СБОРНИК ТЕЗИСОВ · 2016. 11. 4. · 4 vi Международная...

СБОРНИК ТЕЗИСОВ

VI МЕЖДУНАРОДНАЯ КОНФЕРЕНЦИЯ ФИЗТЕХБИО

24-28 мая 2016 годаМосковский физико-технический институт

+7 495 408 42 00

Институтский переулок, дом 9, стр. 7Долгопрудный,

Московская область, Россия, 141700НП «Центр развития БФК «Северный»

www.mipt.ruwww.pharmcluster.ruwww.phystechbio.ru

При поддержке:

Спонсоры:

Информационные партнеры:

Министерство образования и науки Российской Федерации

Инновационный территориальный кластер

Физтех XXI

Правительство Московской области

Проект повышения конкурентоспособности ведущих российских университетов

среди ведущих мировых научно-образовательных центров

Корпорация развитияМосковской области

Научно-исследовательскийинститут молекулярнойбиологии и биофизики

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VI Международная конференция 2016

Оглавление: Итоги VI Международной конференции ФизтехБио 6Тезисы докладов конференции 13Session I: Cancer Cause and Prevention 13Increased lung cancer mortality rates in the Chinese population from 1973-1975 to 2004-2005: an adverse health effect from exposure to smoking Zou X. 13The genome as a record of environmental exposure Phillips D. 14Molecular teranostics Melerzanov A. 14OMICs for precision medicine. Case study of small group of metastatic TNBC patients Serebriyskaya T., Kuzkina N., Kirukhina N., Nikolskaya T. 15Session II: Molecular Carcinogenesis 17Dynamic epigenetic regulation of glioblastoma tumorigenicity through LSD1 modulation of MYC expression Chen C. 17Arsenic induces the highly carcinogenic polyadenylation of canonical histone mRNA by downregulating stem-loop binding protein Costa M. 17Inflammatory and epigenetic regulation in lung cancer Haugen A. 18Global disruption of gene expression in transformed C3H/-10T1/2 mouse embryo cell lines induced by insoluble, carcinogenic nickel compounds Joseph R. Landolph, Jr. Aruni, T. DeSilva-Pehl, and Kazeem A. Akinwumi 19Genotoxicity and сarcinogenicity of engineered nanomaterials Autrup H. 21Enhanced metabolic ROS production by a loss of protein phosphorylation in mitochondria Homma Y. 22Retraction makes more retractions Toshio K., Ukawa A. 22Session III: Cancer Biology 23Regulation of cancer metabolic reprogramming Gao P. 23Role of microRNA in hormonal carcinogenesis Gulyaeva L. 24Genetic disorders in non-small cell lung cancer Demidova I., Gikalo M., Barinov A., Gagarin I., Makhson A. 25Cell signaling and the development of the resistant phenotype of tumor cells Krasil’nikov M. 26The secreted form of heat shock protein-90alpha (HSP90α) defines its tumorigenicity in breast cancer Li W. 27Function of protein kinase CK2 on cell cycle progression Homma M.K,, Ogura M., Homma Y. 27Advancing Oncology research with single-cell approaches Hamilton A. 28Session IV: Cancer Diagnostics 28Roles of a Cell Adhesion Molecule CADM1 in Apoptosis Induction, Cancer Invasion, and Metastasis Ito T., Tsuboi Y., Tsuchiya T., Oba M., Murakami Y. 28Mycoplasma hyorhinis infection conduces toward NFκB-dependent resistance of H292 lung cancer cells to Nutlin3 Boyarskih U., Smetanina M., Zhechev D., Kozlov V., Kel A., Filipenko M. 29Next Generation Sequencing in Hereditary Cancer detection and monitoring Kovalenko S.P., Anisimenko M.S. 30Clinical and molecular-genetic approaches to diagnostics and treatment of thyroid cancer Shevchenko S. 31MicroRNA and somatic mutations in the preoperative diagnostic of thyroid nodules Kolesnikov N., Titov S., Malakhina E., Poloz T., Ivanov M., Shevchenko S., Achmerova L., Veryaskina Y., Zhimulev I. 32

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Modern methods of mathematical modeling in the development of anticancer drugs Yakovlev P. 33Session V. Neuro-Oncology 34Coexpression network analysis of glioma transcriptome Sergeeva M. 34Developing rational medical therapies for von Hippel-Lindau disease: the dilemma of hemangioblastomas in the brain and spinal cord McCutcheon I. 35Radiosurgery for management of brain tumors Chernov M. 36Computerized technologies for cerebral malignant glioma maximum safe resection and postoperative assessment Krivoshapkin A., Gaytan A., Sergeev G., V.V., Maslov N., Mordvinov V., Kanygin, Kurbatov V., Kalneus L. 37Session VI. Breast Cancer 385-Fluorouracil and curcumin inhibit epithelial-mesenchymal transition in breast carcinogenesis Calaf G., Ponce-Cusi R., Gallardо M. 38Selective estrogen mimics for the treatment of tamoxifen-resistant breast cancer using PKCα as a predictive biomarker Tonetti D., Molloy M.E., Zhao H., Pham T.N., Zhao J., Xiong R., Patel H., Gutgesell L., Thatcher G. R. J. 40Heparanase: from basic cancer research to therapeutic applications Vlodavsky I. 41Session VII. Viral Chemotherapy and Cancer Chemotherapy 42Potential drugs to control hazardous viruses including some involved in human malignancies Huberman E. 42Modern surgical oncology in general and visceral surgery (e.g. in the treatment of colon and rectal cancer) Link K.H. 43The role of predictive biomarkers in the treatment of advanced colorectal cancer Kornmann M. 44The tumor microenvironment as a promising target to anti-tumor therapy Cherdyntseva N., Litviakov N., Denisov E., Zavyalova M., Stakheyeva M., Kzhyshkowska J., Perelmuter V. 45Development of novel oral Hedgehog signaling pathway inhibitor for cancer therapy Leonov S. 46

Тезисы постерных докладов 48Somatic mosaic variations in healthy skin fibroblasts Abyzov A., Tomasini L., Zhou B., Vasmatzis N., Mariani J., Amenduni M., Amiri A., Urban A.E., Vaccarinо F. 48Sensing of small molecules using graphene oxide Aftenieva O.A., Stebunov Yu.V., Arsenin A.V. 49Copy number characteristics of the 10q26.3 chromosome region containing the MGMT gene in glioblastoma Alekseeva E., Zaletayev D., Prozorenko E., Zaytsev А., Tanas A., Kirsanova О., Strelnikov V. 50Biopharmaceutical study of oral Ibuprofen complex polymer matrix gel with modified release Antipova I.V., Bakhrushina Е.О., Anurova М.N., Nifontova G.О. 51Frequent mutations in renal angiomyolipoma Anoshkin K., Kuznetsova E., Mosyakova K., Tanas A., Chaplygina M., Alekseeva E., Shpot E., Zaletayev D., Vinarov A., Strelnikov V. 52Content of metalloproteinases in serum of bone sarcoma patients Babkina I.V., Chernomaz I.S., Bondarev A.V., Schupak M.Yu., Soloviev Yu.N. 53Quisinostat Oral HDAC inhibitor for the treatment of non-small cell lung cancer and ovarian cancer Baranovskiy S. 54Gosogliptin — a new innovative drug for the treatment of type 2 diabetes and metabolic syndrome Baranovskiy S. 55Il-16 and VEGF in the blood serum of patients with bone tumors Bondarev A.V., Timofeev Yu.S., Schupak M.Yu., Kuznetsov I.N., Babkina I.V., Alferov A.A., Boulytcheva I.V. 56Using PolyGraphene During Pre-Clinical Tests of the Enterosorption and Safety Assessment Botin A., Buravtsev V., Popova T. 57Sorbtion — Desorbtion of Glycopeptide Antibiotics on the Surface of Oxidized OligoGraphene Buravtsev V., Botin A., Baratova L., Timofeeva A., Katrukha G., Poletaev A. 59Expression of miR-21 and its target genes ACAT1 and ARMCX1 in liver of rats treated with DDT Chanyshev M. 60Profiling of mutations in gastric cancer Cherepanova K.I., Strelnikov V.V., Bykov I.I., Nemtsova M.V., Tanas A.S. 61

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VEGFA isoforms balance shifts towards more angiogenic variants in human hepatocellular carcinoma Chesnokov M., Skovorodnikova P., Shavochkina D., Kustova I., Mugue N., Kudashkin N., Moroz E., Patyutko Y., Lazarevich N. 62

Functional hypermethylation of a number of chromosome 3 genes revealed in breast cancer using noti-microarrays Dmitriev A.A., Krasnov G.S, Beniaminov A.D., Pronina I.V., Puzanov G.A., Kurevlev S.V., Kazubskaya T.P., Loginov V.I., Senchenko V.N., Braga E.A., Kashuba V.I. 64

Storage as a way to improve the efficiency of sequencing data usage Zhegalova I., Petuhova A., Suchkov S. 65

MicroRNAs as biological markers of ovarian cancer Gershtein E.S., Kushlinsky D.N., Tereshkina I.V., Adamyan L.V. 66

RANK/RANKL/OPG system and interleukins 6, 8 16 in blood serum of patients with primary bone neoplasms Gershtein E.S., Zuev A.A., Timofeev Yu.S., Soloviev Yu.N., Kushlinskii N.E. 67

Grammometric labs on a chip and synchronization of the on-chip isolated muscle fiber grammometry with the in situ tissue X-ray analysis and MEMS-assisted excitation time spectroscopy Gradov O.V. 68

Homocysteine plasma level distribution in patients with cancer Ivanov A.V., Nikiforova K.A., Kushlinskii N.E., Kubatiev A.A. 69

Development of plant-produced universal influenza nanovaccines: from “green” biotechnology to medicine Ivanov P.A., Gasanova T.V., Petukhova N.V. 70

Expression of microRNAs and their target genes in various organs of female rats under DDT exposure Kalinina T., Ushakov D., Philippov S. 71

The involvement of methylation in down-regulation of some suppressive miRNA genes and progression of renal carcinoma Khokonova V.V., Pronina I.V., Burdennyy A.M., Dmitriev A.A., Karpukhin A.V., Kazubskaya T.P., Braga E.A., Loginov V.I. 72

Study of PTEN expression in endometrial cancer Kobelev W.S., Geletina N.S. 73

Creating model systems for testing the bioactivity of type I and II IFNs Konyaeva E.P., Kulikova K.V., Gnuchev N.V., Georgiev G.P., Larin S.S. 74

Metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in tumor and plasma of colorectal cancer patients: relationship, clinico-pathologic and prognostic implication Korotkova E.A., Nikolaev A.A., Gershtein E.S., Prorokov V.V. 75

Isolation and characteristic of colon cancer cell with stem cells properties from primary tumors Koshkin S., Raskin G., Petrov N., Bajenova O., Tolkunova E. 76

IGFs and IGF binding proteins as diagnostic and prognostic tumor markers Kostyleva O.I., Maslyaev A.A., Kushlinsky D.N., Mamedov U.R., Korotkova E.A., Adamiyan L.V., Gershtein E.S. 77

Clinical significance of receptor epidermal growth factors in malignant tumors Kostyleva O.I., Gershtein E.S., Azigova R.R., Galdava D.E. 79

Development of technology of manufacturing polymeric microparticles for peptides encapsulation Lyovina T.A., Krechetov S.P., Nifontova G.O. 79

Study of extracellular matrix proteins in metastatic breast cancer Lobanova V., Kononchuk V. 80

Integrins and their ligands osteopontin and thrombospondin-1, can be considered as biomarkers of aggressive papillary thyroid cancer phenotype Logacheva G.P., Mostovich L.A., Svyatchenko S.V., Shevchenko S.P., Gulyaeva L.F. 81

Biochemical markers in neuroendocrine tumors N.V. Lyubimova, M.G. Toms, T.K. Churikova, T.Yu. Kharitidy 82

Neurospecific proteins as biochemical markers of brain tumours Lyubimova N.V., Timofeev Yu.S., Mitrofanov A.A., Bekyashev A.Kh. 83

Study of internalization of PLGA nanoparticles into the intracranial rat C6 glioma: influence of surfactant coating Malinovskaya Y., Gelperina S., Melnikov P., Osipova N., Maksimenko O., Baklaushev V. 84

Normacor® — a new cardioplegic solution to conduct open heart surgeries at normal body temperatureNikolay Merkin 85

Metalloproteinases, MMP tissue inhibitors, and VEGF in blood serum of endometrial cancer patients as potential diagnostic markers Mushtenko S.V., Korotkova E.A., Tereshkina I.V., Dvorova E.K., Gershtein E.S. 86

Сlinical significance of VEGF and uPA evaluation in early stages breast cancer Ovchinnikova L.K., Kostyleva O.I., Alekseeva E.I., Filippova M.A. 87

Protein regulators of epithelial-mesenchymal transition and some components of the VEGF signaling pathway in breast cancerpatients Ovsii O.G., Scherbakov A.M., Guens G.P., Ermilova V.D., Kushlinskii N.E. 88

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Epigenetic mechanisms of expression deregulation of pro-apoptotic APAF1 and DAPK1 genes in breast cancer Pereyaslova E.A., loginov V.I., Pronina I.V., Burdennyy A.M., Kazubskaya T.P., Kushlinskii N.E., Braga E. 89Bioinformatic tools passing through the nigth of neurodegenerative disorders Petuhova А., Zhegalova I., Suchkov S. 91Express way to assess biocompatibility of metal alloys and implants Pilipenko P.N. 92Unsupervised clustering of cancer samples based on expression profiles Rozenwald M. 93Applying NMR spectroscopy to investigation of spider venom’s pharmaceutical potential demonstrated on 3D structure of the two – domain spider toxin OtTx1a Romanovskaya D. 94Peptidoglycan-recognition protein Tag-7/PGLYRP-1 modulate antibiotic-resistance and early steps of innate immune response Slonova D.A., Posvyatenko A.V., Sysolyatina E.V., Ermolaeva S.A., Gnuchev N.V., Georgiev G.P., Kibardin A.V., Larin S.S. 95Lipidome comparison of endometrial tissue by direct mass spectrometry and LC-MS/MS Starodubtseva N.L., Chagovets V.V., Kononikhin A.S., Borisova A.V., Kozachenko A.V., Frankevich V.E., Adamyan L.V. 96Breast cancer molecular classification based on DNA methylation assessed by reduced representation bisulfite sequencing Tanas A., Poddubskaya E. V., Kekeeva T. V., Kerimov R. A., Trotsenko I. D., Kuznetsova E.B., Strelnikov V., Zaletayev D. 98The problem of findingthe cell of origin in glioma Timin G.V., Tolkunova E.N., GulyaevD.A., Lahina Yu.S. 99Insulin-like growth factors (IGF) and IGF-binding proteins in bone sarcoma patients Yu.S. Timofeev, I.V. Boulytcheva, E.S. Gershtein, I.V. Babkina, Yu.N. Soloviev, M.D. Aliev 100PE7/4 Safety and antiviral effect of Elpida (VM-1500), a novel NNRTI (+Truvada) in treatment-naïve HIV-1 infected patients Tyrnova I. 101Tissue-specific expression of microRNAs in rats under DDT and benzo(a)pyrene exposure Ushakov D., Filippov S. 101Using the computational crystallomics to reconstruct full-atomic structure of short fibrin oligomers and protofibrils Zhmurov A., Litvinov R., Zhukov P., Weisel J.W., Barsegov V. 102A molecular dynamics study of the mechanical properties of α-helical coiled-coils Minin K., Zhmurov A., Barsegov V. 103Doclinical research of GZK-111 — dipeptide cognitive enhancer K.N. Kolyasnikova 104Assessment of genetic factors and chronotype for prediction of a bus driver’s road accident risk Dorokhov V.B., Puchkova A.N., Taranov A.O., Tupitsyna T.V., Slominsky P.A., Dementienko V.V., Ermolayev V.V. 104 Natural dipeptide carnosine: a promising neuroprotector and cognitive brain function corrector Fedorova T.N., Stvolinsky S.L., Lopachev A.V., Berezhnoy D.S., Deviatov A.A., Konovalova E.V., Gavrilova S.A., Morozova M.M., Maximova M.Y., Illarioshkin S.N. 106A system for continuous monitoring and control of operator’s and driver’s functional conditions Zakharchenko D.V. 107Efficiency of NGS in molecular diagnostics and CFTR Mutations Spectrum in Russian Cystic Fibrosis Patients Ivanov M.V., Glazova O.V., Matsvay A.D., Krasovskiy S.A., Amelina E.L., Kovalenko S.P., Khafizov K.F. 108Study of life span capacity of new PI3K inhibitors on С. elegans model Marusich E., Ovcharenko D., Lashmanova E., Moskalev A., Leonov S. 109Pollen-based High Content Screening Assay to Discover New Potential Plant Herbicides Chuprov-Netochin R., Neskorodov J., Marusich E., Mishutkina Y., Volynchuk P., Leonov S. 111Multitarget activity profiling predicts in vivo adverse effects of serotonin and norepinephrine reuptake inhibitors Lapushkin G.I., Balakin K.V., Savilova A.G., Voronkov А.E. 112Search of useful side effects of well studied drugs by means of the FDA AERS database Lapushkin G.I. 114The effects of quercetin on lifespan and stress resistance of Drosophilla melanogaster Proshkina E., Lashmanova E., Moskalev A. 115Pharmacoeconomic Analysis of Direct Activity Antiviral Drugs Used for Treatment of Genotype 1 Chronic Hepatitis C Musina N.Z., Savilova A.G., Korzinov O.M. 116Synthesis of Macrocyclic Organo-short\medium peptide Hybrids via CuAAC Vantskul A., Ivanenkov Y., Polyakova M., Sandulenko Y. 117Extra bright fluorescent labels based on organic dyes Rzhevskiy S.A., Shadrin I.A., Babaev E.V., Pakhomov A.A. 118Список докладов круглых столов и питч-сессии стартапов 120Контакты спикеров VI Международной конференции ФизтехБио 128

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Итоги VI Международной конференции

ФизтехБио

ФизтехБио – место для диалога науки и индустрии

Фокусом конференции ФизтехБио стали вопросы передовых онкологических исследований, дискуссии по темам Национальной технологической инициативы и мероприятия для молодых ученых.

VI Международная конференция ФизтехБио проходила в кам-пусе Московского физико-технического института с 24 по 28 мая 2016 г. Насыщенная научная программа и множество сателлитных мероприятий привлекли внимание более чем 400 участников. ФизтехБио в 2016 году посетили представители зарубежных и рос-сийских университетов, научно-исследовательских институтов, медицинских и образовательных центров, фармацевтических и медицинских компаний, ученые, преподаватели, врачи, предпри-ниматели, разработчики.

Организаторами конференции выступили Биофармкластер «Север-ный» и Центр живых систем МФТИ. Основу научной программы Физтех-Био составили сессии 21-го Международного Симпозиума им. Чарльза Хайдельбергера, организованного совместно с НИИ молекулярной био-логии и биофизики (г. Новосибирск).

На открытии ФизтехБио высту-пил заместитель министра образова-ния и науки Российской Федерации Александр Повалко. В своей речи он отметил:

«Важно, что наравне с проекта-ми по запуску ракет в космос, у нас су-ществуют и развиваются проекты, нацеленные на здоровье и долголетие человека, и нам крайне важно продол-жать работу в этом направлении. Это вопросы и фармакологии, и разработки

Заместитель министра образования и науки Российской Федерации Александр Повалко

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медицинской техники, и трансляционной медицины, фундаментальных исследований. И Физтех взялся за решение этой задачи со свойственным ему энтузиазмом. Существует факультет биологической и медицинской физики, создан Биофармкластер «Северный». И мы не собираемся на этом останавливаться. В планах развитие пояса инновационных компаний во-круг Физтеха, создание университетской клиники, интеграция по сетевым программам с другими университетами, имеющими клиническую базу. Но все это возможно, только если будут возникать новые свежие идеи. Мы стараемся обеспечить необходимую инфраструктуру и условия работы для ученых и рассчитываем, что здесь, на Физтехе в Долгопрудном, вам будет комфортно и интересно как учиться, так и работать», — сказал Александр Повалко.

С напутственными словами к участ-никам конференции обратился ректор МФТИ Николай Кудрявцев: «Сначала нас много критиковали, когда мы взяли курс на биологию. Но должен сказать, это было очень мудрое решение, потому что в медицине есть нужда в людях, которые будут улучшать диагностику, развивать новые методы, новые подходы и приме-нять для этого физику». Далее предсе-датель совета директоров ЦВТ ХимРар, член наблюдательного совета МФТИ, президент правления Центра живых си-стем МФТИ Андрей Иващенко сделал до-клад по современной истории развития наук о живом на Физтехе от формиро-вания биофармацевтического кластера «Северный» в 2011 году до постройки но-вого корпуса МФТИ в 2015. Приветствие в адрес ФизтехБио направил также руководитель Росздравнадзора Михаил Мурашко, отметивший особую роль биологического и медицинского на-правления в МФТИ.

Конференция продолжилась выступлениями участников 21-го Меж-дународного Симпозиума имени Чарльза Хайдельбергера. Чарльз Хайдельбергер – бывший президент Американской ассоциации по изуче-нию рака, член Национальной академии наук США, выдающийся ученый в областях химиотерапии рака и канцерогенеза. Значительную часть сво-его состояния Чарльз Хайдельбергер завещал на развитие науки, благо-даря чему онкологические симпозиумы с участием выдающихся мировых

Ректор МФТИ Николай Кудрявцев

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ученых ежегодно проводятся в разных странах мира. Серию докладов открыл профессор Давид Заридзе, заместитель директора Российского онкологическо-го научного центра им. Н. Н. Блохина, который лично был знаком с Чарльзом Хайдельбергом. Профессор Заридзе вы-ступил с сообщением «Эпидемиология и профилактика рака». Далее с докладами на Симпозиуме выступили врачи и ис-следователи из США, Великобритании, Норвегии, Израиля, Германии. Более 50 спикеров конференции представи-ли и обсудили результаты последних исследований в области диагностики и профилактики онкологических забо-леваний, молекулярных механизмов канцерогенеза, нейроонкологии, био-логических аспектов рака, вирусной и раковой химиотерапии. Видеозаписи этих выступлений будут доступны на сайте Биофармкластера «Северный»: www.pharmcluster.ru.

Спикеры 21-го Международного Симпозиума им. Чарльза Хайдельбергера

Давид Заридзе, заместитель директора РОНЦ им. Н. Н. Блохина

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Накануне открытия ФизтехБио более 100 молодых ученых и раз-ных медицинских вузов и центров смогли бесплатно стать участниками Школы молодых ученых, где с образовательными докладами по пер-спективным достижениям клинической и молекулярной онкологии вы-ступили Карл-Генрих Линк, директор хирургического центра лечения опу-холей Asklelpios Rhein/Main (Герма-ния), Кларк Чен, руководитель отделе-ния стереотактики и радиохирургии, профессор подразделения нейрохи-рургии Университета Южной Кали-форнии (США), специалисты из рос-сийских научно-исследовательских институтов.

Для профессионалов отрасли на ФизтехБио были организованы специализированные круглые столы и дискуссии. Так, 25 мая состоялось заседание Рабочей группы Мини-стерства образования и науки Рос-сийской Федерации по рассмотре-нию мероприятий ФЦП «Развитие

Все участники Школы молодых ученых получили сертификаты

Рабочая группа Минобрнауки России Фарма-2020 в Биофармацевтическом корпусе МФТИ

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фармацевтической и медицинской промышленности Российской Фе-дерации на период до 2020 года и дальнейшую перспективу» по про-ведению доклинических исследований инновационных лекарственных средств и разработки новых образовательных программ и образователь-ных модулей для профильных высших и средних специальных учебных заведений.

На круглом столе «Инновационные кластеры Московской об-ласти» представители территориальных кластеров «Пущино», «Дубна», «ФИЗТЕХ-XXI», а также представители Правительства Московской обла-

сти обсудили перспективы сотрудничества и текущие проекты участников кла-стера, в частности вопросы подготовки кадров на базе МФТИ и университетов Мо-сковской области, прове-дение совместных образо-вательных мероприятий и планов по переподготовке кадров в области доклини-ческих и клинических ис-следований, организации производства лекарствен-ных средств по стандартам GLP, GCP и GMP.

Отдельным треком были представлены круглые столы по направле-нию Национальной технологической инициативы НейроНет. Организато-ром трека выступил Отраслевой Союз НейроНет (rusneuro.net). На круглом столе «Научно-исследовательские и технологические проекты Дорожной карты НейроНет» специалисты рас-смотрели различные проекты в рамках проекта «Сobrain» для последующего включения в дорожную карту НейроНет. Формат и структуру НейроНет Центров, создание которых предусмотрено в ре-гионах России, обсудили еще за одной

Встреча представителей кластеров Московской области на круглом столе конференции ФизтехБио

Андрей Иващенко — руководитель рабочей группы НТИ «НейроНет» со вступительным

словом на Круглых столах по НейроНету

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дискуссией. В заключение официальные лица Отрас-левого союза НейроНет на-помнили, что организация открыта к сотрудничеству с представителями крупного и малого бизнеса, научными и производственными органи-зациями, а также частными лицами, работающими в сфе-ре НейроНет.

На круглом столе «ЦНС-Тераностика» экс-перты из ЦВТ «ХимРар», На-учного центра психического

здоровья РАМН, Научного центра неврологии и ИБР РАН рассмотрели дорожную карту мультицентрового проекта по сбору мультимодальных данных для последующей разработки систем диагностики и терапии (те-раностики) заболеваний центральной нервной системы.

В последний день конференции при поддержке ООО «КСИ Вен-чурс» прошла питч-сессия стартапов. Свои проекты в области фар-мацевтики, медицинской техники, агробиотехнологий и нейротехноло-гий представили более двадцати молодых команд из разных городов России – Москва, Санкт-Петербург, Улья-новск, Чебоксары, Владимирская об-ласть, Волгоград, Тюмень. По итогам конкурса были определены три победи-теля, которые получат пакет акселераци-онной поддержки от Биофармкластера «Северный» и «КСИ Венчурс». Награды получили Николай Бушков (МФТИ, «Те-рапевтический аппарат оксигенации»), Ирина Дейген (МГУ им. Ломоносова, «Инновационный препарат для таргет-ной терапии колоректального рака»), Марина Попова (ПСПбГМУ им. И. П. Пав-лова Минздрава России, «Разработка

Круглые столы по направлению НейроНет

Марина Попова, ПСПбГМУ им.И.П. Павлова Минздрава России, один из победителей

питч-сессии

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и организация серийного производства отечественного, «Иммунотера-пия нового поколения в онкологии»).

Питч-сессию также дополнила открытая дискуссия «Междисципли-нарные проекты и технологии в области живых систем». Обсуждение коснулось современных тенденций в инвестировании перспективных проектов для вложений на ранних стадиях — в сельском хозяйстве, вирту-альной реальности и телемедицине.

В течение всех дней конференции в кампусе МФТИ работала экспози-ция фармацевтических, биотехнологический и спе-циализирующихся на ме-дицинском приборострое-нии компаний. В общем сложности свои разработ-ки представили более 30 компаний, среди них: «Хе-ликон», «Биокад», «Рош», «Мерк», «БиоВитрум», «Спектроника», «Диаэм», «Биосан», «Нейроком», «БиоХимМак», «ХимРар», «Сотекс» и другие. В экспозиции конференции также приняли участие стартапы Биофармкластера «Северный» - компании «КардиОН», «Агро-биотех-2020», «ДНК-наследие», «ГенДив», «Брейн-Бит», «Технологии здо-ровья». Научную часть выставки конференции представили 32 постер-ных доклада в области биологии, медицины и кросс-дисциплинарных проектов.

Видеоотчет и фотоотчет о конференции, а также видеозапи-си всех выступлений доступны на официальном сайте конференции физтехбио. рф, а также на сайте БФК «Северный» pharmcluster.ru.

До встречи на VII Международной конференции

ФизтехБио!

Выставочная экспозиция конференции ФизтехБио

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Тезисы VI Международной конференции

ФизтехБиоSession I: Cancer Cause and Prevention

Increased lung cancer mortality rates in the Chinese population from 1973-1975 to 2004-2005:

an adverse health effect from exposure to smoking

Xiaonong Zou, MDProfessor of National Office of Cancer Prevention and Control,

National Cancer Center, Cancer Institute of Chinese Academy of Medical Sciences

Background: Lung cancer incidence and mortality rates have increased substantially in China despite improvements in clinical diagnosis and treatment approaches, as well as significant advances in tobacco control policy implemented in recent decades.

Methods: Age-standardized mortality (ASM) and age-specific rates of lung cancer in China were estimated for the time periods 1973-1975, 1990-1992 and 2004-2005 using data from three National Death Surveys. The percentages of ever smokers among lung cancer cases identified from a hospital-based information system were calculated by histology and demographic variables.

Results: The ASM from lung cancer in China dramatically increased from 7.30 per 100,000 in 1973-1975 to 27.62 per 100,000 in 2004-2005. Increases in lung cancer ASM were consistent in male and female, urban and rural populations. In men of age 75-79, lung cancer mortality increased remarkably to 453.67 per 100,000 in 2004-2005, from 246.78 per 100,000 in 1990-1992 and 53.65 per 100,000 in 1973-1975. Among 6,674 lung cancer cases identified in 2003-2007 from a hospital-based database, 82.97% of men were ever smokers (73.35% with adenocarcinoma, 91.8% with squamous cell carcinoma), and 11.18% of women were ever smokers (6.00% with adeno-squamous carcinoma, 39.02% with squamous cell carcinoma). Differences in numbers of ever smokers were found in age groups but not in year of diagnosis.

Conclusions: The consistent and rapid increases in the lung cancer mortality rate in the Chinese population and high prevalence of exposure to smoking prompt a strong call for the implementation of a comprehensive tobacco control policy and specific public health educational strategies.

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The genome as a record of environmental exposure

David H. PhillipsKing’s College London, Analytical and Environmental Sciences Division,

MRC-PHE Centre for Environment and Health

Whole genome sequencing of human tumours has revealed distinct patterns of mutation that hint at the causative origins of cancer. Experimental investigations of the mutations and mutation spectra induced by environmental mutagens have traditionally focused on single genes. With the advent of faster cheaper sequencing platforms, it is now possible to assess mutation spectra in experimental models across the whole genome. As a proof of principle, we have examined the whole genome mutation profiles of mouse embryo fibroblasts immortalised following exposure to benzo[a]pyrene (BaP), ultraviolet light (UV) and aristolochic acid (AA). The results reveal that each mutagen induces a characteristic mutation signature: predominantly G T mutations for BaP, C T and CC TT for UV and A T for AA. The data are not only consistent with existing knowledge but also provide additional information at higher levels of genomic organisation. The approach holds promise for identifying agents responsible for mutations in human tumours and for shedding light on the aetiology of human cancer. More recently we have begun to investigate mutations in single-cell clones of human stem cells exposed to mutagens to provide whole genome signatures acquired without biological selection.

ReferenceNik-Zainal, S., Kucab, J.E., Morganella, S., Glodzik, D., Alexandrov, L.B., Arlt, V.M., Weningen, A., Hollstein, M., Stratton, M.R. and Phillips, D.H. The genome as a record of environmental exposure. Mutagenesis, 30, 763-770 (2015).

Molecular teranostics

Alexander Melerzanov, MD, PhD, Dean of Department of Biological and Medical Physics

of Moscow Institute of Physics and Technology (State University), Expert of Ministry of Education and Science for Pharma-2020

Most cancer patients die from metastatic disease as the result of circulating tumor cells (CTCs) spreading from a primary tumor through the blood to distant organs. Clinical studies have demonstrated the tremendous potential of using CTC counts as prognostic or predictive markers of metastatic development, cancer recurrence, and therapeutic efficacy.

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Despite significant progress in the development of CTC assays, existing CTC assays in vitro still have many limitations, including time-consuming blood processing and low sensitivity due to the small samples of blood taken.

To overcome these problems, we developed label-free in vivo photoacoustic (PA) flow cytometry (PAFC) and have demonstrated its feasibility in the first pilot trial in melanoma patients. This clinical study revealed the tremendous potential for further improving PAFC technology for label-free detection of weakly pigmented cells by increasing its sensitivity through the safe delivery of a higher level of laser energy to deep vessels. Next generation of the in vivo PAFC platform with advanced parameters by using the principle of nonlinear photoacoustics, fiber delivery of laser energy, sub-nanosecond lasers and a focused ultrasound transducer is under development.

In the course of this study, we will obtain statistically significant data that will demonstrate this innovative technique’s unprecedented sensitivity threshold of 1 CTC/100 mL (with an attempt to achieve 1 CTC/500 mL) for enumerating rare melanoma (as an example) CTCs in vivo without the need for labeling. Application of these technologies may include early melanoma diagnosis, cancer recurrence detection, and rapid evaluation of therapeutic efficacy.

OMICs for precision medicine. Case study of small group of metastatic TNBC patients

Serebriyskaya T., Kuzkina N., Kirukhina N., Nikolskaya T.Moscow Institute of Physics and Technology (State University)

Rationale. One of common trend in personal oncology is search and determination of robust biomarkers which are applicable for groups of patients with similar clinical data. However, number of existing biomarkers is too small to cover all needs of personal oncology. One of the reason is low frequency of occurrence of genetic variations associated with disease or treatment because of tumor heterogeneity that make hard both research process and enrollment of patients in clinical trials. TNBC is characterized by high heterogeneity, up to now there is no special treatment and predictive biomarkers for TNBC. Alternatively, development of OMICs technology provides new opportunity for building of individual models for patient tumor processes that describe his own clinical features with subsequent treatment tailoring.

Research objective. The aim of research was to build individual models of patient tumor processes based on compilation of genomic and expression

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data and to elucidate individual mechanisms of carcinogenesis and of acquired or hereditary drug resistance.

Materials and methods. Small group of 10 patients suffered from metastatic triple-negative breast cancer (TNBC) was enrolled in study. The tumor biopsies were from metastatic lesions from regional lymph nodes. Peripheral blood samples were taken as a control. Whole exome sequencing was performed for the genomic DNA obtained from tumor and blood specimens using Ion Torrent Proton platform. Whole transcriptome analysis was performed using Affymetrix GeneChip Human Gene 2.1 ST Array. DAVID, MetaCore and some other tools were used for functional and integrated analysis.

Results. Whole exome sequence showed high heterogeneity of metastatic samples taken from the same localization - lymph node. Several hereditary risk factors (mutations in TP 53/ Li-Fraumeni syndrome, mutation in MSH6 / Lynch syndrome) and damaging mutations in well- known driver genes were detected in patient samples. Analysis of tumor gene expression data allowed to reveal the concordance with subtypes of Lehmann’s and intrinsic classifications of TNBC. Both classifications were developed based on of primary tumors data, Lehmann’s classification suggest treatment for every subtype. All tumors corresponded to basal type of tumor based on intrinsic classification. Based on Lehmann’s classification basal-like subtype (BL1 and BL2) was detected in 3 patients, an immunomodulatory (IM) — in 2 patients, a mesenchymal (M)- in 3 patients, a mesenchymal stem–like (MSL)- in 2 patients. Expression analysis revealed claudin 7, osteopontin, L-type amino acid transporter 1 (LAT1), MKI67, TOP2A, cyclin A and FOXM1 as the most common upregulated genes for all patients. Overexpression of these genes was shown in many cancers and role for anticancer therapy was discussed. Predictive causative models for individual tumors were built on the basis of integrated analysis of genomic and expression data. Driver characteristics of selected mutations were hypothesized and discussed.

Conclusion. Analysis of genomic and transcriptome data of tumor allows to detect personal features of tumor processes both individually and combined. Classifications developed for primary tumors can be applied for metastatic tumor samples. Integrated analysis allows to build individual models that reflect process of carcinogenesis and in some cases explain the drug resistance. This approach requires further clinical validation.

* Study was conducted in collaboration with FGBU «RONC N.A. N.N. BLOKHIN»

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Session II: Molecular Carcinogenesis

Dynamic epigenetic regulation of glioblastoma tumorigenicity through LSD1 modulation of MYC expression

Dr. Clark Chin-Chun ChenDivision of Neurosurgery, Dept. of Surgery, Univ. of California at San Diego

San Diego, California, USA

The available evidence suggests that the lethality of glioblastoma is driven by small subpopulations of cells that self-renew and exhibit tumorigenicity. It remains unclear whether tumorigenicity exists as a static property of a few cells or as a dynamically acquired property.

We used tumor-sphere and xenograft formation as assays for tumorigenicity and examined subclones isolated from established and primary glioblastoma lines. Our results indicate that glioblastoma tumorigenicity is largely deterministic, yet the property can be acquired spontaneously at low frequencies. Further, these dynamic transitions are governed by epigenetic reprogramming through the lysine-specific demethylase 1 (LSD1). LSD depletion increases trimethylation of histone 3 lysine 4 at the avian myelocytomatosis viral oncogene homolog (MYC) locus, which elevates MYC expression. MYC, in turn, regulates oligodendrocyte

lineage transcription factor 2 (OLIG2), SRY (sex determining region Y)-box 2 (SOX2), and POU class 3 homeobox 2 (POU3F2), a core set of transcription factors required for reprogramming glioblastoma cells into stem-like states. Our model suggests epigenetic regulation of key transcription factors governs transitions between tumorigenic states and provides a framework for glioblastoma therapeutic development.

Arsenic induces the highly carcinogenic polyadenylation of canonical histone mRNA by downregulating stem-loop binding protein

Max Costa Department of Environmental Medicine and Department of Biochemistry and Molecular

Pharmacology NYU School of Medicine

The replication-dependent histone genes are the only metazoan genes whose messenger RNA (mRNA) does not terminate with a poly(A) tail at the 3’ end. Instead, the histone mRNAs display a stem-loop structure at their 3’ end. Stem-loop binding protein (SLBP) binds the stem-loop and regulates canonical

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histone mRNA metabolism. We report that exposure to arsenic, a carcinogenic metal, decreases cellular levels of SLBP by inducing its proteasomal degradation and inhibiting SLBP transcription via epigenetic mechanisms. Notably, arsenic exposure dramatically increases polyadenylation of canonical histone H3.1 mRNA possibly through downregulation of SLBP expression. The polyadenylated H3.1 mRNA induced by arsenic is not susceptible to normal degradation that occurs at the end of S phase, resulting in continued presence into mitosis, increased total H3.1 mRNA, and increased H3 protein levels. Excess expression of canonical histones has been shown to increase sensitivity to DNA damage, as well as increase the frequency of missing chromosomes and induce genomic instability. Our hypothesis is that H3.1 genes are initially transcribed with a normal stem-loop construct at the 3’ end of the mRNA. Arsenic depletes nuclear levels of SLBP resulting in H3.1 transcripts unbound to SLBP. These unbound transcripts lose their stem-loop structure at the 3’ end and acquire a poly(A) tail, which increases the half-life and facilitates translation of the mRNA, two factors that provide for increased H3.1 protein levels. The poly(A) H3.1 mRNA is not susceptible to normal degradation that occurs at the end of S phase allowing for its presence in other phases of the cell cycle. Over-expression of SLBP suppressed the effect of As on H3.1 polyadenylation. Knockdown of SLBP by siRNA induced anchorage independent growth of BEAS2B cells. Transfection of Polyadenlated H3.1 but not H3.3 also induced anchorage independent growth in BEAS2B cells. Anchorage independent growth was also induced in human Urothelial cells by transfection of polyadenylated H3.1 but not H3.3.

Inflammatory and epigenetic regulation in lung cancer

Aage Haugen, Dept. of Toxicology, National Institute of Occuaptional Health, Oslo, Norway

The role of inflammation in lung carcinogenesis has become more evident. Interleukins regulate the expression of several molecules and signaling pathways involved in inflammation. Our earlier studies suggest an association between lung cancer risk and single nucleotide polymorphisms (SNPs) in the regulatory regions of IL1B, IL6 or IL8. We recently reported that SNPs in the regulatory region of the IL1B gene affected the mRNA levels by changing the binding affinity of transcription factors or by creating novel binding sites. IL1B-SNPs at - 3893, -1464, -511 and -31 positions formed a specific risk haplotype (GGCT) with higher frequency in the patients compared to cancer-free controls. The risk GGCT haplotype was correlated with significantly higher IL1B mRNA

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levels in the lung of lung cancer patients. We further characterized the specific transcription factor binding to the IL1B -31 T/C polymorphism which is localized in the TATA box at the core promoter region.

In order to get a better understanding of the regulation of the CYP1A1 gene in lung cancer, we studied CYP1A1 enhancer DNA methylation in normal lung and in adenocarcinomas. We found that in normal lung from lung cancer patients smoking and age affected CYP1A1 DNA methylation (pyrosequencing of bisulfite-treated DNA). We found inverse associations between methylation and CYP1A1 gene expression and DNA adducts in normal lung tissue and TP53 and K-ras mutations in tumor tissue (matched pair). We also found increased methylation in adenocarcinoma compared with normal lung, which is related to significantly reduced gene expression in the tumor. These results indicate overall that the CYP1A1 enhancer methylation may affect the initiation of lung carcinogenesis. Furthermore, our data indicate a role of epigenetic modification in the regulation of expression of key cytokines involved in inflammatory responses and SOX-2 during lung cancer development.

Supported by the Norwegian Cancer Society and the Research Council of Norway.

Global disruption of gene expression in transformed C3H/-10T1/2 mouse embryo cell lines induced by insoluble, carcinogenic nickel

compounds

Joseph R. Landolph, Jr.1, 2, 3, Aruni T. DeSilva-Pehl, and Kazeem A. Akinwumi1, 3

Depts. Mol. Micro./Immun.1 and Path.,2 Cancer Res. Lab.,1303 N. Mission Road, USC Comp. Cancer Ctr.,3 Keck School Medicine, Univ. South. Calif.

Abstract: Nickel (Ni) refinery workers inhaling Ni sulfidic ore dusts and smoking cigarettes in Ni refineries have increased incidences of nasal/lung cancers. Inhalation of Ni3S2 or green NiO induces respiratory cancer in rats. We showed Ni3S2 and green/black NiOs were phagocytosed into C3H/10T1/2 Cl 8 (10T1/2) mouse embryo cells and induced chromosomal aberrations, cytotoxicity, and morphological, A. I., and neoplastic transformation in 10T1/2 cells. Using mRNA differential display to analyze 1/16 of the total mRNA, we found 8 genes were differentially expressed between non-transformed and Ni- and MCA-transformed (Tx) 10T1/2 cells. Extrapolat-ing these results to 100% of the mRNA indicated 144 genes were differentially expressed between non-Tx 10T1/2 cells and 2 MCA and 4 NiO- or NiS-Tx 10T1/2 cell lines. In Ni-Tx and MCA-Tx cell lines, 6 driver genes were over-expressed, leading to a further 52

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genes being over-expressed. In Ni-Tx and MCA-Tx cell lines, 9 suppressor-like driver genes were under-express-ed/not expressed, leading to an additional 77 genes being under-expressed/not expressed. Altera- tion of each driver gene led to alterations in expression of 9 genes downstream. Among these differentially expressed genes, Ni+2-Tx and MCA-Tx cell lines contained a) ect-2 gene amplification / higher levels of ect-2 gene mRNA/protein; b) higher levels of calnexin mRNA /protein and Wdr1 mRNA; and c) no detectable levels of DRIP80 or β-centaurin-2 mRNAs. We hypothesized 1) Ni+2 –induced amplification of the ect-2 gene led to higher levels/aggregation of microtubules (MTs); 2) Ni+2-induced silencing of the β-2-centaurin-2 gene caused higher levels of/aggregation of microfilaments (MFs); and 3) Ni+2-induced silencing of DRIP80 gene altered distributions of Ca+2 ions, in Tx 10T1/2 cells. Testing these hypotheses, we stained cells with fluorescent phall- oidin to decorate MFs, with fluorescent Ab to α-tubulin to decorate MTs, with Fluo 3AM to stain Ca+2 ions, and with DAPI to decorate nuclei, then examined cells by confocal microscopy. In non-Tx 10T1/2 cells, MFs/MTs were arranged homogeneously in long fibers. In NiS/green NiO-Tx cell lines, MFs/MTs were over-expressed and aggregated in some areas/absent in others, changing shapes of Tx cells. In non-Tx cells, Ca+2 ions were found in a) State I, in low density cells, with high concentrations of nuclear Ca+2 ions, lower amounts in the cytoplasm; and b) in State II, in high density cells near confluence, with fewer Ca+2 ions in the nucleus, most cytoplasmic. Non-Tx 10T1/2 cells cycled between States I/II. In six NiO/NiS- or MCA-Tx cell lines, Ca+2 ions were largely cytoplasmic (State II). We conclude mutations/amplifications in 6 target genes lead to over-expression of 52 additional genes. Further, silencing of 9 other target genes led to differ-ential under-expression/silencing of 9 additional genes for each primary gene altered, causing differential down-regulation of expression of 77 additional genes. Together, up-regulation of expression of 52 genes and down-regulation of expression of 77 genes led to differential expression of 129 genes in nickel/MCA Tx cell lines. We further conclude: a) Ni+2 ions amplified the ect-2 gene, leading to expression of higher steady-state levels of MTs, and silenced the β-centaurin-2 gene, leading to expression of higher steady levels of MFs, causing changes in cell shape/gene expression, in Tx cell lines. b) Ni+2 ions silenced the DRIP80 gene, which altered distributions of Ca+2 ions in Tx 10T- 1/2 cell lines. c) These Ni ion-induced events cumulatively caused differen-tial expression of 144 genes, contributing to induction/maintenance of Tx phenotypes in Ni+2 ion-and in MCA-Tx cell lines. Acknowledgements: Res. supported by grants to JRL from: grant R01 ES03341/NIEHS/NIH; funding from M. S. Program, Dept. Micro./Imm. USC; Provost’s Office USC; USC discret. funding; Cancer Ctr. Core Grant 5 P30 CA 09320 from U. S. NCI/NI.

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Genotoxicity and сarcinogenicity of engineered nanomaterials

Herman AutrupProfessor Emeritus, Institute of Public Health, University of Aarhus, Denmark

Nanomaterials are increasingly being used in consumer products

and in many medical applications, e.g., carbon nanotubes. Due to size of nanoparticles, similar to the ultrafine particles in ambient air pollution, and because of the similarities of some multiwall carbon nanotubes MWCNT) to asbestos and asbestos fibers, there is an increasing public health concern for potential carcinogenicity. IARC has recently evaluated a number of carbon nanotubes for their carcinogenicity. Mitsui MWNT-7, was classified as group 2A, due to its ability to induce mesotheliomas.

Testing for genotoxicity is one of the regulatory requirement before compounds can enter the market, and OECD guidelines have been established. A tiered approach using both in vitro and in vivo methods has been developed, but has not been validated for nanomaterials. Literature surveys indicate conflicting results when nanoparticles are tested in the traditional mutation/chromosomal damage assays. Effects are generally only observed at high concentrations and cytotoxicity. The Ames assay can generally not be used for non-soluble nanoparticles. However, the comparison of the published studies is hampered by the poor characterization of the nanomaterial used and by the diversity of cell types – both human and animal. Thus can we use the same methods for nano material that do not react directly with DNA, or should new methods be developed based upon new information on the molecular mechanism of carcinogenesis. One of the general mechanisms of NP toxicity is induction of reactive oxygen species (ROS) and subsequent DNA damage, which can be measured by the comet assay both in vivo and in vitro. DNA adducts induced by ROS has also been detected, e.g., bulky adducts detected by the P32 postlabelling techniques can be seen both in vitro and in vivo, e.g. adducts in brain tissues following intra-tracheal administration of carbon nanotubes.

Taking the large number of potential new nanoparticles and conjugated products, mechanistic based testing or QSAR based models should be considered. The Tox Tracker panel of reporter cell lines is interesting as it can measure both the induction of oxidative stress and p53- associated cellar response. Similarly, transcriptomics could be used to assess the induction.

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Enhanced metabolic ROS production by a loss of protein phosphorylation in mitochondria

Yoshimi HommaInstitute of Biomedical Sciences, Fukushima Medical University School of Medicine,

Fukushima, Japan

Mitochondrial protein phosphorylation is an important mechanism for the modulation of mitochondrial functions. We demonstrated that the phosphorylation of Ser-586 of very long-chain acyl-CoA dehydrogenase (VLCAD) by PKA is significantly reduced in fibroblasts derived from patients with idiopathic pulmonary fibrosis (IPF). Ectopic expression of the S586A mutant of VLCAD produced extremely high amounts of radical oxygen species (ROS) and induced cell death in various cells. On the other hand, some protein tyrosine kinase inhibitors promote ROS production. We identified NDUFV2 (Tyr-193), SDHA (Tyr-215), and FLOT-1 (Tyr-56 and Tyr-149) as c-Src substrates in oxidative phosphorylation systems. Studies with phosphorylation-defective mutants revealed that tyrosine phosphorylation of NDUFV2 is necessary for ATP production and cell survival, and that of SDHA is indispensable for efficient electron transfer and reduced ROS production without any effect on enzyme activity and cell survival. Phosphorylation of FLOT-1 is also important to suppress ROS generation from respiratory complex II. In order to evaluate mitochondrial metabolic ROS are actually involved in carcinogenesis, we have tried to produce Tg mice expressing one of these phosphorylation-defective mutants in B cells or neurons, and so far obtained those expressing the SDHA mutant in B cells. No leukemic change is observed in these Tg mice. Present results suggest that in addition to metabolic ROS, other gene products are required for the development of leukemia.

Retraction makes more retractions

Toshio Kuroki * and Akira Ukawa***Japan Society for the Promotion of Science (JSPS), Tokyo, Japan and ** RIKEN Advanced Institute for Computational Science, Kobe, Japan

Science community as well as the public have been showing great concern over scientific misconduct spurred by its sharp increase from 2000 and repeated media coverage. Retracted publications are an objective measure for gauging scientific misconduct. We approach theoretically the issue of repeated retractions by surveying three databases: the Web of Science (WoS),

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the PubMed and the blog “Retraction Watch (RW)” http://retractionwatch.com by Ivan Oransky and his colleagues.

In both ranking of retracting authors and distribution of retracted publications per authors, we found that a power law is applicable with exponents closed to those in natural and social events, suggesting that retraction is not a random process.

Analysis of retraction distribution suggested that a small fraction of retracting authors with 5 or more retractions, counting only 1-2 %, is responsible to around 10 % of the total retractions. Furthermore, we estimated probability of repeating retractions by retracting authors. Probability to repeat retraction for those with a single retraction is 3-5% in 5 years, while 26-37% for those with 5 or more retractions.

Phenomena under a power law are often called Pareto distribution or preferential attachment. One well known example is a phenomenon of “the rich get richer”: in the same way, one can say that “retraction makes more retractions”. To avoid further retraction, it is therefore important to identify authors even with a single retraction and to remind them research integrity by education.

According to the survey of PubMed by Fang et al (2012), 67.4% of retracted articles were attributable to misconduct, including fraud or suspected fraud (43.4%), duplicate publications (14.2%) and plagiarism (9.8%), while 21.3% were due to errors. Although retractions represent a minuscule percentage, 0.02% of total publications (Fanelli, 2013), a series of surveys demonstrated that a sizable fraction of retractions was attributable to a minor fraction of scientists who have repeatedly retracted their publications (Grieneisen et al, 2012; Steen et al, 2011; Fang et al, 2013).

Session III: Cancer Biology

Regulation of cancer metabolic reprogramming

Ping Gao University of Science & Technology of China

One of the emerging hallmarks of cancer has been the deregulated cellular metabolism, which is well beyond the Warburg Effect, or the aerobic glycolysis, as Otto Warburg described some 90 years ago. In this presentation, I will discuss our recent evidence regarding regulation of glucose,lipid and amino acid in cancer cells. In particular, I will discuss how these metabolic alterations may ultimately contribute to cancer cells’s survival and proliferation under stressed conditions.

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Role of microRNA in hormonal carcinogenesis

Lyudmila Gulyaeva The Institute of Molecular Biology and Biophysics, Novosibirsk, Russia

Novosibirsk State University, Novosibirsk, Russia

Epigenetic alterations affecting gene expression are increasingly being recognized for their roles in mechanisms of carcinogenesis. In recent years a number of studies have shown that disregulation of microRNAs (miRs) expression plays a fundamental role in cancer development. Our study confirmed that expression of several oncogenic miRs such as miR-21,-155, -221, -222 increases considerably in breast cancer whereas tumor suppressor miR-205 decreases. The in silico analysis has shown that oncogenic miRs can bind mRNAs of ERα, PR, CYP19 which are key markers of breast cancer. It is related to the fact that the studied miRs are involved in hormonal carcinogenesis. Moreover, miR expression profile can change under neoadjuvant radio- and chemotherapy. These results triggered our study aimed at understanding the reasons for the changes in miR profile. Using the in silico approach we have chosen 13 miRs: miR-21,-155, -221, -222, -205,-429 and a set of “novel” miRs: miR-326,-483, -3573, -126a, -1843a,-190a, -6327, potentially regulated by nuclear receptors CAR or AhR. We determined the expression levels of microRNAs by stem-loop RT followed by TaqMan PCR as well as by RT-PCR their “host” genes and target mRNAs involved in cell proliferation and differentiation in the liver, breast, ovaries and uterus of rat treated with DDT and benzo(a)pyrene once a week or within 3 months. We also studied the morphological changes in rat tissues under the long-term xenobiotic exposure. It was shown that the tissue-specific expression of studied miRs might be explained by the tissue-specific activation of CAR/AhR by xenobiotics. Thus, the results testify in favor of the role of microRNAs in carcinogenesis when a change in their expression is accompanied with affected expression of the target genes and emergence of morphological pathological changes.

This work was supported by Russian Science Foundation № 15-15-30012

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Genetic disorders in non-small cell lung cancer

Demidova I, Gikalo M, Barinov A, Gagarin I, Makhson A Moscow Oncological hospital 62, Moscow, Russia

Introduction and Aim of the studyIdentification of some specific genetic lesions, known as drivers of

neoplastic process, can improve survival in a subset of patients with non-small lung cancer (NSCLC), eligible for modern targeted therapies. Detection of mutations in EGFR gene and ALK rearrangements became standard of diagnostic, but some new genetic aberrations are promising as potential targets for new or known medications. Our study is aimed on revealing of rare genetic abnormalities: mutations of BRAF and MET genes, MET amplification, rearrangements of ROS1, RET, BRAF, NTRK and NRG genes.

Matherials and Methods Ninety six planned 200 FFPE samples of NSCLC patients, negative for

EGFR mutations and ALK translocations, were investigated to date. Mutations in 11 and 15 exons of BRAF gene and mutations/deletions in exon 14 of MET gene were investigated using PCR and fragment length analysis, amplifications and rearrangements were tested by FISH. Eight cases not tested for any genetic anomalies were investigated by targeted resequencing on Illumina MySeq V2.0 device using universal custom panel, including exons of interest in EGFR, KRAS, NRAS, BRAF, MET, PI3K, KIT genes.

Results At the first step, all 96 samples were investigated for KRAS ex2 mutations

by PCR and 16 cases, positive for KRAS were excluded from the study. In 80 “triple negative” cases (for EGFR, ALK and KRAS) we revealed two samples positive for BRAF mutations (one – V600E), one ROS1-rearranged and one MET-amplified samples. One deletion and one mutation in MET exon 14 were detected and proved by Sanger sequencing. One rearrangement of NTRK gene is under investigation for discovering of second partner now. Targeted resequencing of 8 cases find out one EGFR and one KRAS mutation, also proved by other methods.

Conclusion Using additional investigation, we used to reveal potentially targetable

aberrations in 7 patients (7,3% of total group). However, new approaches based on next generation sequencing are promising in simplifying and fastening of testing procedure.

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Cell signaling and the development of the resistant phenotype of tumor cells

Mikhail Krasil’nikovBlokhin Russian Cancer Research Center, Moscow, Russia

The main goal of this study is the analysis of the role of cell-cell interactions in the formation of the tumor cell resistance to hormonal drugs. The efficiency of endocrine therapy of tumors, including breast cancer, is limited by development of hormone-independent tumors which are resistant to antiestrogens initially or acquire resistance under prolonged therapy with antiestrogens (tamoxifen, raloxifene). The most frequent mechanism of hormonal resistance is based on the blockage of estrogen receptor signaling (estrogen ablation, antiestrogen treatment) and activation of hormone-independent mitogenic signaling, including receptor tyrosine kinases, cell cycle-regulating proteins, PI3K/Akt cascade etc.

Here we have hypothesized that the formation of the hormone resistance of tumors may be based, at least in part, on the transferring of the resistant phenotype from the resistant to hormone-sensitive cells – as a result of the secretion of the specific factors acting in the paracrine manner or via the direct cell-cell contacts.

Using the estrogen-dependent breast cancer cells MCF-7 and the resistant subline MCF-7/T, we investigated the possible changes in the hormonal sensitivity of these cells caused by the co-cultivation in vitro. To distinguish the cell cultures, the MCF-7/T cells were previously transfected with the plasmid containing the gene of the green fluorescent protein (GFP), and GFP-positive hormone-resistant subline MCF-7/T/ GFP+ was developed.

We found that the co-cultivation of the parent and resistant breast cancer cells leads to development of the tamoxifen resistance in the parent cells, whereas PI3K inhibitor wortmannin prevents the progression of the resistance. Furthermore, both the resistant MCF-7/T subline and new-generated resistant cells were characterized with the common features: partial loss of estrogen receptor and activation of PI3K/Akt/ Snail1 pathway.

Totally, we conclude that: • The acquired hormonal resistance of breast cancer cells may be

transferred, at least in a part, via horizontal connection – from cell to cell;

• The horizontal transfer of hormonal resistance is mediated via PI3K/Akt/ Snail1 signaling, indicating the significant role of this pathway in the development of hormone resistance.

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The secreted form of heat shock protein-90alpha (HSP90α) defines its tumorigenicity in breast cancer

Wei Li Department of Dermatology and the Norris Comprehensive Cancer Center, University of

Southern California Keck Medical Center, Los Angeles, CA 90033, USA

Secreted heat shock protein-90 (Hsp90) supports tumorigenesis and is potentially a new therapeutic target. However, there is a paucity of evidence demonstrating which Hsp90 isoform to target and where in the molecule to target for development of new anti-tumor agents. Here, we report that knockout of Hsp90β causes tumor cell death, whereas knockout of Hsp90α specifically nullifies tumorigenesis of the cells. Interestingly, the Hsp90α knockout-caused tumorigenic defects are corrected by supplementation with extracellular Hsp90α, but not Hsp90β, protein. Consistently, monoclonal antibodies, 1G6-D7 and 5C4-D4, targeting tumor cell–secreted Hsp90α eliminate invasiveness of the cells. Furthermore, we identify lys-270 and lys-277 in Hsp90α critical for its tumorigenic activity and sufficient to convert Hsp90β to Hsp90α-like protein to promote tumor cell invasion. These findings suggest that inhibitors targeting the dual lysine motif of tumor-secreted Hsp90α would be effective and safer, due to lack of interference with the intracellular chaperone function of Hsp90β.

Function of protein kinase CK2 on cell cycle progression

Miwako Kato Homma, Masato Ogura, Yoshimi Homma Department of Biomolecular Sciences, Fukushima Medical University School of Medicine,

Fukushima 960-1295, Japan

Protein kinase CK2 is a highly conserved serine/threonine kinase that functions in multiple cellular processes including cell growth and development. We have reported the cell-cycle dependent association of cellular CK2 with adenomatous polyposis coli (APC) protein that regulates CK2 activity, and have identified its downstream target as eukaryotic translation initiation factor 5 (eIF5). In this study, we demonstrate that CK2α is phosphorylated following stimulation of G0-arrested cells by growth signal and provide evidence for phosphorylation sites in that occurred in human normal fibroblasts. Results obtained from those phosphorylation site mutants support the involvement of CK2 phosphorylation for its nuclear localization and for its role in the nucleus that linked to the progression of the cell cycle. FRAP experiments on living

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stable cell lines expressing CK2α protein indicated that the α subunit was a highly mobile protein, although a passive cytoplasmic–nuclear translocation was not observed. Proteomics studies identified components of the CK2 immune complexes and elucidated downstream target molecules for CK2 by functional classification based on the Gene Ontology information. These data may provide the broad biochemical framework for understanding the function of CK2 in complex with cellular partners during the progression of cell cycle.

Advancing Oncology research with single-cell approaches

Amy HamiltonFluidigm Europe B.V.(Official partner of Helicon Company),

Senior Field Applications Specialist, EMEA

Being able to examine individual cells and their molecular contents stands to improve our capacity to unravel the complex and personalized genetics of human cancers. We will review several examples of how single-cell techniques can bring insight into different areas of cancer research, from whole transcriptome and whole genome sequencing to multi-parametric protein analysis.

Session IV: Cancer Diagnostics

Roles of a Cell Adhesion Molecule CADM1 in Apoptosis Induction, Cancer Invasion and Metastasis

Ito T, Tsuboi Y, Tsuchiya T, Oba M, Murakami Y.Division of Molecular Pathology, Institute of Medical Science, The University of Tokyo

Disruption of cell adhesion is a crucial step in invasion and metastasis of human cancer. An immunoglobulin superfamily cell adhesion molecule, CADM1, is involved in the formation of an epithelial cell structure. CADM1 is frequently inactivated in various cancers in advanced stages, suggesting its involvement in invasion and/or metastasis. This is supported by spontaneous development of lung tumors in Cadm1-/- mice and by development of more invasive lung tumors when crossed with mutant K-ras knock-in mice. We demonstrated

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that trans-homophilic interaction of CADM1 activated PI3K to reorganize actin cytoskeleton. On the other hand, when cell adhesion by CADM1 was disrupted, cells were induced to apoptosis, which was dependent on digestion of the cytoplasmic fragment of CADM1 by caspase-7 as was observed in dependence receptors. Essential roles of CADM1 in cell adhesion might be caused at least in part by its stable characters on the cell membrane demonstrated by FRAP analysis. In contrast, CADM1 is overexpressed in ATL or small cell lung cancer. Tissue- or cell context-dependent function of CADM1 in tumor progression will be discussed.

Mycoplasma hyorhinis infection conduces toward NFκB-dependent resistance of H292 lung cancer cells to Nutlin3

Uljana A. Boyarskih1, Mariia A. Smetanina1*, Dmitry A. Zhechev1, Vadim V. Kozlov2, Alexander E. Kel1,3, Maxim L. Filipenko1,4

1Laboratory of Pharmacogenomics, Institute of Chemical Biology and Fundamental Medicine, 8 Lavrentiev Avenue, Novosibirsk 630090, Russia

[email protected]; [email protected]; [email protected] department, Novosibirsk Regional Clinical Oncological Center, 2 Plakhotnogo Str.,

Novosibirsk 630108, Russia [email protected]

3Department of Research and Development, geneXplain GmbH, Am Exer 10b, Wolfenbüttel D-38302, Germany

[email protected] of natural sciences, Novosibirsk State University, 2 Pirogova Str., Novosibirsk

630090, Russia [email protected]

Background: Nutlin3 is an antitumor compound that selectively induces cell cycle arrest and death of tumor cells through p53 reactivation. We examined the effect of Mycoplasma hyorhinis on sensitivity of H292 lung cancer cells to Nutlin3 and a possible mechanism mediating this effect.

Methods: Determination of sensitivity of Н292 and Н292Myc.h cells to Nutlin-3 was performed using viability test with resazurin. Epithelial–mesenchymal transition (EMT) was assessed using a test for the capability of forming colonies in soft agar. Genome-wide transcriptional profiles of H292 and Н292Myc.h cell lines were determined using Human HT-12 v3 Expression BeadChips (Ilumina). Bioinformatics analysis was performed using R/Bioconductor software package integrated into geneXplain/BioUML platform. Mycoplasma species in human lung tumor DNAs was tested by real-time PCR.

Results: Mycoplasma infection reduces the cytotoxic effect of Nutlin3 on H292 cells (in cell culture infected with M. hyorhinis the percentage of viable cells after incubation with 30 µM Nutlin3 was significantly higher than

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in uninfected cell culture; 11.5 ± 5.2 vs. 59.5 ± 11.4; p<0.001). Additionally, epithelial-mesenchymal transition of cell culture was a concomitant effect of mycoplasma infection. Microarray-based transcriptome analysis of infected and uninfected cells revealed the activation of IL-1/NFκB signaling pathway in H292 cells infected with M. hyorhinis. We showed that up to 10% human lung tumor could be carrier of Mycoplasma species.

Conclusions: This finding suggests that activation of IL-1/NFκB signaling pathway by M. hyorhinis is the most likely mechanism of H292 cells resistance to Nutlin3. Mycoplasma infection may therefore bias treatment of lung cancer patients with Nutlin3 or other antitumor drugs. Presence of Mycoplasma species in human tumors could potentially lead to increased drug resistance during chemotherapy.

Next Generation Sequencing in Hereditary Cancer detection and monitoring

Kovalenko S.P,1,2., Anisimenko M.S.11Institute of Molecular Biology and Biophysics, Novosibirsk, Russia

2Moscow Institute of Physics and Technology, Russia

Breast cancer is the most frequent malignancy in women. 5-10% of breast cancer cases are connected with germ-line mutations, mainly in BRCA1 and BRCA2 genes. Disease-causing mutations in BRCA1 and BRCA2 genes have a very high penetrance, so detection of those mutations is widely used for the targeting of high cancer risk group of patients.

Meanwhile, the analysis of mutations by complete sequencing of coding regions of BRCA1 and BRCA2 genes is time-consuming and expensive procedure. We calculated the economical benefit of genetic testing for all breast cancer patients taking into consideration the difference in expenses for treatment of patients on early (1-2) and late (3-4) stages of breast cancer in Russia. Markov model was constructed for the analysis. The model demonstrated a very high economical benefit of genetic testing for all breast cancer patients with subsequent follow-up of mutation carriers if 2-3 founder mutations are analyzed. Nevertheless, an economic benefit disappears if complete sequencing of BRCA1/2 is applied.

Complete sequencing of BRCA1/2 genes can be performed by NGS. Simultaneous loading of 96 samples per machine run with relevant bar-coding can dramatically reduce the cost of sequencing. To reduce the cost even more we developed a special economic procedure for processing of patient’s DNA

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samples. The procedure presumes simple construction of specific sticky ends for each “service” DNA fragment which supposed to be connected with template DNA. We called this procedure LEGO-like connection.

We analyzed the most frequent mutations (BRCA1 5382insC, T300G, 4153delA, 185delAG, BRCA2 6173 and CHEK2 1100delC) by AS-PCR in 3543 samples of genomic DNA of non-selected breast cancer patients. 4,2% of all tested patients were mutation carriers. 350 samples of DNA samples of cancer patients were analyzed by targeted sequencing of BRCA1 gene on PGM IonTorrent machine, 3 mutations among 350 were found in addition to those detected earlier by AS-PCR. Meanwhile, NGS can be considered as economically effective tool for the analysis of hereditary cancer mutations by direct screening of all breast cancer patients. A possibility for monitoring of BRCA1 mutation carriers by NGS of liquid biopsy will be discussed.

Clinical and molecular-genetic approaches to diagnostics and treatment of thyroid cancer

Shevchenko Sergey Head and Neck Surgery Department, Novosibirsk Clinical Hospital No. 1,

Novosibirsk State University, Novosibirsk, Russia

The growing incidence of thyroid cancer in the world makes the diagnosis and treatment of this pathology an important issue. Currently new molecular markers to distinguish between benign and malignant thyroid tumors are being searched. One way is to investigate intra- and extracellular signaling pathways, the deregulation of which leads to thyroid cell transformation. The report examines the use of modern diagnostic methods, such as ultrasonography, sonoelatography, hormonal status, fine-needle aspiration biopsy, and computer-aided analysis, as well as up-to-date molecular and genetic methods in Novosibirsk Municipal Clinical Hospital No.1. Differential diagnostics of nodular thyroid neoplasms is one of the most acute problems in thyroidology. We determined somatic mutations in BRAF gene, expression and activity of integrins, angiogenic factors, GSTP and microRNA in benign and malignant tumors for their differential diagnostics. In this study 120 samples of benign nodular neoplasms and 112 samples of papillary thyroid cancer (PTC) were analyzed. The expression profile of ITGA3, ITGAV, ITGA6, ITGA9, ITGB1 integrin genes and their ligands OPNa, OPNb, TSP1 in papillary carcinoma with various status of BRAF V600E mutation was detected. The revealed changes in studied gene expression testify to their potential role in the tumor progression, as well as

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possible effect of mutant BRAF on gene expression levels. The gene expression and activity of the GSTP enzyme involved in hormone metabolism and c-Jun signaling pathways were different in PTC and benign nodular neoplasms. The results showed that GSTP can be used as cancer-specific marker for thyroid neoplasms with 83-88% sensitivity, 70-80% specificity and 76-84% diagnostic accuracy. Thus, the studied molecular markers could be used for differential diagnostics of benign and malignant thyroid neoplasms and the treatment approach.

MicroRNA and somatic mutations in the preoperative diagnostic of thyroid nodules

Nikolay Kolesnikov1, Sergei Titov1,2, Ekaterina Malakhina3, Tatyana Poloz4, Mikhail Ivanov 1,2 Sergei Shevchenko5, Larisa Achmerova1, Yulia Veryaskina1, Igor Zhimulev1.

1The Institute of Molecular and Cell Biology, SB RAS, Novosibirsk, Russia. 2JSC «Vector-Best», Koltsovo, Russia.

3Institute of Chemical Biology and Fundamental Medicine, SB RAS, Novosibirsk, Russia. 4Non-governmental Healthcare Institution

«Railroad Clinical Hospital on the Station Novosibirsk-Glavny», JSC Russian Railways, Novosibirsk, Russia.

5Novosibirsk Municipal Budgetary Healthcare Institution «Municipal Clinical Hospital №1», Novosibirsk, Russia.

Context: The major method used to preoperatively subtype and grade thyroid nodules is cytological analysis of material collected during ultrasound-guided fine-needle aspiration biopsy (FNAB). The performance of this method critically depends on pathologist’s qualification and experience; therefore, it is very vulnerable to errors. Moreover, up to 30% of FNA samples classified as indeterminate thyroid lesion. Identification of specific molecular markers such as microRNA and somatic mutations should help to increase the robustness and objectivity of nodules subtyping. Micro RNAs (miRNAs) can serve as such markers, since recent studies showed that expression of many of these is subject to profound changes in thyroid neoplasms.

Objective: The goal of our study was to develop a new approach to extract nucleic acids from the cytological samples (stained FNA smears) and perform microRNA profiling and detection of BRAF, RET/PTC mutations using RT-qPCR.

Setting: Preoperative FNA material from cytology stained smears (n=435) and surgical specimens (n=208) were evaluated for 9-13 miRNA expression classifier and testing BRAF, RET/PTC mutations. All cytological slides were subdivided on six groups according to the Bethesda classification. Molecular results were compared with cytological conclusion from the same analyzed preparations and with surgical histopathology diagnosis.

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Results: In FNA smears the high level of expression of miR-146b and mir-221 discriminate between the malignant and benign lesions and mostly are common for papillary carcinoma. Together with BRAF mutation it increases the accuracy of diagnostic classification. Mir-375 is a prominent marker for medullary thyroid carcinoma in cytological preparation on preoperative stage as well as in the case of postoperative specimen. Using our developed microRNA platform we determined 16 malignant lesions in 99 cases of samples with indeterminate characteristics (I, III and IV classes of Bethesda), including 5 BRAF mutations. Also miRNA expression analyses have shown the existence of 19 malignant lesions among 221 samples classified by the cytological evaluation as benign (class II, Bethesda).

Conclusions: The results confirm practical feasibility of usage of molecular markers for improving the diagnostic accuracy especially in the case of indeterminate thyroid nodules in preoperative diagnostic and for typing of thyroid neoplasm in surgical specimens. Hopefully this approach will become the important part of molecular cytology, clinical oncology in the nearest future.

Keywords: Thyroid nodules, FNAB, cytological analysis, microRNA, BRAF, RET/PTC, qRT-PCR.

This work was supported by fundamental scientific research on the project 0310-2015-0007.

Modern methods of mathematical modeling in the development of anticancer drugs

Yakovlev PavelBIOCAD, Russia

Modern anti-tumor drugs R&D mostly depend on high-throughput screening methods. In case of small molecules design the dominant way is fragment-based screening. This approach shows great results, but requires long and expensive post-screening analysis procedures like crystallography. Also screening libraries are limited by fragment diversity and reactions set that are often very simple.

Source library is often determines the success of antibody discovery process too. That is why considerable efforts are applied to the methods of creating new high-throughput display methods and construction such libraries from human donors, immunized animals or just random maturated genes.

This research trend leads to the fact that post-screening candidates often do not fit the target molecule profile by affinity and immunogenicity for

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antibodies or ADME(T) for small molecules. This candidates need significant tweaking, that can affect a number of molecule parameters including the specific activity, as the epitope can be moved. This become especially important with immune checkpoints inhibitors, as many of them need to be agonists. As a evident way of success probability magnification is libraries size increase, there is another way.

The alternative is structural-based computer-aided (or in silico) design of molecules. Modern methods of protein folding and small molecule conformational search, docking and molecular dynamics show a cheap and accurate way of leads optimization. Combined with effective gene synthesis platform or reactions design, mathematic methods may be used to create small and directed maturation libraries from lead candidates. In silico predictions also can save development budget and time by replacing in vitro (or «wet-lab») analyzes.

In this talk I will present the next step of drug discovery evolution. Our department works on novel methods of target representation for directed drug candidates selection in the space of non-immunogenic high-affinity proteins and high-selectivity ADME(T)-optimal small molecules, so I will introduce first promising results of this work.

Session V. Neuro-Oncology

Coexpression network analysis of glioma transcriptome

Dr. Marina G. SergeevaA.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia

The available evidence suggests that the lethality of glioblastoma is driven by small subpopulations of cells that self-renew and exhibit tumorigenicity. It remains unclear whether tumorigenicity exists as a static property of a few cells or as a dynamically acquired property.

We used tumor-sphere and xenograft formation as assays for tumorigenicity and examined subclones isolated from established and primary glioblastoma lines. Our results indicate that glioblastoma tumorigenicity is largely deterministic, yet the property can be acquired spontaneously at low frequencies. Further, these dynamic transitions are governed by epigenetic reprogramming through the lysine-specific demethylase 1 (LSD1). LSD depletion increases trimethylation of histone 3 lysine 4 at the avian myelocytomatosis

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viral oncogene homolog (MYC) locus, which elevates MYC expression. MYC, in turn, regulates oligodendrocyte

lineage transcription factor 2 (OLIG2), SRY (sex determining region Y)-box 2 (SOX2), and POU class 3 homeobox 2 (POU3F2), a core set of transcription factors required for reprogramming glioblastoma cells into stem-like states. Our model suggests epigenetic regulation of key transcription factors governs transitions between tumorigenic states and provides a framework for glioblastoma therapeutic development.

Developing rational medical therapies for von Hippel-Lindau disease: the dilemma of hemangioblastomas in the brain and spinal cord

Ian McCutcheon, MDUniversity of Texas M D Anderson Cancer Center, Houston, Texas USA

Von Hippel-Lindau disease (VHL) causes tumors of kidney, brain, and spinal cord, pancreas, adrenal glands, and endolymphatic sac, and retinal hemangiomas. Given the variety of tumor types within a single individual with VHL disease, treatment necessitates a personalized, multidisciplinary approach; and, given that the most frequent alterations in sporadic clear cell renal cell carcinoma involve loss of the 3p chromosomal arm including the VHL gene, treatment discoveries for this rare, heritable disease have implications for a much wider patient population. In VHL, hypoxia-inducible factors decrease leading to angiogenesis in hypervascular tumors. Although standard treatment for VHL is selective surgical removal of these neoplasms of whatever type, tyrosine kinase inhibitors used systemically affect renal tumors more than the hemangioblastomas seen in the brain and spinal cord. Through differential analysis of cell proteins in VHL-associated tumors, drugs have been chosen based on their angiogenic profile. We have completed two clinical trials of medical therapy for VHL and are soon to embark on a third. This talk will discuss the broader clinical issues and landscape of VHL, what medical strategies have been tested, and what the future holds for controlling the tumors associated with this disease, particularly those arising in the brain and spinal cord. Although no medical therapy has yet been identified that works in all patients with VHL, selective successes with some multi-kinase inhibitors in some patients offer proof-of-principle that drug therapy in VHL can achieve tumor control when designed intelligently.

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Radiosurgery for management of brain tumors

Mikhail F. ChernovFaculty of Advanced Techno-Surgery and Department of Neurosurgery

Tokyo Women`s Medical University, Tokyo, Japan

The idea of sterotactic radiosurgery as precise delivery of a single fraction of high-dose ionizing radiation to an imaging-defined target was originally developed by the Swedish neurosurgeon professor Lars Leksell. In 1951 he performed the first radiosurgical procedure to treat a patient with trigeminal neuralgia. The initial technique involved focusing multiple beams of ionizing radiation in such a way that all of the beams collided at the same point in three-dimensional space. Each individual beam contained a dose low enough to minimize the risk of damaging normal brain. At the point of intersection of all of the beams, however, a destructive dose of radiation was delivered.

The new technique was designated as Gamma Knife radiosurgery (GKR), implying the administration of high energy radiation with surgical precision. At present GKR has evolved into multidisciplinary field and utilized for treatment of patients with a wide variety of intracranial lesions, including benign and malignant brain tumors. Around 50.000 patients undergo GKR each year worldwide.

Contemporary Gamma Knife employs robotized computer-controlled device, which provides extremely high precision of irradiation within 0.1 mm in any coordinate direction. Conformal and selective dose planning is based on high-resolution MRI.

Modern treatment strategies permit to attend not only stabilization of the tumor growth, but shrinkage of the neoplasm, which in many cases accompanied by definite regress of the neurological symptoms. Sharp dose fall outside the treated volume provides minimal irradiation of the adjacent structures within the recommended safe range, thus treatment-associated complications are extremely rare and usually transient. In conclusion, modern Gamma Knife radiosurgery seems to be beneficial for patients with brain tumors and in many cases serves as a reasonable alternative to open microsurgical procedures.

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Computerized technologies for cerebral malignant glioma maximum safe resection and postoperative assessment

A.L. Krivoshapkin, А. S. Gaytan., G.S. Sergeev, V.V., N.A. Maslov, V.A. Mordvinov, Kanygin, V P. Kurbatov, L.E. Kalneus.

European Medical Center, Moscow, Novosibirsk State Medical University, Institute of Theoretical &Applied Mechanics, Institute of Cytology &Genetics, Meshalkin Institute of Circulation

Pathology, Novosibirsk

IntroductionNumerous clinical trials of the past decade have shown that the extent of

malignant glioma resection determination based on the results of postoperative contrast enhanced MRI data is of utmost importance. Nowadays, computerized technologies are used to achieve gross total resection. 5-ALA fluorescence-guided resection of malignant brain gliomas is one of those current trends which significantly increase the extent of tumor resection. The disadvantage of the technology is that the tumor fluorescence can best be seen in the dark which often makes resection difficult. We developed a new attachment computerized device for operative microscope for clear 5- ALA induced tumor visualization in the daylight. It should be noted that evaluation of tumor removal extent based on postoperative enhanced MRI is difficult as much as the residual malignant glioma interferes with haemostatic tissue, blood remnants and edema. New software for objective assessment of GBM resection based on mathematical analysis of postoperative MRI data was proposed.

MethodsThe device consists of the fluorescence excitation source (light-emitting

diode wavelength 400-410 nm), image receiving unit (set of portative HD cameras with filters), microcomputer with original data processing software and HD display. The fluorescence excitation source and image receiving unit are attached to the Carl Zeiss®Pentero 900 microscope and connected by cable to the control computer. Excitation source is used to expose the target with white light and fluorescence emission. The data is collected in the receiving unit and then processed and displayed on the monitor with the fluorescent area being marked contrast green. The new device was used to activate fluoresce of the Carl Zeiss® blue400 conventional target in the daylight. In addition to experiment with conventional target, ten patients with glioblastomas underwent microsurgical resection. 5-ALA was given to patients orally 3 hours before inductions of anesthesia in dose of 25 mg/kg body weight. Tumor samples and nearby tissue were obtained during surgery. The microscope attachment was used to detect both tumor samples and adjacent tissue fluorescence in the daylight. Five independent specialists (3 neurosurgeons and

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2 radiologists) examined those 10 patients postoperative MRI data and tested the program specificity and sensitivity for the tumor enhanced part (TEP). In 7 cases Surgicel® and in the other 3 - Tachocomb® was used.

ResultsThe developed device showed HD real-time video of the Carl Zeiss®

blue 400 fluorescent target in the daylight overlapped by digitally processed image of the fluorescent area. There was clear difference between malignant tumor and adjacent tissue in the daylight during all the ex vivo experiments with glioblastoma samples. In all cases software visualizes the volume of tumor remnants after subtotal and partial resection. 100% sensitivity and specificity of the program for Surgicel® were demonstrated. High resectability of results for TEP volume assessment on average 0,14 ± 0,02 (± m) ccm was shown. The program-assisted data evaluation for a patient took 2,21 ± 0,14 (± m) min only.

ConclusionThe developed device might be useful for detection of malignant brain

tumours fluorescence in the daylight. This device can also enhance the ability of microscopes not initially provided with fluorescent modules. Further research is required to assess efficiency and safety of the proposed device. The new software for objective assessment of GBM resection based upon mathematical analysis of postoperative MRI is accurate and easy to use for both daily neurosurgical practice and research in malignant glioma management.

Session VI. Breast Cancer

5-Fluorouracil and curcumin inhibit epithelial-mesenchymal transition in breast carcinogenesis

Gloria M. Calaf1, 2, Richard Ponce-Cusi1 and Marcela Gallardo1

1Instituto de Alta Investigación, Universidad de Tarapacá, Arica, Chile; and 2Center for Radiological Research, Columbia University Medical Center, New York

Breast cancer remains the major cause of global female mortality despite substantial progress in prevention, diagnosis and treatment. 5-Fluorouracil (5-FU) is a widely used anticancer drug, a heterocyclic aromatic organic compound with a structure similar to that of the pyrimidine molecules of nucleic acids that interferes with nucleoside metabolism. 5-FU is a chemotherapeutic agent for the treatment of a variety of solid cancers that arrest cell cycle and induce apoptosis in cancer cells.

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Curcumin is an antioxidant known as a dietary natural yellow pigment derived from the rhizome of the herb Curcuma longa. Curcumin has demonstrated antioxidant and antiproliferative properties in breast cancer. The aim of this study was to evaluate genes that could be targeted by 5FU and curcumin in a breast carcinogenesis in vitro model induced by radiation and estrogen. Evidence strongly implicate that Epithelial-Mesenchymal Transition (EMT) is involved in malignant progression inducing genes such as Axl, Twist and Slug.

Since such genes are aberrantly expressed in multiple tumor types and are known to favor the metastatic dissemination process the effect of 5FU and curcumin was evaluated in breast cancer cell lines by qRT-PCR. An established breast cancer in vitro model was used. It was developed with the normal immortalized breast epithelial cell line, MCF-10F that was exposed to low doses of high LET (linear energy transfer) alpha particles (150keV/um) of radiation, and cultured in presence of 17B-estradiol. This model consisted of the following cell lines: i) MCF-10F, ii) Alpha5, pre-tumorigenic, and iii) Tumor2, derived from Alpha5 injected in nude mice.

Curcumin effect in breast cancer cell lines was studied with 10 and 30uM for 48 hrs. The expression of genes implicated in EMT was evaluated by qRT-PCR. Results showed that 5 FU and curcumin decreased expression of genes related with EMT as Axl, Slug, Twist, E-Cadherin, considered epithelial markers and increased mesenchymal-related genes as N-Cadherin, Vimentin, Fibronectin, as well as those related with metastasis such as c-Ha-ras, Rho-A, p53, Caveolin 1 and others. MicroRNA expression was analyzed in breast cell lines by the same method.

Curcumin increased miR-34a expression, which in turn repressed several genes that are situated at the core of several signaling pathways known to mediate EMT. The migratory capacity was measured by wound healing assay and it was observed that 5FU and curcumin decreased the migratory and invasive capabilities. Thus, our studies show that 5 FU and curcumin may prevent or delay cancer progression and dissemination through its ability to disrupt EMT It can be concluded that 5FU and curcumin influenced biochemical changes associated with EMT, and among micro RNAs the miR-34a and its downstream genes Axl, Slug and Twist seem to promote such transition. Supported by FONDECYT 1120006 (GMC).

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Selective estrogen mimics for the treatment of tamoxifen-resistant breast cancer using PKCα

as a predictive biomarker

Debra A. Tonetti1*, Mary Ellen Molloy1, Huiping Zhao1, Thao N. Pham1, Jiong Zhao2, Rui Xiong2, Hitisha Patel2, Lauren Gutgesell2, Gregory R. J. Thatcher2

1 Department of Biopharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, 833 S. Wood St., Chicago, IL, USA

2 Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, 833 S. Wood St., Chicago, IL, USA

Endocrine-resistant breast cancer is a major clinical obstacle. Although recent clinical trials have demonstrated the efficacy of 17β-estradiol (E2) or diethylstilbestrol (DES) following exhaustive use of antiestrogens, E2 treatment is associated with major side effects such as an increased risk of uterine cancer, deterring the clinical community from adopting it as a treatment strategy. Our laboratory previously reported that protein kinase C alpha (PKCα) overexpression predicts tamoxifen (TAM) resistance in the clinic and may also predict a positive response to estrogenic therapy.

Ectopic overexpression of PKCα in T47D breast cancer cells results in a TAM-resistant, E2-inhibited phenotype in vivo accompanied by the translocation of estrogen receptor alpha (ERα) to extranuclear sites. We synthesized and characterized novel selective estrogen mimics (SEMs), with the goal of achieving the positive therapeutic effects of E2 treatment, while minimizing the side effects.

We identified two SEMs, BTC and TTC-352, which display partial estrogenic activity in breast cancer cell lines. Treatment with SEMs results in significant tumor regression in two xenograft models of TAM-resistant, PKCα-overexpressing breast cancer. Similar to E2, tumor regression is accompanied by translocation of ERα to extranuclear sites, possibly defining mechanism of action. SEMs are unable to support the growth of TAM-sensitive xenografts indicating safety and efficacy may be achieved in heterogeneous tumors. E2 and TAM both increase the risk of uterine cancer. Interestingly, SEMs do not increase uterine weight in mice suggesting reduced risk of uterine cancer. We are in the process of developing patient-derived xenograft (PDX) models to further validate our findings.

These data suggest that further development of SEMs targeted to TAM-resistant breast cancer is a feasible therapeutic strategy.

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Heparanase: from basic cancer research to therapeutic applications

Israel VlodavskyCancer and Vascular Biology Research Center, Rappaport Faculty of Medicine, Technion,

Haifa 31096, Israel

Heparan sulfate proteoglycans (HSPGs) are primary components at the interface between virtually every eukaryotic cell and its extracellular matrix (ECM). HSPGs not only provide a storage depot for heparin-binding molecules (i.e., growth factors, chemokines, enzymes) in the tumor microenvironment, but also decisively regulate their accessibility, function and mode of action. As such, HSPGs are intimately involved in modulating cell invasion and signaling loops that are critical for tumor growth. Heparanase, the sole heparan sulfate degrading endoglycosidase, regulates multiple biological activities that enhance tumor growth, angiogenesis and metastasis. Much of the impact of heparanase on tumor progression is related to its function in mediating tumor-host crosstalk, priming the tumor microenvironment to better support tumor take and growth. Immunohistochemistry, in situ hybridization, qPCR and western blotting demonstrate that heparanase expression is enhanced in almost all cancers examined including various carcinomas, sarcomas and hematological malignancies. It has been demonstrated that heparanase stimulates gene expression, autophagy, exosome formation, inflammatory responses and signal transduction via enzymatic and non-enzymatic activities that dynamically impact multiple regulatory pathways that together drive tumor survival, growth, dissemination and drug resistance. Numerous clinical association studies have consistently demonstrated that upregulated heparanase expression correlates with increased tumor size, tumor angiogenesis, enhanced metastasis and poor prognosis. Knockdown of heparanase expression or treatments of tumor bearing mice with compounds that inhibit heparanase enzyme activity markedly inhibit tumor progression further underscoring the potential of anti-heparanase therapy for multiple types of cancer. Importantly, there is only a single, enzymatically active form of heparanase in humans, it is expressed in very low levels in normal tissues and heparanase knock-out animals exhibit no obvious deficits. These characteristics imply that inhibition of heparanase will cause minimal side effects in cancer patients, together stirring heparanase as a highly desirable and druggable target for anti-cancer therapy. Development of heparanase inhibitors focused on carbohydrate-based, heparin-like compounds of which four are being evaluated in clinical trials for various types of cancer, including myeloma, pancreatic carcinoma and hepatocellular carcinoma. Heparanase neutralizing monoclonal antibodies were recently

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found to inhibit myeloma and lymphoma tumor growth and dissemination in preclinical models. Heparanase-inhibiting small molecules are being developed based on the recently resolved crystal structure of the heparanase protein.

Collectively, the overarching premise guiding our work is that heparanase is a master regulator of the aggressive phenotype of cancer, an important contributor to the poor outcome of cancer patients and a prime target for therapy.

Session VII. Viral Chemotherapy and Cancer Chemotherapy

Potential drugs to control hazardous viruses including some involvedin human malignancies

Eliezer Huberman Novadrug, LLC and University of Illinois at Chicago, USA

Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) are two RNA viruses implicated in human cancer causation. HCV causes liver tumors as an outcome of viral-induced chronic tissue damage while HIV triggers malignancies as a result of viral-evoked immunodeficiency. A substantial number of people worldwide are infected with these viruses. There is therefore an ongoing effort to develop drugs to control these infections since an early intervention can impede malignancies evoked by these viruses.

By means of a bacterial construct and appropriate mammalian cell systems, we identified three distinct classes of small molecules with anti-HCV activity. At low doses, these drugs reduced HCV load in infected cells with little to no cytotoxicity. Compounds of these classes also impacted other unrelated viruses. They inhibited the replication of HIV in infected human peripheral blood monocytic cells (PBMC) stimulated to grow by CD3/CD28 and Il-2, corona virus in infected human MRC-5 fibroblasts and Ebola virus in infected monkey Vero cells. In spite of this broad spectrum activity, NovaDrug’s compounds display virus specificity because they fail to inhibit the replication of other RNA viruses such as Coxsackie or influenza virus.

NovaDrug’s effective compounds do not display genotoxicity in the mouse micronuclei test nor in the Ames or cell culture mutagenesis assays. At antiviral doses, these compounds are within a safe dose range in the hERG assay. The hERG gene product, a potassium ion channel, contributes to the heart’s electrical activity that coordinates its beating. Inhibition of this channel can result in sudden death.

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To identify the mode of action of NovaDrug’s compounds, we used the HIV/PBMC model because it is well characterized and has useful tools for mechanistic studies. To this end, we examined the ability of a group of compounds to affect different steps of the HIV life cycle including virus cell entry, reverse transcription, genomic integration, translation and viral release. The results indicated that that the compounds operate after virus entry but prior to reverse transcription.

A comparative structure analysis of a subgroup suggested that it may affect a cellular protein kinase, which we termed PK. This subgroup inhibited the catalytic activity of the PK. Moreover, inhibiting its expression by specific siRNAs restrained HCV replication, thus suggesting that the subgroup blocks HCV replication by inhibiting PK activity. Based on its cellular function, this kinase can be involved an early stage of the virus life cycle. Unlike the subgroup’s compounds, the other ones do not appear to be protein kinase inhibitors because they failed to inhibit the catalytic activity of 359 currently known wild type protein kinases. Also, all three classes did not consistently affect the production of 30 known cytokines. Studies are ongoing to pinpoint the relevant targets of NovaDrug’s antivirals.

Currently, there are a limited number of clinical drugs with a wide range of antiviral specificity. NovaDrug’s compounds display such a range since they affect unrelated viruses including HCV and HIV as well as corona and Ebola viruses.

Modern surgical oncology in general and visceral surgery (e.g. in the treatment of colon and rectal cancer)

Karl Heinrich Link, M. D., Ph. D.Professor Dr. Medicine and Director of the Surgical Center and Asklepios Tumor Center Rhein/

Main Asklepios Paulinen Klinik (APK), Wiesbaden, Germany

Modern Developments of the Ulm University Surgical Oncological Group FOGT in Colon- and Rectal Cancer(C+RC) (Head: K.H.L.) are presented.

1. Successful public measures for Secondary Prevention of C+RC (“Carcinogenesis”).

2. Achievements in Multimodal Therapy with 3 Phase III -Trials involving more than 1800 C+RCPts and 65 Hospitals (“5-FU and Modulation”).

3. Inaugural establishment of Molecular Prognostic Factors for Individualization of Multimodal Treatment of C+RC Primary Tumors (“Translational Research”).

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4. Inaugural proof that Treatment Individualization involving Molecular Biology + In Vitro Chemosensitivity Testing significantly improve response and survival in sensitive vs. resistant patients with isolated C+RC Liver Metastases (CRLM) when drugs were individually selected in a prospective decision aiding trial (“Individualization”).

5. First large scale series to report benefit of 4. in “Downstaging and Resection of previously nonresectable CRLM”; one of two groups with large patient numbers to publish “Downstaging-Resection”. First report on Split Time Liver Resection. Help to establish Cytoreductive Surgery+Hyperthermic Abdominal Perfusion (CRS/HIPEC) for treatment of Peritoneal Carcinosis of C+RC in Germany/Europe (“Expand limits of surgery”).

6. Representation of various aspects of C+RC in National/International Societies, -Meetings, -Working Groups.

1.-5. were inspired in part by Charles Heidelberger in 1980, achieved in mutual cooperation with USC CRL (P.V. and K.Danenberg: “Prognostic Factors and Treatment Individualization”. J.Landolph: “Continuous Translational Communication between Basic Research – Surgical Oncology”). Results were continuously presented on the most prestigious meetings and published in highest rated journals. 4. was elected as “Best Paper of the Year” in “Cancer”, 2000. Thanks to Charles Heidelberger as mentor at my time USC-CRL and the subsequent cooperations with his former laboratory coworkers (P.V. and K.Danenberg)!

The role of predictive biomarkers in the treatment of advanced colorectal cancer

Marko KornmannDepartment of General and Visceral Surgery, University of Ulm, Germany

Multimodal treatment of advanced colorectal cancer has dramatically improved within the last decades. In parallel, several subtypes of colon and rectal cancer have been characterized based on their molecular profile. In order to more effectively navigate treatment of these patients, individual tumor tailored approaches need to be established. In this review an update about the influence of thymidylate synthase (TS), epidermal growth factor receptor (EGFR) and the Ras/Raf mitogen-activated kinase pathways on the outcome of treatment of metastasized colon and rectal cancer are presented. High TS expression is associated with poor response to fluoropyrimidine monotherapy. Patients with colorectal liver metastases and high intratumoral TS receiving

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5-FU alone displayed response rates below 20%, whereas 5-FU in combination with irinotecan (CPT-11) achieved a response greater 50% (53%). In contrast, in patients with low TS response rates were comparable (41% vs. 46%). Selection of KRAS, NRAS, HRAS, and BRAF wild-type tumors in initially unresectable metastasized disease followed by intensive treatment (5-FU, oxaliplatin, and irinotecan) in combination with EGFR inhibition resulted in tumor response allowing R0 surgical resections in 35% of the patients. Individual molecular tumor profiling is going to direct multimodal treatment of colon and rectal cancer in the future resulting in a reduction of treatment induced toxicity and an increase in response and survival.

The tumor microenvironment as a promising target to anti-tumor therapy

1,2Cherdyntseva N., 1,2Litviakov N., 1,2Denisov E., 1,3Zavyalova M., 1Stakheyeva M., 2Kzhyshkowska J., 1,3Perelmuter V.

1 Tomsk Cancer Research Institute, Tomsk, Russian Federation2 Laboratory for translational cellular and molecular biomedicine, Tomsk State University, Tomsk,

Russian Federation3Siberian State Medical University, Tomsk, Russian Federation

Only 30% of cancer patients, on average, benefit from cytotoxic chemotherapy. Even in cases of complete clinical response the tumor progression is event of high level expectation. The main reasons for tumor progression are: intratumor heterogeneity resulted from clonal evolution, drug resistance, tumor-promoting microenvironment. Risk of metastasis formation is provided by both tumor cell, biological characteristics and the microenvironment features within the primary tumor along with local and systemic conditions for metastatic niche formation. Reprogramming of microenvironmental stromal-inflammatory components, as expected, could allow tumor phenotype reversion (Rosenfeld, 2013). L.Zitvogel et al. anticipate that the comprehension of the mechanisms governing the immunogenicity of cell death will have a profound impact on the design of anticancer therapies. It is necessary to take into account the phenomenon of intra-tumor heterogeneity and inflammatory infiltration heterogeneity in order to improve the strategy for tumor-associated immune biomarkers and therapeutic agents. The heterogeneity is responsible for the patient-specific tumor-stroma interactions and formation of individual tumor microenvironment. It is important to perform systematic and multicomponent analysis of the biological properties of tumor cell affecting their metastatic potentials as well as the analysis of

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the specific local (tumor microenvironment) and systemic conditions, including immune system, responsible for mobilization of niche-forming component and supporting the metastasis in cancer patients.

Identification of microenvironmental factors and mechanisms promoting tumor dissemination is necessary to identify new prognostic markers to predict metastatic process, and optimize anti-metastatic therapeutic strategy, as well as new targets to create anticancer drug.

The work was supported by Russian Science Foundation grant 14-15-00318.

Development of novel oral Hedgehog signaling pathway inhibitor for cancer therapy

Sergey LeonovMD, PhD, Chief Science Officer, Life Science Center, Head of Laboratory of Innovative Medicine,

Moscow Institute of Physics & Technology

An alteration in the Hedgehog (Hh) pathway, either by misexpression of components of that pathway or by changes in the expression of other cellular components that interfere with the Hh signaling system, may trigger the development of several types of cancer. Hh plays a central role in the proliferative control and differentiation of both embryonic stem cells and adult stem cells. The theory suggests that conventional chemotherapies kill differentiated or differentiating cells, which form the bulk of the tumor but are unable to generate new cells. A population of cancer stem cells (CSCs) remains untouched and contributes to cancer cell invasion, relapse of the disease, metastasis and drug resistance. Development of specific therapies targeted at cancer stem cells holds hope for improvement of survival and quality of life of cancer patients, especially for sufferers of metastatic disease, where little progress has been made in recent years. The cellular signal crosstalk and deregulation of multiple cellular signaling pathways including Hh play critical role in self-renewal of CSCs, sphere formation, epithelial-to-mesenchymal transition (EMT) and tumorigenesis in pancreatic cancer (PC). Several previous studies demonstrated that Sonic hedgehog expression was detected in 70% of PC tissues and up-regulation of sonic hedgehog is observed in pancreatic CSCs, whereas down-regulation of Hh signaling by several inhibitors of Hh results in the inhibition of pancreatic CSCs and EMT-type cell growth. The Hh pathway is clinically validated target with GDC-0449 (Erivedge) as the first and only FDA-approved (Jan 2012) therapy for advanced and metastatic forms of

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basal cell carcinoma. Therefore, targeting this important signaling pathway could eliminate pancreatic CSCs and EMT-type cells.

The goal of current work was to develop an innovative orally available new drug that targets Hedgehog signaling pathway with pancreatic cancer as primary indication for mono- or complex therapy. Total 8 series have been identified and patented during the course of the project. Robust SAR (3 logs potency range, approximately 1000 analogs synthesized) resulted in identification of Preclinical candidate CD-005-0080-01 with high specific (against Ambit 96 Kinases panel and MDS LeadProfiling screen 68 receptors panel) activity (IC50 (alkaline phosphatase assay)= 1.5 nM) and low cytotoxicity (TC50 (AlamarBlue assay)=>0,1 mM) m in vitro. In vivo (AsPC-1 cells as a xenograft model of human pancreatic cancer) CD-005-0080-01 treatment 100 mg/kg PO twice daily for 3 weeks not only inhibited tumor growth (comparable to Gemcitabine: 100 mg/kg IP twice weekly for 3 weeks) but also significantly reduced mGli1 level without body weight loss, while body weight loss observed by the end of treatment in Gemcitabine received group. ADME studies of CD-005-0080-01 revealed high oral bioavailability (60-100%) in mice and rats, dose-dependent exposure increasing in rats at doses up to 30 mg/kg and favorable PK parmeters (T1/2 =3 h (mice) and 7 h (rats)). In concentrations up to 10 uM CD-005-0080-01 didn’t inhibit any CYP`s isoforms, being metabolized predominantly by CYP3A4 in human liver microsomes. High permeability and absence of efflux in Caco-2 cells suggested that CD-005-0080-01 is not a Pgp substrate. Safety studies of CD-005-0080-01 demonstrated no mortality in mice and rats after single oral dose 5 g/kg in acute toxicity, being not mutagenic in Ames test and chromosomal abberation (at doses up to 500 mg/kg) assay. CD-005-0080-01 was not immunotoxic with no signs of allerginicity. Chronic toxicity studies of CD-005-0080-01 in rats and dogs after 3 months of administration verified: no clinical signs of toxicity with no affection of respiratory and cardiovascular systems in rats, slightly increasing heart rate and decreasing R-R interval in dogs, with some changes in hematology and biochemistry of blood that were not confirmed by results of histopathology analysis. In conclusion, the CD-005-0080-01 can be an excellent orally available, highly selective, safe and efficient candidate for further clinical studies.

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Тезисы постерных докладов

Somatic mosaic variations in healthy skin fibroblasts

Alexej Abyzov1, Livia Tomasini2, Bo Zhou3, Nikolaos Vasmatzis1, Jessica Mariani2, Mariangela Amenduni2, Anahita Amiri2, Alexander E Urban3, Flora M Vaccarino2

1Mayo Clinic, Department of Health Sciences Research, Rochester, MN 55905, 2Yale University, Child Study Center, 3Stanford University, Departments of Psychiatry and

Genetics, Palo Alto, CA, 94305

Multiple studies have been performed on the analysis of somatic genomic alterations in cancer, but only a few have been conducted to understand natural somatic mosaicism, that is post-zygotic accumulation of mutations in cells of the human body. Fundamental knowledge about somatic mosaicism is not only crucial for finding determinants of cancer development and progression, but also for an understanding of various diseases and aging. We have compared genomes of 32 clonally derived human induced pluripotent stem cell (hiPSC) lines to the genomes of 11 (5 children and 6 adults) primary skin fibroblast samples, parental to the hiPSC lines. The clonal nature of hiPSC lines allows the discovery of somatic genomic variants present in the founder cell, but not in all fibroblast cells, thereby providing a mean for a high-resolution (and not compromised by amplification) analysis of single cell genomes. Adjusted for discovery sensitivity we estimated that on average, an iPSC line/a single fibroblast cell in children has 1,035 single nucleotide variants (SNVs) of which 181 could be directly confirmed as somatic by an in-depth re-analysis of fibroblast bulk genomic data with ultra-deep sequencing and digital droplet PCR, down to an allele frequency of 0.02%. Progressive increase in re-analysis’ sensitivity confirmed additional SNVs as somatic, suggesting that the estimated numbers are true counts of somatic SNVs per fibroblast cell. Similar analyses in adults revealed on average ~30% increase in the count of SNVs per cell (counts ranged was 900 to 2,000) as compared to children, suggesting that a large fraction of somatic SNVs in human fibroblasts occurs during prenatal and early childhood development. Except for a few SNVs occurring at neighboring nucleotides (likely results of exposure to UV light) SNVs were distributed randomly across the genome and their mutation spectrum was an almost perfect match to a previously uncharacterized mutation signature observed in cancers (Alexandrov et al., Nature, 2013; Lawrence et al., Nature, 2013). We, thus, propose that this cancer signature reflects normal development. Finally, in four children, allele frequency distribution for somatic SNVs had distinct narrow peaks, which, we hypothesize, are either the results of cell clonal

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selection or bursts of mutations during development. These new discoveries reveal a large degree of somatic mosaicism existing in healthy human tissues, link the mosaicism with development, and explain a mutational signature observed in cancers.

Sensing of small molecules using graphene oxide

O.A. Aftenieva*, Yu.V. Stebunov, A.V. ArseninMoscow Institute of Physics and Technology, Laboratory of Nanooptics and

Plasmonics, Dolgoprudny, Russian Federation

Surface plasmon resonance (SPR) spectroscopy has been approved as a precise and reliable method for investigation of biochemical interactions in pharmacological research. However nowadays, most of developing drugs are based on compounds with low molecular weight and already existed methods of SPR sensing could not provide sufficient sensitivity to detect interactions of small molecules.

Recently, we suggested a new type of linking layer for SPR biosensing based on graphene oxide (GO), that increases sensitivity and enables small molecules investigation. Overall, numerous biosensing applications of graphene and grapheme oxide (GO) have been demonstrated during the last years. These nanomaterials possess unique physical and chemical properties including high surface area, possibility of pi-stacking interaction with wide range of biological objects, rich availability ofoxygen-containing functional groups in GO, and excellent optical properties, which make it an ideal candidate for use as a universal immobilization platform in SPR biosensing.

Here, we propose a new SPR biosensing assay for sensitive and selective detection of low-molecular- weight compaunds. This assay exploits GO linking layers deposited on gold surfaces of SPR sensor chips and aptamers as a recognition element. Aptamers are short artificial oligonucleotides showing a high affinity toward the desired targets and similar in function to antibodies and enzymes with several advantages including small size, thermal and chemical stability, and low cost. These properties are useful for a multitude of applications, especially for development of molecular recognition elements. For instance, biosensor application of aptamers offers the possibility for fast and easy detection of environmental relevant substances or determining the drugs efficacy and toxicity at the preclinical stage.

In our experiments, aptamers with terminal amino groups and biotin-TEGs having high binding affinity to adenosine triphosphate (ATP) were

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immobilized on the surface of SPR chips via carboxyl groups of GO layer using carbodiimide crosslinking chemistry and biotin-streptavidin site-specific interactions respectively.

This procedure performed in the flow cell of SPR instrument led to the modification of carboxyl groups to N-Hydroxysuccinimide esters. Analysis of the changes in SPR signal during the activation of carboxyl groups demonstrated that GO linking layers have 6 times more activated carboxyl groups comparing to the commercial sensor chips based on carboxymethylated dextran. After immobilization, injections of an ATP solutions produced different signals, depending on aptamers immobilisation techniques: using amino-modified aptamers led to high level of non-specific binding, while with biotin-TEG aptamers shows the maximum level of response without non-specifity.

In conclusion, using such techniques we developed the SPR biosensing assay for highly sensitive detection of low-molecular- weight ligands using aptamers as a selective recognition layer immobilized on the surface of graphene oxide linking layer.

Copy number characteristics of the 10q26.3 chromosome region containing the MGMT gene in glioblastoma

E.Alekseeva1, D.Zaletayev1,2,5, E.Prozorenko3, A.Zaytsev4, A.Tanas1,5, O.Kirsanova4, V.Strelnikov1,2,5

1Research Centre for Medical Genetics, Moscow, Russian Federation. 2I.M. Sechenov First Moscow State Medical University, Moscow, Russian Federation.

3N.N. Blokhin Russian Research Center for Oncology, Moscow, Russian Federation. 4P.A. Hertsen Moscow Oncology Research Institute, Moscow, Russian Federation.

5N.I. Pirogov Russian National Research Medical University, Moscow, Russian Federation.

Inactivation of the MGMT gene located at 10q26.3 predicts glioblastoma’s sensitivity to alkylating agents. Methylation of the MGMT gene promoter is considered to be the key mechanism for this gene silencing and the marker of a favorable response to alkylating drugs. Deletion can present the alternative mechanism of MGMT gene inactivation. Previously we were the first to conduct a targeted analysis of loss of heterozygosity (LOH) at 10q26.3 and we showed LOH at the MGMT region in 63.2% (74/117) glioblastoma samples. However, LOH merely reflects allelic imbalance in the area without detailed information on the gene copy number. In order to assess copy number alterations at the 10q26.3 region in glioblastoma samples with identified LOH, we have developed a system for quantitative microsatellite analysis (QuMA). QuMA is based on amplification of microsatellite loci that contain (CA)n repeats where

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the repeat itself is the target for hybridization by the fluorescently labeled probe. The reference pool contains primer pairs for six genomic regions located on different chromosomes in which copy number violations are not typical for glioblastoma. In 51.5% (34/66) of the samples only one copy of the tested locus was found (deletion), while in 48.5% (34/66) two copies were detected (acquired uniparental disomy, aUPD). Thus, we have shown that MGMT LOH in glioblastoma can reflect either a deletion or an aUPD. The deletion of MGMT gene requires detailed study as a potential marker of glioblastoma sensitivity to alkylating agents.

Biopharmaceutical study of oral Ibuprofen complex polymer matrixgel with modified release

I.V. Antipova1, Е.О. Bakhrushina1, М.N. Anurova1, G.О. Nifontova1,2

1 I.M. Sechenov First Moscow State Medical University, Moscow, Russian2 MIPT Life Sciences Centre, Laboratory of drug formulation, Dolgoprudny, Russia

Investigation of innovative modified-release dosage form is one of the main tasks for pharmaceutical technology. Oral gels have high bioavailability in comparison with solid dosage forms, what is explained by dissolving of active pharmaceutical ingredient. The objective of this study was to develop oral modified release Ibuprofen gel.

The gel matrix was obtained by codissolution of composite polymeric carrier (CPC, interpolymer complex of polymethacrylic acid and polyethyl glycol) and hydroxypropyl methylcellulose Benecel K 100 M (Ashand). To achieve aggregative stability and improve viscosity modification polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer - Soluplus® (BASF) was added in two modifications: as native powder and as product of hot melt extrusion with Ibuprofen. Rheological measurements were performed using rotational viscometer. Dissolution profiles were evaluated accordingly USP using basket apparatus at pH 1,2 or 6,8. Ibuprofen quantative assay was performed by UV spectrophotometry at 220 nm.

As a result of the investigation the optimal gel matrix composition in three modifications: containing Ibuprofen (Comp.1), Ibuprofen and Soluplus (Comp.2), extrudate of Soluplus and Ibuprofen (Comp.3) was developed. Comp.2 and Comp.3 have thixotropic properties, Comp.1 was destroyed after yield value. The dissolution test showed the most sustained release of Ibuprofen from extrudate at pH 6,8 (71±8% per 5 hour). Ibuprofen release at pH 1,2 was smaller than 10% because of composite polymeric carrier

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pH-depended dissolving. Thus Ibuprofen and Soluplus extrudate application in oral CPC and Benecel gels improves rheological properties and allows to obtain Ibuprofen modified release.

Frequent mutations in renal angiomyolipoma

Kirill I. Anoshkin1,2, E.B. Kuznetsova1,3, K.M. Mosyakova3, A.S. Tanas1,2, M.S. Chaplygina1 E.A. Alekseeva1, E.V. Shpot3, D.V. Zaletayev1,2,3, A.Z. Vinarov3, V.V. Strelnikov1,2

1Research Centre of Medical Genetics of the Russian Academy of Medical Sciences, Moscow2Pirogov Russian National Research Medical University, Moscow

3I.M. Sechenov First Moscow State Medical University, Moscow

Kidney damage in tuberous sclerosis (TS) is the third most common symptom, occurring in about 85% of patients. The most common kidney injury is angiomyolipoma (AML), which is detected in approximately 70% of patients with kidney disease in TS. Mutation profiling of AML is traditionally limited to two genes, TSC1 and TSC2, because of association with TS. The aim of this study was to investigate mutations with a wide panel of cancer related genes, in blood and surgical material of patients with AML.

By using new genomic sequencing method on the Ion Torrent PGM, we have studied 7 of 35 patients with renal angiomyolipoma diagnosis.

We have identified mutations in a total 45 of 409 sequenced genes. Most frequently mutations were detected in the TSC2, ROS1, LAMP1, RAD50.

Mutations in ROS1 (c. A176G: p.Q59R, c.G2677T: p.G893C, c.G2187A: p.W729*, c.C1958T: p.S729F) and in TSC2 (c. C1111T:pQ371X, c.С3275T:pP1092L, c.C320A: p. A107D, c.5260-15C> T) genes are mostly pathogenic in agreement with bioinformatic analysis, and affect PIK3/AKT/mTOR signaling pathway.

Less common mutations were in LAMP1 and RAD50 genes (c.A389G: p.N130S, c.G118A: p.A40TX, and c.T2840C:p.I947T,c.G943T:p.V315L respectively), where mutations in RAD50 gene may affect the pathway of initiation of repair processes of double strand DNA breaks. Mutations in LAMP1 gene may affect the normal function of autophagy, similarly to the violation in mTORC1 pathways. Seven of the listed mutations have never previously been reported.

It was also noticed that six examined patients were carriers of the gene polymorphism (rs1058172) in cytochrome p450 CYP2D6. CYP2D6 protein is involved in the metabolism of 25% of all drugs metabolizable by cytochromes. This observation draws attention to the role of this protein in the etiopathogenesis of AML, and also requires to consider the genetic status of CYP2D6 in drug therapy.

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In conclusion, further search and characterization of mutations seems to be promising for the development of effective AML therapy, including targeted therapy, and will contribute to the prevention of disease progression by allowing its early detection.

Content of metalloproteinases in serum of bone sarcoma patients

Babkina I.V., Chernomaz I.S., Bondarev A.V., Schupak M.Yu., Soloviev Yu.N.N.N. Blokhin Russian Cancer Research Center, Moscow

Background: bone sarcomas belong to malignant tumors due to development hematogenous metastasis. Tumors markers expression study is impotent to detection new potential chemotherapy targets and evaluation disease prognosis.

Aim of the study: a comparative study of the content of metalloproteinases - MMP-2, -7, -9 and tissue inhibitor TIMP-1 in the serum of patients with primary malignant bone tumors and healthy people to identify relationship with the tumor histological structure and the disease prognosis.

Materials and methods: a comparative study of the content of metalloproteinases-2, -7, -9 and their inhibitor TIMP-1were performed in the serum of 54 patients with primary bone tumors (malignant - 45: central osteosarcoma 21, periosteal osteosarcoma - 4, Ewing’s sarcoma - 11, primary chondrosarcoma - 6, undifferentiated pleomorphic sarcoma - 3 and borderline giant cell tumors - 9 and healthy individuals - 26 was conducted using «Biosource» USA for TIMP-1 and «RαD» USA for MMP-2, -7 and -9.

Results: TIMP-1 levels in the serum of patients with central osteosarcoma and periosteal osteosarcoma was significantly higher, then in the serum of healthy persons. A direct correlation of TIMP-1 and MMP-9 was evaluated in the serum of patients with central osteosarcoma, periosteal sarcoma and Ewing’s sarcoma. Significant differences in 5 year survival rate and levels of serum TIMP-1, MMP-2, -7, -9 were not evaluated. But, 5-year survival rate with the level of MMP-2 >160 ng/ml in the serum was 1.6 times higher, then if the levels of MMP-2 were lower. If the levels of ММP-9 were <377 ng/ml – the 5-year survival rate was 1.4 times higher, then if ММP-9 was >377 ng/ml. Minimal 5-year survival rates (33%) were evaluated in the serum of patients with ММP-2 <160 ng/ml и ММP-9 >377 ng/ml.

Conclusion: the expression of MMP-2, MMP-9 and TIMP-1 can be linked to pathogenetic changes associated with the growth and metastatic sarcomas of bone and, in particular, osteosarcoma, and may be the subject of further studies to determine the levels of these indicators and their value in prognosis.

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Quisinostat Oral HDAC inhibitor for the treatment of non-small cell lung cancer and ovarian cancer

Sergey Baranovskiy, ChemRar RnD, Khimki, Moscow region, Russia

Indications: Non-small cell lung cancer (NSCLC):NSCLC accounts for 85% of all lung cancer casesThe American Cancer Society’s estimates for lung cancer in the United States for 2015 are: • About 221 200 new cases of lung cancer (115,610 in men and 105 590 in

women) • An estimated 158 040 deaths from lung cancer (86 380 in men and 71 660

among women) Lung cancer accounts for about 27% of all cancer deaths

Indications: Ovarian cancerThe American Cancer Society estimates for ovarian cancer in the United States for 2015 are:• About 21 290 women will receive a new diagnosis of ovarian cancer.• About 14 180 women will die from ovarian cancerOvarian cancer ranks 5-th in cancer deaths among women

Quisinostat – novel HDACi• Potent HDAC inhibitor status (HDAC1: IC50=0,16nM)• Inhibition of cell proliferation in tested tumor lines• Potent antitumor efficacy in a number of human xenograft models (ovarian,

colon, lung breast, prostate cancer)• No significant activity against large panel of receptors, kinases or zinc-

requiring enzymes• Oral administration• Favourable PK and safety profiles according to current clinical results• High efficacy according to interim clinical data

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Gosogliptin — a new innovative drug for the treatment of type 2 diabetes and metabolic syndrome

Sergey Baranovskiy, ChemRar RnD, Khimki, Moscow region, Russia

SatRx, LLC is an innovative biotech ChemRar group company formed in 2010.

The company develops new compounds starting from medical chemistry and high-throughput screening up to preclinical studies, full cycle of clinical trials and registration of new drugs for the treatment of type 2 diabetes and metabolic syndrome.

Gosogliptin — a DPP-4 inhibitor for the treatment of type 2 diabetesLike other gliptins, gosogliptin prevents the inactivation of incretins such

as glucagon-like peptide 1 (GLP-1) and therefore regulates the glucose levelAdvantages of Gosogliptin comparing to the marketed products:1. High affinity and high selectivity to DPP-42. PK profile enabling once a day dosage3. Favorable safety profile and reliable glucose-lowering action — mean

decrease of HbA1c by 0,92%The phase III clincal trial comparing Gosogliptin and Vildagliptin as both

monotherpies and combinations with metformin in newly diagnosed treatment-naive patients with type 2 diabetes has been successfully completed. Safety and non-inferiority of Gosogliptin comparing to Vildagliptin has been proved. Gosogliptin has been registered in Russia

The development has been conducted in terms of program «Pharma 2020» with the support of The Ministry of Industry and Trade of Russian Federation.

Fixed dose combination of Gosogliptin and MetforminOptimal combination of compounds in one dosage form for pathogenetic

correction of metabolic abnormalities in type 2 diabetesThe current technological process of development of fixed dose

combination will be completed with a comparative clinical trial to prove the advantages of this dosage form comparing to the administration of single drugs in equivalent dosages. The registration of fixed dose combination is expected in 2017.

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Il-16 and VEGF in the blood serum of patients with bone tumors

Bondarev A.V., TimofeevYu.S., SchupakM.Yu., Kuznetsov I.N., Babkina I.V., Alferov A.A., Boulytcheva I.V.

N.N. Blokhin Russian Cancer Research Center, Moscow

Aim: A comparative study of the initial levels of IL-16 and VEGF in the serum of patients with malignant, borderline and benign bone tumors.

Materials and methods: A comparative study of serum IL-16 and VEGF levels was perfomed in 138 patients with primary bone tumors: benign (10); borderline — giant cell tumor of bone (22); malignant (106), aged 14-50 years, immunoassay reagents «Biosource» (USA) — IL-16 and — «R&D» (USA) — VEGF prior to a particular treatment. Sarcomas were presented with osteosarcoma (45) (typical — 35, parosteal — 6, periosteal — 4), chondrosarcoma (24), Ewing’s sarcoma (27), an undifferentiated pleomorphic sarcoma (7) and chordoma (3).

Results: Frequency of IL-16 determination in the serum bone tumors patients was 93%, no significant differences in the IL-16 levels in taking into account the histological structure of the tumor is not revealed. There were no relationship between the size of the primary tumor and IL-16 content in serum. Overall 3- and 5-year survival of patients with malignant bone tumors with IL-16 serum levels >33.0 pg/ml was significantly lower than patients with levels of IL-16 ≤33,0 pg/ml. Osteosarcoma overall 5-year survival rate among patients with high IL-16 in serum was 1.6 times at Ewing sarcoma 1.7 times at chondrosarcoma 1.8 times lower than those containing IL- 16 serum ≤33,0 pg/ml. VEGF levels in bone sarcomas patients were significantly higher than in the borderline and benign tumors, while statistical analysis of significant difference in the levels of VEGF in view of the primary tumor histological structure is not revealed. Maximum VEGF levels marked with periosteal osteosarcoma, the minimum - at parosteal osteosarcoma. When the VEGF concentration in the serum above the average for the group (>493 pg/ml) the total 3-and 5-year survival in patients with malignant bone tumors was higher than that at low levels of this indicator. Similar results were obtained in osteosarcoma, whereas in Ewing sarcoma and chondrosarcoma high levels of 3 and 5-year survival rates were observed in patients with serum VEGF content less than 493 pg/ml.

Conclusion: These data suggest that the expression of IL-16 and VEGF could be associated with pathophysiological changes related to growth and metastatic process of bone sarcomas, and may be an object for further studies to determine the levels of these biomarkers and their value in the prediction of bone malignancies

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Using PolyGraphene During Pre-Clinical Tests of the Enterosorption and Safety Assessment

Alexander S. Botin1,2, Vladimir N. Buravtsev1, Tamara S. Popova2

1 Institute of Chemical Physics, Russian Acad. of Sciences, Kosygin Str.4, 119991, Moscow, Russia2 Sklifosovsky Institute of Emergency Medicine, Big Sukharevskaya Sq.3, Build. 10, Moscow, Russia

During experiments it was studied a new form of an oxygen-containing expanded graphite, which after repeated thermal activation and chemical modification with using ultrasound results in a material with stacks of carbon layers with higher multiplicity (10-40), but containing both single sheets of graphene. It is possible to include this material in classification of nanocomposite sorbents and the graphene-containing carbon forms. The authors of this work introduce a new classification of this type of material and see it as an PolyGraphene (PG).

Enterosorption — the method based on linkng and removal from the digestive tract (DT) with the medical or preventive purpose of endogenous or exogenous substances, metabolites, various products of a microbic origin.

The last achievements in the field of physiology and pathology of digestion allow to consider enterosorption mechanisms from mass exchange positions between the internal and enteral medium. In this regard, enterosorbents can be estimated not only as effective remedies of a detoxication of an organism, but also as a factor, in itself having essential impact on activity of the digestive and transport conveyor and an exchange of the main nutrients.

Though materials and manufacturing techniques of sorbents significantly differ, the main medical requirements to enterosorbents remain rather constant:

1) convenient pharmaceutical form and lack of unpleasant organoleptic properties of a preparation;

2) not toxicity — preparations in the course of passing up through a gastrointestinal tract shouldn’t collapse to fragments which can be soaked up and make negative impact on bodies and systems;

3) preparations shouldn’t injure the mucous; 4) there has to be a good evacuation warning a sorbent congestion in an

intestines gleam; 5) high sorption ability to the deleted components; 6) at not selective sorbents possibility of sorption of useful nutrients has

to be minimum; 7) lack of a desorption in process of advance through a gastrointestinal

tract, lack of dependence from рН of medium.

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Objective. The study of the interaction of PG with the structure of the mucous of small intestine to determine possibility of using the PG for detoxification general and selective action.

Conditions of experiments. Histologic research of a small intestine. For 10 rats through a probe was entered PoliGraphen’s suspension into initial department of a duodenum. After 2.5 hours for receiving samples of biomaterial to rats was done euthanasia according to the European bio-ethical standards of manipulations with laboratory animals. Further was opened an abdominal cavity of rats, was cuted sites of medial department of a duodenum, initial department of lean gut and distal department of ileum gut. Samples placed in the cooled fixating solution and processed according to the standard scheme for histologic research.

The results. PolyGraphene remains as a part of a himus and in the field of near wall of a mucous membrane of a gut. PG doesn’t get directly to a surface of cages of an epithelium that treats as large poly-particles of PG (100-500 microns), and smaller poly-particles (10-50 microns). At one-time introduction of PG, it goes as transit goods through a small intestine, without being late and without getting directly to a surface of an intestinal epithelium which is closed by a continuous dense mucous bed, and also in space between intestinal fibers. PG works, mainly in a gleam of intestines and at a surface of a mucous layer, without having direct negative destructive effect on cells of an intestinal epithelium and PG has a large capacity.

Conclusions. The results of tests PG as acting basis for the enterosorbents of new generation indicate a good promising potential for wide medical applications of PolyGraphene.

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Sorbtion - Desorbtion of Glycopeptide Antibiotics on the Surface of Oxidized OligoGraphene

Vladimir N. Buravtsev1, Alexander S. Botin1,2, Lyudmila A. Baratova3, Alla V. Timofeeva3, Genry S. Katrukha4, Andrey I. Poletaev1

1 Institute of Chemical Physics, Russian Acad. of Sciences, Kosygin Str.4, 119991, Moscow, Russia2 Sklifosovsky Institute of Emergency Medicine, Big Sukharevskaya Sq.3, Build. 10,

Moscow, Russia3 Belozersky Institute of Physico-Chemical Biology, Moscow State University,

119992, Leninskye gory, house 1, building 40, Moscow, Russia4 Gause Research Institute of New Antibiotics, Russian Academy of Medical Sciences,

Bolshaya Pirogovskaya Street, house 11, 119021, Moscow, Russia

Important phenomena were discovered that oxygen-containing expanded graphite — an Oxidized OligoGraphene (OOG) obtained after hydrothermic treatment of modified graphite became to be able to interact as sorbent OOG with glycopeptide antibiotics.

Introduction. In this work was studied the sorption properties of carbon material as an example of Oxidized OligoGraphene (OOG) concerning glycopeptide antibiotics clinically important groups, as well as study conditions desorption from OOG of antibiotics of this group.

OOG - version of ultrafine carbon sorbent, which was developed on the basis of the modified oxygen-containing expanded graphite (OCEG).

OOG is a potential sorbent for the separation of the antibiotic from the culture medium producing strain after the fermentation process. Originally studied sorption characteristics of OOG with respect to antibiotics-standards as an example of clinically important glycopeptide (Vancomycin-Ristomycin) group to which are known antibiotics vancomycin, ristomycin, teicoplanin A2, ehremomitcin. The interest to antibiotics of this group is steadily increasing, it is associated with a sufficiently high their stability with respect to MDR Gram-positive microorganisms such as: Methicillin Resistant Staphylococus Aureus (MRSA), Methicillin Resistant Staphylococcus epidermidis (MRSE), Clostridium difficile, enterococcs, pneumococcs, and others, so antibiotics -glikopeptids become indispensable in the treatment of infections caused by these microorganisms.

Sorption of antibiotics–glykopeptides on OOG. Technological feature of OOG as sorbent is need of filling of its openwork structure with solution in which tests were carried out it.

Follows from the obtained data that anti-biotics-glikopeptides possess ability to be oc-cluded on OOG in the chosen conditions. And the quantity of a sorbed antibiotic increases with increase in time of sorption.

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For an eremomitsin, a vankomitsin and a teykoplanin the liquid phase of a sorbent served in all experiments as «transit area» - the antibiotic was immobilized on a carbon matrix and its share in a liquid phase of a sorbent didn’t exceed 23% of all mass of the antibiotic accumulated on a sorbent. Thus, during the conducted research high sorption ability of a new sorbent of OOG concerning a number of antibiotics-glikopeptides for the first time is established.

Desorption of antibiotics from OOG. It is established that degree of a desorption of antibiotics from OOG depends on properties of organic solvent and also on structure of the analyzed antibiotic. It is shown that antibiotics, stripped from OOG, keep the physical and chemical properties and antibacterial activity. The obtained data by results of researches on sorption and a desorption allow to claim that OOG can be used in practice of allocation of antibiotics — polypeptides from solutions.

Conclusions. Researches have the theoretical and practical importance, the received results can be applied both in screening new antibiotics, and at allocation and cleaning of already known antibiotics in production.

Expression of miR-21 and its target genes ACAT1 and ARMCX1 in liver of rats treated with DDT

Michael Chanyshev Institute of Molecular Biology and Biophysics, Novosibirsk, Russia

MicroRNAs (miRs) are small, non-coding, single-stranded RNAs of 19–23 nucleotides in length that may complementarily bind appropriate mRNAs (targets) at the 3′-untranslated regions, leading to mRNA degradation and translation repression. In this manner miRs regulate expression of most of genes and thereby participate in biological processes, such as cell proliferation, differentiation and apoptosis. Understanding of gene expression regulation is crucial for treatment and diagnostics of many diseases including cancer. One of the most important miRs is miR-21 known as oncogenic miR. It’s known that treatment with some chemicals is accompanied by changes in miR-21 expression level.

In present study we assessed the regulation of genes by miR-21 and its possible outcomes. Female Wistar rats (n = 15) were treated weekly i.p. for three months with a single dose of 10 mg/kg («low-dose», n = 5) or 50 mg/kg («high-dose» n = 5) body weight of DDT in corn oil, rats of control group (n = 5) were treated with oil. We measured miR-21 level in the liver by qPCR with U6 as

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a reference gene, reverse transcription was performed using specific stem-loop primer. DDT treatment led to increase of miR-21 level and this effect was dose-specific (approximately 1.5 and 3 times, respectively).

Using in silico approach we found targets for miR-21. Since most of mRNAs are regulated by different miRs, we focused on those mRNAs that can be targeted by miR-21 only. From list of them we chose two targets: ACAT1 (acetyl-CoA acetyltransferase 1) and ARMCX1 (member of the ALEX family of proteins, expressed at very low level in liver). Relative mRNA level and protein content of ACAT1 and ARMCX1 genes in the liver was measured by qPCR and Western-blot analysis respectively, GAPDH was used as a reference gene. DDT treatment significantly decreased mRNA and protein content of ACAT1 in dose-specific manner, higher dose led to a greater effect. Level of ARMCX1 mRNA was also decreased whereas ARMCX1 protein content was decreased by treatment with the high dose of DDT only. Obtained results fit with hypothesis that selected genes might be regulated by miR-21.

ACAT1 is important enzyme in metabolism of ketone bodies in the liver. Several studies have shown the decrease of ACAT1 expression with fibrosis. Interestingly, that analysis of morphological changes in the liver of DDT treated rats displayed state of moderate fibrosis. Thereby high expression of miR-21 may be cause of this cell outcome. The reasons of miR-21 overexpression under DDT exposure remain unknown and require the additional studies.

Study was supported by the Russian Science Foundation (grant # 15-15-30012)

Profiling of mutations in gastric cancer

K.I. Cherepanova1,2, V.V. Strelnikov1,2, I.I. Bykov3, M.V. Nemtsova1,4, A.S. Tanas1,2;1Research Centre for Medical Genetics, Moscow, Russian Federation,

2Pirogov Russian National Research Medical University, Moscow, Russian Federation,3I.M.Sechenov First Moscow State Medical University, Moscow, Russian Federation,

4Russian Medical Academy for Postgraduate Education, Moscow, Russian Federation

Introduction: Gastric cancer (GC) is one of the most common oncological diseases with high morbidity and mortality. Genetic factors may increase the risk of developing GC. The aims of this study are to identify new genes involved in the development of GC and to characterize mutations in these genes.

Materials and methods: The study included GC 50 patients aged 35-59 years, who had given the tumor material, normal gastric tissue and venous

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blood. Search for mutations in 52 cancer related genes was carried out by high-throughput parallel DNA sequencing. For the validation of the detected mutations we used Sanger sequencing. Determination of germline or somatic nature of the identified mutations was carried out by DNA sequencing of normal tissues of the same patients.

Results: Mutations were found in 22 out of 52 genes. We showed that diffuse and intestinal GC types were distinguished by the spectra of somatic mutations: somatic mutations in the CDH1 gene were more common in the diffuse type (p=0.04) and mutations in the TP53 gene are more common in the intestinal type of GC (p=0.04). Germline mutations in patients with GC were for the first time detected in the genes MET (p.N375S, p.R970C), RB1 (p.H686N), EGFR (p.V292M), JAK3 (p.V722I), APC (p.I1289K), ATM (p.F858L), CDKN2A (p.D86N). New mutations were discovered in the CDH1 gene (c.-71C>G, p.K182N, p.T303P, p.S838G, c.1320+2T>G).

Conclusions: Our results suggest that germline mutations in genes other than CDH1 might contribute to CG development.

VEGFA isoforms balance shifts towards more angiogenic variants in human hepatocellular carcinoma

Mikhail S. Chesnokova*, Polina A. Skovorodnikovac,d, Darya A. Shavochkinaa, Inna F. Kustovaa, Nikolai S. Muguec,e, Nikolay E. Kudashkinb, Ekaterian A. Morozb, Yuri I. Patyutkob, Natalia L. Lazarevicha,d

aInstitute of Carcinogenesis andbInstitute of Clinical Oncology, N.N. Blokhin Russian Cancer Research Center,

cA.N. Belozersky Institute of Physico-Chemical Biology and dBiological Faculty, Lomonosov Moscow State University,

eN.K. Koltsov Institute of Developmental Biology.* Corresponding author: Mikhail S. Chesnokov, Ph.D., research assistant,

Institute of Carcinogenesis, N.N. Blokhin Russian Cancer Research Center, 115478, Kashirskoye shosse 24-15, Moscow, Russian Federation

Hepatocellular carcinoma (HCC) is the most common form of liver tumors that is characterized by high aggressiveness, low efficiency of chemotherapy and lack of sensitive diagnostic markers. One of the essential processes involved in HCC progression is angiogenesis that is mostly regulated via VEGFA signaling pathway. Inhibition of VEGFA pathway is considered to be a promising approach for HCC-targeted therapy.

Due to alternative splicing, VEGFA is expressed as a set of isoforms with different functional properties and physiological relevance. Changes in VEGFA isoforms expression pattern may affect both properties of tumor and effectiveness of VEGFA-targeting treatment.

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The aim of present study is to quantitatively characterize underinvestigated pattern of VEGFA isoforms expression in HCC and evaluate possible associations between VEGFA expression changes and clinicopathological characteristics of the tumor. We used 50 paired samples of HCC and non-tumorous (NT) tissue to study the expression of VEGFA isoforms using conventional and quantitative RT-PCR with total-VEGFA- and isoform-specific primers.

The predominant VEGFA isoforms expressed in examined specimens were VEGFA-189, VEGFA-165 and VEGFA-121. The more potent stimulators of angiogenesis VEGFA-165 and VEGFA-121 were upregulated in 38% and 44% of cases and downregulated in 26% and 24% of cases, respectively. Expression of less active membrane-bound VEGFA-189 isoform was repressed in 66% and upregulated in 14% of cases. 80% cases displayed statistically significant decrease in VEGFA-189 fraction of total VEGFA amount and parallel increase in fraction of VEGFA-165 and VEGFA-121.

Decrease in VEGFA-189 and increase in VEGFA-165 fractions correlated with advanced tumor stage. Changes in VEGFA-189 and VEGFA-165 expression level negatively correlated with tumor capsule presence. Upregulation of VEGFA-165 and VEGFA-121 expression was most pronounced in ascite-positive cases, while higher fraction of VEGFA-165 and VEGFA-121 corresponded to increased serum level of onco-fetal marker α-fetoprotein.

Thus, using quantitative approach, we have revealed HCC-specific pattern of VEGFA isoforms expression that consisted in prevalent downregulation of less active VEGFA-189 isoform and activation of more angiogenic VEGFA-165 and VEGFA-121 ones. Shift in balance of VEGFA isoforms expression was associated with prominent clinicopathological properties of tumors. The clinical significance of presented data consists in their potent impact on optimization of HCC treatment since VEGFA isoforms ratio may be a promising factor for prediction of anti-angiogenic therapy efficacy. Further studies of functional properties of VEGFA isoforms in HCC are necessary to evaluate its possible association with other processes determining tumor progression, survival and recurrence.

The work was partly supported by grant from Russian Ministry of Education and Science (contract 14.607.21.0049, RFMEFI60714X0049).

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Functional hypermethylation of a number of chromosome 3 genes revealed in breast cancer using noti-microarrays

A.A. Dmitriev1, G.S. Krasnov1, A.D. Beniaminov1, I.V. Pronina2, G.A. Puzanov1, S.V. Kurevlev2, T.P. Kazubskaya3, V.I. Loginov2, V.N. Senchenko1, E.A. Braga2, V.I. Kashuba4

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia2Institute of General Pathology and Pathophysiology, Moscow, Russia

3N.N. Blokhin Russian Cancer Research Center, Moscow, Russia4Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institute,

Stockholm, Sweden

Introduction. Genetic and epigenetic alterations can lead to the activation of oncogenes and inactivation of tumor-suppressor genes (TSGs) followed by the development of malignant tumors. NotI-microarrays designed by prof. E.R. Zabarovsky is a unique tool that allows us to simultaneously detect hypermethylation of CpG islands and deletions of DNA — two major reasons of TSG silencing.

Aim of the study. The aim of the present research was to evaluate the frequency of chromosome 3 genetic and epigenetic aberrations in breast cancer and search for novel TSG-candidates.

Material and methods. In total, 47 paired tumor/normal breast cancer samples were obtained from the N.N. Blokhin Russian Cancer Research Center (Moscow, Russia). Frequency of CpG island methylation and DNA copy number aberrations was assessed using comparative (tumor/normal) DNA hybridization on NotI-microarrays. The microarrays contained 180 NotI-clones (in 6 replicates each) associated with 188 chromosome 3 genes. Data analysis was performed using original NIMAN (NotI-microarray analysis) software. Expression alterations were evaluated with the use of qPCR technique, ΔΔCt method and original ATG (Analysis of transcription of genes) software. Non-parametric tests were used for statistical analysis.

Results. With the use of NotI-microarrays 35 NotI-sites with high (15-38% of cases) hypermethylation/deletion (HM/D) frequency were revealed in breast cancer (BC). Among genes associated with this sites, there are both known TSGs and TSG-candidates (EPHB1, ITGA9, ALDH1L1, VHL, CTDSPL etc.) as well as genes, which involvement in breast oncogenesis was shown for the first time (TOPAZ1, LRRN1, GORASP1, FOXP1, PRICKLE2 etc.). NotI-microarray data were verified selectively using bisulfite sequencing for VHL, NKIRAS1, ITGA9, LRRC3B, and CTDSPL genes. Several genes with high HM/D frequency (ALDH1L1, EPHB1, ITGA9, and ROPN1) were tested for expression alterations using qPCR. Frequent (57-90% of cases) and significant (> 2-fold) down-regulation was shown for all of them in BC. The most significant expression loss was observed for ALDH1L1 gene – 50-fold mRNA level decrease on the average in 90% of BC samples.

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Obtained data allowed us to suggest the set of potential markers based on DNA methylation status, for BC diagnostics. Moreover, in 21% of BC samples HM/D were revealed in 16 or more NotI-sites simultaneously. Such samples most likely represent CIMP+ (CpG-island methylator phenotype) tumors characterized by dense methylation of promoter CpG islands of a number of genes.

Conclusions. We revealed 35 NotI-sites associated with 40 chromosome 3 genes that were hypermethylated/deleted in more than 15% of examined BC samples. The involvement of the majority of the genes in breast oncogenesis was shown for the first time. These genes are novel TSG-candidates in BC. Significant expression decrease associated with DNA hypermethylation was observed for ALDH1L1, EPHB1, ITGA9, and ROPN1 genes. Besides, 10 out of 47 investigated BC samples most likely represents CIMP+ tumors. Detection of CIMP+ tumors is important both for diagnostics and prognostics as well as for the choice of treatment strategy.

Funding. The methylation and expression analysis of genes, revealed by Not-microarrays, was financially supported by the Russian Science Foundation (grant 14-15-00654). Participants of this grant thank Engelhardt Institute of Molecular Biology, Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institute and N.N. Blokhin Russian Cancer Research Center for collaborative studies.

Storage as a way to improve the efficiency of sequencing data usage

Irina Zhegalova, Anna Petuhova, Sergey Suchkov, I.M. Sechenov First Moscow State Medical University

In January 2015, the US government announced a new program on Precision Medicine, during which electronic medical records and sequencing data of million volunteers will be re-ceived. Framed with Personalized medicine and translated into the practice, such data becomes the basis for the field of early (including predictive and pre-clinical) diagnostics.

Each sequencer generates nearly one and a half terabytes of «raw» data. After cleaning and mapping it is reduced at times, but continues to contain important information about indi-vidual features of patients. But in clinical practice of our country it is more often for genomic da-ta to become an object of study only for physician-geneticist, and later such data is sent to «ar-chive» and is not used further. Top-local sources of scientific information become practically «disposable». For large university hospitals such information flows can reach several tens of en-tries per day.

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Even if we are talking about the impossibility of fully functioning genetics services in our country at the present time, we can increase the effectiveness of the effort through the collection of information in a single hospital database.

Already, an increased number of people suffered from orphan diseases, the growing field of pharmacogenomics and the rapid expansion of the market segment in targeted therapies make it possible to talk about the need for Uniform Hospital Database to form a flow of patients under clinical trials of highly specialized products. Furthermore, such data will be the basis for long-term projects on personalized.

MicroRNAs as biological markers of ovarian cancer

E.S. Gershtein1,3, D.N. Kushlinsky2, I.V. Tereshkina3, L.V. Adamyan2,3

1N.N. Blokhin Russian Cancer Research Center; 2Scientific Center for Obstetrics, Gynecology and Perinatology named after V.I. Kulakov;

3Moscow State University of Medicine and Dentistry name after A.I. Evdokimov

Introduction. MicroRNAs (miRNAs) - small non-coding RNAs targeting multiple mRNAs - are posttranscriptional regulators of gene expression involved in carcinogenesis, metastasis, and invasion. MiRNAs expression is altered in various human tumors including ovarian cancer.

Aim of the study. Analysis of published data describing miRNAs expression pattern and its changes in ovarian cancer tissue and in peripheral blood with the accent on the possibility of their application as diagnostic, prognostic, and predictive biomarkers in this disease.

Materials and Methods. The results of the most valuable published clinical laboratory studies found in PubMed database in the period from 2007 to 2016 are summarized and critically evaluated.

Results. Expression of several tens of miRNAs was shown to be both up- and downregulated in ovarian cancer compared to normal ovarian tissues. Similar changes were revealed in peripheral blood of ovarian cancer patients as compared to healthy persons. Substantial number of evidence indicating to good prospects of miRNAs application both in diagnostics, including non-invasive serological approach, and in general prognosis and, more important, in prediction of resistance to standard chemotherapy (platinum derivatives, taxans) has accumulated. However, the diversity and inconsistence of these data prevents marking out of accurate and reliable candidate clinically valuable biomarkers at the present stage.

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Conclusion. MicroRNAs are promising biological markers for ovarian cancer, but further data accumulation, unification of methods and results’ evaluation criteria are needed for successive practical

RANK/RANKL/OPG system and interleukins 6, 8 16 in blood serum of patients with primary bone neoplasms

E.S. Gershtein, A.A. Zuev, Yu.S. Timofeev, Yu.N. Soloviev, N.E. Kushlinskii N.N. Blokhin Russian Cancer Research Center, Moscow

Introduction. RANK/RANKL/OPG ligand-receptor system is a key player in bone tissue homeostasis directly regulating osteoclast differentiation and osteolysis. Many pathologic processes imposed by impairments in bone remodeling including tumor growth and metastasizing are associated with misbalancing of this system.

Aim of the study. Comparative evaluation of RANK/RANKL/OPG and some proinflammatory cytokines (IL-6, 8, 16) serum levels in primary bone neoplasms’ patients and practically healthy persons and analysis of associations between these markers and principal clinico-pathologic characteristics of bone tumors.

Material and methods. 101 malignant bone tumor (37 osteosarcoma, 41 chondrosarcoma, 12 chordoma, 7 Ewing sarcoma, 2 pleomorhic undifferentiated sarcoma, 2 fibrosarcoma) 32 borderline giant cell bone tumor (GCBT), and 30 benign bone tumor patients were involved. Control group comprised 71 persons. OPG, sRANKL, sRANK, IL-6, 8, 16, and Receptor for Advanced Glycation End product (sRAGE) serum levels were measured by standard ELISA kits.

Results. Measurable RANK levels were detected in blood serum of less than 50% of malignant and benign bone tumor patients and control persons; the median of RANK concentration in GCBT patients comprised 57 pg/ml. Serum sRANKL and OPG levels in GCBT patients’ were increased both in relation to control, and to malignant bone tumor patients. OPG level is also markedly increased in patients with malignant and benign neoplasms as compared to control, while IL-6 is significantly increased in benign and borderline tumor patients as compared to those with benign lesions and control. The degree and direction of changes in RANK/sRANKL/OPG system depends also on histologic type of malignant bone tumors.

Conclusion. The changes revealed in this study evidence in favor of disturbances in the balance of osteolysis activators and inhibitors in patients with primary bone tumor depending on the character of neoplasm (malignant, borderline or benign) and histological structure of malignant tumors.

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Grammometric labs on a chip and synchronization of the on-chip isolated muscle fiber grammometry with the in situ

tissue X-ray analysis and MEMS-assisted excitation time spectroscopy

Gradov O.V.Talrose Institute for Energy Problems of Chemical Physics, RAS, Moscow, Russia

This poster presents the main concept of our studies performed in 2011 on the structural and functional changes monitoring in single muscle fibers on a chip, allowing multivariate measurements. We propose here the synchronization of the following analytical techniques: micromechanical measurements performed using a grammometer-type dynamometric / tensometric system with digital recording; real-time on-chip morphometric microscopy; X-ray tissue analysis with the discrete identification («barcoding») of the structural data from the matrix registration array detector; stimulation electromyography at various excitation parameters using the chip providing the response spectra of the sample at each excitation signal type.

The graphical representation of the above measurements correlation provides useful information on the relationship between the structure and function of the sample at both cytophysiological and supramolecular levels and can be compared with the database.

Correlation analysis of the biochemical response of the isolated muscle fiber to either electrophysical or electrochemical stimulation allows to detect teinochemical effects resulting from the conformational changes both at macromolecular and supramolecular scale, since the deformation value and, hence, the fiber contraction, depends on the variable medium characteristics or the external effect parameters and can be corresponded to them in a special database. Furthermore, the technique proposed allows a direct on-chip study of the effects of pharmacological and physiotherapeutic agents causing similar conformational changes in situ.

The using of X-ray methods of colloid and tissue analysis provides identification of the results of the following processes: dehydration, thermal effects, electrophysiological excitation and chloroform-caused anesthesia. The design of organ-specific or tissue-specific descriptors depending on the fiber orientation would allow to distinguish between the various muscle fiber types and sources within the basis of comparative histological interpretation.

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Homocysteine plasma level distribution in patients with cancer

Ivanov A.V.1, Nikiforova K.A.1, Kushlinskii N.E.2 and Kubatiev A.A.11FSBSI “Institute of General Pathology and Pathophysiology”, 125315, Moscow, Baltiiskaya str., 8,

phone +79653725301, e-mail: [email protected]. Blokhin Russian Cancer Research Center, Russia, Moscow, 115478, Kashirskoe highway, 23,

phone +74993242604

Homocysteine (Hcy) is a product of methionine demethylation. Various genetic, nutritional and hormonal factors are the prime cause of increase of plasma homocysteine level (hyperhomocysteinemia or Hhcy). Numerous clinical and epidemiological data confirm the association of Hhcy and vitamins B9, B6 and B12 status, and SNP of key enzymes of methionine cycle. Therefore Hcy imbalance is involved in a DNA synthesis and methylation, and hence it has been hypothesized to be associated with carcinogenesis. But only in some cases (for example lung, colorectal cancer) the association of Hcy with cancer was found [Tastekin D. et al. Exp. Oncol. 2015,37(3):218-22; Keshteli A.H. et al. World J. Gastroenterol. 2015,21(4):1081-90], whereas it is absent obvious impact of Hhcy in many cases cancer. The aim of this work was a detailed study of the distribution of Hcy and cysteine in blood plasma in patients with cancer of the internal organs.

We have tested the normal distribution of the blood plasma samples of patients with renal cell carcinoma (N=41, Age 60±11), ovarian (N=21, Age 48±14), colorectal (N=74, Age 59±11) and breast (N=45, Age 56±11) cancer using Shapiro-Wilk test at α=0.05. Age-related distribution was normal for all patients groups. Hcy and cysteine analysis was performing by capillary electrophoresis method [Ivanov A.V. et al. J. Chrom. B 2015, 1004:30-6]. The distribution of Hcy is not normal in all cases, while for the samples of patients with ovarian, colorectal and breast cancer ratio Cysteine / Hcy was normal. Thus, the internal heterogeneity of the Hcy distribution in studied groups was demonstrated, and that causes the search and study of Hhcy cancer-associated factors.

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Development of plant-produced universal influenza nanovaccines: from “green” biotechnology to medicine

Ivanov P.A., Gasanova T.V., Petukhova N.V.Lomonosov Moscow State University, Department of Virology, Moscow, Russia

A new approach for super-expression of conserved M2e epitope (23 amino acid residues, aa) in plants has been developed on the basis of a recombinant tobacco mosaic virus (TMV-U1, wild type, wt) genome designed for Agrobacterium-mediated delivery into the plant cell nucleus. The TMV coat protein (CP) served as a carrier and three versions of human consensus M2e sequence were inserted between amino acid residues 155 and 156. Cysteine (Cys) residues in the foreign peptide were thought likely to impede efficient assembly of chimeric particles. Therefore, in addition to the initial sequence, we introduced mutations by the substitution of cysteines at the positions 17 and 19 with either serine (Ser) or alanine (Ala) residues. Agroinfiltration experiments proved that all the recombinant viruses were capable of spreading via vascular system of Nicotiana benthamiana, N. excelsior, N. sylvestris, N. clevelandii and N. tabacum. The chimeric particles were stable in plant extracts and during purification; influenza antigens were exposed on their surface as shown by immunoelectron microscopy. Preparations of particles carrying M2e-ser/ala epitopes can be sterilized through nitrocellulose filters (0.45 mkm) that enables one to store them at +4°C for at least 4-6 months. Reverse transcription of genomic RNA and total RNA from systemic leaves proved genetic stability of TMV-M2e recombinant viruses as well as repeated inoculations using extracts from symptomatic upper leaves. Nicotiana benthamiana plants produced as much as 5 g of TMV-M2e-ala or 1 g of TMV-M2e-ser CPs per 1 kg of fresh weight of non-inoculated leaves in 2 weeks under typical greenhouse conditions. Following the purification, particles incorporated up to 90% of CP-M2e fusion protein. During vaccination, experimental mice did not lose weight comparing with the control animals. Antisera to chimeric virions contained far more antibodies specific to influenza virus than those specific to TMV particle itself (up to 5/1 ratio). IgG1/IgG2a ratio varied from 0.7 (Ser) to 3.2 (Ala). The longevity of M2e-specific immune response declined insignificantly during the 7 month period. A whole cell ELISA assay confirmed that the antisera to TMV-M2e-ala and TMV-M2e-ser are capable of recognizing M2 protein exposed on the surface of epithelial cells infected with influenza A/PR/8/34 (H1N1). Mice immunized with the chimeric virions were resistant to five lethal doses (LD50) of the homologous influenza virus strain, A/PR/8/34 (H1N1) and TMV-M2e-ala conferred protection (5LD50, 70% of survival rate) against a heterologous strain

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(4 amino acid changes in M2e plus cysteine substitutions) A/California/07/2009 (H1N1). Thus, TMV-M2e-based nanovaccine may be called “universal”. It should be noted that amino acid sequence of M2e antigen from avian influenza usually differs from human consensus by 3-4 substitutions. Similarly, we constructed another vector designed for expression of hydrophobic fusion peptide (fp, 14 aa) from hemagglutinin. Systemic infection of N. benthamiana with TMV-fp vector caused severe symptoms and genetic stability of TMV-fp vector was demonstrated as well. Electron microscopy of TMV-fp preparation represented the rod-shaped particles similar in morphology to TMV-wt virions. It may imply that rigid helical epitope geometry mimics the native tetrameric structure of M2 protein ion channel and provides a high level of immune response with a promising antigen/carrier correlation and protection against lethal heterologous infection. Therefore, a new generation plant-derived technologically efficient universal influenza A nanovaccines has been obtained.

Expression of microRNAs and their target genes in various organs of female rats under DDT exposure

1,2Tatyana Kalinina, 2Dmitry Ushakov, 2Sergey Philippov1Novosibirsk State University

2The Institute of Molecular Biology and Biophysics, Novosibirsk, Russia

All living organisms are in constant interaction with the chemical environmental factors that aren’t natural components of physiological processes in cells. Among these foreign substances dichlorodiphenyltrichloroethane (DDT) is a potent insecticide that was used worldwide for agricultural and public health purposes until the 1970. DDT possesses various mechanisms of toxic action: it changes microRNA expression or activate nuclear receptors CAR, PXR, ERα. As a result, DDT affects the expression of many genes that encode proteins involved in many cellular processes. In our study the expression level of oncogenic microRNAs (miR-21, -221, -222, -429, - 205) in liver, uterus and ovaries of female rats treated with DDT was determined. Using in silico approach we also found miR-190a, which contain in their promoter putative PBREM and can be regulated by CAR. Changes in the expression profile of miR-21, -429, -190a depending on rat’s organ and DDT doses (10 and 50 mg/kg) in acute and chronic experiments have been revealed. Elevated expression of miR-205 in all studied organs after a single dose of DDT has been shown. Also, changes in the expression of miR-190a, -21 target genes (Soat1, Ago1, Tp53inp1, Phlpp1, Armcx1) after DDT exposure were shown. Using obtained results the

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scheme of interactions: xenobiotic — target (receptor) — microRNAs — target gene – changes in cellular processes (proliferation, apoptosis, differentiation) has been constructed. It is important to note that all changes identified in our experiments resulted in the changes in gene expression including those participate in carcinogenesis.

This work is supported by the Russian Science Foundation № 15-15-30012.

The involvement of methylation in down-regulation of some suppressive miRNA genes and progression of renal carcinoma

V.V. Khokonova1,2, I.V. Pronina1,2, A.M. Burdennyy1, A.A. Dmitriev3, A.V. Karpukhin2, T.P. Kazubskaya4, E.A. Braga1,2, V.I. Loginov1,2

1Institute of General Pathology and Pathophysiology, Moscow, Russia 2Research Center of Medical Genetics, Moscow, Russia

3Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia4N.N. Blokhin Russian Cancer Research Center, Moscow, Russia

Introduction. Epigenetic mechanisms play a great role in expression deregulation of miRNA genes and downstream signaling pathways in cancer. The hypermethylation may lead to down-regulation of suppressive miRNA genes and activation of their pro-tumor target genes. These events may serve as drivers for malignant transformation of cells and cancer progression.

Aim of the study. The goal of our study was to reveal miRNA genes regulated by methylation and assess the contribution of methylation of miRNA genes to clear cell renal cell carcinoma development and progression.

Material and methods. The group of tumor suppressive miRNAs was selected using databases, in particular, miRWalk 2.0. The set of 60 paired (tumor/normal) samples of clear cell renal cell carcinoma (ccRCC) from patients not exposed to prior radiation or chemotherapy was used. Only samples with tumor cell content of at least 70% were selected. The methylation profiles of CpG islands of miRNA genes were assessed by methylation specific real-time PCR. We used the certified real-time PCR kits of Applied Biosystems to analyze the expression level of miRNAs. RNU6B and RNU48 were applied as references. Fisher’s exact test and Spearman’s rank correlation coefficient were used for statistical analysis.

Results. We have shown that the frequency of methylation of MIR-124a-1/-2/-3, MIR-129-2, MIR-34b/c, MIR-193a, MIR-9-1/-3 genes in ccRCC samples was significantly higher than in histologically normal tissues of the

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same patients (P ≤ 0.05 by Fisher’s exact test). The frequencies of methylation of the listed miRNAs in renal tissues of post-mortal patients without cancer history were in the range from 5% to 9%. We have identified statistically significant positive correlations between the expression down-regulation and hypermethylation for the following pairs of mature miRNAs / encoding genes: miR-124a-5p / MIR-124a-3 (Spearman’s correlation coefficient, Rs = 0.74, P < 0.05); miR-129-5p / MIR-129-2 (Rs = 0.90, P < 0.05); miR-34b-3p / MIR-34b/c (Rs = 0.82, P < 0.05); mi1R-34c-3p / MIR-34b/c (Rs = 0.70, P < 0.05) and miR-9-5p / MIR-9-1/-3) (Rs = 0.61, P < 0.05). In addition, we revealed statistically significant correlations of the frequency of methylation of MIR-124a-2 and MIR-129-2 genes with the size and the stage of tumor development (P ≤ 0.001 by Fisher’s exact test), methylation of MIR-124a-3 gene with the degree of tumor differentiation (P = 0.0001), and methylation of MIR-129-2 gene with the presence of metastases in adjacent lymph nodes (P = 0.01).

Conclusion. Our data extend the range of suppressive miRNAs deregulated by methylation in kidney tumors. Data on the relationship of methylation of some miRNA genes with progression and metastasis are relevant to clinical oncology.

Study of PTEN expression in endometrial cancer

Kobelev W.S.1,2, Geletina N.S.2

1The Institute of Molecular Biology and Biophysics, Novosibirsk, Russia2Novosibirsk State University, Novosibirsk, Russia

Endometrial cancer (EC) represents the most common gynaecological malignancy in many industrialized countries. About 70–80% of sporadic endometrial carcinomas designated as type I carcinomas have estrogen-related pathway. At present, endometrial carcinomas are characterized by distinctive types of genetic instability and molecular alterations. The most frequently altered gene in EC is PTEN, which codes for a protein with tyrosine kinase activity. Up to 70% of EC and 50 % of precancerous lesions reveal altered PTEN, characterized by loss of function. The underlying genetic alteration with lost PTEN function is mostly mutation and, less frequently, loss of heterozygosity or promoter methylation. MicroRNAs (miRNAs) are short noncoding RNAs also can regulate PTEN expression negatively. The goal of our study was to determine PTEN gene expression in EC and reveal the possible mechanisms of its inactivation. We use the laser micro-dissection capture of tumor tissue to collect EC cells. PTEN gene expression level in 14 samples of EC is determined.

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It is shown that in 43% of cases its expression didn’t change, in 57% - has been lowered by 4-6 times. In two samples with the lowered expression of PTEN rs 398123319 mutation in intron between the 4th and the 5th exon is registered. In other samples there were no mutations in the “hot spot” region of 5th exon. We evaluated the expression of miRNA (miR-1908, miR-181a, and miR-186) selected by in silico analysis as a potential negative regulators of PTEN in the same samples of EC. Expression of miR-1908 didn’t change, whereas miRNA-181a expression was increased in 35% and miRNA-186 was decreased in 57%.

Thus, our results indicated that microRNAs may play the important role in the regulation of PTEN gene expression and function.

This study is supported by the Russian Science Foundation (grant # 15-15-30012)

Creating model systems for testing the bioactivity of type I and II IFNs

E.P. Konyaeva1,2, K.V. Kulikova1,2,3, N.V. Gnuchev3, G.P. Georgiev3, S.S.Larin1,2,3

1Moscow Institute of Physics and Technology (State University)2FNKC «Centre of Pediatric Hematology, Oncology and Immunology» named after D. Rogachev

3Institute of gene biology RAS

Interferons (IFNs) are crucial for the control of immune response. They belong to a group of signaling molecules. All IFNs have been classified into three groups: I, II and III types. IFN-γ is IFN type II, which activates STAT-signaling pathway (Signal transducer and activator of transcription) in the cell. The recruitment of IFN-γ to its receptor causes phosphorylation of transcriptional factor STAT-1. Activated STAT-1 forms homodimers, which translocate to the nucleus and bind to GAS (Interferon Gamma Activated Site) sequences. IFN-α or IFN-β (IFNs type I) binding to its receptors leads to phosphorylation of STAT-1 and STAT-2. STAT-1/2-heterodimers enter the nucleus and bind to ISRE (Interferon-Stimulated Response Element) sequences. Thus, IFN-γ activates GAS-dependent genes and IFN-α/IFN-β activates ISRE-dependent genes.

Currently, the major way to determine the bioactivity of the IFNs is to measure the interferon-induced inhibition of the viral cytopathic effect on indicator cells. The assay is quite complicated and requires special precautions. Therefore, there is a necessity to develop new approaches for determination of the bioactivity of the IFNs. The one possiblily is to create a model system based on the ability of IFNs to activate STAT-signaling pathways.

To define the IFNs bioactivity two different model systems were established. The first model system is represented by a stable cell line with

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genome integrated GAS-reporter cassette. This reporter cassette includes six tandem repeats of GAS-element, minimal promoter and firefly luciferase gene (FFly). It allows to detect activity of IFN-g/STAT-1 signaling pathway. The second model system is designed for monitoring of activity of IFN-α(IFN-β)/STAT-signaling pathway. It’s a stable cell line with genome integrated reporter cassette consisting of a luciferase gene under control of ISRE-containing promotor.

For the both reporter systems the detectable IFNs concentration range was detemined. Applicability of the model systems in the IFN antibodies testing was also demonstrated.

Metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in tumor and plasma of colorectal cancer patients: relationship,

clinico-pathologic and prognostic implication

Korotkova E.A., Nikolaev A.A., Gershtein E.S., Prorokov V.V., N.N. Blokhin Russian Cancer Research Center, Moscow

MMPs with their tissue inhibitors TIMPs play a crucial role in cancer invasion and metastasizing, and are regarded as helpful prognostic and predictive markers in colorectal cancer (CRC).

Study Aim: To evaluate associations between MMP-2, 3, 7, 9, TIMP-1, 2 tumor and plasma levels in colorectal cancer (CRC) patients with the main clinico-pathologic features and disease prognosis in order to reveal most promising biological markers.

Method: The study enclosed 93 colorectal cancer patients operated in 2005-2008. MMPs 2, 3, 7, 9, and TIMPs 1, 2 in plasma and in tumor and adjacent unchanged tissue extracts were determined by standard ELISA kits.

Results: Detectable levels of all proteins were found in all tissue samples. MMPs content was significantly increased as compared to adjacent mucosa in 70-80% colorectal cancers, TIMP-1 - in 87%, and TIMP-2 - in 63%. No meaningful correlations were revealed between the levels of individual proteins in tumors and adjacent tissues, but MMP-2 and MMP-3 levels were positively associated both in tumors and in unchanged colon mucosa. All proteins were represented in blood plasma, but no significant differences from control levels and no unambiguous associations between intra-tumor and circulating concentrations of any of the markers were discovered. Multivariate analysis demonstrated independent unfavorable prognostic role of high plasma MMP-7 and TIMP-1 in CRC with cut-off levels of 4.0 and 347 ng/ml respectively. Tumor MMPs/TIMPs

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content did not significantly affect the survival of patients without distant metastases, but univariate analysis revealed unfavorable prognostic value of high (>7.8 ng/mg protein) tumor MMP-7 in patients with disseminated process.

Conclusion: In 70-90% of colorectal cancers a significant increase of MMPs representing 4 subfamilies (collagenases, gelatinases, stromelysins, and matrilysin) expression was found. Plasma MMP-7 and TIMP-1 were shown to be independent prognostic factors in CRC and can be considered as possible biological marker characterizing colorectal cancer metastatic and invasive potential.

Isolation and characteristic of colon cancer cell with stem cells properties from primary tumors

Sergey Koshkin1, Grigory Raskin2, Nikolai Petrov1,Olga Bajenova3,4, Elena Tolkunova1

1Institute of Cytology, Russian Academy of Sciences, St. Petersburg 194064, Russia;2Russian Research Centre for Radiology and Surgical Technologies. St. Petersburg, 197758;

3Theodosius Dobzhansky Center for Genome Bioinformatics, St. Petersburg State University, St. Petersburg 199034, Russia;

4Department of Genetics and Biotechnology, St. Petersburg State University, St. Petersburg 199034, Russia

Colorectal cancer is the fourth most frequently diagnosed cancer and the second leading cause of cancer death in the USA. According to the data of the American Cancer Society, in 2015, 93000 new cases of colon cancer were estimated.

According to British Cancer Research, at the moment of diagnosis, near half of the patients are diagnosed with the I and II stages, which can be radically treated by surgical operations. Another part of the patients have regional or distant metastasis. For effective treatment of this group, we require drugs which are able to destroy different populations of cancer cells.

According to hypothesis of cancer stem cells (CSC), tumors represent a mixed-cell population that consists of healthy and cancerous cells with varying degrees of differentiation. The most differentiated cancer cells in the primary tumor have the features of the epithelial tissue from which the tumor originated, enabling its identification. The least differentiated cells have the characteristics of stem cells that confer tumor the ability to de- and re-differentiate, metastasize and to acquire drug resistance.

The aim of this study is to search for genes that are involved in the development and progression of colorectal cancer and their CSC properties. We made an attempt to extract a population of cancer stem cells from the

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primary cell culture of colorectal cancer. We have also assessed the role of Oct4 transcription factor in tumorigenic and drug resistance properties of colorectal cancer cells.

In this work we selected and then characterized colon cancer stem-like cells from primary tumors. From 6 endoscopic tissue samples, 2 cell lines were obtained. For selecting stem-like cells, two methods were used. First – Transfection of a bicistronic DNA construct 2A2B-TKiresPur that features the thymidine kinase and puromycin resistant genes under the control of 2A2B enhancer from the pluripotency-associated gene Oct4. Second - MACS for CD133+ cells. Via injections into immune-deficient NUDE mice, we obtained cells, that had caused the tumors. Mice tumors expressed the same as original tumor IHC markers (CK-20, beta-catenin, CDX2 and ALDH1 positive).

Sensitivity to cytostatic was measured by MTT method with 5-FU and Irinotecan. The selected cell populations had shown similar response to Irinotecan treatment. The cells enriched in endogenous Oct4 expression have significantly greater resistance to 5-FU, that is consistent with the observations that CSCs have higher resistance to modern chemotherapeutic agents. The subpopulation of cells expressing the CD133 marker (CD133+) shows the same resistance to the 5-FU as the original cells and lower than the CD133- subpopulation.

Altogether, this approach can be used in developing of new therapeutic strategies aimed at eradicating the tumorigenic subpopulation of CSCs within colorectal malignant tumors. To attain maximal clinical results, we consider it necessary to use the Oct4 positive cancer cells as therapeutic targets.

The work was supported by the Russian Science Foundation grant14-50-00068.

IGFs and IGF binding proteins as diagnostic and prognostic tumor markers

Kostyleva O.I.1, Maslyaev A.A.1, Kushlinsky D.N.2, Mamedov U.R.1,Korotkova E.A.1, Adamiyan L.V.2, Gershtein E.S.1

1N.N.Blokhin Russian Cancer Research Center, Moscow; 2Scientific Center for Obstetrics, Gynecology and Perinatology named after V.I. Kulakov, Moscow

Aim of the study: To evaluate the relationship between insulin-like growth factors (IGF-I and II) and key IGF binding proteins (IGFBP-1, 2, 3) in tumors and peripheral blood of cancer patients with major clinical and morphological features and prognosis of diseases in order to reveal clinically important biological markers.

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Material and Methods: The study involved 104 patients with ovarian tumors (74 malignant, 14 benign, and 16 borderline); 95 – with colorectal cancer (CRC), 79 – with breast cancer, 64 patients with cervical cancer and intraepithelial hyperplasia (CIN), and 113 - with bone tumors and tumor-like lesions. The control group consisted of 77 healthy women and 17 men. The markers were determined by standard direct ELISA kits («Mediagnost», Germany).

Results: Significant IGF/IGFBP imbalances were demonstrated indicating to an increase of IGF bioavailability for tumor cells. The direction and extent of these changes depended on the type and clinical and morphological characteristics of the tumors. Thus, IGF-I decreased in serum of patients with tumors of female reproductive system, but was elevated in patients with bone neoplasms and CRC.

IGFBP-2 was found to be a promising marker for the differential diagnosis of ovarian cancer: its diagnostic sensitivity at the specificity of about 80% exceeded 85% from the earliest stages of the disease. IGFBP-2 was also significantly elevatedin CRC patients’ sera, buta complex of 3 tests - IGFBP-2, IGFBP-1 and IGF-I – allows to make apreliminary differential diagnosis between these diseases in female patients. These diagnostic markers were also associated with the prognosis of overall survival in ovarian cancer, and low IGF-I serum levels retained its independent unfavorable prognostic value in multivariate analysis.

Ovarian cancer prognosis wasalso influenced by IGF-II and IGFBP-1 content in the tumor tissue. Marked reduction of IGF-I, II and IGFBP-3 serum levels and an increase ofserum IGFBP-1 was found in cervical cancer patients indicating the involvement of these proteins in cervical carcinogenesis and making them prospective markers of latent invasion in severe forms of CIN. Significant prognostic value of serum IGFs and IGFBPs content prior to specific treatment was demonstrated for patients with bone sarcomas.

Conclusion: Determination of IGFs and IGFBPsserum levels can be applied for diagnostics and prognosis of various cancers. And since IGF signaling system components are potential targets for molecular-targeted therapy, these markers can also be considered as possible predictors of the sensitivity to such agents.

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Clinical significance of receptor epidermal growth factors in malignant tumors

O.I. Kostyleva, E.S. Gershtein, R.R. Azigova, D.E. GaldavaN.N.Blokhin Russian Cancer Research Center, Moscow

The review of literature data and the own results of investigations of EGFR and their ligand significance in malignancies pathogenesis and metastasizing and of their relationships with clinico-morphological characteristics, and also of their influence on general and relapse free survival in patients with different cancers are presented. 598 patients with malignant and benign tumours were investigated: 291 breast cancer patients, 44 ovarian cancer and 12 benign ovarian tumours, 58 endometrial cancers and 31 patients with hyperplastic endometrial processes, and also 115 patients with bone tumours and tumour-like bone lesions. A general tendency to increasing of EGFR+ tumours phenotype with the growth of malignancy grade was found in different cancers. Complex investigation of EGFR and estrogen and progesterone receptors was recommended to breast cancer patients, because auto/paracrine tumour phenotype is reliable factor of poor prognosis for breast cancer (p<0.03). And also EGFR and their ligands are significant prognosis factors in non small cell lung cancer. EGFR investigation is important for successful antiEGFR target therapy.

Development of technology of manufacturing polymeric microparticles for peptides encapsulation

T.A. Lyovina, Ph.D. S.P. Krechetov, G.O. NifontovaMoscow Institute of Physics and Technology (State University), Dolgoprudny, Russia

A large number of drugs wildly used in clinical practice are proteins (hormones, antibodies, vaccines) and their biological activity strongly depends on the stability of the secondary, tertiary and quaternary structures. One of the promising directions in drug delivery system development is polymeric microparticles usage. Due to their structure microparticles provide a modified (prolonged) drug release and drug protection from pH, temperature and other aggressive microenvironment factors. Therefore, it is reasonably to use polymeric microparticles to achieve prolonged release and peptides stability.

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The aim of this study was to evaluate polymer structure influence on peptides encapsulation and release characteristics from microparticles.

Copolymers of lactic and glycolic acid (PLGA) 50:50, 65:45, 75:35 were used in microparticles preparation process. Bovine serum albumin (BCA) was used as a model of peptide. Microparticles were prepared by double emulsion technique w/o/w. Copolymer solution in methylene chloride was used as primary disperse medium, as secondary disperse medium – water solution of polyvinyl alcohol and methyl cellulose. Primary emulsion was obtained by dispersing the solution of BSA in the primary disperse medium. Water cooled to 4ºС was used for particles maturation. Washed and precipitated microparticles were lyophilized.

Microparticles size and morphology were studied with optical and confocal microscopy. For protein encapsulation evaluation polymeric microparticles were dissolved in dimethylsulfoxide with subsequent polymer hydrolysis in sodium hydroxide solution. BSA release was carried out in PBS buffer medium at room temperature. The samples were taken up to 40 days. Lowry assay was used for determining the protein quantity in the sample solutions.

The prepared microparticles size range was approximately 15-50 µm, they characterized by round shape and granular structure accordingly to the optical microscopy assay. The maximal encapsulation (about 30%) of BSA was revealed in PLGA 50:50 based microparticles. The other PLGA based microparticles showed lower BSA encapsulation and release rate. High protein release rate from the obtained microparticles was observed in the first days (up to 40%). Further constant-rate protein release without complete release was determined within 40 days.

The obtained results showed that PLGA 50:50 based microparticles to be effective for achieve prolonged release of BSA and other peptides.

Study of extracellular matrix proteins in metastatic breast cancer

Valeria Lobanova1, Vladislav Kononchuk2

1Novosibirsk State University, Novosibirsk, Russia2The Institute of Molecular Biology and Biophysics, Novosibirsk, Russia

The ability of breast cancer (BC) to develop metastases is the major cause of the high mortality from this disease. To date there is no clear explanation why positive for estrogen and progesterone receptors tumors metastasize predominantly to bone, negative for these receptors tumors metastasize predominantly to lungs, positive for Her2/neu tumors metastasize to the liver

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and brain. The possibility of predicting the development of metastases in distant organs is of great clinical importance due to the emergence of effective therapies. In recent times, there is an active search for molecular markers of metastasis. This process directly depends on the interaction of tumor cells with extracellular matrix proteins. Integrins are one of those proteins. The purpose of this study was to evaluate the role of extracellular matrix proteins (integrins) in the metastasis development in BC patients. Subunits of integrins αν, α2, α6, β1 were investigated by immunohistochemical method in samples of tissue of the primary tumor, normal tissue and tissue of lymph nodes with metastases. It is shown that the overexpression of these integrins in breast cancer results in statistically significant development of metastases in the lymph nodes. Thus, the opportunity is shown to highlight a group at high risk of developing metastases. This study is supported by the Russian Science Foundation (grant №15-15-30012)

Integrins and their ligands osteopontin and thrombospondin-1, can be considered as biomarkers

of aggressive papillary thyroid cancer phenotype

G.P. Logacheva1,3, L.A. Mostovich2, S.V. Svyatchenko1, S.P. Shevchenko1,3, L.F. Gulyaeva1,2

1Novosibirsk State University2Institute of Molecular Biology and Biophysics

3Novosibirsk Clinical Hospital № 1

IntroduсtionPapillary thyroid cancer (PTC) is the most common malignancy of the

endocrine system. The most frequent genetic alteration in PTC is V600E mutation in BRAF gene which leads to changes in the activity of intracellular signaling pathways. As a result, expression levels of cell membrane integrin receptors and their ligands - extracellular matrix proteins - osteopontin (OPN) and thrombospondin-1 (TSP1) change. Such changes promote migration, invasion and metastasis of tumor cells. Thus, integrin receptors and their ligands are potential biomarkers of an aggressive PTC phenotype. The aim of our study was to compare the gene expression profile of integrins ITGA2, ITGA3, ITGAV, ITGA6, ITGA9, ITGB1, ITGB3 and their ligands OPNa, OPNb, and TSP1 in PTC with different BRAF V600E mutation status.

MethodsIntraoperative thyroid tissue samples from 41 patients diagnosed with

papillary thyroid carcinoma (n = 26), diffuse nodular nontoxic goiter (n = 10) and follicular adenoma (n = 5) were analyzed to evaluate the expression levels

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of the investigated genes that had been determined by real time RT-PCR. Fluorescent immunohistochemistry (IHC) was used to confirm the PCR results and to estimate the amount of protein products. For IHC frozen sections were used. The presence of BRAF V600E mutation was identified using allele-specific amplification. Nonparametric criteria (Kruskal Wallis, Wilcoxon and Mann-Whitney tests) were used to evaluate group differences. P values of less than 0,05 were considered as statistically significant.

ResultsAn increase of gene expression level of ITGA3 (2,9-fold, p = 0,014), ITGAV

(1,9-fold, p = 0,038), ITGB1 (1,7-fold, p = 0,026), OPNb (2,5-fold, p = 0,0001) and TSP1 (3,2-fold, p = 0,017) was identified in PTC tissue, and high gene expression level of OPNb (5,9-fold, p = 0,003) and TSP1 (12,1 раз, p = 0,005) was identified in tissue samples of lymph node metastases compared with conventionally normal tissue. In samples with a widespread process (T3, T4, TNM) expression levels of ITGA3, ITGA6 and ITGA9 were higher compared to samples T1. Expression levels of ITGA3 and ITGAV were significantly higher in PTC BRAF V600E positive samples than in samples that were BRAF V600E negative. In compare with benign tumors increased levels of gene expression of OPNa (11,4-fold, p= 0,0112), OPNb (10,2-fold, p= 0,0216) and TSP1 (33,5-fold, p= 0,0005) were identified in follicular adenoma compared with PTC tissue. For ITGA2 and ITGB3 there was a significant increase of expression in PTC tissue compared with benign thyroid tumors (8,9-fold, p=0,019 and 38,4-fold, p=0,014, respectively.

Identified changes in expression levels of the studied genes indicate their potential role in tumor progression, and the possible impact on their expression of the mutant product of gene BRAF. Integrins and their ligands OPN and TSP1 can be considered as potential markers in determining prognosis and treatment of PTC.

Biochemical markers in neuroendocrine tumors

N.V. Lyubimova, M.G. Toms, T.K. Churikova, T.Yu. KharitidyN.N. Blokhin Russian Cancer Research Center, Moscow

Introduction. Current diagnosis of neuroendocrine tumors (NETs) is based on a determination of substances produced by the tumor cells (peptides, amines, hormones).

Purpose of the study. Estimation of diagnostic efficiency of chromogranin A (CgA), serotonin and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) as biochemical markers of NETs.

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Materials and methods. Determination of CgA and serotonin in blood serum and 5-HIAA in the 24h urine was performed in 330 patients with NETs of lung, pancreas, stomach, small bowel, colon, and in 115 healthy men and women with enzyme immunoassay test systems «Chromogranin A ELISA kit» (Dako A/S, Denmark), «Serotonin ELISA» and 5-HIAA ELISA» (IBL International GMBH, Germany).

Results and discussion. CgA levels in all groups of NETs were significantly (p<0.000001) higher than the corresponding value of control group, the elevation of serotonin and 5-HIAA were significant in all groups except gastric NETs. The secretion of biomarkers were significantly higher in patients with hepatic metastases and carcinoid syndrome compared to patients without these clinical signs. The diagnostic significance of CgA, serotonin and 5-HIAA was evaluated according to the cutoff values (33 ng/ml, 320 ng/ml, 60 µmol/day, respectively). Diagnostic sensitivity of CgA in whole group of patients was 80.6% (specificity 98.5%). Serotonin and 5-HIAA shows comparable diagnostic sensitivity only in patients with carcinoid syndrome (72.5% and 60.3%).

Conclusion. The data confirm the high efficiency of CgA as a marker of NETs, which improves the accuracy of diagnosis and tumor progression. Serotonin and 5-HIAA are markers of the carcinoid syndrome.

Neurospecific proteins as biochemical markers of brain tumours

N.V. Lyubimova, Yu.S. Timofeev, A.A. Mitrofanov, A.Kh. BekyashevN.N. Blokhin Russian Cancer Research Center, Moscow

Introduction. Differential diagnosis of primary and metastatic brain tumors remain one of the actual problem of neurooncology. Magnetic resonance imaging and computed tomography are not enough sensitive and specific for the differentiation of brain tumors of different etiology. Thereby there is a necessity to make a search for the additional noninvasive biochemical markers, such as neurospecific proteins (GFAP, S-100).

Purpose of the study. Comparative evaluation of GFAP and S100 in blood serum of patients with primary and metastatic brain tumors, those with neurological nontumor diseases, and practically healthy persons.

Materials and methods. Neurospecific proteins were measured in blood serum of neurooncologic patients (n=175), patients with brain pathology of non-oncologic origin (neurodegenerative and cerebrovascular diseases; n=38) and healthy people (n=69). The evaluation of GFAP and S100 concentrations

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were performed by plate format ELISA kits («BioVendor» and «Fujirebio») based on highly specific monoclonal antibodies to this protein.

Results and discussion. The significant elevation of GFAP and S-100 was revealed in glioblastoma (G IV) patients compare to the patients with anaplastic astrocytoma (G III), benign meningioma (G I), cerebral metastasis and healthy controls. The concentration of S-100 in blood serum of patients with anaplastic astrocytoma, benign meningioma, and cerebral metastasis did not significantly differ among themselves, and in relation to the control group there was a significantly increase only in patients with cerebral metastasis. GFAP was characterized by high frequency of its detection in patients with glioblastoma (83%) compare to other groups of patients and healthy donors, in which it was practically undetectable. Our results may suggest the correlation between increased GFAP serum levels and disturbance of blood-brain barrier during the development of malignant brain neoplasms.

Conclusion. These data suggest the possibility of using GFAP as a marker of glioblastoma and S-100 – as an additional biochemical criteria of cerebral lesions in oncology patients.

Study of internalization of PLGA nanoparticles into the intracranial rat C6 glioma: influence of surfactant coating

Yulia Malinovskaya1, Svetlana Gelperina1, Pavel Melnikov2, Nadezhda Osipova1, Olga Maksimenko1, Vladimir Baklaushev2,

1 Drugs Technology Ltd, Moscow, Russia2 Pirogov Medical University, Moscow, Russia

Introduction: Our previous results have shown that PLGA nanoparticles (PLGA NPs) coated with poloxamer 188 (P188) enable the delivery of drugs across the blood–brain barrier (BBB) after intravenous injection. Doxorubicin loaded PLGA NPs (Dox-PLGA) coated with P188 produced a considerable anti-tumour effect against the intracranial glioblastoma in rats. The objective of the present study was to evaluate the uptake of the P188-coated PLGA NP in the intracranial C6 glioma in rats.

Experimental methods: Doxorubicin-loaded PLGA NPs were prepared by a double emulsion solvent evaporation technique. For visualization using scanning laser confocal microscopy (SLCM) and an intravital fluorescence imaging system Ivis®Spectrum CT (Perkin-Elmer), the NP were labeled with DiI (DiI-PLGA NP). The DiI-PLGA NP were administered i.v. into rats with intracranial C6 glioma on day 15 after tumour inoculation. The presence of mass lesion

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was verified by previous MRI. Two hours after administration of the NP, the rats were perfused transcardially with p-formaldehyde, organs were recovered, and the fluorescence intensity was assessed using an Ivis Spectrum CT system. To assess NP localization in brain sections immunohistochemical staining with antibodies against GFAP (astroglial marker), beta-III Tubulin (neuronal marker), was performed.

Results: The fluorescence intensity of the hemisphere with the implanted glioma was > 4-fold higher for the P188-coated NP (DiI-PLGA/P188 NP), as compared to the uncoated NP (45.1×106 vs 9.5×106 photons/sec/cm2, respectively. The quantitative fluorescence analysis of the tumor sections using SLCM also showed a significantly higher accumulation of the DiI-PLGA/P188 NP, as compared to the uncoated DiI-PLGA NP. Mean fluorescence intensity values in the tumor were 1698.9±536.6 and 558.9±181.0 CU for the P188-coated and uncoated NP, respectively. The intensity values in the contralateral hemispheres for the same preparations were 293.4 ± 32.3 and 203.2 ± 22.9 CU, respectively. Thus, according to the SLCM data, the penetration of the DiI-PLGA/P188 NP into the tumor was 3 times more effective than that of the uncoated NP. The analysis of the magnified fluorescence images showed considerable accumulation of the DiI-PLGA/P188 NP both in the tumor interstitial fluid and inside the C6 glioma cells. At the same time, DiI-PLGA-NPs were mainly localized in epithelial cells of cerebral microvessels of the contralateral hemisphere. relatively intense DiI fluorescence was also observed on Purkinje cells of cerebellar cortex.

Conclusion: Together with the data obtained previously, the results of the present study demonstrate that coating of the PLGA nanoparticles with poloxamer 188 considerably enhances their delivery to the brain tumor.

Normacor® — a new cardioplegic solution to conduct open heart surgeries at normal body temperature

Nikolay Merkin, ChemRar RnD, Khimki, Moscow region, Russia

CardioSystemPharma, a ChemRar group company, has developed a new

cardioplegic solution Normacor® to conduct open heart surgeries at normal body temperature. Normacor® is expected to be launched to the Russian pharmaceutical market soon. Normacor® solution allows to prolong time of the open heart surgeries require stopping the heart during blood cardioplegia under normothermic conditions. Normacor® is the most effective myocardial

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protection system in the world according to the results of efficiency, safety and convenient of the practical use. The composition of cardioplegic solution Normacor® is protected by the patent of the Russian Federation.

Cardioplegia is temporary cardiac arrest caused by an injection of pharmacological drug, electric current or subsequent selective hypothermia. It is used in open heart surgeries in conditions of cardiopulmonary bypass. Cardioplegia can be classified into two types as hypothermia (with decrease of temperature) or normothermia (at normal temperature). Normothermic blood cardioplegia can reduce the mortality rate among patients during open heart surgeries and in postoperative period without changing tactical approach. Also normothermic cardioplegia can reduce the risk of postoperative myocardial infarction and brain damages and can expand indications for surgery. This simplified technique of open heart surgeries is safer and easily reproducible. Use of cardioplegia in recent years has undoubtedly improved the prognosis of a number of patients undergoing surgical correction of complex cardiac lesions. Normacor® excludes myocardial ischemia-reperfusion injury that allows rendering the greater number of patients and improves the quality of treatment.

Metalloproteinases, MMP tissue inhibitors, and VEGF in blood serum of endometrial cancer patients as potential diagnostic markers

Mushtenko S.V.1, Korotkova E.A.2, Tereshkina I.V.1, Dvorova E.K.2, Gershtein E.S.1,2

1A.I.Evdokimov Moscow State University of Medicine and Dentistry2N.N.Blokhin Russian Cancer Research Center, Moscow

Introduction: Metalloproteinases (MMPs) and MMPs’ tissue inhibitors (TIMPs) play a crucial role in cancer invasion and metastasizing. MMPs and TIMPs were also shown to actively interact with VEGF in the regulation of neoangiogenesis playing an important role in human endometrium both in normal conditions, and pathology.

Study Aim: To evaluate the relationship of MMP-2, 7, 9, TIMP-1, 2, and VEGF concentrations in blood serum of endometrial cancer patients with major clinical and morphological features in order to reveal clinically important biological markers.

Method: The study group enclosed 103 endometrial cancer patients, and 104 age-matched practically healthy women. MMPs 2, 7, 9, TIMPs 1, 2 and VEGF levels were measured in pre-treatment blood serum by standard ELISA kits (R&D, USA).

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Results: Highly significant (p<0.0001) increase of serum MMP-7, MMP-9 and TIMP-1 was revealed in blood sera of endometrial cancer patients as compared to control healthy women. There was practically no difference in serum VEGF between endometrial cancer patients and control group, and TIMP-2 level was only slightly increased (p<0.05). Most prominent changes were found for MMP-2, but its serum level, on the contrary to other markers, was drastically decreased (medians 31.3 and 586 ng/ml respectively; p<0.0001). Thus, decrease of MMP-2 in blood serum proved to be the most informative diagnostic marker for endometrial cancer: its sensitivity at cut-off level 90 ng/ml determined by ROC curve comprised 98.7%, specificity in relation to healthy control – 95.4%. For comparison, sensitivity of MMP-7 at cut-off 4.0 ng/ml corresponded to 82.5%, specificity – to 76.8%; MMP-9 and TIMP-1 had even worse diagnostic characteristics. The levels and diagnostic characteristics of the markers did not significantly depend on disease stage. Meanwhile, significantly lowest MMP-2 and highest MMP-9 levels were found in patients with poorly differentiated endometrial cancer.

Conclusion: MMP-2 can be regarded as a promising serological marker of endometrial cancer requiring further thorough evaluation.

Сlinical significance of VEGF and uPA evaluation in early stages breast cancer

Ovchinnikova L.K., Kostyleva O.I., Alekseeva E.I., Filippova M.A.N.N. Blokhin Russian Cancer Research Center, Moscow

Introduction. The tumor neoangogenesis is an attractive topic of oncology because it is known that tumor growth and development is associated with concomitant forming of mycrovessels, providing cellular breathing and feeding. Vasculoendothelial growth factor (VEGF) is one of the important neoangiogenesis activators, stimulating endotheliocytes proliferation and migration. Urokinase type plasminogen activator (uPA) plays a key role in process of tumor invasion, contributing to the extracellular matrix degradation and to easier endotheliocytes migration.

The aim of study – Evaluation of serum VEGF levels and tumor cytosols uPA levels in early stages breast cancer patients, and the investigation of their associations with basic clinico-morphological characteristics of breast cancer.

Materials and methods. In the study 248 breast cancer patients and 55 healthy women were accepted. The blood serum content of VEGF was determined by EIA standarts kits “Quantikine human VAGF” (“R&D Systems

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Inc.”, USA). The uPA level in cytosol fraction of tumor was evaluated by EIA reactives, created by Dr. T.Benraad (Nymegen, Netherlands).

Results. The reliable association between VEGF and uPA levels and stage of disease or malignancies degree was found by the multivariate analysis. Also, association between serum VEGF and tumor cytosols uPA levels in T1 and T2 stages was shown in general group of patients (r=0,67; p=0,001), this correlation was more expressed in lobular infiltrative breast cancer (r=0,92;p=0,001, and significantly less expressed in ductal infiltrative breast cancer (r=0,49; p=0,01).

Conclusions. The data obtained support the hypothesis about engagement of VEGF-depending neoangiogenesis and also about its association with urokinase type system plasminogen activation in early stages breast cancer.

Protein regulators of epithelial-mesenchymal transition and some components of the VEGF signaling pathway

in breast cancerpatients

O.G. Ovsii1, A.M. Scherbakov2, G.P. Guens1, V.D. Ermilova2, N.E. Kushlinskii21Moscow State University of Medicine and Dentistry named after A.I. Evdokimov

2N.N. Blokhin Russian Cancer Research Center, Moscow

Introduction. Epithelial–mesenchymal transition (EMT) – complex process of changing of cell phenotype from epithelial to mesenchymal. EMT is a sequence of programmed morphogenetic events, that starts in response to stimulation of a number of signal pathways, including activation of Snail transcription factors - SNAI1 and SNAI2/SLUG, and also including Zeb (ZEB1 иZEB2) proteins, E47 proteins of bHLH and KLF-8 protein. EMT is considered as a key stage of tumor progression; and capacity of tumor cellsfor invasion and metastatic propensity depend largely on EMT. Tests in vitro, carried out on breast cancer cell lines with different receptor status, revealed inverse relationship between SNAI1 content and ERα level; SNAI1 has protective properties and protects breast cancer cells against hypoxic conditions.

Aim of the study. Complex immunohistochemical evaluation of the level of SNAI1, SNAI2, NF-κB (one of Snail coactivators)proteins expression, VEGF and its receptors of the first and second orders in tumors of breast cancer patients.

Material and methods. 157 breast cancer patients were examined (112 of them have ductal infiltrative cancer, 23 – lobular breast cancer, 11- tubular breast cancer; other breast cancer types are presented as single observations). Most of tumors (111 specimens) had II G., 25 specimens had III G., 19 specimens

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had I G. All the patients had treatment in N.N. Blokhin Russian Cancer Research Centersince 2008 till 2013. Expression of Snail proteins-regulators, their NF-κBcoactivator and VEGF-signal pathway components (VEGF and its 2 receptors) of EMT was analyzed by immunohistochemical method. Statistical data processing was done in Statistica 6.0 - “StatSoft” programm.

Results. In most tumors (54%) SNAI1 expression is not detected. There are about 65% of high and middle SNAI1 expression tumors. The expression level of SNAI1 and SNAI2 and their NF-κBcoactivator have a significant positive correlation(p<0.05). The expression level of SNAI1 and SNAI2 has significant and positive relationship with the expression level of VEGFR1 (p<0.05). Correlation relationship of VEGFR2 expression level and SNAI1, SNAI2 expression level has expressed positive character (p<0.05). VEGFR2 and SNAI2 coexpression is detectedin 105 cases. SNAI1- andSNAI2-positivetumor incidenceis approximately the same in A and B luminal types of breast cancer. Incidence of SNAI1-positive tumors, NF-κB expression, VEGF and its receptors is decreasing to the complete absence in the case of triple negative breast cancer(p<0.01). There are 94% of cases of SNAI2-positive breast cancer tumors of SNAI2 expression detected in the case of triple negative breast cancer.

Conclusion. Thus, coordinated activation of proteins-regulators of EMT and of main components of VEGF-signal pathways is noticed in breast cancer tumors. SNAI2 protein is one of the main factors, supporting the growth and progression of breast cancer, including triple negative breast cancer. Investigations and development of methods of therapy, aimed to suppression of SNAI1 and SNAI2 proteins in malignant tumors of breast cancer, are very promising.

Epigenetic mechanisms of expression deregulation of pro-apoptotic APAF1 and DAPK1 genes in breast cancer

E.A. Pereyaslova1, V.I. loginov1, I.V. pronina1,2, A.M. burdennyy1, T.P. Kazubskaya3, N.E. Kushlinskii3, E.A. Braga1

1Institute of General Pathology and Pathophysiology, Moscow, Russia 2Research Center of Medical Genetics, Moscow, Russia

3N.N. Blokhin Russian Cancer Research Center, Moscow, Russia

Introduction. The epigenetic mechanisms including methylation of promoter CpG islands and microRNA (miRNA) interaction with mRNA of target genes are involved in deregulation of genes in tumors. APAF1 and DAPK1 genes are known as positive and BCL2 as negative regulators of apoptosis. The involvement of epigenetic mechanisms in expression deregulation of these genes in tumors needs to be clarified.

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Aim of the study. Our study was aimed to reveal a possible role of methylation and miRNAs in expression deregulation of pro-apoptotic APAF1, DAPK1 and anti-apoptotic BCL2 genes in breast carcinomas.

Material and methods. A number of databases, for example: miRWalk 2.0 (http://www.ma.uni-heidelberg.de/apps/zmf/mirwalk/), TargetScan 7.0 (http://www.targetscan.org/) etc. were applied to search for miRNAs which are potentially involved in the expression deregulation of the DAPK1, APAF1, and BCL2 genes. The set of 30 paired (tumor/normal) samples of breast carcinoma from patients not exposed to prior radiation or chemotherapy was used. Only samples with tumor cell content of at least 70% were selected. The methylation profiles of CpG islands of APAF1, DAPK1, and BCL2 genes were analyzed using methylation specific PCR. The mRNA level alterations were evaluated using quantitative SYBR Green real-time PCR with B2M as a reference gene. We used the certified real-time PCR kits of Applied Biosystems to analyze the expression level of miRNAs. RNU6B and RNU48 were applied as references. Fisher’s exact test and Spearman’s rank correlation coefficient were used for statistical analysis.

Results. We have shown that the frequency of methylation of APAF1 and DAPK1 genes was significantly higher in breast cancer tissues than in histologically normal tissues of the same patients (P ≤ 0.05 by Fisher’s exact test). We have identified statistically significant positive correlations between the expression down-regulation and hypermethylation for APAF1 and DAPK1 genes (Spearman’s correlation coefficients: Rs = 0.55, P = 0.04 and Rs = 0.57, P = 0.03, respectively). Comparative analysis of the data on expression alterations of 3 protein-coding genes (DAPK1, APAF1, and BCL2) and a group of miRNA genes, which were predicted as potential regulators by miRWalk 2.0, revealed the negative correlation between expression levels for miR-203а and APAF1 gene (Rs = -0.50, P = 0.05).

Conclusion. Our data suggest an important role of methylation in down-regulation of pro-apoptotic APAF1 and DAPK1 genes in breast cancer. In addition, we suggest APAF1 mRNA as a potential target of miR-203а. This knowledge is important to clarify the mechanisms of deregulation of pro-apoptotic APAF1 and DAPK1 genes in breast cancer.

This work was financially supported by grant 14-15-00654 from the Russian Science Foundation.

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Bioinformatic tools passing through the nigth of neurodegenerative disorders

Anna Petuhova, Irina Zhegalova, Sergey SuchkovI.M.Sechenov First Moscow State Medical University

Due to the development of science and technology medicine has made a big step forward, but it is still too early to talk about success in some areas. One of these is the area of neurodegenerative disorders. According to the World Health Organization and Alzheimer’s Disease International, over 46 million people worldwide are living with dementia, and this number will have risen to 131.5 million by 2050. In just three years’ time, the global economic cost of dementia will reach US $ 1 trillion.

Every three seconds, someone in the world develops dementia and the most of the patients with dementia are people over 60 years old. Because of increasing number of eldery people in the world (currently, 16% of the European population is over 65, and this figure is expected to reach 25% by 2030), searching for methods of diagnostics and curing of neurodegenerative diseases is an actual social and economic problem standing behind medical and research community and is also a problem of scientific interest according to researching of aging mechanisms.

To solve this problem we have to extend our understanding of the whole range of reasons of neurodegenerative diseases. This approach suggests running of process of collecting of large amounts of patients biological data, data processing and interpretation that means using a powerful tool of bioinformatics. Variety of purposes can be achieved whith this multifunctional tool. We can validate genes that play the main role in the disease development or we can build a gene regulstory network which shows the gene expression products influence on the metabolic processes.

We can design the 3D-structure of potentional targets of the therapy or we can choose the molecule which is the most appropriative for the role of drug from the library of well-known compounds. Bioinformatic programs are becoming more comfortable, demonstrative and accurate, and such useful instruments like full genome and exome sequensing that were very expensive recently are becoming cheaper and cheaper year by year. All of these processes make bioinformatics, its decisions and achievements more available for a wide range of professionals and encourage the development of bioinformatics as a tool an as a science.

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Express way to assess biocompatibility of metal alloys and implants

Pilipenko P.N.Moscow Institute of Physics and Technology, Dolgoprudny

Metal alloys are among the most commonly used materials for making

various kinds of implants. The most important selection criterion of implants is the biocompatibility of implantable materials – absence of negative effects on the body. One implant biocompatibility parameters is the absence of inflammatory reactions caused by the activation of immune cells in experiments in vivo and in vitro.

It is known that an universal inducer of inflammatory reactions of the body, as well as signal chemotactic molecule for immune cell is hydrogen peroxide (H2O2). Peroxide is generated by the body’s cells under the influence of various physical and chemical factors, including, probably, contact with the surface of the foreign material. Molecular mechanisms of biological reactions of peroxide generation are poorly understood. In this work, we studied the H2O2 formation rate in contact with the surface of various metals in aqueous solutions containing molecular oxygen. We detected peroxide by the enhanced chemiluminescence system (luminol-p-iodophenol plus horseradish peroxidase). It is found, that when in contact with titanium a minimum quantity of H2O2 was generated, while in contact with copper the surface speed of H2O2 formation was about 300 times more. Titanium is well known for its biocompatibility, and copper when sutured under laboratory animals skin can simulate chronic inflammation.

In experiments on cultures of luminous bacteria E. coli strain is shown that the toxicity of copper metal indeed due to the formation of H2O2, because survival of these bacteria was significantly higher in the presence of horseradish peroxidase. This allowed us to conclude that the mechanism of the biocide (oligodynamic) action of copper and silver discovered by the Swiss botanist Carl Naegeli in 1893 is in the proportion to its ability to generate peroxide.

It is found that the rate of formation of H2O2 depends on the oxygen concentration and pH, as well as on the degree of passivation of a metal surface oxide film. The rate of formation of H2O2 in presence of phosphate ions in solution has a peak at physiological pH values.

We proposed mechanism of H2O2 and other reactive oxygen species formation at the interface metal-solution. An experimental method of estimating the biocompatibility of implants and metal alloys as well as the device for realization of this method were described. Proposed solutions are easy to implement and should greatly accelerate research in search of new metal alloys for medicine, including work to improve their surface modification technologies, as well as fabricated metal implants testing procedures.

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Unsupervised clustering of cancer samples based on expression profiles

Rozenwald MichalStudent of Applied Mathematics and Informatics Faculty

of Computer Science Higher School of Economics

BackgroundTranscriptomic gene signatures are a powerful tool of modern

personalized medicine that is utilized for diagnostic, prognostic and predictive purposes. In particular, several gene signatures (e.g., OncotypeDX [1], MammaPrint [2]) are already routinely used in clinics for the prognosis of recurrence for ER-positive breast cancer patients and for the making a decision about chemotherapy prescription.

Prognostic and predictive gene signatures are standardly designed using methods of computer science. Generally these are supervised methods of machine learning: centroid methods [3], support vector machines [4], analysis correlations with the outcomes [1,2], etc. We decided to check whether unsupervised clustering can bring insights into the construction of prognostic gene signatures.

MethodsPublically available microarray dataset GSE17705 (“Validation dataset of

298 ER-positive patients treated with tamoxifen for 5 years” [5]) was utilized. K-means clustering [6] was selected as a method of unsupervised clustering.

Clustering pipeline used a combination of R, Python and C++ scripts.Clustering was applied to the whole list of transcripts (more than 20 000)

and to a list of key transcripts of OncotypeDX gene signature. In both cases four clusters were constructed.

For these clusters proportions of two phenotypic characteristics were analyzed: 5-years recurrence and nodal status.

ResultsFor the whole list of transcripts no correlation between clusters and

analyzed phenotypic characteristics was observed. Instead, strong correlation between these clusters and a lab that processed and profiled tissue samples was detected. For a list of key transcripts of OncotypeDX this correlation was not detected. However, correlation with the analyzed phenotypic characteristics was still poor in spite of the fact that supervised methods provided a construction of a reliable recurrence prediction classifier for this gene signature.

ConclusionUnsupervised clustering of samples can be efficiently utilized for the

analysis of biological samples based on expression profiles: it is capable

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of revealing outliers, heterogeneities induced by certain phenotypical differences and discrepancies in processing and profiling procedure. However, unsupervised clustering can hardly be used for the construction of prognostic classifiers even in case of consideration of a list of pre-selected transcripts that definitely can be used for the construction of such a classifier.

References[1]Paik S. et al. A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N. Engl. J. Med., 2004, 351, 2817–2826.[2] Van ’t Veer L. J. et al. Gene expression profiling predicts clinical outcome of breast cancer. Nature, 2002, 415, 530–536.[3] Parker J. S. et al. Supervised risk predictor of breast cancer based on intrinsic subtypes. J. Clin. Oncol., 209, 27, 1160–1167.[4] Galatenko V.V. et al. Highly informative marker sets consisting of genes with low individual degree of differential expression. Sci Rep., 2015, 5: Article 14967.[5] Symmans, W.F. et al. Genomic index of sensitivity to endocrine therapy for breast cancer. J. Clin. Oncol., 2010, 28, 4111–4119.[6] Hartigan J. A., Wong J. A. Algorithm AS 136: a K-means clustering algorithm. Journal of the Royal Statistical Society. Series C (Applied Statistics),1979, 28(1), 100–108.

Applying NMR spectroscopy to investigation of spider venom’s pharmaceutical potential demonstrated

on 3D structure of the two – domain spider toxin OtTx1a

Romanovskaya Daria DmitrievnaMoscow Institute of Physics and Technology (State University)

Spider toxins are complicated mixtures of different compounds with a large differentiation of biological activity. Knowledge of toxin’s structure and molecular mechanisms of activity allows us to use their unique capacity in creating new antibiotics and analgesics.

In this work we studied OtTx1a — a protein from Oxyopes takobius spider venom. It consists of two distinct modules, separated by a short linker. N-terminal module (residues 1–40) possesses cytolitic activity against a large number of pathogens, whereas C - terminal domain (residues 50–100) exhibit neurotoxic activity.

We produced N-terminal fragment on a Syro I peptide synthesizer using Fmoc/t-butyl chemistry, C-terminal segment we expressed in E.coli with the use of Novagen pET system protocol with addition of 15N-labeled media. The structures of both N- and C-terminal modules were solved by the liquid-state NMR spectroscopy.

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In water OtTx1a — AMP, the N — terminal fragment, is disordered. After adding DPC micelles, which mimic membrane environment, the peptide changed its conformation to α-helix. In DPC micelles, N -terminal module consists of two amphiphilic α-helixes, connected by a short unstructured linker.

In addition, we determined the structure of a C-terminal domain, OtTx1a-ICK. Five S-S bonds of this module form classical cysteine knot motif. Two antiparallel β — sheets and sophisticated system of hydrogen bonds stabilize hydrophobic core of the molecule.

Our results prove the data received by circular dichroism spectroscopy and theoretical analysis of amino acid sequence of the protein and give an opportunity to speculate on molecular mechanisms of activity of modular antimicrobial peptides.

Peptidoglycan-recognition protein Tag-7/PGLYRP-1 modulate antibiotic-resistance and early steps of innate immune response

Slonova D.A.1,2*, Posvyatenko A.V.1,2,3, Sysolyatina E.V.4, Ermolaeva S.A.4, Gnuchev N.V.3, Georgiev G.P.3, Kibardin A.V.2,3, Larin S.S.1,2,3

1 Moscow Institute of Physics and Technology (State University); 2 Federal Research Center of Pediatric Hematology, Oncology and Immunology name of Dmitry

Rogachev, The Ministry of Health of the Russian Federation; 3 Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia;

4 Gamaleya Research Institute of Epidemiology and Microbiology, Moscow, Russia

Wide spreading of antimicrobial resistance becomes serious problem because of mortality caused by infection especially in hospitals. Peptidoglycan recognition proteins recognize conservative peptidoglycan pattern of bacteria cell wall and participate in induction of innate immune response. Here we propose, that Tag-7/PGLYRP-1 can protect host during early steps of infection.

We demonstrated the ability of the recombinant human Tag-7/PGLYRP-1 protein binds to cell wall of Gram-positive and Gram-negative bacteria. Tag-7/PGLYRP-1 did not display direct bactericidal and bacteriostatic activity and did not change growth dynamics of E. Coli at the used concentrations. However, increased sensitivity of bacteria to the antibiotics was observed at the presence of Tag-7/PGLYRP-1. Stimulation of phagocytosis of the intracellular pathogen L. monocytogenes by macrophage like cell line ANA-1 was demonstrated in the presence of Tag- 7/PGLYRP-1. Using in vitro infection model we showed that the presence of Tag-7/PGLYRP-1 inhibits the intracellular survival of L. monocytogenes, protecting eukaryotic cells from infection.

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Thus our results supports hypothesis concerning participation of Tag-7/PGLYRP-1 in host protection by decreasing intracellular infection and down regulation of antibiotic resistance.

This work is supported RFBR grant №15-04-07649.

Lipidome comparison of endometrial tissue by direct mass spectrometry and LC-MS/MS

N.L. Starodubtseva, V.V. Chagovets, A.S. Kononikhin, A.V. Borisova, A.V. Kozachenko, V.E. Frankevich, L.V. Adamyan

A novel aspectFirst performed a comprehensive lipidome comparison of endometrial

tissues using the method of direct mass spectrometry (MS) without prior sample preparation and the classical approach with a preliminary extraction of lipids and subsequent MS analysis with high resolution.

AbstractThe study aimed at developing strategy for early diagnosis and prognosis of

hyperproliferative gynecological pathologies (particularly endometriosis) based on lipidomic approaches and methods of direct MS profiling of the molecular composition of the samples. According to the World research foundation of endometriosis (WERF), one in ten women of reproductive age worldwide suffer from endometriosis (about 176 million women aged 17 to 49 years). While there has recently been a steady increase in the frequency of this pathology.

Comparison of lipid composition and ectopic and eutopic endometrium can not only help to clarify the reasons for the development of the disease and different severity of its course, but also in the future to be useful in the development of new methods and treatment strategy. In this regard, the determination of the differences in the lipid profile of endometriotic and endometrial heterotopias in combination with histological examination of these tissues, is one of the urgent tasks in clinical gynecology.

Created in the framework of the project analytical methodology will form a methodological basis of the technology platform for new molecular methods of assessing the proliferative potential of tissues with altered functionality on the basis of comparative mass spectrometric analysis of lipid profiles of tissues at different stages of pathological processes.

Materials and MethodsSamples of endometrial tissue (ectopic and eutopic endometrium) were

obtained during the laparoscopic surgery from 30 patients with endometriosis

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(III or IV endometriosis stage) in the Operative Gynecology department at the V. I. Kulakov Research Center for Obstetrics, Gynecology and Perinatology. Diagnosis (external genital endometriosis) was confirmed histologically. Each tissue sample was divided into three parts for pathological studies, direct mass spectrometry based on method EESI (liquid-lipid extraction of tissues with simultaneous ionization) and lipid extraction by Folch followed by HPLC-MS/MS analysis at Maxis Impact (Bruker, Germany).

Preliminary resultsTypical mass spectra of endometrial tissue were obtained by the direct

mass-spectrometry: a piece of tissue was placed directly into the ion source and tissue molecules extraction, desorption and ionization was performed at the same time. Since the main peaks in the mass spectra, obtained by direct mass spectrometry, correspond to lipids, we compared our direct method of analysis with standard analysis of lipid extracts (lipid extraction by Folch followed by HPLC-MS/MS analysis). It turned out that the resulting mass spectra correlated with each other.

Ectopic endometrium in comparison to eutopic was shown to contain the elevated levels of certain lipids belonging to four classes: phosphatidylcholine, phosphoethanolamine, sphingomyelin and triglycerides. Phosphatidylcholines and sphingomyelins can be proposed as potential biomarkers for endometriosis since these lipids are closely associated with the suppression of apoptosis by oxidative stress and cell malignancy.

Lipidomic analysis of endometrial tissue can give new information about the cellular mechanisms that cause the increased cellular growth being a main feature of aggressive form of endometriosis. This can give the opportunity to develop new treatments aimed at pathophysiological mechanisms.

This work was supported by project No. 16-14-00029 sponsored by the Russian Science Foundation, N.S. acknowledges the support of lipid identification part of the research by the grant project № МК-8484.2016.7 of the Ministry of Education of Russian Federation.

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Breast cancer molecular classification based on DNA methylation assessed by reduced representation bisulfite sequencing

A. Tanas1,5, E. V. Poddubskaya3, T. V. Kekeeva1, R. A. Kerimov3, I. D. Trotsenko4, E.B. Kuznetsova1,2, V. Strelnikov1,5, D. Zaletayev1,2,5

1Research Centre for Medical Genetics, Moscow, Russian Federation, 2I.M. Sechenov First Moscow State Medical University, Moscow, Russian Federation,

3N.N. Blokhin Russian Research Center for Oncology, Moscow, Russian Federation

We have developed a modification of the RRBS method applicable to assess methylation in large collections of DNA samples within biomedical research. Briefly, our modification further reduces the size of the representation to be sequenced simultaneously increasing the inner fraction of CpG islands and shores. By use of this approach, we have obtained RRBS results for 80 breast cancer (BC) samples, 6 BC cell lines and 10 samples of normal breast tissue. To date, this is the most comprehensive collection of RRBS data for a set of tumors of the same organ. Cluster analysis of this data distinguishes at least five molecular subtypes of breast tissues (cells) based on the CpG methylation. Specific subtypes are assigned to normal breast tissues, BC cell lines, triple negative and HER positive (indistinguishable based on the present dataset), and two independent groups of luminal BC regardless of their HER status. The most pronounced differences of the DNA methylation patterns are observed between normal breast tissues and BC cell lines, the latter presenting the highest CpG methylation levels within the samples, notwithstanding different immunoprofiles (LumA, LumB and triple negative).

One of the most striking findings is separation of the luminal tumors into two clusters differing by the density of CpG methylation, independently on HER status (LumA or LumB). This separation may reflect differences in the tumor biology between groups and requires further research in terms of utility for prognostication and treatment that may arise from the DNA methylation markers.

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The problem of findingthe cell of origin in glioma

Timin G.V.1, Tolkunova E.N.1, GulyaevD.A.2, Lahina Yu.S.2

1. Laboratory of molecular mechanisms of cell differentiation, Institute of Cytology, RAS, St. Petersburg, Russian Federation

2. Neurosurgery department, Almazov federal north-west medical research centre, St. Petersburg, Russian Federation

Gliomas are brain tumors of glial origin. They are divided into four groups due to histological similarity of tumor tissue with gliaependymomas, astrocytomas, oligodendrogliomas and mixed gliomas. However, this classification is not strongly related to the glioma origin because of glioma cells anaplasia, heterogeneity of tumor tissue and the presence of stem cells in adult brain.The cell of origin is the cell type that initiates tumor formation. There is mixed evidence as to whether or not the cells of origin are neural stem cells, progenitor cells or differentiated progeny.Therefore, the origins of glioma still remain not well understood. We approach the topic by analyzing tumor specimens and primary glioma cell culturesin the assumption thatcells of the mature tumor retain the molecular markersexpression profile of the cell of origin. Twenty specimens of human glioma tissue have been assayed with RT-PCR for the expression of molecular markers specific for various types of glial cells and neural stem cells (NeuN, MOG, MBP, NG2, Olig2, Vimentin, GFAP, Aldh1L1), stem cells markers (Oct4, C-Kit) and cancer stem cells marker (CD133). We found that the expression profiles of these markers are overlapped for different types of glioma. From three of these glioma specimens primary cell cultures were obtained and cultured in serum-containing media. They were enriched with the stem component using puromycin resistance vector under control of the Oct4 gene enhancer 2A2B. Enriched cell cultures were compared with primary cell culturesand tumor specimens via RT-PCR. To examine tumarigenic potential of the cells we stereotactically injected the cells into the ratus brains. We concluded that culturing of primary tumor cells in serum conditions leads to serious changes that make serum-based cultures inappropriate for the search of cells of origin in glioma.

The work was supported by the Russian Science Foundation grant14-50-00068.

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Insulin-like growth factors (IGF) and IGF-binding proteins in bone sarcoma patients

Yu.S. Timofeev, I.V. Boulytcheva, E.S. Gershtein, I.V. Babkina, Yu.N. Soloviev, M.D. AlievN.N.Blokhin Russian Cancer Research Center, Moscow

Introduction. Insulin-like growth factors (IGF) signaling system plays a major role in the development and progression of various sarcomas and other solid tumors, and therefore its components are regarded as promising diagnostic and prognostic tumor markers and targets for molecular oriented therapy.

Aim of the study. Evaluation of basal IGF-I and II, and IGF-binding proteins (IGFBP) serum levels prognostic significance in patients with various malignant bone tumors.

Materials and Methods. 25 osteosarcoma, 21 chondrosarcoma and 18 Ewing sarcoma patients followed up for 15-61 (median – 35) months with known pretreatment IGF-I, II, and type 1 and 3 IGFBPs serum levels measured with DSL Inc. (USA) ELISA kits were enclosed in the study.

Results. Increased IGF-I levels (>243 ng/ml) were associated with low relapse-free survival (RFS) in all bone sarcoma patients, and in those with chondrosarcoma. Overall survival (OS) depends on IGF-I level only in the total group. Patients with serum IGF-II exceeding 1.97 µg/ml also had less favorable RFS prognosis than those with lower levels of this marker. This association was most pronounced in Ewing sarcoma patients. IFG-II level also affected on OS indices in osteosarcoma. Significant RFS and OS decrease were found at high (>31 нг/мл) IGFBP-1 serum levels in the total group, osteosarcoma and Ewing sarcoma patients. And IGFBP-3 level >5.85 µg/ml was associated with worse RFS in osteosarcoma, and decreased OS both in the total group, and in osteosarcoma and Ewing sarcoma patients.

Conclusion. Unfavorable prognostic significance of high serum IGFs and IGFBPs concentrations both in the total group of bone sarcoma patients, and in various histological types of bone tumors was demonstrated. The value of individual IGF system components may change depending on tumor histological type.

Key words: IGF-I, IGF-II, IGFBP-1, IGFBP-3, osteosarcoma, chondrosarcoma, Ewing sarcoma, prognosis

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PE7/4 Safety and antiviral effect of Elpida (VM-1500), a novel NNRTI (+Truvada) in treatment-naïve HIV-1 infected patients

Irina Tyrnova, ChemRar RnD, Khimki, Moscow region, Russia

Viriom presented the results of a randomized, placebo-controlled, double-blind dose-finding clinical trial of Elpida® (VM-1500) in patients with HIV infection who are antiretroviral therapy-naïve at the EACS-2015 conference. Elpida is the pro-drug of the active compound VM-1500A, a potent, highly selective NNRTI.

The purpose of this clinical trials was to evaluate the impact of different dosage regimens of Elpida (VM-1500) in combination with drugs used for standard antiretroviral therapy (ART) versus a combination containing efavirenz (EFV) and standard ART (2 nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs/HtRTs)) on the efficiency of treatment in view of a reduction in viral load to an undetectable level (< 50 copies/ml) at 24 weeks in previously untreated HIV-1-infected patients. At this study stage (II), an interim analysis of the efficiency and safety of therapy was performed after 12-week therapy.

In treatment-naïve patients, Elpida 20 and 40 mg QD (with TDF/FTC) at week 12 demonstrated potent antiviral activity, comparable to EFV, and favourable safety/tolerability profile. Fewer drug-related AEs were observed for Elpida compared with EFV. The safety of 12-week ART regimens incorporating Elpida (VM-1500) was higher than that of the regimen containing EFV. Elpida (VM-1500) 20 mg daily was chosen to be further investigated.

Tissue-specific expression of microRNAs in rats under DDT and benzo(a)pyrene exposure

Dmitry Ushakov, Sergey FilippovThe Institute of Molecular Biology and Biophysics, Timakov 2/12, Novosibirsk, 630117, Russia

The information that was recently made available indicates that genotoxic carcinogens can take a toxic effect, not only directly destroying the DNA structure, but also affecting the mRNA expression by activating the corresponding CAR and AhR receptors, as well as estrogen receptors (Era and ERb). Benz(o)pyrene, a widely occurring genotoxic pollutant, can cause carcinogenesis, immunosuppression, teratogenicity, and endocrine-related

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effects. On the contrary, DDT, a high efficiency insecticide, does not show high toxicity, but can be accumulated both in tissues of animals and plants.

An in silico analysis of the rat genome was carried out to find the microRNA that can be regulated by АhR, CAR, and ER. Several microRNAs were found: miR-3577,-208a,-193b, expected to be CAR-activated. Thee miRs were found: miR-336, -207, 208b that were part of other genes with a DRE-element in promoter, that can be a coupling element of AhR benz(o)pyrene. The results have shown that all miRs, but for miR-208b, are expressed in the liver and the ovarian, to a lesser extent in the uterus and the mamma. These tissue-specific variations may be connected with specific expression characteristics of the “principal gene” (“host-gene”) that hosts the miR. In these organs the expression of target genes was determined for receptors ER (Cyclin D1), CAR (CYP2B/3A), and AhR (CYP1A/1B) when subjected to the induction of the xenobiotics under study. BP caused an increase in the expression of CYP1A and CYP1B in the liver 17.5 and 46.6 times, respectively. Correspondingly the DDT-induced expression of CAR-regulated genes CYP2B and CYP3A increased 992- and 57-fold. In case of BP induction an up-regulation of genes CYP2B1 and CYP3A1 was also observed by a factor of 12.5 and 34, respectively. These results justify that the considered xenobiotics activate AhR and CAR receptors, thus, considerably increasing the expression of their target genes. The expression of Cyclin D1 regulated by ER change neither under BP, nor under DDT, which validates the hypothesis of no effect low effect of xenobiotics on ER, whose target is Cyclin D1.

This work was supported by Russian Foundation for Basic Research № 15-03-01700 А.

Using the computational crystallomics to reconstruct full-atomic structure of short fibrin oligomers and protofibrils

Artem Zhmurov1, Rustem I. Litvinov2,3 Pavel Zhukov1, John W. Weisel2, and Valeri Barsegov1,4

1 Moscow Institute of Physics & Technology, Moscow region, 141700, Russian Federation 2Department of Cell & Developmental Biology, Perelman School of Medicine,

University of Pennsylvania, Philadelphia, PA 19104, USA 3Institute of Fundamental Medicine and Biology, Kazan Federal University,

Kazan 420012, Russian Federation 4Department of Chemistry, University of Massachusetts, Lowell, MA 01854, USА

Fibrin is a filamentous network formed in blood to stem bleeding. Polymerization starts when activated fibrinogen (fibrin), a rod-like plasma protein, self-associates to make protofibrils, critically important soluble intermediate oligomers with largely unknown structure and properties.

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Protofibrils self-associate laterally to form fibers arranged into an insoluble three-dimensional network. To study the mechanisms of fibrin formation, we developed a “computational crystallomics” approach to molecular modeling and employed 27 crystal structures of human fibrinogen and its fragments resolved to date to reconstruct, for the first time, double-stranded half- staggered fibrin oligomers up to a 19-mer protofobril. We incorporated missing flexible portions of fibrin and performed stepwise elongation beginning with end-to-end single-stranded constructs reinforced by covalent γ-γ crosslinking, which were then connected laterally via non-covalent A- a and B-b knob-hole bonds. The full-atomic structures of fibrin protofibrils with the αC regions, which were validated by quantitative comparison with experimental ultramicroscopic images of fibrin oligomers and protofibrils, reveal intermolecular interactions that stabilize a twisted arrangement. Quantitative physical characteristics of fibrin polymers are calculated and profiled for oligomers and protofibrils of different length.

A molecular dynamics study of the mechanical properties of α-helical coiled-coils

Kirill Minin1, Artem Zhmurov1 and Valeri Barsegov 1,2

1 Laboratory of Computer and Mathematical Modelling of Biological Systems, Moscow Institute of Physics and Technology, Dolgoprudny, Russia

2 Department of Chemistry, University of Massachusetts at Lowell, Lowell, MA, United States

Coiled coil is a very common motif of protein structure. It contains several (usually from two to five) α-helices, wrapped around one another to form a supercoil. Much experimental and computational efforts were made to investigate the mechanical properties of coiled coils, whose response to external tension has unique footprint: elastic at low strains, plastic at moderate and non-linear when the stress is high. We used molecular dynamics simulations to investigate the origin of the mechanical properties of the coiled coils and to compare these properties for coiled coils from several different proteins. We found that all the system show similar force-extension profiles with three distinct regimes. The Young modulus in the initial (elastic) regime is system dependent, has higher values for more regular systems and lower values for those having distortions in the coiled-coil motif, the height of the force plateau in the plastic regime does not depend on the micromolecular composition but only on the number of α-helices in the structure. We provide detailed mechanical characteristics of each system and show how the atomistic structure of the coiled coils is related to those properties.

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Doclinical research of GZK-111 – dipeptide cognitive enhancer

K.N. KolyasnikovaInstitute of Pharmacology named after V.V. Zakusov

Cyclo-(prolyl-glycine) (CPG) was discovered as an endogenous cyclic dipeptide in rat brain in 1996 (Gudasheva et al., 1996). It posesses nootropic (Gudasheva et al., 1999), anxiolytic (Gudasheva et al., 2001), antihypoxic (Koliasnikova et al., 2012) and neuroprotective (Povarnina et al., 2015) activities. Previous studies have demonstrated that CPG positively modulates AMPA receptor function and increases level of brain-derived neurotrophic factor (BDNF) in neuronal cell cultures SH-SY5Y and HT-22 in normal and pathological conditions (Gudasheva et al., 2016, in press). We suppose that CPG is an endogenous positive modulator of AMPA receptors. It is well known that positive AMPA receptor modulators (ampakines) affect brain physiology, networks and neurotrophins.

We designed and synthesized linear substituted dipeptide N-phenylacetyl-glycyl-L-proline ethyl ester (GZK-111) which can convert into CPG (Russian Patent Application 2016100425). CPG formation from GZK-111 in the presence of plasma and brain enzymes was shown in vitro. It was shown that GZK-111 possesses nootropic, neuroprotective, antihypoxic and anxiolytic activities inherent to CPG. Thus, GZK-111 can be considered as a prodrug of endogenous ampakine, and therefore can be the basis for development of a novel cognitive enhancer.

Assessment of genetic factors and chronotype for prediction of a bus driver’s road accident risk

Dorokhov V.B. 1, Puchkova A.N. 1, Taranov A.O. 1, Tupitsyna T.V. 2, Slominsky P.A. 2, Dementienko V.V. 3, Ermolayev V.V. 4

1– IHNA&NPh RAS, 2 – IMG RAS, 3- Neurocom , 4- MSPU (Moscow, Russia)e-mail: [email protected]

High working performance and low risk of mistakes is important for any occupation but especially crucial for professional drivers, who are responsible for their passengers’ and other members’ of traffic safety. Their alertness and level of attention depend on many factors, amongst which homeostatic mechanisms, circadian rhythms and sleep quality play an important role. All people differ in the

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preferred timing of sleep and activity, e.g. chronotype. “Owls” – late chronotypes, and “larks” – early chronotypes have different optimal performance periods. These differences have a significant genetic component. Due to individual differences in sleep, biological clock functions and sleep deprivation resistance a person may be better suited to work on a particular schedule, but this fact is often neglected. Thus appears the need to assess objective performance and sleep habits data and search for their genetic correlates.

The objective of this study is to assess the sleep parameters, chronotype, performance of professional bus drivers and search for their genetic correlates. This study has looked for possible connections between single nucleotide polymorphisms (SNP) in sleep, cognition and circadian-related genes, chronotypes and road accident history.

300 professional bus drivers (all male, ages 21-68, average 45,7) participated in the study, 188 of them had traffic accident records. The records included all the accidents in which the driver was involved for the period of his work in the coach company. The drivers worked on a rolling schedule with shifts starting at 3:30, 6:30, 9:30, 12:30, 15:30, 17:30. To assess the chronotype we have used a Russian version of Munich Chronotype Questionnaire (MCTQ) and a Sleep-Wake Pattern Assessment Questionnaire (SWPAQ). MCTQ gives the measures of average sleep length, midsleep time (chronotypes) and social jetlag (difference between sleeptimes on workdays and holidays). SWPAQ measures the scales of morning lateness (M, high score means low activation in the morning) and evening lateness (E, high score means high activation in the evening) and compliments the MCTQ data. SNPs were tested in: CLOCK, RORA, NPAS2, NPSR1, PER2, DRD3, SLC6A3, DBH, DRD2 genes.

Chronotype assessment of the drivers with MCTQ (6,9±1,2 h), and prominent social jetlag (average 1,6±1,4 h) caused by the shift work. As expected, midsleep time and social jetlag parameters were correlated. Unlike the general population, drivers exhibited both high negative and positive scores in social jetlag. SWPAQ has shown correlation between M and E scores and predomination of a mixed “owlark” type. There was a small but significant correlation between SNPs in CLOCK, RORA, and NPSR genes and chronotype parameters (mainly with social jetlag); and significant connection between SNPs in CLOCK, PER3, NPSR1, NPAS2, SLC6A3 and “guilty in the accident” parameter of road accident history.

Conclusions.Based on our results we can say that the interplay of genetic influences

of SNPs in chronotypes- and sleep-related genes and chronotypes themselves results in differences in such a high-order behavioral characteristic as driving safety and that assessment of these individual characteristics may be useful for making a safer and more optimized working schedule and better hiring decisions.

The study is supported by RFH grant 14-06-00963.

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Natural dipeptide carnosine: a promising neuroprotector and cognitive brain function corrector

Fedorova T.N., Stvolinsky S.L., Lopachev A.V., Berezhnoy D.S., Deviatov A.A., Konovalova E.V., Gavrilova S.A., Morozova M.M., Maximova M.Y., Illarioshkin S.N.

Research Center of Neurology, Volokolamskoye Shosse 80, Moscow, 125367, Russia

Oxidative stress (OS) is one of the leading pathogenic factors of neurodegenerative and vascular diseases of the brain, resulting in damage to the cell structure of the nervous tissue, the development of neurological deficits and reduced brain cognitive functions. Natural protectors of cells and tissues from OS are antioxidants, however currently the spectrum of antioxidant drugs with proven effectiveness used in complex treatment and prevention of CNS diseases is limited. In this regard, development of new antioxidant drugs with neuroprotective effects is an urgent task. The development of OS leads to functional and structural disorders in various biological components of the brain tissue, making drugs with antioxidant effects advisable in these conditions.

Among promising compounds is a natural neuropeptide carnosine (β-alanyl-L-histidine), with a wide range of biological properties, including those of a direct antioxidant, proton buffer, chelator for metals of variable valence, antiglycation agent, and immunostimulator. In various experimental models of CNS diseases characterized by oxidative brain damage such as immobilization and electrical-pain stress, acute hypobaric hypoxia, cerebral ischemia, hyperhomocysteinemia, and MPTP-induced parkinsonism it has been shown that carnosine prevents the development of oxidative damage to brain tissue, increasing the efficiency of the endogenous antioxidant system.

Neuroprotective effect of carnosine has been shown to reduce the ischemic brain damage zone in Wistar rats in a model of 72-hour irreversible focal ischemia and to preserve motor activity and long-term memory in animal subjected to testing in the open field, hole board, and Morris water maze tests. Potentiating effect of carnosine on rat memory and learning has been revealed during the formation of the conditioned reflex of active avoidance of negative reinforcement, paired with electrical-pain stress. Protective effect of carnosine has been shown in the original model of MPTP-induced parkinsonism in senescence-accelerated mice SAMP1, characterized by impaired antioxidant status. Carnosine prevented a decrease in motor and exploratory activity, the development of rigidity, as well as oxidative damage to the brain.

In pilot clinical studies it was shown that adding carnosine to the treatment regimen of patients with chronic cerebral ischemia and Parkinson’s

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disease enhanced the effectiveness of the treatment and was accompanied by a steady decline in neurologic symptoms, improvement of cognitive function of the brain, and restoration of antioxidant status.

Overall, ongoing research is aimed at creating new promising drugs with neuroprotective effect based on carnosine, characterized by the ability to cross the blood-brain barrier and demonstrate desired properties during targeted delivery to the brain.

A system for continuous monitoring and control of operator’s and driver’s functional conditions

Zakharchenko D.VNeurocognitive technologies, Moscow, Russia

The presentation shows the results of the first phase of the project to develop a system for continuous monitoring functional condition of operators, dispatchers, drivers and locomotive machinists. At the moment we have developed the main element of the diagnostic system – high-precision algorithm for the detection of falling asleep in real time.

According to the official statistics, annually about 25 thousand people dies in a car accidents in Russian Federation alone. Cause about 20% of all fatal accidents is the driver falling asleep at the wheel. In addition to direct losses associated with the death of workers, the companies-owners of vehicle have a significant economic cost: repair costs of vehicles, compensation for the value of the goods, legal and insurance expenses (including payment of treatment, medical insurance and disability pensions, a different kind of compensation). Such costs can be substantial and many times exceed the cost of the in-vehicle operator’s (driver’s) monitoring systems – especially in air transportation, railroad, and hazardous industries (chemical, energy, aerospace, or nuclear).

The system, developing by us, will monitor the functional status and level of human performance in real-time and, if necessary, can automatically maintain the required level through timely signal for the test of vigilance. This system will allow to the driver/dispatcher/operator to avoid falling asleep, using the automatically tracking condition, which cannot arbitrarily be controlled by a human.

The system includes:1. The device for registration of electroencephalographic information

(ergonomic EEG headset)2. Computing module (smartphone, tablet PC or car navigator)

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3. Software for the detection of falling asleep in real time.Our system is focused to the mass market. Having a low cost (about

$100 per device), it can be used both as a tool of personal security driver (for example, private passenger cars) and as part of a corporate system of personnel management (in transport, industry or energy companies).

The result of the project will be commercially security system for driver/dispatcher/locomotive machinist. This system will have a good prospects in both domestic and foreign markets. The implementation of this system will significantly reduce road deaths, improve the safety of transportation and reduce the costs, caused by accidents on transportation and hazardous industries.

This project received a high appraisal by experts of Fund «SKOLKOVO», as well as working groups «Neuronet» and «CoBrain». Currently filed applications for venture funding. We invite to cooperation partners and investors from Russia and foreign countries.

Efficiency of NGS in molecular diagnostics and CFTR Mutations Spectrum in Russian Cystic Fibrosis Patients

Ivanov MV1,*, Glazova OV1, Matsvay AD1, Krasovskiy SA2, Amelina EL2, Kovalenko SP1, Khafizov KF1,3

1 Moscow Institute of Physics and Technology2 Research Institute of Pulmonology

3 Central Research Institute of Epidemiology FBUNcontact e-mail: [email protected]

BackgroundCystic fibrosis (CF) is one of the most common, life threatening, autosomal

recessive genetic disorders. Remarkable developments in molecular analysis techniques have resulted in the identification of more than 1,500 disease-causing CFTR mutations, which complicates molecular diagnosis using conventional techniques.

MethodsWe conducted Next Generation Sequencing (NGS) on the set of 89 adult

patients that were prescreened for the CFTR mutations with “gold standard” techniques. CNV were detected employing MLPA. Clinical information was available for 82 patients.

ResultsAcross 89 patients diagnosed with CF, 48 unique mutations were

identified and 25 of them were detected in single cases. All detected with NGS

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pathogenic mutations were successfully validated with direct sequencing. Patients with mild genotype (n=36) were associated with lung form of the disease, while ones with severe genotype (n=46) - with mixed (p-value < 0.01). Diabetes mellitus was seen only in patients with severe genotype (22% vs 0%; p <0.01). At least two pathogenic mutations were detected for each patient. Nine patients harbored 3 potentially pathogenic mutations. Three of them had genotype [3849+10kbC>T, R668C; Phe508del], which was associated with lower patient weight (p=0.004). Complex genotypes with R0170Q and S466X in cis were associated with milder pulmonary disease (FVC 108% vs 67%). Four patients had F508del and L467F co-located on the same chromosome. Exone 2,3 deletions as well as 6,10 duplications could not be detected with NGS.

ConclusionNGS was more cost-efficient, scalable, workload reducing and information

gaining technology as compared to standard routine methods. Combining with the equivalent diagnostic performance compared to Sanger sequencing, we conclude that it can be implemented in clinical practice, although careful validation is required. Observed complex genotypes and genotype-phenotype correlations demonstrates opportunities of using NGS in patient prognosis. Though role of mutation in modifier genes remains NGS was more cost-efficient, scalable, workload reducing and information gaining technology as compared to standard routine methods. Combining with the equivalent diagnostic performance compared to Sanger sequencing, we conclude that it can be implemented in clinical practice, although careful validation is required. Observed complex genotypes and genotype-phenotype correlations demonstrates opportunities of using NGS in patient prognosis. Though role of mutation in modifier genes remains unknown. Study was funded by RFBR (№15-04-04730)

Study of life span capacity of new PI3K inhibitors on С. Elegans model

E. Marusich1, D. Ovcharenko1, E. Lashmanova1, A. Moskalev1, 2, S. Leonov1

1MIPT, Moscow Institute of Physics and Technology (State University), Moscow, Russia2 Institute of Biology, Komi Scientific Center, Russian Academy of Science, Syktyvkar, Russia

Every day 150 thousand people die on the earth, 75% among them die from the age-related diseases. Even in developing countries, these type of diseases causes up to 90% of death. Search of the novel drugs with high capacity of life extension plays a pivotal role in improvement of quality of

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human life ((healthcare). Caenorhabditis elegans nematode is very suitable and widely used biological model for screening of new chemical compounds with geroprotective properties.

In this study we tested the effect of specific inhibitors of phosphatidylinositol 3-kinase (PI3K) on lifespan of Caenorhabditis elegans. Considering the key regulatory role of PI3K in cell function, we also focused on demonstration the ability of PI3K inhibitors to interfere with cell aging. In order to assess the biological activity of the chemical compounds library we used a phenotypic screening based on high-tech platform analysis of the ImageXpress Micro XL High Content Screening System (Molecular Devices, USA).

We tested 62 new chemical compounds previously selected from the focused library composed by 3D Molecular docking ICM-Pro v. 7.2 program analysis of PI3K binding site.

Lifespan assay performed in liquid S-medium, at 20 °C in 96-well plates with synchronized worms in presence of E.Coli OP-50 and FUDR in presence of tested chemical at 10 μM concentration. Then, selected hits were tested in second round to determine dose-response curve using range of concentrations from 0.001 μM up to 10 μM. The number of live and dead animals monitored nematodes were monitored three time a week. The assay revealed two hits G5 and E7 that were able to extend nematodes’ lifespan by 28.3% and 20.8%, respectively.

Geroprotective properties of selected hits were retested in culture of senescent human embryonic lung fibroblasts cultured in the presence of these hits. The level of beta-galactosidase activity identified by staining with X-Gal (β-Galactosidase Staining Kit # 9860) was chosen as an evaluative criterion for aging. We demonstrated that 0.1 μM of G5 compound significantly reduced beta-galactosidase activity in human fibroblasts.

G5 and E7 hits were also tested for their impact on the accumulation of lipofuscin, a well-known marker of oxidative stress during aging, in the body of the nematodes. The experiment conducted during 30 days in 384-well plates, in liquid S-medium in the presence of tested compounds at 0.1 μM and 10 μM concentrations. These studies demonstrated the ability of G5 compound (at 0.1 μM) to decrease of lipofuscin accumulation in nematode’s body by 33 % (in average) compared with controls.

The presence of G5 compound (at 0.1 μM) extended the nematode’s lifespan by 32 % compared with controls in the stress-resistance test during 9 hours of incubation at 37 0C.

Thus, we have found a new chemical compound G5, which can be considered as a good candidate for further optimization step of potential drug candidate with strong geroprotector properties.

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Pollen-based High Content Screening Assay to Discover New Potential Plant Herbicides

Roman Chuprov – Netochin1, Jaroslav Neskorodov2, Elena Marusich1, Yana Mishutkina2, Polina Volynchuk1, Sergei Leonov1

1 Life Sciences Center, Moscow Institute of Physics and Technology, Dolgoprudny, Moscow region, Russia,

2 Center «Bioengineering», Russian Academy of Sciences, Moscow, Russia.

The resistance to existing herbicides among the weeds populations increased dramatically over past years and became a major threat to the sustainability and profitability of crop systems. Therefore, the discoveries of new herbicides turn out to be essential task for weed management. Due to technical difficulties and high costs involved in discovery and development of new herbicides, conventional old-fashion methods happen to be extremely inefficient, leading to discovery of one novel herbicide per screening of at least 10,000 chemical compounds. High Content Screening (HCS) is one of the innovative new approaches to assess biological effect of large number of new chemical compounds reducing the assay time by 4-5 times. This approach, based on a multi-parametric analysis of digital images obtained by automated optical systems allows quantitative and qualitative phenotypic screening of tested compounds in real time.

The goal of present study was to develop the biological assay model for screening and discovery of new herbicides using HCS platform in 384-well plates format, based on ImageXpress XL system. Mature tobacco (Nicotiana tabacum L.) pollen was chosen as a model system for initial screening of new herbicides from extended library of chemical compounds. This model has certain unique features, making it an ideal system to discover plant modulators: a) unlimited number of uniform population of pollen grains; b) high speed of germination (assay could be completed within 2 hours); c) experiments can be done at non-sterile condition and finally, d) almost 70% of plant genes are expressed in developing pollen. The last feature of the pollen system implies that any inhibitor, found to inhibit pollen germination, theoretically could also inhibit root germination, growth and elongation.

To obtain good reproducibility of the assay the harvesting time and density of pollen grain per well were optimized. It was found, that the pollen grains should be collected from freshly harvested mature, open tobacco flower buds, whereas the optimal density can be achieved at final concentration 0,2 mg/ml. Basic media for this pollen germination assay was GVH media.

Suspension of pollen in GVH medium and compounds were dispensed using a robotic system Biomek. To evaluate the effect of 1000 compounds on

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tobacco pollen germination and tube growth the HCS screening and analysis in 384-well plates was performed after 120 minutes of incubation of pollen at room temperature. We developed our own HCS analysis algorithm based on the assessment of the area of all pollen grain population per well, which allows measuring the inhibition efficacy of chemical compounds. Our assay demonstrated meaningful results (Z factor >0,4) with good robustness and reproducibility. The pollen suspension in GVH media containing 0.1% DMSO was used as positive control. Pollen suspended in GVH media containing 0.1% DMSO and 1 µM salicylic acid was served as a negative control.

The primary screening (10 µM) revealed 24 potential inhibitors (hits) of pollen germination and pollen tube. The dose-dependent (in the range of 0.01 – 50 µM) effect of hit compounds was further evaluated, and the IC50 values of inhibitory activity were revealed for only 7 compounds. The inhibitory potential of those 7 compounds was verified by their effect on Arabidopsis seed germination and root growth and they were confirmed as hits.

Our pollen-based HCS assay demonstrated several benefits. Firstly, it enabled screening of medium-large libraries of compounds within a few hours. Secondly, the assay showed acceptable levels of robustness and reproducibility. Thirdly, the Mature tobacco pollen can be used as an ideal model system to discover new plant modulators in HCS assays. Finally, the hits selected via first stage can be further tested for dose dependence and reconfirmed by testing of their biological activity on different plant models. In conclusion, we developed effective medium to high throughput screening assay based on advanced HCS of biological effects on pollen tube growth, which can be used to discover new compounds that can effectively inhibit the growth of crop pests (weeds, fungal diseases) without causing any adverse effects on crop plants and humans.

Multitarget activity profiling predicts in vivo adverse effects of serotonin and norepinephrine reuptake inhibitors

Georgy I. Lapushkin1, Konstantin V. Balakin 2, 1, Anastasia G. Savilova 1, Andrey E. Voronkov 1

1Moscow Institute of physics and technology (State University), Dolgoprudnyi, Moscow region.2Kazan Federal University, Kazan.

[email protected]

Early identification and control of drug adverse effects (AEs) is an important and very difficult issue in modern drug discovery and development. Even multicenter long-term clinical trials are not always able to provide an exhaustive understanding of the potential pitfalls of a drug’s effects on health,

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which represents a serious problem. Such critical knowledge usually appears only after multi-year post-marketing surveilance, which allows the uncovering of side-effects that remained unnoticed during the clinical trials. Often, these effects are so serious that they lead to the recall of the products from the market due to their harmful side-effects. This raises the need for approaches accurately estimating the negative side effects as early as possible in the drug discovery process.

The early detection of such undesirable side-effects of drugs would enable a reduction in the risks for the patients during clinical trials and to reduce the costs of drug development process.

In this study, we have demonstrated the ability to evaluate potential adverse effects profile for a series of antidepressants, namely serotonin and norepinephrine reuptake inhibitors (SNRIs), based on analysis of their in vitro multitarget activity data. Our approach is based on the use of computational data mining methods. We have proposed a method for analysis of topology of the adverse effects space based on nonlinear Sammon mapping approach. The map can be used for visual analysis of differences in the individual adverse effects profiles of drugs and groups of drugs beyond SNRIs. Other data visualization and mapping approaches may also be applicable.

We have found relationships between the complex profiles of adverse effects, corresponding to the 1st level of the MedDRA classification, and multi-target activity profile of 10 SNRIs. This demonstrates the principal ability to predict and classify the potential adverse effects profile for compounds using results of in vitro experiments based on a relatively small panel of targets. It is likely in future such results could be entirely in silico derived.

Taking into consideration the fact that profiling a molecule or new drugs clinical adverse effects can take years and typically many years of clinical and post-marketing surveillance studies, our approach using results of in vitro or in silico experiments could be very insightful to propose further experiments and refine safety studies. At the post-marketing stage such information could warn of possible safety problems and thus reduce possible health risks.

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Search of useful side effects of well studied drugs by means of the FDA AERS database

Lapushkin G.I., Moscow Institute of Physics and Technology (MIPT), 9, Institutskiy per., Dolgoprudny,

Moscow region, 141700, Russia. [email protected]

A study relies on input data from the public release of the FDA’s Adverse Event Reporting System (AERS) are widely used for searching of Adverse Effects (AEs) using statistical methods. Now only the amounts of PRR much more then one are used to make conclusions.

But analyzing a formula PRR it is possible to make the conclusion, that low amounts of PRR probably can approve the effect of treatment by particular drug for particular disease. For the purpose of verification of this assumption statistical analysis was made for some drugs (ISOTRETINOIN, METHYLPHENIDATE HYDROCHLORIDE, IBUPROFEN, ESOMEPRAZOLE MAGNESIUM, ROSUVASTATIN CALCIUM, QUETIAPINE FUMARATE). For this set of drugs a number of side effects with low values PRR has been chosen, and it has turned out that value PRR at combined use of couples of these drugs for this set of AEs considerably decreases, that can mean existence of positive side medical effect. Such effect can be in case two drugs with the different mechanism of impact on an illness are at the same time used.

Also this method has been applied to search of side medical effects against Psoriasis, in the course of work it has become clear that reliable useful side effect is rendered by ATORVASTATIN CALCIUM and PREGABALIN.

Taking into consideration the fact that search and check of safety new drugs for adverse effects can take years and typically many years of clinical and post-marketing surveillance studies, and also is very expensive, this approach to use positive side effects of well-known drugs and the combinations of such drugs can save a lot of money, reduce medicine conclusion time for the market, reduce risk of new dangerous AEs.

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The effects of quercetin on lifespan and stress resistance of Drosophilla melanogaster

Ekaterina Proshkina1, Ekaterina Lashmanova2, Alexey Moskalev1,2

1 Institute of Biology, Komi Science Center, Ural Branch of RAS, Syktyvkar, Russia2 Moscow Institute of Physics and Technology (State University), Dolgoprudny

E-mail: [email protected]

Flavonoids are important phytochemicals with revealed antioxidant, antidiabetic, anticancer, cardioprotective and neuroprotective properties. The goal of the present study was to investigate the effects of long- and short-term consumption of quercetin, a most abundant and well-studied flavonoid in human diet, on the lifespan of Drosophilla melanogaster. In addition, effects of this flavonoid on the resistance of flies to thermal (35°С), oxidative (20 mM paraquat), genotoxic (1000 Gy γ-radiation) stresses, and starvation were studied.

In our experiments we used the wild-type Canton-S line. The flies were kept on standard nutrient medium at 25°С in 12/12 light/dark regime. In experiments on long-term quercetin consumption and stress resistance, the studied flavonoid was applied on the surface of nutrient medium in final concentrations 0.3, 0.5 and 1 µМ. In experiments on short-term consumption, quercetin in concentration of 1 µМ was given to flies during the first 10 days of their life. All experiments on stress resistance were performed on the 10th day of flies’ life.

It was revealed that the long-term consumption of the flavonoid decreased the median and maximum lifespans of males by 7-13% (P <0.05). For females only a decrease in maximum lifespan by 4-6% was observed (P <0.05). In contrast, the short-term consumption of quercetin increased the median and maximum lifespans of females by 6-13% (P <0.001). There were no statistically significant changes in parameters of males’ lifespan after short-term consumption of quercetin. The addition of quercetin had no or even negative effects on resistance of flies to starvation, thermal and oxidative stresses. However, it increased males’ resistance to γ-radiation.

Thus, short-term consumption of quercetin increased lifespan of Drosophilla melanogaster. However, long-term consumption of this flavonoid had opposite effect. The hermetic response is a possible explanation of quercetin’s positive effects. The negative effects could have been a result of its toxic properties. The further investigations are needed to be done to confirm these theories.

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Pharmacoeconomic Analysis of Direct Activity Antiviral Drugs Used for Treatment of Genotype 1 Chronic Hepatitis C

N.Z. Musina, A.G. Savilova, O.M. KorzinovMoscow Institute of physics and technology (State University)

E-mail: [email protected]

Hepatitis C is one of the most important global health care problems. There is a high level of hepatitis C virus (HCV) infection in Russian Federation, and the morbidity is still increasing. Genotype 1 hepatitis C virus is a dominant form (80%) in Russian Federation, and it is highly resistant to therapy.

Hepatitis С is socially important disease and its dissemination leads to serous economic losses. Timely diagnostics and treatment of hepatitis C allows preventing serious complications and even achieving complete liver regeneration.

The goal of this study was to perform a comparative pharmacoeconomic analysis of therapeutic schemes used in treatment of patients with genotype 1 chronic HCV by using direct-acting antiviral agents (boceprevir, simeprevir and telaprevir) in combination with pegylated interferons and ribavirin.

Initially, a systematic survey of the literature was performed to evaluate efficacy and safety of preparations used to treat patients with genotype 1 HCV, both treatment-naive and unsuccessfully treated with pegylated interferons and ribavirin. For this purpose, Clinicaltrials.gov and Medline databases were used. Results of completed efficacy studies were selected, in which the preparations were used in doses recommended by special instructions for a certain cohort of patients. Patients with combined HIV/HCV infection were not taken into consideration.

In the current study Markov decision process model was built in order to assess long-term costs and efficacy of preparations under study. Comparison of therapeutic schemes for chronic HCV infection treatment was performed with the use of cost-effectiveness and cost-utility analysis. In costs-effectiveness analysis, therapeutic schemes for CHC treatment that include telaprevir, boceprevir and simeprevir, life years gained (LYG) and quality adjusted life years (QALY) served as a criterion of benefit in costs-effectiveness and cost-utility analysis respectively.

Pharmacoeconomic analysis of treatment of chronic hepatitis C viral infection showed that the best combination of costs and effectiveness was observed in combined treatment with the use of pegylated interferon, ribavirin and boceprevir in treatment-naïve patients as well as patients with unsuccessful treatment experience.

Our results showed that combined therapy with the use of boceprevir is the most pharmacoeconomically justified in comparison with telaprevir and simeprevir-based treatment.

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Synthesis of Macrocyclic Organo-short\medium peptide Hybrids via CuAAC

Vantskul Aa. Ivanenkov Yb. Polyakova Ma. Sandulenko Ya.a Moscow Institute of Physics and Technology,

9 Institutskiy per., Dolgoprudny, Moscow Region, 141700, Russian Federationb Moscow State University

Leninskie Gory, Moscow, 119991, Russian Federation [email protected]

The main aim of our work is investigation of possibilities of the formation of macrocycles, which include short/medium peptide fragment.

It’s well known, macrocyclic peptides have attracted increasing attention as a potential new source of chemical probes and therapeutics [1]. In particular, their conformationally restricted structure combined with a high degree of functional and stereochemical complexity allows considering them as promising scaffolds for targeting biomolecules with high affinity and selectivity [2].

Up to this time, we have developed a methodology for the synthesis of hybrid organo-peptide macrocycles via the cyclization with non-peptidic organic linkers which contain azide-alkyne fragments as our ‘synthetic precursors’. This strategy relies on the copper (I)-catalyzed alkyne azide 1,3-dipolar cycloaddition (CuAAC) or ‘click’ reaction. It’s a highly versatile reaction that can be performed under a variety of reaction conditions including various

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solvents, mild conditions of pH and temperature, and using different copper catalysts, with or without additional ligands or reducing agents. All these features of CuAAC are very important, when we work with structures, which include peptidic fragments; moreover, the reaction is highly selective and can be performed in the presence of other functional groups [3]. So, this approach to a macrocyclization was found to proceed with high efficiency among a range of different sizes of cycles (14-23 ring members) which can include different natural and synthetic fragments of chains.

This versatility combined with the possibility to integrate non-proteinogenic fragments into peptide sequences and fragments results in methodology of particularly high value for the creation and screening of highly diverse libraries of peptide-based macrocycles [4].

1. Robinson JA, Demarco S, Gombert F, Moehle K, Obrecht D. The design, structures and therapeutic potential of protein epitope mimetics.Drug Discov Today.2008;13:944–951.[PubMed]2. Smith JM, Frost JR, Fasan R. Emerging strategies to access peptide macrocycles from genetically encoded polypeptides.J Org Chem.2013;78:3525–31.[PubMed]3. Contemporary strategies for peptide macrocyclization.Nat Chem.2011;3:509–524.[PubMed]4. Frost JR, Vitali F, Jacob NT, Brown MD, Fasan R. Macrocyclization of Organo-Peptide Hybrids through a Dual Bio-orthogonal Ligation: Insights from Structure-Reactivity Studies.Chembiochem.2013;14:147–160.[PubMed].

Extra bright fluorescent labels based on organic dyes

Rzhevskiy S.A.a, Shadrin I. A.a, Babaev E.V.b, Pakhomov A.A.c a Moscow Institute of Physics and Technology,

9 Institutskiy per., Dolgoprudny, Moscow Region, 141700, Russian Federation;b Moscow State University, Faculty of Chemistry; c-M.M. Shemyakin and Yu.A. Ovchinnikov

Institute of bioorganic chemistryof the Russian Academy of [email protected]

Fluorescent tags enable visualization of biochemical processes in vivo. For wide use at organismic level such tags should be characterized by water solubility, bioavailability and high quantum output without short UV light. That’s why fluorescent tags based on organic dyes integrated into a single molecule by means of dendritic matrix are objects of high interest. Synthetic approaches for obtaining and functionalization of dendrimers developed in our laboratory will help to control the size and topology of fluorescent probes containing several coloring agents and reactive bioorthogonal groups (azides and acetylenes).

The «click» – promoted tetra methyl BODIPY was synthesized.

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It was used for optimization of introduction conditions of fluorescent labels to biomolecule1.

Extra bright fluorescent labels based on tetra methyl BODIPY-substituted

acetylene promoted 4th generation bis-MPA dendrones were prepared. 4 fold increase in brightness was observed compared to single molecule of tetra methyl BODIPY.

[1] Russian Journal of Bioorganic Chemistry, 2015, vol 41, № 4, p. 505–508, 2015

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Список докладов круглых столов и питч-сессии стартапов

Круглый стол «Научно-исследовательские и технологические проекты Дорожной карты НейроНет» Семячкина-Глушковская Оксана Валерьевна, зав. кафедрой физиологии человека и животных Биологического факультета Саратовского государственного университета им. Н.Г. Чернышевского«Разработка компьютерных диагностических комплексов для прогноза риска развития неонатального инсульта и восстановления интеллектуальных зон мозга»

Галкин Алексей Петрович, Санкт-Петербургский государственный университет«Восстановление и сохранение ресурсов мозга через регуляцию амилоидогенеза»Крылов Владимир Владимирович, руководитель лаборатории технологий больших данных, Нижегородский государственный технический университет им. Р.Е. Алексеева«Технология аудиовизуального представления запаховых полей и навигации в них»

Шишкин Сергей Львович, начальник лаборатории, НИЦ «Курчатовский институт»«Контроллер с открытым кодом для скорейшей реализации намерений пользователя на основе технологий интерфейсов глаз-мозг-компьютер (Мышь исполнения желаний – The WishMouse)»

Иванюк Наталья Михайловна, генеральный директор ООО «Бионик Натали», Каримов Владимир Русланович, коммерческий директор ООО «Бионик Натали»«Создание бионического протеза руки»

Попов Сергей Валентинович, НИИ кардиологии, Национальный исследовательский Томский политехнический университет, Томский государственный университет систем управления и радиоэлектроники«Разработка единого инструментально-методического комплекса ранней МР-томографической диагностики атеросклероза сосудов мозга и риска нарушений мозгового кровообращения»

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Милов Юрий, Шкотин Александр, Private R&D «Контролируемый Русский Язык»«Язык логического программирования для представления знаний в семантическом интернете по-русски»

Дорохов Владимир Борисович, ФГБУН Институт Высшей Нервной Деятельности и Нейрофизиологии РАН«Нейротехнология нефармакологического улучшения качества сна, путем неивазивной стимуляции во время сна, вызывающей активацию естественных восстановительных и репаративных процессов»

Захарова Олеся Владимировна, Межрегиональное общественное движение «Сахаджа Йога»«ECO-meditation»

Александрова Евгения Владимировна, невролог НИИ нейрохирургии им. Н.Н. Бурденко Минздрава РФ«Комплексный многоуровневый мониторинг патологической пластичности мозга человека в процессах нарушения и восстановления сознания»

Цукерман Валерий Давидович, Академия биологии и биотехнологий им. Д.И. Ивановского Южного федерального университета«Создание Лаборатории моделирования нейродинамики когнитивных функций мозга и разработки гибридной (аналого-цифровой) технологии решения прикладных задач»

Козунов Владимир Вячеславович, Московский МЭГ Центр, Московский Городской Психолого-Педагогический Университет«Проект создания инструментария функционального картирования мозга»

Копейкин Кирилл Владимирович, проректор СПбДА, директор Научно-богословского центра междисциплинарных исследований СПбГУ«Проект ТЕОС (ТЕологическое Осмысление Сознания)»

Воронежская Елена Евгеньевна, Институт биологии развития им. Н.К.Кольцова РАН (ИБР РАН)«Трансляционная модификация белков нейрональными моноами-нами как новый механизм направленного изменения поведения и активизации адаптивных ресурсов организма»

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Овчинникова Валентина Андреевна, ФГАОУ ВПО «Уральский федеральный университет имени первого Президента России Б.Н. Ельцина»«Разработка комплексной системы мониторинга и коррекции психофизиологического состояния оператора в процессе высокоинтенсивного взаимодействия в интерфейсах человек-машина с повышенным риском»

Надежда Павлова Валерьевна, ФГАОУ ВПО «Уральский федеральный университет имени первого Президента России Б.Н. Ельцина»«Лонгитюдное междисциплинарное исследование детей с риском развития аутизма и СДВГ»

Краснощекова Елена Ивановна, д.б.н., профессор кафедры цитологии и гистологии, биологический факультет СПбГУ «Разработка новых технологий картирования мозга и предиктивной диагностики патологий нейровирусного генеза»

Миланич Александр Иванович, Институт Общей Физики им. А.М. Прохорова РАН (ИОФ РАН)«Объективное тестирование состояния мозга по саккадам»

Иванов Антон Андреевич, Арлашкин Михаил Николаевич, ООО “Нейроменеджмент”Проект «TreeBCI»

Намазова-Баранова Лейла Сеймуровна, Каркашадзе Георгий Арчилович, Савостьянов Кирилл Викторович, ФГАУ «Научный центр здоровья детей» Минздрава«Инновационная технология нейромаркирования вариативной типологии когнитивной и психофизиологической деятельности человека»

Кичигина Валентина Федоровна, ФГБУН Институт теоретической и экспериментальной биофизики РАН (ИТЭБ РАН), ФГБУН Институт математических проблем биологии РАН«Исследование механизмов когнитивной деятельности мозга и их нарушений при височной эпилепсии»

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Ратушняк Александр Савельевич, КТ Институт вычислительной техники СО РАН, лаборатория биомедицинской информатики«Клеточно-молекулярная основа базовой негэнтропийной функции мозга и перспективы исследований и разработок по устранению дефицита нейроресурсов»

Осадчий Алексей Евгеньевич, Шестакова Анна, Высшая Школа Экономики (Центр нейроэкономики и когнитивных исследований) МГППУ (Московский МЭГ центр)«Динамическая коннектомика головного мозга: алгоритмы, экспериментальные парадигмы и инструменты, Магнитоэнцефалография высокого пространственного разрешения на основе атомарных магнетометров. Универсальный нейроинтерфейс с обратной связью для индивидуального и коллективного пользования»

Васильев Николай Сергеевич, руководитель отдела исследований и разработок компании «Интеллект Хауз» «Визуальный интерфейс мозг-компьютер»

Крыжановский Эдвард Владимирович, ООО «БрейнБит»«Технология опторитмографии»

Павлова Галина Валериевна, Гельперина Светлана Эммануиловна, ФГБУН Институт биологии гена РАН«Разработка наноразмерной формы GDNF для направленной терапии нейродегенеративных заболеваний на модели ишемического инсульта»

Мнацаканян Елена Владимировна, ФГБУН Институт высшей нервной деятельности и нейрофизиологии РАН«Мультидисциплинарные исследования социальной перцепции в норме и патологии» Сафоничева Ольга Георгиевна, д.м.н., профессора кафедры нелекарственных методов лечения и клинической физиологии ИПО ГБОУ ВПО «Первый МГМУ им. И.М. Сеченова«Научное обоснование новых подходов к исследованию ресурсов мозга, разработка оптимальных условий и технологий для повышения успешности обучения и сохранения пластичности»

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Аведьян Эдуард Дзеронович, Центр Нейросетевых технологий Международного центра по информатике и электронике«Технология создания перспективной программно-аппаратной платформы для моделирования структур разделов мозга большой размерности»

Дорохов Владимир Борисович, Лигун Наталья Владимировна, Блохин Илья Сергеевич«Нейротехнология нефармакологического улучшения качество сна, путем неивазивной стимуляции во время сна»

Пучкова Александра Николаевна, Таранов Антон Олегович, Сломинский Петр Андреевич, лаборатория нейробиологии сна и бодрствования ФГБУН Института высшей нервной деятельности и нейрофизиологии РАН«Технология генетической экспресс-диагностики индивидуального хронотипа, как способ повышения работоспособности при круглосуточном и сменном режиме работы»

Круглый стол «Создание НейроНет Центров в России»

Луковникова Наталья Михайловна, Борщева Ольга Игоревна, Центр научно-технологического форсайта«Северо-Западный центр разработки и коммерциализации нейротехнологий»

Зайцев Андрей Станиславович, ООО «Интеллект Хауз»«Нейронет центр Кубани»

Любимов Игорь Иванович, ген. директор, партнер ООО «Гурус БиоФарм»«Научно-внедренческий центр перспективных фармакологически препаратов для лечения Болезни Паркинсона (БП) и родственных заболеваний ЦНС. (НВЦ «Нейрофармтех»)»

Узденский Анатолий Борисович, д.б.н, профессор, Академия биологии и биотехнологии Южного федерального университета«НейроНет-центр ЮФУ»

Пятин Василий Федорович, кафедра нормальной физиологии СамГМУ«Поволжский НейроНет Центр (ПННЦ)»

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Питч-сессия стартапов

Астапенко Елена, начальник Управления организации государственного контроля и регистрации медицинских изделий Росздравнадзора «Государственное регулирование обращения медицинских изделий на территории Российской Федерации. Новеллы законодательства и результаты контроля»

Воробьёв Константин Александрович, ООО «НеоМед»«Гамма-Зонд» Радионуклидная диагностика в онкологии

Жмырко Екатерина Викторовна, ООО «Джинэкст»«Mens Diagnostics». Комплексное решение для диагностики рака предстательной железы»

Антипов Артем, Пыльца Club«Пыльца Club» — краудсорсинговая платформа помощи людям, страдающим от аллергии»

Олейник Сергей Викторович, ООО «Ксеникс» «Многофункциональная система для комплексной неинвазивной диагностики органов и тканей организма в коротковолновом инфракрасном спектральном диапазоне»

Бушков Николай Алексеевич, МФТИ«Разработка и организация серийного производства отечественного Терапевтического аппарата оксигенации (ТАО)»

Павшинцев Всеволод Вячеславович, ООО «АДГ-Фарм»Препарат для терапии алкогольной зависимости

Сидельников Георгий Дмитриевич, Международный биотехнологический центр «Генериум»«Разработка инновационного препарата против широкого спектра онкопатологий на основе антигенов Trichinella spiralis»

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Моисеева Ольга Владимировна, Международный биотехнологический центр «Генериум»«Новый подход в терапии раковых заболеваний: метод полимерного капсулирования»

Коновалова Евгения Викторовна, ФГБНУ «Научный центр неврологии»«Разработка лекарственного препарата на основе нанокомплекса карнозина с адресной доставкой к очагу поражения»

Воронков Андрей, МФТИDrug discovery at home

Попова Марина Олеговна, Первый Санкт-Петербургский государственный медицинский университет им. акад. И.П. Павлова«Next generation immunotherapy for cancer»«Fighting with HIV»

Дейген Ирина Михайловна, МГУ им. М.В. Ломоносова«Инновационный препарат для таргетной терапии колеректального рака»

Иванов Антон Андреевич, ООО «Нейроменеджмент» «TreeBCI». Нейроуправляемая виртуальная реальность»

Захарченко Дмитрий, ООО «Нейрокогнитивные технологии»«Система непрерывного мониторинга и контроля работоспособности водителей и машинистов локомотивов»

Кислов Андрей Александрович, BrainRay«BrainRay — технология объективной оценки марочного капитала»

Морозова Татьяна Игоревна, МФТИ«Динамические каблуки или как наука спасет красоту»

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Панихин Григорий Игоревич, НИТУ МИСиС«Разработка мобильного аэропонного контейнера для овощей»

Живалина Юлия Андреевна, Тюменский государственный университет«Разработка биотехнологического метода утилизации отходов свиноводческих компаний»

Илья Ильяшенко, ООО «Триэл»«Изготовление модульных цехов по переработке плодов сок содержащих растений и строительство завода по производству бикомпонентных экологически чистых и полезных для здоровья напитков, молочных продуктов»

Открытая дискуссия «Междисциплинарные проекты и технологии в области живых систем»

Сергей Шумский, председатель совета директоров ЗАО «Айкумен – информационныебизнес-системы»«Роботы и машинный интеллект в сельском хозяйстве: зачем, как, когда?»

Екатерина Филатова, исполнительный директор Ассоциации дополненной и виртуальной реальности«Технологии дополненной и виртуальной реальности на основе существующих кейсов»

Александр Мелерзанов, декан факультета биологической и медицинской физики МФТИ«Развитие телемедицины в Российской Федерации. Российский телемедицинский консорциум»

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Контакты спикеров VI Международной конференции

ФизтехБиоChinese SpeakersDr. Xiao Nong Zou, Ph. D. Professor, Department of Epidemiology, Office of Cancer Prevention and Control National Cancer Center, Cancer Institute of the Chinese Academy of Medical SciencesSouth 17 Panjiayuan, Chaoyang DistrictBeijing, 100021, The People’s Republic of ChinaTelephone: (86) 10 8778 8656 FACS: (86) 10 6771 8227E-mail: [email protected]

Dr. Ping Gao, Ph. D. Professor, School of Life Science, University of Science and Technology of China, 443 HuangshaHeifei, The People’s Republic of ChinaTelephone: 86-551-6367033E-mail: [email protected]

European Union SpeakersDr. Herman Autrup, Ph. D, Professor, Aarhus Universitetet, Bygning 1260, Vennelyst Boulevard 6, 8000 Arhus CTelephone: 8942 6180E-mail: [email protected]

Dr. Robert Ladner, Ph. D., Associate Professor/Reader, School of Medicine and the Cancer Center, Queen’s University, Belfast Health Science Building , 97 Lisburn Road Belfast, Ireland, BT9 7BL BT9 7BL United Kingdom, CEO and Founder, CV6 Therapeutics, Inc.Telephone: 1-(323)-381-8745 in U. S. A. or 44(0)2890 973091 in the U. K.E-mail: [email protected]

Dr. Marko Kornmann. M. D., Department of General, Visceral, and Transplantation SurgeryUniversity Hospital of Ulm, Steinhovelstrasse 9, 89075 Ulm, University of Ulm, Ulm, GermanyTelephone: 49-731/500-53 560FACS: 49-731/600-53 561 E-mail: [email protected]

Prof. Dr. Med. Dr. H. C. Karl-Heinrich Link, M. D., Ph. D., Director Surgical Center and Asklelpios Tumor Treatment Center Rhein/Main (ATC), Asklepios Paulinen Klinik (APK)Geisenheimer Strasse 10, D 65197 Wiesbaden, Germany Telephone: 0611-8472431; 0177-3357542FACS: 0611-8472459E-mail: [email protected]

Dr. David H. Phillips, Ph. D, Professor of Environmental Carcinogenesis, Research Group of Genetic and Environmental Toxicology, Dept. of Analytical and Environmental Sciences, MRC-HPA Centre for Environmental and Health, 3.86, 3rd Floor Franklin –Wilkins BuildingKing’s College, London, 150 Stamford Street, London, SE1 9NH, United KingdomTelephone: 44 (0) 2078484569E-mail: [email protected]

Amy Hamilton, Senior Field Applications Specialist, EMEA, Fluidigm Europe B.V. (Official partner of Helicon Company)Tel: +33 1 60 92 42 40E-mail: [email protected]

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Israeli Speakers Dr. Israel Vlodavsky, Ph. D., Cancer and Vascular Biology Research Center, Rapaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 31096, IsraelTelephone: 972-4-8295410FACS: 972-4-8523947E-mail: [email protected]

Japanese Speakers Dr. Miwako Homma, M. D., Associate Professor, Institute of Biomedical Sciences, Department of Biomedical Research, School of Medicine, Fukushima University, Hikarigo-oka, Fukushima 960-1295, JapanTelephone: 81-24-547-1659FACS: 81-24-548-3041

Dr. Yoshimi Homma, M. D., Director Institute of Biomedical Sciences, Department of Biomolecular Research, School of Medicine, Fukushima University, Hikarigo-oka, Fukushima 960-1295, JapanTelephone: 81-24-547-1659FACS: 81-24-548-3041E-mail: [email protected]. Toshio Kuroki, M. D., Ph. D., Japan Society for the Promotion of Science (JSPS), University of Tokyo and Gifu University, 5-3-1, Kojo-machi, Chiyoda-ku, Tokyo, 102-0083, JapanTelephone: 81-3-3263-2130E-mail: [email protected] or [email protected]

Dr. Yoshinori Murakami, M. D., Ph. D., Professor and Director Division of Molecular PathologyInstitute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 JapanTelephone: 81-3-5449-5260FACS: 81-3-5449-5407E-mail: [email protected]

Dr. Mikhail F. Chernov, M. D., D. Med.Sci Assistant Professor, Faculty of Advanced Techno-Surgery and Department of Neurosurgery, Tokyo Women’s Medical University; 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, JapanTelephone: +81-3-3353-8111 (Ext. 66003)FACS: +81-3-5312-1844E-mail: [email protected] or [email protected]

Norwegian SpeakersDr. Aage Haugen, Ph. D., Senior Scientist and Director, Department of Toxicology National Institute of Occupational Health, P. O. Box 8149 Dep. N-0033, Oslo, NorwayAdjunct Professor, Norwegian Institute of Science and Technology, Oslo, NorwayTelephone: 47 2319-5270FACS: 47 2319-5203E-mail: [email protected]

Dr. Arne Klungland, Ph. D., Professor, Laboratory for Genome Repair and Regulation Centre for Molecular Biology and Neuroscience, Institute of Medical Microbiology, BIG CAS-OSLO Genome Research Corporation, Oslo University Hospital, Rikshospitalet, Oslo 0027 NorwayTelephone: 47-2307-4072FACS: 47-2307-5061E-mail: [email protected] or [email protected]

Russian SpeakersDr. Nadezhda V. Cherdyntseva, Ph. D., Professor Department of Molecular Oncology and Immunology, Deputy Director for Sciences Tomsk Cancer Research Institute, Kooperativny per., 5 Tomsk, 634009, RussiaE-mail: [email protected]

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Dr. Irina Demidova, M. D., Molecular Diagnostic Laboratory, Moscow Oncological Hospital №62 Russia, 143423, Istra settl/Stepanovskoe, Krasnogorsky District, Moscow Region, Moscow, RussiaTelephone: 8-916-159-7481E-mail: [email protected] [email protected]

Dr. Maxim Filipenko, Ph. D., Laboratory of Pharmacogenomics, Institute of Chemical Biology and Fundamental Medicine, Lavrentiev Str. 8, Novosibirsk, 630090, RussiaTelephone: +7-913-921-7392E-Mail: [email protected]

Dr. Lyudmila Gulyaeva, Ph. D, Professor and Senior Scientist, Head of the Laboratory of Molecular Carcinogenesis, Institute of Molecular Biology and Biophysics, Academy of Medical Sciences, Siberian Division, Timakov Street 2/12, Novosibirsk 630117, RussiaVice Dean, Medical Department, Novosibirsk State UniversityTelephone: 7-383-334-8840 FACS: 7-383-335-9847E-mail: [email protected] or [email protected]: lyudmilla.gulyaeva

Dr. Evgeny Imyanitov, Department of Tumor Growth Biology, N. N. Petrovinstitute of OncologyPesochny str. 2,, St. Petersburg, 197758, RussiaDepartment of Medical Genetics, St. Petersburg Pediatric Medical Academy, Litovskaya str. 2, St. Petersburg, 19410, RussiaDepartment of Oncology, I. I. Mechnikov North-Western Medical University Kirochnaya str. 41, Saint-Petersburg, 191015, RussiaTelephone: 7-812-596-89-51E-mail: [email protected] or [email protected]

Dr. Nikolay Kolesnikov, Ph. D. Institute of Molecular and Cell Biology, Novosibirsk, RussiaTelephone: +8- 913-463-4544E-mail: [email protected]

Dr. Sergei Kovalenko, Ph. D., Senior Scientist and Deputy Director, Institute of Molecular Biology and Biophysics, 630117 Novosibirsk, RussiaSenior Scientist, Life Science Center, Moscow Institute of Physics and Technology, Moscow, Russia Telephone: +8 905-952-1411E-mail: [email protected]

Dr. Michael Krasilnikov, Blokhin Russian Cancer Research Center, Moscow, RussiaTelephone: +8 916-303-0412E-mail: [email protected]

Dr. Alexei L. Krivoshapkin, M. D., Professor Neurosurgery Department Novosibirsk State Medical University, Krasny Prospect 52, Office 105, Novosibirsk, 630091, RussiaNeurosurgery Department, European Medical Center, Schepkina str. 35, Moscow, 129090, RussiaPrincipal Research Officer, Meshalkin State Research Institute of Circulation Pathology, Rechkunovskaya str. 15, Novosibirsk, 630055, RussiaTelephone: 7 (383)-347-60-06 E-mail: [email protected]

Dr. Aleksandr Melarzanov, MD, PhD, Dean of Department of Biological and Medical Physics of Moscow Institute of Physics and Technology (State University), Expert of Ministry of Education and Science for Pharma-2020 Moscow, RussiaTelephone: +7(915)155-55-50E-mail: [email protected]

Dr. Marina G. Sergeeva, Belozevsky Institute of Physico-Chemical Biology, Moscow State University, Moscow, RussiaTelephone: +8-916-657-93-77E-mail: [email protected]

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Dr. Tatyana Serebriyskaya, Ph. D., Moscow Institute of Physics and Technology, Moscow, RussiaTelephone:+7(916)401-65-22E-mail: [email protected]

Dr. Sergey Shevchenko, M. D., Surgical Pathology Unit Novosibirsk State University Pirogova str. 2, Novosiribrsk, 630090, Russia Head and Neck Surgery Department, Novosibirsk Clinical Hospital No. 1, Zalesskogo str. 6, Building 5Novosibirsk, 630047, RussiaTelephone: +8 913-913-1077E-mail: [email protected]

Dr. David Zaridze, M. D., Professor and Deputy Director, Blokhim Russian Cancer Research Centre24 Kashirskoye Shossa 115478, Moscow, RussiaTelephone: +7-499-324-1824E-mail: [email protected]

Sergey Leonov, MD, PhD, Chief Science Officer, Life Science Center, Head of Laboratory of Innovative Medicine, Moscow Institute of Physics & Technology Telephone:+7(915)055-56-43E-mail: [email protected]

Korzinov Oleg, «Northern» Biopharmaceutical Cluster Development Center, Nonprofit Partnership, Executive Director Tel. +7 495 408 42 00E-mail: [email protected]

Dr. Kudryavtsev N.N., Member of the Russian Academy of Sciences, Rector of MIPTTel: +7(495)408-57-00E-mail: [email protected]

Yakovlev Pavel, BIOCAD, Saint-Petersburg, RussiaTel:+7 (812) 380 49 33E-mail: [email protected]

South American SpeakersDr. Gloria M. Calaf, Professor, Universidad de Tarapaca, Instituto De Alta InvestigacionCalle Antofagasta 1520, Arica, Chile South AmericaTelephone: (56 58) 2255 371, FACS: (56 58) 2230 334Center for Radiological Research, School of Medicine and Medical Center, Columbia University530 W. 168th Street, VC11-238, New York, United States of America, Telephone: (212)-305-9981E-mail: [email protected]

U. S. A. SpeakersDr. Clark Chi-Chung Chen, M. D., Associate Professor, Department of Surgery, University of California at San Diego, 3855 Health Science Drive #0987, La Jolla, California 92093-0987, United States of AmericaTelephone: 619-246-0674 FACS: 858-822-4715E-Mail: [email protected]

Dr. Max Costa, Ph. D., Professor and Chairman, Dept. of Environmental Medicine, School of MedicineNew York University, 57 Old Forge Road, Tuxedo, New York 10987, United States of AmericaTelephone: (845)-731-3515 (office)Telephone: (646)-469-5298FACS: (201)-418-8596E-mail: [email protected]

Dr. Curtis C. Harris, M. D., Chief, Laboratory of Human Carcinogenesis, CCR, NCI, NIH37 Convent Drive, Room 3068A, MSC 4258, Bethesda, Maryland 29892, United States of AmericaTelephone: (301)-496-2048FACS: (301)-496-0497E-mail: [email protected]

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Dr. Eliezer Huberman, Ph. D., CEO, NovaDrug, LLC, 500 W. Superior, #2802, Chicago, Illinois 60654Adjunct Professor, School of Pharmacy, University of Illinois in ChicagoTelephone: (312)-644-4622E-mail: [email protected] or [email protected]

Dr. Joseph R. Landolph, Jr., Ph. D., Associate Professor of Molecular Microbiology, Immunol., and Pathology, Member, USC/Norris Comprehensive Cancer Center, Keck School of Medicine, Associate Professor of Molecular Pharmacology/Pharmaceutical Science, School of Pharmacy Member, Free Radical Institute, Laboratory of Chemical Carcinogenesis and Molecular Oncology Cancer Research Laboratory, Room #218, 1303 N. Mission Road, University of Southern California Keck School of Medicine, Health Sciences Campus, Los Angeles, California 90033, United States of AmericaTelephone: (323)-224-7781 FACS: (323)-224-7679E-mail: [email protected] or [email protected]

Dr. Wei Li, Ph. D., Professor, Department of Dermatology, 6320 Norris Cancer ResearchUSC/Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, California 90089-9176, United States of AmericaTelephone: (323)-865-0618FACS: (323)-865-0105E-mail: [email protected]

Dr. Ian E. McCutcheon, M. D., Professor of Neurosurgery, Director, Neuroendocrine ProgramDept. of Neurosurgery, Affiliate Faculty, Program in Clinical Cancer Genetics University of Texas M. D. Anderson Cancer Center, Department of Neurosurgery, Baylor College of Medicine, Houston, TexasTelephone: 713-563-8706FACS: 713-794-4950E-mail: [email protected]

Dr. Debra A. Tonetti, Ph. D., Associate Professor, Department of Pharmaceutics and Pharmacodynamics, College of Pharmacy, University of Illinois, 833 Wood Street, Room 335, Chicago, Illinois 60612 United States of AmericaTelephone: (312)-249-1448FACS: (312) - 996-0098E-mail: [email protected]

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Тираж: 125 экз.«Сборник тезисов VI Международной конференции ФизтехБио»

24-28 мая 2016 г., 141700, Россия, Московская область, г. Долгопрудный, Институтский пер., д. 9

Московский физико-технический институтНП «Центр развития БФК «Северный»,

+7 495 408 42 [email protected]

www.mipt.ruwww.pharmcluster.ru

СБОРНИК ТЕЗИСОВ

VI МЕЖДУНАРОДНАЯ КОНФЕРЕНЦИЯ ФИЗТЕХБИО

24-28 мая 2016 годаМосковский физико-технический институт

+7 495 408 42 00

Институтский переулок, дом 9, стр. 7Долгопрудный,

Московская область, Россия, 141700НП «Центр развития БФК «Северный»

www.mipt.ruwww.pharmcluster.ruwww.phystechbio.ru

При поддержке:

Спонсоры:

Информационные партнеры:

Министерство образования и науки Российской Федерации

Инновационный территориальный кластер

Физтех XXI

Правительство Московской области

Проект повышения конкурентоспособности ведущих российских университетов

среди ведущих мировых научно-образовательных центров

Корпорация развитияМосковской области

Научно-исследовательскийинститут молекулярнойбиологии и биофизики

СБОРНИК ТЕЗИСОВ · 2016. 11. 4. · 4 vi Международная...

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