肿瘤生物学与转化医学 cancer biology & translation medicine 第四军医大学...

肿瘤生物学与转化医学Cancer Biology & Translation Medicine

第四军医大学[email protected]

陈志南Zhi-Nan Chen

第四军医大学 Fourth Military Medical University

Global Action Against Cancer

Update Edition 2005

Global (Year)

7.6 millionDeaths

10.9 millionNew Cases

5Liver

1Lung

7Esopha-

geal

3Colon

2Breast

4Gastric

6Cervical

Cancer


Disease Control Division,Ministry of Health of PRC, 2008

Cancer

6Cervical

5Colon

8Nasopharyn-

geal

1Lung

2Liver

7Breast

3Gastric

4Esopha-

geal

City 24.41 (2nd)

Incidence 2,000,000Mortality 1,500,000

Cancer

Countryside 26.93 (1st)

Liver CancerMortality/100,000

Against Cancer in China


The third time investigation of death causes: The third time investigation of death causes:

cancer ranks second with a mortality rate of 22.32%.cancer ranks second with a mortality rate of 22.32%.

————Ministry of Health, April 2008Ministry of Health, April 2008


增生

异常

• Inherited susceptibility

• Chemical carcinogens

• Radiation

• Infectious agents

• DNA repair system

• General health (diet, stress, etc)

• Immune system

Cancer Cell: Contributing Factors

Cancer EtiologyCancer Etiology

Multiple CarcinogenMultiple Carcinogen

Cancer: Three CharacteristicsCancer: Three Characteristics


TumorTumor

InitiationInitiation

DevelopmentDevelopment

InvasionInvasion

MetastasisMetastasis

TumorTumor

InitiationInitiation

DevelopmentDevelopment

InvasionInvasion

MetastasisMetastasis

Dysplasia

Carcinomain situ

Cell

Primary cancer

Metastasis(2nd Cancer)

genomic DNA

transcription

translation

post-transcription

post translation

Multiple StagesMultiple Stages


EMBO reports 1, 2, 115-119, 2000

Loss of genomic integrity and imbalance of Loss of genomic integrity and imbalance of molecules are mechanism for the cancer incidencemolecules are mechanism for the cancer incidenceLoss of genomic integrity and imbalance of Loss of genomic integrity and imbalance of molecules are mechanism for the cancer incidencemolecules are mechanism for the cancer incidence

Multiple Gene MutationMultiple Gene Mutation


Cancer Initiation & Invasion

Four Mechanisms

Cancer Initiation & Invasion

Four Mechanisms


Mining the cancer genome

Tobias Sjoblom, et al. Science 314: 268-274, 2006

Cancer Genome AtlasCancer Genome Atlas

Sequencing: 13,023 genes

removing errors, normal variants

Breast cancer 11 Samples Colon cancer 11 Samples

1149 mutation genes

(Individual tumors: average 90)

(Significant frequency: 189 genes)

(Significant frequency: average 11 per tumor)

Cancer MolecularBalance & Mutation


Sohrab P. Shah, et al. Nature 461: 809-813, 2009

Mutational evolution in a lobular breast tumour profiled at single nucleotide resolutionMutational evolution in a lobular breast tumour profiled at single nucleotide resolution

Recent advances in next generation sequencing 1, 2, 3, 4 have made it possible to precisely characterize all somatic coding mutations that occur during the development and progression of individual cancers. We found 32 somatic non-synonymous coding mutations present in the metastasis. Five of the 32 mutations (in ABCB11, HAUS3, SLC24A4, SNX4 and PALB2) were prevalent in the DNA of the primary tumour removed at diagnosis 9 years earlier, six (in KIF1C, USP28, MYH8, MORC1, KIAA1468 and RNASEH2A) were present at lower frequencies (1–13%), 19 were not detected in the primary tumour, and two were undetermined.

Recent advances in next generation sequencing 1, 2, 3, 4 have made it possible to precisely characterize all somatic coding mutations that occur during the development and progression of individual cancers. We found 32 somatic non-synonymous coding mutations present in the metastasis. Five of the 32 mutations (in ABCB11, HAUS3, SLC24A4, SNX4 and PALB2) were prevalent in the DNA of the primary tumour removed at diagnosis 9 years earlier, six (in KIF1C, USP28, MYH8, MORC1, KIAA1468 and RNASEH2A) were present at lower frequencies (1–13%), 19 were not detected in the primary tumour, and two were undetermined.

Sohrab P. Shah, Ryan D. Morin, Jaswinder Khattra, Leah Prentice, Trevor Pugh, Angela Burleigh, Allen Delaney, Karen Gelmon, Ryan Guliany, Janine Senz, Christian Steidl, Robert A. Holt, Steven Jones, Mark Sun, Gillian Leung, Richard Moore, Tesa Severson, Greg A. Taylor, Andrew E. Teschendorff, Kane Tse, Gulisa Turashvili, Richard Varhol, René L. Warren, Peter Watson, Yongjun Zhao, Carlos Caldas, David Huntsman, Martin Hirst, Marco A. Marra & Samuel Aparicio

Sohrab P. Shah, Ryan D. Morin, Jaswinder Khattra, Leah Prentice, Trevor Pugh, Angela Burleigh, Allen Delaney, Karen Gelmon, Ryan Guliany, Janine Senz, Christian Steidl, Robert A. Holt, Steven Jones, Mark Sun, Gillian Leung, Richard Moore, Tesa Severson, Greg A. Taylor, Andrew E. Teschendorff, Kane Tse, Gulisa Turashvili, Richard Varhol, René L. Warren, Peter Watson, Yongjun Zhao, Carlos Caldas, David Huntsman, Martin Hirst, Marco A. Marra & Samuel Aparicio

Cancer Genome AtlasCancer Genome Atlas


Travel

Intravasation

Primary tumor (Cell escape)

Invasion

Extravasation

Transport

Growth(2nd tumor)

Migration

Invasion & Metastasis— Main Death Causes


Mesenchymal cell-like movement

Amoeba-like movement

A little long cell

Need protease

Form of pseudopodium

A little rely onprotease

Myosin strongContraction

A little circle cell

Rac/WAVE2 signal

Rho/ROCK signal

Cell. 2008 Oct 31; 135 (3): 510-523

Adhesion MovementAdhesion Movement


G1→S→G2→MR

Control of cell cycleControl of cell cycle

G1: DNA pre-synthesis

S: DNA synthesis

G2: DNA post- synthesis

M: Cell division

GO: Oncogenes

STOP: Tumor superessor


Epithelial lining cellsTransformed epithelial cells

Tumor fbTissueMartrixTransformed Transformed

epithelial cellsepithelial cellsMMP-7, 13MMP-7, 13 (9)(9)

Epithelial cells ofEpithelial cells oftumor angiogenesistumor angiogenesis

MMP-1, 2, 14MMP-1, 2, 14

Tumor fbTumor fbMMP-MMP-1, 2, 3, 11, 141, 2, 3, 11, 14

Matrix degradation: Matrix degradation: Structural base of tumor invasion & metastasisStructural base of tumor invasion & metastasisMatrix degradation: Matrix degradation: Structural base of tumor invasion & metastasisStructural base of tumor invasion & metastasis

Matrix Degradation —— MMPsMatrix Degradation —— MMPs


Tumor>2-3mm: Need vessels

Key Molecular: MMPs, VEGF, bFGF, PDGF

Tumor vessels density is a marker for early

diagnosis and prognosis Nature Rev Cancer, 4, 2004

AngiogenesisAngiogenesis


Cancer: Wounds that fail to heal

Nature, 420, 2002

The Chain of Inflammation & Cancer

Tumor Development


CancerCancerChronic inflammation

Chronic inflammation

BacterialH pylori幽门螺杆菌

BacterialH pylori幽门螺杆菌

VirusEB, HCV

VirusEB, HCV

Parasiteflukes, schistosomes

吸虫，血吸虫

Parasiteflukes, schistosomes

吸虫，血吸虫

Chemial irritisPMA 佛波酯

Chemial irritisPMA 佛波酯

Nondigestible Particles

asbestos, silica石棉纤维，矽

Nondigestible Particles

asbestos, silica石棉纤维，矽

Strong association: inflammation and cancer

Strong association: inflammation and cancer


Oxidative Stress 氧化应激 Oxidative Stress 氧化应激

Aldehydes 醛 Aldehydes 醛 Oxidized DNA nucleosides

氧化脱氧核苷酸

Oxidized DNA nucleosides

氧化脱氧核苷酸

DNA mutation DNA mutation

Peroxynitrite过氧亚硝基

Peroxynitrite过氧亚硝基

ROS活性氧ROS

活性氧

DNA damage DNA damage

P53 、 Rb P53 、 Rb

DNA repair DNA repair

RNS活性氮RNS

活性氮

Chronic inflammationChronic inflammation


Inflammatory cytokines & OncogenesInflammatory cytokines & Oncogenes

OncogenesInflammatory

cytokines

RASIL-1a, IL-1b, IL-6, CXCL8, IL-11

MYC

IL-1b, CCL2, CCL5, CCL7, CXCL1, CXCL2

RET tyrosine kinase

CXCL8, CXCR4, CCL2, MCP-1, GM-CSF

EGFR CXCL8

proto-oncogene Tpl2 NF-kB

OncomiRs, miR-155

Inflammatory mediators in macrophages and in monocytesCancer Letters, 267, 2008

Tumor inflammation environment


ROS

HIF

Blood vessel

Cancer cell

Macrophage

Cytokines Chemokines

CD147

CypA

Erk1/2 P38Proliferation

Anti-apoptosis

Angiogenesis

CypA

Hypoxia

Our study

Not confirmed

Reported

PI3K

Signal TransductionSignal Transduction

The Seven Hallmarks of Cancer and Their Links to Tumor Metabolism

Cancer Cell. 13: 472-482, 2008

凋亡

血管生成

无限增殖

侵袭转移抵抗抑瘤

免疫逃避

增生环路

Tumor Metabolism


J Clin Invest. 118 (12): 3835-3837, 2008

Intratumoral hypoxia and metabolic symbiosis

or Aerobic stromal cell

Anaerobic glycolysisAnaerobic glycolysisAnaerobic glycolysisAnaerobic glycolysis


Molecular Mechanisms of Cancer-Specific Metabolic Reprogramming

葡萄糖转运

糖原分解

乳酸产生

氧化磷酸化降低

脂类合成

氧化抑制


Leland H. Hartwell, et al. Nat Biotech, 24 (8), 2006

高危评估 Risk assessment

早期诊断 Noninvasive screening for early-stage disease

检测定位 Detection and localization

预后判断 Disease stratification and prognosis

治疗反应 Response to therapy

复发监测 Screening for disease recurrence

Cancer Biomarker -- A Systems ApproachCancer Biomarker -- A Systems Approach


• Biomarker Discovery:

Expression Mapping (Modification mapping)

Functional proteomics: interaction of the proteins

Epitope mapping (active core)

• Biomarker Discovery:

Expression Mapping (Modification mapping)

Functional proteomics: interaction of the proteins

Epitope mapping (active core)

Seperation and identification of mixture sample Ab array, Cell array, Tissue array, Co-IP (pull-down), Biosence, etc. Comparative proteomics 2DE, 2DELC-MS

Validation peptides sequence of protein MALDI-MS, SELDI-MS, LC-MSMS, ESI-MS (m/z) Analysis of the databases

Seperation and identification of mixture sample Ab array, Cell array, Tissue array, Co-IP (pull-down), Biosence, etc. Comparative proteomics 2DE, 2DELC-MS

Validation peptides sequence of protein MALDI-MS, SELDI-MS, LC-MSMS, ESI-MS (m/z) Analysis of the databases

How Identified Cancer Biomarker?How Identified Cancer Biomarker?


System Biology Approaches “-omic” Technologies (Preclinical or Clinical Utilization)System Biology Approaches “-omic” Technologies (Preclinical or Clinical Utilization)

Trends Biotechnol. 23 (11), 544-546, 2005

Amplichip Cyp 450 TestAmplichip Cyp 450 TestGlobal SNP ArraysGlobal SNP Arrays

MicroarrayMicroarraySAGESAGE

MALDI-MS/MSMALDI-MS/MS2D Gels- MS2D Gels- MS

NMRNMRGC-MSGC-MSLC-MSLC-MSFT-IRFT-IR


What is an Ideal Cancer Biomarker?What is an Ideal Cancer Biomarker?

Screening a healthy population or a high risk population for the presence of cancer

Making a diagnosis of cancer or of a specific type of cancer

Determining the prognosis in a patient

Monitoring the course in a patient in remission or while receiving surgery, radiation, chemotherapy, or biotherapy

Screening a healthy population or a high risk population for the presence of cancer

Making a diagnosis of cancer or of a specific type of cancer

Determining the prognosis in a patient

Monitoring the course in a patient in remission or while receiving surgery, radiation, chemotherapy, or biotherapy


Application of Cancer BiomarkerApplication of Cancer Biomarker

Identification and diagnosis

Individuals affected with disease

People who may be at risk but do not yet exhibit symptoms

Monitor progress of disease

Monitor effects of treatment

Remission

Follow-up

Cancers found in early stage: low morbidity and recurrence rates

Cancers identified in late stage: high recurrence and mortality rates

Identification and diagnosis

Individuals affected with disease

People who may be at risk but do not yet exhibit symptoms

Monitor progress of disease

Monitor effects of treatment

Remission

Follow-up

Cancers found in early stage: low morbidity and recurrence rates

Cancers identified in late stage: high recurrence and mortality rates


Cancer Biomarker and Types of Cancer: statistically significant association between a particular cancer and the associated cancer marker (s)

AFP = alpha fetoprotein

CEA = carcinogenic embryonic antigen

CA 15-3 = carbohydrate antigen 15-3

CA 19-9 = carbohydrate antigen 19-9

CA 125 = carbohydrate antigen 125

PSA = free prostate specific antigen + prostate specific antigen - alpha(1)antichymotrypsin complex

PSAF = free prostate specific antigen

PSAC = prostate specific antigen - alpha(1)antichymotrypsin complex

PAP = prostatic acid phosphatase

hTG = human thyroglobulin

hCGb = human chorionic gonadotropin beta

Ferr = Ferritin

NSE = neuron specific enolase

IL-2 = interleukin 2

IL-6 = interleukin 6

A2M = alpha 2 macroglobulin

B2M = beta 2 microglobulin


Problems of Cancer BiomarkerProblems of Cancer Biomarker

No cancer biomarker is absolutely specific

No cancer biomarker test is free of false negatives

No cancer biomarker test is free of false positives

No cancer biomarker is absolutely specific

No cancer biomarker test is free of false negatives

No cancer biomarker test is free of false positives


Antibody Based Cancer BiomarkersAntibody Based Cancer Biomarkers

Cancer Marker Antibody Cancer

CA125 OC 125 ovarian cancer

CA 15-3 DF3, 115D8 breast cancer, ovarian cancer

CA 549 BC4E549, BC4N154 breast cancer, ovarian cancer

CA 27-29 B27.29 breast cancer

MCA b-12 breast cancer, ovarian cancer

DU-PAN-2 Du-PAN-2 pancreatic cancer, ovarian cancer,gastrointestinal cancers, lung cancer

CA 19-9 19-9 pancreatic cancer, liver cancer,gastrointestinal cancers

CA 19-5 19-5 pancreatic cancer, ovarian cancer,gastrointestinal cancers

CA 50 C50 pancreatic cancer, rectal cancer,gastrointestinal cancers

CA 72-4 B27.3, cc49 gastric carcinoma, pancreatic cancer,ovarian cancer

CA 242 C242 gastrointestinal cancerspancreatic cancer

Her-2/neu Herceptin breast cancer

HAb18G/CD147HAb18G/CD147 HAb18HAb18 hepatocellularhepatocellular carcinoma, ectcarcinoma, ect



Human genome is 2.91-billion base pairs in length (1990)

There are about 25,000 genes exist in the human genome (42% have an unknown

function)

Approximately 12,000 genes that appear to have the capacity to make secreted

proteins, all the genes have been determined the entire nucleotide sequences

(3141 genes locus on the first chromosome)

At May 2006, the first chromosome sequences were completed, at least 1,000

new genes were found. This is the end of 16 year’s Human Genomic Plan.

This has empowered more direct means of target identification,

e.g. by expanding protein databases and enabling the mapping of

novel cancer-associated genes (Venter et al. 2001).



Complete genome sequences have provided a plethora of

potential drug targets. But the hard task of finding their weak

spots is just beginning (about 5000 genes can be used as drug

target)

A challenging new development in the field of drug-target

discovery is systems biology, or the recognition that genes, or

better the gene products, are part of, and function, in large

complex networks.

Complete genome sequences have provided a plethora of

potential drug targets. But the hard task of finding their weak

spots is just beginning (about 5000 genes can be used as drug

target)

A challenging new development in the field of drug-target

discovery is systems biology, or the recognition that genes, or

better the gene products, are part of, and function, in large

complex networks.

Nature, 428, 225-231, 2004.


转化医学与公众健康转化医学与公众健康转化医学与公众健康转化医学与公众健康


• EA. Zerhouni ， NIH 路线图计划（ NIH Roadmap ）， 2003

• 将医学生物学基础研究成果迅速有效的转化为可在临床实际应用的理论、技术、方法和药物

• → →基础研究临床应用，实验室成果产业化• 在实验室到病房（ Bench to Bedside, B2B ）之间架起一

条快速通道• 双向、开放：

• 基础研究提供新疗法、新药物• 临床研究者对疾病的进程和特性提供反馈意见

• “ ”驱动临床研究引擎的激发器

转化医学概念转化医学概念转化医学概念转化医学概念


• 美国已经在 38 所大学建立了转化医学研究中心，在 2012 年以前将会达到 60 个以上， NIH 每年资助经费达 5 亿美元

• 英国已投入 4.5 亿英镑用于转化医学研究，并启动世界上首个转化医学合作研究中心

• 欧洲共同体为转化医学计划投入 60 亿欧元

• Science Translational Medicine 、 Journal of

Translational Medicine 和 Translational Research 三本国际性专业杂志

转化医学发展现状转化医学发展现状转化医学发展现状转化医学发展现状


• 转化医学战略研讨会• 2009 ，中国工程院• 2010 ，中国科学院

• 转化医学中心• 中南大学• 上海交通大学• 同济大学

• 成果转化率： 25% ，商品化： <15%

中国的转化医学中国的转化医学中国的转化医学中国的转化医学


转化医学与转化医学与 4P4P 医学医学转化医学与转化医学与 4P4P 医学医学

Predictive Medicine – 预测医学

Preventive Medicine – 预防医学

Personalized Medicine – 个体化医学

Participate Medicine – 参与医学


Personalized Prevention & Early Detection Personalized Prevention & Early Detection

Lifestyle changes

Screening

Chemoprevention

Prophylactic Surgery

Average Moderate High Very High

RISK


Personalized Medicine – 个体化治疗Personalized Medicine – 个体化治疗• No “one size fits all” drug• Most drugs work for 30% to 70% of patients• Multiple factors determine drug responses• Phamacogenetics is essential for individualized therapy


Herceptin, the first

marketed personalized

medicine, was approved

using a coordinated

drug/diagnostic approval

process that will become

more common.

Personalized Medicine – 个体化治疗Personalized Medicine – 个体化治疗


Our StudyMolecular Docking

HAb18G & Its Antibodies

HAb18G/CD147 & HAb18, 6H8, 5A12 Antibodies

HAb18G/CD147 I Set


Our Study Crystal Structure of HAb18G/CD147

C2

I

Xiao-Ling Yu, Zhi-Nan Chen, J Biol Chem, 283 (26), 2008National patent: 200710018514.X

PCT patent: PCT/CN2007/003034 PDB ID: 3B5H

HA

b18G

/C

D147

Our Study Tissue Atlas - HAb18G/CD147



Shao-Hui Hu, Zhi-Nan Chen, et al. Proteomics, 7 (13), 2007

Lung

Liver

Fetal Normal Cancer

Case Positive Positive Rate(%) P

Fetal 67 1 1.49 <0.0001

Normal 196 16 8.16 <0.0001

Cancer 1565 1040 66.4566.45


Yu Li, Zhi-Nan Chen, et al. Histopathology, 2009


组织类型病例数阳性率灵敏度（真阳性率）

特异度（真阴性率）

肝癌 147 例 77.66%77.66% 100%良性病变肝组织 44 例 0.00%

肺癌 186 例 95.04%95.04% 98%良性肺疾病 100 例 2.00%

乳腺癌 200 例 65.35%65.35% 91%良性乳腺疾病 100 例 9.00%

食管癌 199 例 86.22%86.22% 97.5%良性食管疾病 80 例 2.77%

卵巢癌 101 例 76.23%76.23% 83.67%良性卵巢疾病 52 例 16.33%

胃癌 193例 76.68%76.68% 83.37%良性胃疾病 98例 16.63%

宫颈癌 120例 71.84%71.84% 88.46%良性宫颈疾病 55例 11.54%

直肠癌 195 例 77.68%77.68% 93.93%良性直肠疾病 100 例 6.07%

平均值 78.75% 91.73%

第四军医大学 Fourth Military Medical UniversityCancer Biol Ther 5 (7), 2006

Our Study Iodine (131I) Metuximab Injection(LICARTINTM)


Anti-recurrence Treatment after Liver Transplantation

recurrence rate30.42↓ survival rate

20.62↑

AFP44.08%

57.09%

26.67%

82.50%

61.88%

P=0.0174 P=0.0289

P=0.0016

Control groupTreatment group

60 cases HCC (III, IV stage)

87.82%


Hepatology, 45 (2): 269-276, 2007



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肿瘤生物学与转化医学 cancer biology & translation medicine 第四军医大学...

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