cikk-referáló

Génátírás szabályozása: transzkripciós mechanizmusok

Dr. Marcsek Zoltán

GénexpresszióGénexpresszió

Courtesy The Protein Society

eukaryoták promóter elemei

• a promóter elemei többé-kevésbé jól definiáltak

• a transzkripció start helyétől különböző távolságra vannak

általában van

gyakran hiányzik

egy tipikus promóter

az eukaryota transzkripció szabályozása

szabályozás transzkripciós faktorokkal

• egy transzkripciós faktor (vagy transzkripciós faktorok egy csoportja) egy géncsoport expresszióját szabályozhatja (pl. heat shock fehérjék, G1/S restrikciós pont fehérjék, stb)

• egy gén átírását egy vagy több, egymástól független transzkripciós faktor szabályozhatja (pl. metallothionine)

• magas expresszió kiváltásához gyakran több, különböző 'response' elemhez kötő transzkripciós faktor lehet szükséges

Nuclear Factor-κBNuclear Factor-κB: A Holy Grail in Cancer Prevention and TherapyA Holy Grail in Cancer Prevention and Therapy

Bharat B. Aggarwal, Bharat B. Aggarwal, Gautam Sethi, Gautam Sethi,

Asha Nair, Asha Nair, Haruyo IchikawaHaruyo Ichikawa

Cytokine Res. Lab, Dept. Exp. Therapeutics, Cytokine Res. Lab, Dept. Exp. Therapeutics, Univ. Texas M.D. Anderson Cancer Center, Houston, TxUniv. Texas M.D. Anderson Cancer Center, Houston, Tx

’’invited review’ invited review’ Current Signal Transduction and TherapeuticsCurrent Signal Transduction and Therapeutics

2006.2006.

NFkBmi az, mit csinál,

miért jó, ha ismerjük?

transzkripciós faktor

NFB• Baltimore & Sen fedezték fel mint egy B-sejt-specifikus

magi proteint amely az immunoglobulin könnyű láncának meghatározott szekvenciájához kötődik és a gén átírását serkenti (enhancer) (Cell 47:921-928, 19861986)

• az NFB a Rel transzkripciós faktor családba tartozik amely nagyszámú, az immunválasszal, a gyulladással, a sejtosztódással és apoptózissal összefüggő gén expresszióját szabályozza

• az emlős sejtekben öt eltérő NFkB (rel) alegységet ismerünk amelyek kb. 300 nagyon konzervált aminosat tartalmaznak

• a citoplazmában az NFkB inaktív formában több, különböző kombinációban található (heterodimer p50/p65 (Rel A) és az inhibítor IB alegység)

Family of NFB and Inhibitory B (IB) proteins

Nat.Rev.Cancer 2:301-310, 2002

p105 (contains p50 NFkB1)

• Name: p105 (contains p50 subunit, a.k.a. NFkB1)• Descr: Once in the nucleus initiates transcription of

hundreds of genes• Length: 968

p100 (contains p52 NFkB2)

• Name: p100 (contains p52 subunit, a.k.a. NFkB2)• Descr: Once in the nucleus initiates transcription of

hundreds of genes• Length: 899

IkB• Name: Inhibitor of NF kappaB beta• Length: 356

• Consists mostly of ankyrin repeats that serve inhibitory function

IkB

• Name: IkappaB epsilon• Length: 361

• Consists mostly of ankyrin repeats that serve inhibitory function

IKK

Name: IKappaB kinase alpha

Descr: Forms complexes with IKKLength: 745 aa

Conserved Serine/Threonine protein kinase Domain detected at 10 – 300 aa.

IKK

• Name: IKappaB kinase beta• Descr: Forms complexes with IKK and destroys IkB’s• Length: 756

Similarly to IKK a slightly less conserved Serine/Threonine protein kinase Domain detected at 10 - 290 aa.

IKK

• Name: IKappaB kinase gamma• Length: 487 aa

Contains Chromosome segregation ATPases domain

az NFB aktiválása• NFB a citoplazmában található inaktív formában mint

az IB inhibítorához kötött p50/65 heterodimer • extracelluláris indukciós hatásra az IB-kináz komplex

(IKK) aktiválódik és foszforilálja az IBtszerin 32 és 36) amely az IB lizin21 és lizin22 ubiquin-kötését okozza

• ez az IB alegység lebotását eredményezi (26S proteasome)

• az inhibitor alegység lebomlása a p50/p65 komplex aktivációját eredményezi: az NFkB a citoplazmából a sejtmagba kerül (transzlokáció)

• a sejtmagban az NFkB transzkripciós faktor a ‘consensus szekvenciához’ (5’-GGGACTTTC-3’) kötődik és aktiválja a specifikus gének expresszióját

Aktiváló Stimulus(kfl)

IB-kináz/NEMO/IKAP komplex

IB

p65p50

IB

p65p50

PIB

degradáció

p65p50CITOPLAZMA

p65p50

NUKLEUSZ

mRNS

gyulladási-/immun-stb. fehérjék

transzlokáció

transzkripció

Target genes

Leukemia 16:1053-1068, 2002

Factors that induceinduce NFB

Reactive Oxygen Species (ROS)

Free Radic Biol Med25:346-361

kérdés:kérdés:

mely szignálok aktiválják az NFkB aktivitást?

CytokinesCytokinesTNFα , TNFβRANKL, TRAlLCD40 ligandCD30 ligandTWEAKTHANKBAFFLTβIL-1, IL-2, IL-4IL-8, IL-12, IL-15IL-17, IL-18CD11b/CD18 ligandCD2 ligandCD35 ligandCD3 ligandCD28 ligandCD43 ligandCD4 ligandCD66a ligandLlFS100BAgonist antibodies to AT1 receptorIgM ligandBeta-D-glucan ligandIgG2aFlt-1 ligandLy6A/E ligand

Growth factors and Growth factors and hormoneshormones

GMCSFM-CSFNGFEGFInsulin-like growth factor 1InsulinHepatocyte growth factorGastrinPEDFPlatelet-derived growth factorBone morphogenic protein 2Bone morphogenic protein 4Cortical releasing hormone Follicle-stimulating hormoneMullerian inhibiting substancePAFPlant steroids (diosgenin, hecogenin, tigogenin)Prostratin

OncogenesOncogenesRasMycBcr-ablAll-trans retinoic acidRET/PTC3 fusion oncoprotein S100BSerumSulphatideTGF-αTGF-β

Table 1. A list of inducers of NF-kB activationinducers of NF-kB activationChemical AgentsChemical Agents L-NMA, Ethanol, MonensinNicotine, Adriamycin2-DeoxyglucoseAnisomycinBenzyl isothiocyanateBisperoxovanadium phosphotyrosylBrefeldin A, CadmiumCalyculin A, ColchicineCalcium lonophore plus phorbol esterCeramide-beta-galactose2-chloroethyl ethyl sulfide Con A, Cycloheximide Cyclopiazonic acidDiquat, 2,4-DinitrofluorobenzeneFerrocene, ForskolinGadolinium chlorideGlass fibersSodium butyrate trichostatin AMalondialdehydeMDMA, MEN 17055N-methyl-D-aspartateMycophenolic acidNafenopin, N-NitrosomorphineNocodazol, Okadaic acidPeplomycin, PhytohemagglutininPhorbol ester, Podophy/lotoxin Polychlorinated biophenyl-77Prostratin, PyrogallolStaurosporine, TunicamycinTransglutaminase 2 WF10, WY-14 643, CFTR, GPR56Hematopoietic progenitor kinase 1Ig heavy chain, MHC Class I

Table 1. A list of inducers of NF-kB activationinducers of NF-kB activation (ctd)VirusesVirusesAdenovirusCytomegalovirusEBVHammarskjöld and hepatitis B virusHVSHuman herpes virus 6HIV-1Herpes simplex virus-1HTLV-1Influenza virusMeasies virusMoloney murine leukemia virusNewcastle disease virusRespiratory syncytial virusRhinovirusSARS coronavirusSendai paramyxovirusSindbis virusVaccinia virus akaraWest Nile flavavirus

Viral productsViral productsAdenovírus 5: E1AAdenovirus: E3/19KAfrican swine fever virus: IAPAntibody to dengue virus nonstructural protein 1CMV: ie1EBNA-2EBV: LMP1HBV: HBxHBV: LHBsHBV: MHBstHCV: Core proteinHVS13HVS: StpCHIV-1: gp160HIV-1: NefHIV-1: p9 (9 aa peptide)HIV-1:TatHTLV-I: TaxHTLV-II: TaxInfluenza vírus: hemagglutininKSHV: K1Parvovirus B19: NSIRespiratory syncytial virus: M2-1SV40

Bacteria and fungiBacteria and fungiAnaplasma phagocytophilumAngiostrongylus cantonensis Bacteroides forsythus Bartonel/a henselae Bordetel/a pertussisBorrelia burgdorferiChlamydia pneumoniaeCryptococcus neoformans Ehrlichia chaffeensisEnteropathogenic Escherichia coliFusobacterium nucleatum Gardnerella vaginalis Helicobacter pyloriLactobacilliListeria monocytogenesMycoplasma fermentans Mycobacteria tuberculosis Neisseria gonorrhoeae Neisseria meningitidisPorphyromonas gingivalis Prevote/la intermedia Pseudomonas aerogenosa Rhodococcus equiRickettsia rickettsiiSalmonella dublinSalmonella typhimurium Shigella fIexneri Staphy/lococcus aureus StreptococcusStreptomyces californicus Trichomonas vaginalisUreaplasma urealyticum Yersinia enterocolitica

Bacterial/fungal productsBacterial/fungal productsLPSWogoninEnterotoxinGIycosylphosphatidylinositolsFumonisin B1Staphylococcus enterotoxins A and BApicularen ACpGCytolysinDiphosphoryl lipid AExotoxin 8 Lipoprotein LpKLipoprotein LpKLipoteichoic acidMembrane lipoproteins(Mycoplasma fermentans,Mycoplasma penetrans)MannoproteinsMuramyl peptides Mycobacterium species lipoarabinomannan PicA, PlcB Porin 1BToxic shock syndrome toxin 1

Eukariotic parasietisEukariotic parasietisEimeria tenella and necatrix Leishmania PhospholipomannanPneumocystisTheileria parvaTrypanoplasma borreli

Table 1. A list of inducers of NF-kB activationinducers of NF-kB activation (ctd)Physiologic Physiologic (stress)(stress) cond. cond.Asthma, Rheumatoid arthritisCrohn's disease/ulcerative colitisCorneal epidemic keratoconjunctivitisCoronary artery bypass Hypercholesterolemia Hyperglycemia Hyperhomocysteinemia Hyperosmotic shock, Shear stressHypoxia, ReoxygenationAcute jung injuryAcute respiratory distress syndromeAdhesion, HemorrhageAngina pectoris Antiphospholipid antibodies AppendicitisDepolarizationIschemia (transient, foGaI) Ischemic preconditioning Liver regenerationBiotin deficiencyButter and walnut diet Cecalligatin and puncture Mechanical ventilation Memory retrievalMiddie cerebral artery occlusionMuscle disuse, Muscular dystrophy Neuronal firing Overventilation pancreatitis ProteinuriaSenescenceNeuroonal trimethyltin injury Uniaxial cyclic cell stretching T -cell selection

Physical stressPhysical stress UV A, B and C radiation Bile duct ligationCyclic mechanical muscle strainExercice γ radiationHeavy ion irradiationHigh-fat dietLaminar shear stressDietary lipid hydroperoxidePPME photosensitizationMechanical lung ventilationObesityWounding combined with He-Ne irradiationWounding combened with thermal irradiathion

OxldativeOxldative stressstressGlutathione Hydrogen peroxide Catalase Nitric oxide OzonePeroxiynitriteButyl peroxidePervanadateReoxygenation

Environmental hazardsEnvironmental hazardsCigarette smokeArsenicPolychlorinated biphenyl Benzo[a]pyrene diol epoxide Crocidolite asbestos fibers DicambaDiesel exhaust particlesGliadinHouse dust miteDust particlesFear-potentiated startie responseLead Lead chromateNoiseOlly fly ashSilica partlclesWood smokeZymosan

Chemotherapeutic drugsChemotherapeutic drugsCisplatin, PaclitaxelDoxorubicin, TamoxifenVinblastine, VincristineEtoposideAra-CBryostatin-1CamptothecinDaunomycinDaunorubicinMitoxantroneGemcitabine

Therapeutic drugsTherapeutic drugsCelecoxib, AZT, CiprofibrateBleomycinABR-25757 SN38SA 981, WR1065AdAMP, AnthralinCycloprodigiosinDacarbasineOrengedeokutoDiazoxide5,6-dimethylxanthenone-4-acetic acidFlavone-8-acetic acidHaloperidolCunbi-BoshinHangam-TangMethamphetamineOltiprazPhenobarbitalProtocatechuic acid

Table 1. A list of inducers of NF-kB activationinducers of NF-kB activation (ctd)Fatty acids Fatty acids Palmitate Leukotriene B4Free fatty acids Llnolelc acidOleic acidSaturated fattx acidsLysophosphatldlc acid

Modified Proteins Modified Proteins LDL AGEs Amyloid protein fragment Glycosylated oxyhemaglobin Maleylated bovine serum albumin Neurotrophin receptor proteolytic fragmentsNonamyloid beta component of Alzheimer's diseaseATM

Heat shock proteinHeat shock proteinHSP60 HSP71

Apoptotlc MediatorsApoptotlc Mediators Anti-Fas/Apo-1PARP

MetalsMetalsLithiumTitanium and copper implantsNickel sulfateIronManganaseNickelChromiumCobaltAluminium

Natural productsNatural productsAreca nut extractBaicalinBeta-caroteneQuercetinQuinolinic acidSafflower polysaccharidesSanglifehrin A

Physiological MediatorsPhysiological MediatorsAlloxanAngiotensin IIBradykininFibrinogenHemoglobinHyaluronanThioredoxinKainan acidL-GlutamateMelaninViolaceinVitamin D3AdesonineActivin AAlbuminAllerginAcrp30/adiponectinAdrenomedullinAeginetia indica L (AIL)b-ABronchoalveolar lavage fluidBrusatolHomocysteineCryptdinsDeoxicholic acid12-hydroxyeicosatrienoic acid6-HydroxydopamineHuman peptide transporter 1LTPLyso PCNeuomelaninNS-398OsteopontinPAFPCSCProlactin

PIF12-LipoxygenaseThrombinStreptozotocinSubstance PPotassiumBum patient serumAnaphyloxin C3a and C5aAngiotensin II type 1 receptoractivating antibodiesAntiphospholipid antibodiesBasicCalciumPhosphate crystalsC2-ceramide ceruleinChelidonium majus extractCollagen laticeCollagen type 1Cysteinyl leukotrinesDes-Arg 10-kallidinMixed meal ingestionMonosudium urate crystalsNeutrophil elastasePhellinus linteus proteoglycanPlatelet type arachidonatePolysachcharides of Poria cacosRegulatory RNAS100BSleep deprivationTUDCATuberous sclerosis complexUremic toxins, VLDL

IK P

NFBIB

P

NFB

IB Degraded

IB geneNFB

IB

IL-6 geneNFB NFB

IB

PNFBIB

Nucleus

Cytoplasm

Constitutive NF-Constitutive NF-B Signaling B Signaling ModuleModule

IK

Negativefeedback

Input signal

NLS

kérdés:kérdés:

mely szignálutak vesznek részt az NFkB aktivitásában?

Table 2. protein kinases implicated in NF-kB activationNF-kB activation

KinaseKinase LigandLigand

IKKα Proinflammatory cytokines (eg TNF, IL-1, IL6), LPS, IkBIKKβ Proinflammatory cytokines (eg TNF, IL-1, IL6), LPS, IkBIKKγ Proinflammatory cytokines (eg TNF, IL-1, IL6), LPS, IkBIKKi/IKKε TNF, PMA, IkBNIK TNF, CD95, and IL-1NAK Phorbol esters, growth factorsPI-3K IL-1Akt1 TNF, TCR/CD28HPK-1 UnknownPKC-α TPA, TNF, Bimp1, Bcl1 O/MAL T1PKC-β IgM receptor, Bimp1, BcI10/MAL T1PKC-τ CD3-CD28 (TCR/CD28), Bimp 1, Bcl1 O/MAL T1PKC-δ TNFαPKC-ε Phorbol ester, Bimp1, Bcl1 O/ MAL T1aPKC TNF, ras p21MEKK1/2 Proinflammatory cytokinesMEKK3 TNF, other proinflammatory cytokinesTPL-2/Cot kinase TNFpp90rsk Pseu omonas aeruglnosaRaf-1 kinase Serum ~rowth factors, phorbol ester, and PTK oncogenesPKR dsRNA TAK1/MEKK XIAP, TGF-βMLK3 CD3/CD28PAK1 p21, LPSBTK B-cell antigen receptorJAK2 ErythropoietinPKA IL-1, LPS, many othersIRAK-1 IL-1IRAK-2 IL-1, LPsIRAK-M IL-1, LPSP56 Ick CeramideSyk TNF, H2O2

min. 32 ismert kinázmin. 32 ismert kináz

kérdés:kérdés:

mely betegségekben mutatható ki abberáns (állandóan magas) NFkB aktivitás?

többet tudunk, ha ismerjük a rendszer hibáit...

Table 3. association of NF-kB activationNF-kB activation with diseases

CancerCancerB-cell lymphomaBreast cancerHodgkin's diseaseLiver cancerMantle cell lymphomaMultiple myelomaNon-Hodgkin's lymphomaOvarian cancerProstate cancerT -cell lymphomaThyroid cancerBladdercancerColon cancerEsophagealcancerHead and neck squamous cell carcinomaLaryngealcancerLung cancerPancreatic cancerPharyngealcancerRenal carcinomaAcute lymphoblastic leukemiaAdult T-cell leukemiaCervical cancerNasopharyngeal carcinomaMelanoma

Cardiovascular diseasesCardiovascular diseasesAtherosclerosisCardiac hypertrophyHeart failureHypercholesterolemia

Diabetes mellitusDiabetes mellitusType IType II

Inflammatory diseasesInflammatory diseasesAlzheimer's diseaseArthritisAsthmaBone resorptionChronic obstructive pulmonary disease (COPD)Crohn's diseaseHuntington's diseaseInflammatory bowel diseaseInflammatory response syndromeMultiple sclerosisOcular allergyParkinson's disease

Other diseasesOther diseasesAgingAIDSCatabolic disordersEctodermal dysplasiaGut diseasesHeadacheHelicobacter pylori-associated gastritisInontinentia pigmenti Ischemia/reperfusion injurySystemic lupus erythematosus Muscular dystrophyNeuropathologic diseasesPsychosocial stress diseasesRenal diseasesSepsisSkin diseasesSleep apnea Viral infection

kérdés:kérdés:

mely gének aktiválódnak az NFkB hatására? mi a hiba okozója (fenotípus)

Table 4. list of genes regulated by NF-kBregulated by NF-kB CytokinesCytokinesTNF-α, TNF-βIL-1-α, IL-1βIFN-α, IFN-γTRAlL, M-CSFRANTESFAS ligand Lymphotoxin α, βG-CSF, GM-CSFIL- 2, IL- 6, IL -8, IL- 9, IL-10, IL-11, IL-12, IL-13, IL-15NK4, GCP-2 (CXCL6) Gro α, Gro γ, Gro-1ICOSIL-1 receptor antagonistIP-10, KC, BMP-2CCL 15, CCL 22, CCL 28, CCL 5CD40 ligandCINC-1, CXCL 11ENA-78 (CXCL5) Eotaxin Erythropoietin MCP-1/JE, MIP-1 α βMIP-2, MIP-3 α, Mob-1Neutrophil- activating peptide 78NK-1RStem cell factorAngiotensinogenTCA3TFF3TSP-1TSP-2

ReceptorsReceptorsbeta2 microglobulin µ-opioid receptor2AA1 Adenosine receptor Amiloride-sensitive Na channelAndrogen receptor B 7.1Bradykinin B-1 receptorBRL-1, CCR 5, CCR 7, CD 137CD 154, CD 23, CD 40, CD 48CD 69, CD 83, CD 95EGFRGal 1 receptor, GFBP-2Glucocorticoid receptor IGFBP-1IL-1 receptor antagonistIL-2 receptor a-chainIg γ4, Ig Cγ1, Ig ε heavy chain Ig к light chain, invariant chain IILox-1, Mdr 1MHC class (HLA-B7) MHC class I (H-2Kb) Neuropeptode YY1- receptorNMDA receptor subunint NR-1NMDA receptor subunit Nod 2Polymeric Ig receptorRAF receptor 1,RAGET -cell receptor β chainT -cell receptor CD3γTNF-Receptor, p75/80 DR4, DR5

Early response genesEarly response genesp62, p22/RPG1, B 94Egr-1, TIEG

Transcription factors Transcription factors NF-КB1, NF-КB2, c-Rel, RelBIkBαc-fos, p53, c-myb, JunBE2F3a, Elf3 , ELYSETR 1001IRF-1, IRF-2, IRF-4, IRF-7Mail, Nurr1, Stat5a, WT1

Metastasis Metastasis ICAM-1, VCAM-1 ELAM-1, E-selectin Endoglin, FibronectinMadCAM-1, P-selectin Tenascin-C, DC-SIGNMMP-3, MMP-9, CXCR4KA11/CD82, uPA

AntiapoptosisAntiapoptosisBcl-2, Bcl-XL, Bfl1/A1TRAF-1, TRAF-2, TRAF6c-FLIP, IAPsSurvivin, A20, BaxCaspase-11FAP-1, Fas-ligandIEX-1 L, Nr13

AngiogenesisAngiogenesisVEGF

Cell proliferationCell proliferationCOX-2, c-myc

Cell cycleCell cycleCyclin D1, P21, GADD 45Cyclin D2, Cyclin D3

VirusesVirusesHIV-1EBV wp promoterHBV pregenomic promoterHSV, SV-40Adenovirus (E3 region)Avian leukosis virusBovine leukemia virus CytomegalovirusJC virus, HPV 16, SIV

Proteins involved in Proteins involved in antigen presentationantigen presentation TapasinLMP2TAP 1Complement BComplement receptor 2Complement component 3

Table 4. list of genes regulated by NF-kBregulated by NF-kB (ctd)

KinasesKinasesPI3KPIK3PKC8MAP 4K1

P450P450CYP2C11CYP2E1

Oxidative stress Oxidative stress XODGSTMn SOD INOS

EnzymesEnzymesγ-GCS11bGSD212-LOX5-LOXABC transporters ADHARFRP1AromataseBACECatepsin B and ceramide glycosyl transferase Collagenase 1CRAD1 and 2 Dihydrodiol dehydrogenaseDT –diaphorase

Enzymes Enzymes (ctd)Gelatinase B GSTP1-1Guanylyl cyclase αH+-K+-ATPase a2 HeparanaseHO-1Hyaluronan synthase lodothyronine deiodinaseLysozymeMKP-1N- Acethylglucosaminyl transferase 1NGALNQO1PDE 7A1PGES\L-PGDS Phospholipase A2Phospholipase C δ1PIM-1PTGISRACK1REV3Seprin 2ASNARKTERTTransglutaminase

MiscellaneousMiscellaneousα-1 acid glycoprotein a1-Antitrypsin,a2(1)-Collagenα-FetproteinAMHApolipoprotein C IIIβ-AmyloidBiglycanCaveolin-1Claudin-2Clone 156Clone 330Clone 68Connexin 32Epsilon-globinFactor VIIIGadd45βGalectin 3GIFGS 3686HMG-14K15 keratinK3 keratin K3 keratin K6 keratin

Laminin B2 chainmCGM3Mts1MUC-2Mx1Neutrophil gelatinaseassosiated lipocalinNlF 1P11PAI-1Pax 8PCBDPerforinPGKPSARICKS100A6 (calcyclin) Spergen-1 Syndecan-4TFPI-2TransferrinUrokin 16UBE 2MUCP-2VimentinWilms' tumor suppressor gene

kérdés:kérdés:

mivel gátolható az abberáns NFkB aktivitás?

Table 5. list of agents that inhibit NF-KB activationinhibit NF-KB activationAnti-inflammatoryAnti-inflammatoryagentsagentsAspirin, IndomethacinDiclofenac, NaproxenDexamethasonePhenylbutazoneTamoxifen, SulfasalazineSulindac, IbuprofenLeflunomide Trichodion Acetaminophen Gabexate mesilate Glucosamine sulfate Petrosaspongiolide M Dexanabinol, BSASMCloricromene, RolipramPranlukast

ImmuInosupressive ImmuInosupressive agentsagentsCyclosporin, LF15-019515-Deoxyspergualin6(5H)-Phenanthridinone and benzamide

Chemopreventive Chemopreventive agentsagentsCarboplatin, VesnarinoneCycloprodigiosin(- )-Cycloepoxydon Campthothecin, GemcitabineLevamisole, Epoxomicin

AngiogenesisAngiogenesis2-Methoxyestradiol TNP-470

Cell proliferation 6-Aminoquinazoline derivatives Raxofelast

VirusVirusK1L, HIV-1 Vpu protein Canine distemper virus

AntioxidantsAntioxidantsADT, Apocynin, BenidipineBHA, CarvedilolCatechol derivativesDHEAS, DMDTCDMSO, DisulfiramEbselen, EPC-K1 21Ethyl pyruvateEthylene glycol tetraacetic acidIron tetrakis, Mn-SODMelatonin, NAL, NDGAOrthophenanthrolineHydroquinone and tert-butyl hydroquinoneRef-1, Tempol, TepoxalineN-Acetylcysteine

Reactive oxygen speciesReactive oxygen speciesNitric oxide

Antibacterial Antibacterial agentsagentsFosfomycin 5-Aminosalicylic acid ClarithromycinRifampicinRoxithromycin

Antidiabetes Antidiabetes agentsagentsAdiponectinGlimepiride

Tyrosine Tyrosine phosphatase phosphatase inhibitorsinhibitorsDiamidesPervanadatePhenylarsine oxide

Serine protease Serine protease inhibitorinhibitorPefabloc

Phosphathidyl-Phosphathidyl-choline - choline - phospholipase C phospholipase C inhibitorinhibitorD609

HIV-1 protease HIV-1 protease

inhibitorsinhibitors NelfinavirRitonavirSaquinavir

ACE inhibitorACE inhibitor Ouinadril

COX-2 inhibitorCOX-2 inhibitorCelecoxib

Interferon Interferon inducible proteininducible proteinp202a

MethanesulfonilidMethanesulfonilide antiarthritis e antiarthritis inhibitor inhibitor T -614

PTK inhibitorPTK inhibitorHerbimycin A

Farnesyl protein Farnesyl protein transferase transferase inhibitor inhibitor SCH66336

Table 5. list of agents that inhibit NF-KB activationinhibit NF-KB activation (ctd)Protease inhibitor Protease inhibitor Nafamostat mesilate

PPARγ ligandPPARγ ligandPioglitazone

Heat shock Heat shock proteinsproteinsHSP70, HSP72

StatinsStatinsAtorvastatinErbstatinFluvastatinLovastatin

MetalsMetalsChromium Cadmium GoldLeadMercuryZinkArsenicTitanium

IKB-like proteinIKB-like proteinEncoded by ASFV

Acetylation Acetylation inhibitorinhibitorHDAC

OthersOthersβ-Amyloid protein, β –Cateninγ-Glutamylcysteinesynthetase1,2,3,4,6-Penta-O-galloyl- beta-D-glucose1,2,4- Thiadiazolidine derivatives 15-Deoxy-prostaglandin J(2)17 -Allylamino-17- demethoxygeldanamycin 2-Acetylaminofluorene3,4,5- Trimethoxy-4'- fluorochalcone4-Hydroxy-2 -nonenal5'-MethylthioadenosineAcrolein, AdenosineAMP-activated protein kinaseAmrinone, AnandamideAngiopoietin-1, ANPAntihrombin, Antithrombin IIIAPCO576, ArseniteAuranofin, AvrA proteinAZT, Bcl-Ab1, BenfotiamineBiliverdinBiotinylated isopanepoxydoneBisphenol A, BMTBovine serum albuminBZLF1, C5aCarbon monoxideCyclopentonesCYL-19s and CYL-26zCYLD, DesloratadineDisulfiram, DobutamineDa 65-79, DTD

E3330, E-73, Ecabet sodiumHydroquinone, HypochloriteInhaled isobutil nitriteJesterone dimer, KT-90 APCLosartin, Calcitriol, CaMKKCampthothecin, CaprofinDecoy oligonucleotidesDehydroxymethylepoxy quinomicinDiethyldithiocarbamic acid DiethylmaleateDiltiazem, DMF, EpoxyquinolErythromycinEstrogen (E2)Ethacrynic acidEthyl 2-[(3-methyl-2,5- dioxo(3-pyrrolinyl))Ethyl pyruvate, Evans blueFibrates, Fish oil feedingFlunixin meglumineFlurbiprofen, Fox1jG-120 3032, GangliosidesGemfibrozil, GhrelinGILZ, GlucocorticoidsHydroquinone, HSCOHuman breast milk HycrochlorideHyperosmolarity Hypothermia, IMD-O354Intravenous immunoglobulin Jesterone, JSH-21, JSH-23Kaposi's sarcoma-associated herpesvirus K1 protein

Ketamine, KL-1156, LMBMacro/ide antibioticsMAST205, MEB, MelatoninMercaptopyrazineMesalamine, MethotrexateMonochloramineMonomethylfumarateMTA, Murr1, MX781, EM, Myxoma Virus MNF N-(p-coumaroyl) serotonin Neurofibromatosis-2 protein nicotinamide 3- aminobenzamideNilvadipine, Nitrosoglutathione Nitrosylcobalamin mild hypothermia, N-OctylcaffeateN-OleoyldopamineNPM-ALK oncoprotein, NRFNucling, ox-LDL and HNEOXPAPC, pB, p53, PACAPPAN1, Panepoxidone, PBNPDTC, PEDF, PentoxifyllinePhoto oxidative stress PIAS, Pirfenidone, PPM-1BProbioticsPg J2, Pg A1, Pg E2, PSKPsychosine, PTEN , PTX-BPyrithione, Quinazolinespyrrolidinedithiocarbamic -acid Rapamycin, RebamipideRelA peptides, Retinoic acid receptor-related orphan receptor-alpha

Table 5. list of agents that inhibit NF-KB activationinhibit NF-KB activation (ctd)OthersOthers (ctd)RibavirinRifamides ,RitonavirRKIPRocaglamideSalineSelenomethionineSiah2, SiRNASLPI, SOCS1Sodium salicylateSUN C8079Surfactant protein ASurvantaTAT -SR-IКBαTetrathiomobdateThalidomide THI 52ThiopentalTNAPTom1TranilastTriflusalTriglyceride-rich lipoproteinsTroglitazone Tyrphostin Ursodeoxycholic acid Uteroglobin Vasoactive intestinal peptidevlRF3YopJZAS3 proteinZUD protein

kérdés:kérdés:

hol találhatók olyan anyagok, amelyek csökentik a kóros

NFkB aktivitást?

Table 6. natural products that suppress NF-kB activationsuppress NF-kB activation α-Lipoic acid α-Pineneα-Tocopherol α-Torphryl succinate β-Lapachone1’-Acetoxychavicole acetate20(S)-Protopanaxatriol23-Hydroxyursolic acid7-Amino-4-methy- coumarinAcetyl-boswellic acidsAllyl-cysteineMentoflavoneAndalusolAndrographolideAnetholeApigeninArtemisia sylvatcaArtemisininAtaxanthinAstragaloside IVAucubinAvicinBaicaleinBambara groundnutBee venonBenzyl isothiocynateBetulinic acidBis-eugenolBupleurum fruticosum phenylpropanoidsCacospongionolide BCalagualineCancer bush

CAPECapsaicinCapsiateCamosolCatalposideCinnamaldehyde/2-methoxycinnamaldehydeCycloepoxydonCelastrolCepharantinCytochalasin DConophylineCheongyeolsaseuplangChromene derivativesChicory rootChitosanCurcuminCyclolinteinoneDanshenshuDDCDehydroascorbic acidDiarylheptanoidDibenzylbutyrolactone lignansDiferoxamineDigitoxinDihydroisoeugenolDihydrolipoic acidDIM/I3CEbselenEGCGEmodinEnt-kaurene diterpenoidsErgothioneineEvodiamine

Epoxyquinone AFalcarindolFlavenoidsFlavopiridolFungal gliotoxinGanoderma lucidum polysaccharidesGarcinolGenisteinGaldenamycinGenipinGinkgolide BGlabridinGlitoxinGlossogyne tenuifoliaGlycyrrhizinGuggulsteroneHelenarinHemateinHerbal compound861Hydroxyethyl starchHypericin10Z-hymenialdisineHypoestoxideIndirubin-3’-oximeIRFI 042Isomallotochromanol isomallotochromeneIsorhapontigeninKahweol Quinic acidKamebakurinLacidipineL-Ascorbic acidLazaroids

LupeolLuteolinMagnololγ-MangostinManassantisMangiferinNicotineOleandrinOnconasePanduratin APalthenolidePC-SPESPhenethylisothiocyanatePhomolPhytic acidPhallacidinPiceatonnolPiperineQingkailingShuanghuanglianQuercetinQuercetinResiniferatoxinResveratrolRheinRocaglamideRotenoneSanggenon CSanguinarineSaucerneolSauchinoneSesaminSesquiterpene lactonesSilymarinSinomenine

Solona nigrum LSphondinStaurosporineTansinonesTetrandineTheaflavinTrilinoleinTriptolideUrsolic acidVitamin CWedalolactoneWithaferin AWogoninWortmanninYakuchinone A and BZerumbone

Table 6. natural products that suppress NF-kB activationsuppress NF-kB activationOthersOthers1,2,4-Thiadiazolidine derivativesAged garlic extract, Apple juiceArtemisia capillaris extractBlueberry and berry mixBlack raspberry extractsCat’sclaw bark, Extract of the stem bark of Mangifera indica LExtracts of Ochna macroccalyxFomes fomentarius methanol extractsFructus benincasae recens extractGinkgo biloba extractHarpagophytum procumbens extractsKochia scoparia fruit, Omega 3 fatty acidsPhyllanthus amarus extractsPlatycodi radix extract, Red wineSophorae radix extractStinging nettle plant extrats Tanacetum larvatum extractUncaria tomentosum plant extractTrichomonas vaginalisXanthium strumarium LYucca schidigera extract

kérdés:kérdés:

praktikusan - mely kis-molekula- tömegű természetes vagy szintetikus anyagok csökkentik az NFkB aktivitását?

Table 7. proteins and peptides that inhibit NF-kB activationinhibit NF-kB activation Interleukin Interleukin IL-4IL-11IL-10IL-13

InterferonInterferonIFN-α

Growth factors and hormonesGrowth factors and hormonesVEGFHepatocyte growth factor MSH HB-EGFChorionic gonadotropin

PeptidesPeptidesTRAF6 inhibitory peptideNEMO binding peptidep65 binding peptideBoronic acid peptideNLS cell permeable peptides Peptide nucleic acid-DNA decoys Peptide YYTIRAP inhibitor peptide

Table 8. synthetic compounds that inhibit NF-kB activationinhibit NF-kB activation

IKK inhibitors IKK inhibitors BAY-11-7082 BAY-11-7085 BMS-345541 SPC839 AS602868 PS1145MNL 120SC-514

Polyubiquitination inhibitorPolyubiquitination inhibitor Ro106-9920

Ubiquitin Ligase Inhibitors Ubiquitin Ligase Inhibitors Z-LLL, Z-LLnV

Proteosome inhibitorsProteosome inhibitorsALLnL PS-341Lactacystine, β-lactone SN-50FK506Ro106-9920PS5190TLCK ,TPCKCyclosporin ADCICDeoxyspergualinDFPLLMBTEEMG115 MG120APNE

P42 MAPK inhibitorP42 MAPK inhibitor AG 126

P38 MAPK inhibitorP38 MAPK inhibitor SB203580

PI-3K-kinase inhibitorPI-3K-kinase inhibitor LY294002

PKC inhibitorPKC inhibitor RO31-8220

MEK1/2 inhibitorMEK1/2 inhibitor UO126

Protein kinase inhibitorProtein kinase inhibitor A771726

kérdés:kérdés:

mely pontokon lehet/kellszerkezeti változtatást létrehozni (pl. rekombináns technikákkal)

az NFkB aktivitásának specifikus csökkentéséhez?

Table 9. inhibition of NF-kBinhibition of NF-kB through the critical cysteincystein residue

IKKα/β IKKα/β Cys179Cys179

IKKβ IKKβ Cys59Cys59

IKKγ IKKγ Cys417Cys417

P65 P65 Cys38Cys38

p50 p50 Cys62 Cys62

kérdés:kérdés:

milyen célmolekulákat tervezzünkcélmolekulákat tervezzünk az NFkB aktivitásának csökkentsére? a szignálút melyik szakaszára?

kérdés:kérdés:

az aktivációs mechanizmus gátlásával változik-e az NFkB aktivitása?

gátló stimulusok használhatók-e? hát az IkB gátlása?

Aktiváló Stimulus(kfl)

IIB kinB kináz áz /NEMO/IKAP

IB

p65p50

IB

p65p50

PIB

degradáció

p65p50CITOPLAZMA

p65p50

NUKLEUSZ

mRNS

transzlokáció

transzkripció

Target genes

az NF-kB NF-kB gátlás kehetőségei 1.

inaktív forma maradinaktív forma marad

gyulladási-/immun-stb. fehérjék

kérdés:kérdés:

az IkB kináz ativitásának vagyaktiválásának gátlásával lehet

csökkenteni az NFkB aktivitását? mi az ubiquitináció gátlás hatása?

Aktiváló Stimulus(kfl)

IB kináz/NEMO/IKAP

IB

p65p50

IB

p65p50

PIB

degradációdegradáció

p65p50CITOPLAZMA

p65p50

NUKLEUSZ

mRNS

transzlokáció

transzkripció

Target genes

az NF-kB NF-kB gátlás kehetőségei 2.

inaktív forma maradinaktív forma marad

gyulladási-/immun-stb. fehérjék

Aktiváló Stimulus(kfl)

IB kináz/NEMO/IKAP

IB

p65p50

IB

p65p50

PIB

degradáció

p65p50CITOPLAZMA

p65p50

NUKLEUSZ

mRNS

transzlokáció

transzkripció

Target genes

az NF-kB NF-kB gátlás kehetőségei 3.

gyulladási-/immun-stb. fehérjék

nem megoldott:

- sejt-specificitás

- alacsony toxicitás

sok fejleszteni való van hátra

screen – screen – ~220eFt/kísérlet~220eFt/kísérletHuman NFκB Signaling Pathway RT² Profiler™ PCR Array Functional Gene Grouping (more detailed information is provided in the gene table) Activation of NFκB pathwayLigands and receptors: IL1B, IL8, MYD88, TLR1, TLR2, TLR3, TLR4, TLR6, TLR8, TNF, TNFRSF10A, TNFRSF10B. Membrane molecules: IRAK1, IRAK2. Kinases: CHUK, IKBKB, IKBKG, IRAK1, IRAK2. I-kappa-B kinase/NF-kappa-B cascade: EDARADD, IKBKG, IRAK2, STAT1, TLR8. Cytoplasmic sequestering/releasing of NF-kappa-B: BCL3, IL10, NALP12, NFKBIA, TNFRSF7, TNFSF14. Transcription factors: IKBKB, IKBKG, IRAK1, NFKB1, NFKBIA, RELA, STAT1, TNF. Inflammatory response: IL1B, IL8, IRAK2, MYD88, NFKB1, TLR1, TLR2, TLR3, TLR4, TLR6, TLR8, TNF. Positive regulation of I-kappa-B kinase/NF-kappa-B cascadeLigands and receptors: CD40 (TNFRSF5), F2R, FASLG (TNFSF6), HTR2B, LTBR, SLC20A1, TICAM2, TNFRSF1A, TNFSF10, TRIF. Membrane molecules: EDG2, GJA1, HMOX1, RHOA. Kinases: IKBKE, RIPK1, TBK1. Other: BCL10, BIRC2, CARD4, CASP1, CASP8, CFLAR, CTL2, FADD, MALT1, PPM1A, REL, RFP2, TMED4 (HNLF), TRADD. NFκB Responsive GenesAcute phase response proteins: AGT, BF. Adhesion molecules: ICAM1. Extracellular molecules: BF, CSF2, CSF3, IFNA1, IFNB1, IFNG, IL6. Ligands and receptors: AGT, CCL2, ICAM1, LTA. Cytokines: CCL2, CSF2, CSF3, IFNA1, IFNB1, IFNG, IL6, IL8, LTA, TNF. Inflammatory response: CCL2. Other factors involved in NF-kappa-B pathwayExtracellular molecules: IL1A. Ligands and receptors: IL1R1, RAF1, TLR7, TLR9. Kinases: AKT1, MAP3K1, RAF1. Transcription factors: ATF1, EGR1, ELK1, FOS, JUN, NFKB2, RELB, TNFAIP3. Inflammatory response: FOS, IL1A, IL1R1, TLR7, TLR9  

köszönöm a figyelmetköszönöm a figyelmet

Leukemia 16:1053-1068, 2002

(konstitutívan) aktív NFB-t expresszáló tumorok (pl.)

• B cell lymphoma• Hodgkin’s disease• T-cell lymphoma• Acute lymphoblastic

leukemia• Breast• Liver• Thyroid

• Prostate• Melanoma• Head and neck SCC• Colon• Multiple myeloma• Ovarian• Bladder • Lung