Post on 31-Mar-2015
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