肿瘤生物学与转化医学

肿瘤生物学与转化医学CancerBiology&TranslationMedicine第四军医大学znchen@陈志南Zhi-NanChenGlobalActionAgainstCancerUpdateEdition2005Global(Year)7.6millionDeaths10.9millionNewCases5Liver1Lung7Esopha-geal3Colon2Breast4Gastric6CervicalCancerDiseaseControlDivision,MinistryofHealthofPRC,2008Cancer6Cervical5Colon8Nasopharyn-geal1Lung2Liver7Breast3Gastric4Esopha-gealCity24.41(2nd)Incidence2,000,000Mortality1,500,000CancerCountryside26.93

(1st)LiverCancerMortality/100,000AgainstCancerinChinaThethirdtimeinvestigationofdeathcauses:cancerrankssecondwithamortalityrateof22.32%.——MinistryofHealth,April2008增生

异常InheritedsusceptibilityChemicalcarcinogensRadiationInfectiousagentsDNArepairsystemGeneralhealth(diet,stress,etc)ImmunesystemCancerCell:ContributingFactorsCancerEtiologyMultipleCarcinogenCancer:ThreeCharacteristicsTumorInitiationDevelopmentInvasionMetastasisDysplasiaCarcinomainsituCellPrimarycancerMetastasis(2ndCancer)genomicDNAtranscriptiontranslationpost-transcriptionposttranslationMultipleStagesEMBOreports1,2,115-119,2000LossofgenomicintegrityandimbalanceofmoleculesaremechanismforthecancerincidenceMultipleGeneMutationCancerInitiation&InvasionFourMechanismsMiningthecancergenomeTobiasSjoblom,etal.Science314:268-274,2006CancerGenomeAtlasSequencing:13,023genesremovingerrors,normalvariantsBreastcancer11SamplesColoncancer11Samples1149mutationgenes(Individualtumors:average90)(Significantfrequency:189genes)(Significantfrequency:

average11pertumor)CancerMolecularBalance&MutationSohrabP.Shah,etal.Nature461:809-813,2009MutationalevolutioninalobularbreasttumourprofiledatsinglenucleotideresolutionRecentadvancesinnextgenerationsequencing1,2,3,4havemadeitpossibletopreciselycharacterizeallsomaticcodingmutationsthatoccurduringthedevelopmentandprogressionofindividualcancers.Wefound32somaticnon-synonymouscodingmutationspresentinthemetastasis.Fiveofthe32mutations(inABCB11,HAUS3,SLC24A4,SNX4andPALB2)wereprevalentintheDNAoftheprimarytumourremovedatdiagnosis9yearsearlier,six(inKIF1C,USP28,MYH8,MORC1,KIAA1468andRNASEH2A)werepresentatlowerfrequencies(1–13%),19werenotdetectedintheprimarytumour,andtwowereundetermined.SohrabP.Shah,RyanD.Morin,JaswinderKhattra,LeahPrentice,TrevorPugh,AngelaBurleigh,AllenDelaney,KarenGelmon,RyanGuliany,JanineSenz,ChristianSteidl,RobertA.Holt,StevenJones,MarkSun,GillianLeung,RichardMoore,TesaSeverson,GregA.Taylor,AndrewE.Teschendorff,KaneTse,GulisaTurashvili,RichardVarhol,RenéL.Warren,PeterWatson,YongjunZhao,CarlosCaldas,DavidHuntsman,MartinHirst,MarcoA.Marra&SamuelAparicioCancerGenomeAtlasTravelIntravasationPrimarytumor(Cellescape)InvasionExtravasationTransportGrowth(2ndtumor)MigrationInvasion&Metastasis—MainDeathCausesMesenchymalcell-likemovementAmoeba-likemovementAlittlelongcellNeedproteaseFormofpseudopodiumAlittlerelyonproteaseMyosinstrongContractionAlittlecirclecellRac/WAVE2signalRho/ROCK

signalCell.2008Oct31;135(3):510-523AdhesionMovementG1→S→G2→MRControlofcellcycleG1:DNApre-synthesisS:DNA

synthesisG2:DNA

post-synthesisM:CelldivisionGO:OncogenesSTOP:TumorsuperessorEpithelialliningcellsTransformedepithelialcellsTumorfbTissueMartrixTransformedepithelialcellsMMP-7,13

(9)EpithelialcellsoftumorangiogenesisMMP-1,2,14TumorfbMMP-1,2,3,11,14Matrixdegradation:Structuralbaseoftumorinvasion&metastasisMatrixDegradation——MMPsTumor>2-3mm:NeedvesselsKeyMolecular:MMPs,VEGF,bFGF,PDGFTumorvesselsdensityisamarkerforearlydiagnosisandprognosisNatureRevCancer,4,

2004AngiogenesisCancer:WoundsthatfailtohealNature,420,2002TheChainofInflammation&CancerTumorDevelopmentCancerChronicinflammationBacterialHpylori幽门螺杆菌VirusEB,HCVParasiteflukes,schistosomes吸虫，血吸虫ChemialirritisPMA佛波酯NondigestibleParticlesasbestos,silica石棉纤维，矽Strongassociation:inflammationandcancerOxidativeStress氧化应激

Aldehydes醛OxidizedDNAnucleosides氧化脱氧核苷酸DNAmutationPeroxynitrite过氧亚硝基ROS活性氧DNAdamageP53、RbDNArepairDNArepairRNS活性氮ChronicinflammationInflammatorycytokines&OncogenesOncogenesInflammatorycytokinesRASIL-1a,IL-1b,IL-6,CXCL8,IL-11MYCIL-1b,CCL2,CCL5,CCL7,CXCL1,CXCL2RETtyrosinekinaseCXCL8,CXCR4,CCL2,MCP-1,GM-CSFEGFRCXCL8proto-oncogeneTpl2NF-kBOncomiRs,miR-155InflammatorymediatorsinmacrophagesandinmonocytesCancerLetters,267,2008TumorinflammationenvironmentROSHIFBloodvesselCancercellMacrophageCytokinesChemokinesCD147CypAErk1/2P38ProliferationAnti-apoptosisAngiogenesisCypAHypoxiaOurstudyNotconfirmedReportedPI3KSignalTransductionTheSevenHallmarksofCancerandTheirLinkstoTumorMetabolismCancerCell.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TumorMetabolismJClinInvest.118(12):3835-3837,2008IntratumoralhypoxiaandmetabolicsymbiosisorAerobicstromalcellAnaerobicglycolysisMolecularMechanismsofCancer-SpecificMetabolicReprogramming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LelandH.Hartwell,etal.NatBiotech,

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(8),2006高危评估Riskassessment早期诊断Noninvasivescreeningforearly-stagedisease检测定位Detectionandlocalization预后判断Diseasestratificationandprognosis治疗反应Responsetotherapy复发监测ScreeningfordiseaserecurrenceCancerBiomarker--ASystemsApproachBiomarkerDiscovery:ExpressionMapping(Modificationmapping)Functionalproteomics:interactionoftheproteinsEpitopemapping(activecore)

SeperationandidentificationofmixturesampleAbarray,Cellarray,Tissuearray,Co-IP(pull-down),Biosence,etc.

Comparativeproteomics2DE,2DELC-MS

ValidationpeptidessequenceofproteinMALDI-MS,SELDI-MS,LC-MSMS,ESI-MS(m/z)

AnalysisofthedatabasesHowIdentifiedCancerBiomarker?SystemBiologyApproaches“-omic”Technologies(PreclinicalorClinicalUtilization)TrendsBiotechnol.23(11),544-546,2005AmplichipCyp450TestGlobalSNPArraysMicroarraySAGEMALDI-MS/MS2DGels-MSNMRGC-MSLC-MSFT-IRWhatisanIdealCancerBiomarker?ScreeningahealthypopulationorahighriskpopulationforthepresenceofcancerMakingadiagnosisofcancerorofaspecifictypeofcancerDeterminingtheprognosisinapatientMonitoringthecourseinapatientinremissionorwhilereceivingsurgery,radiation,chemotherapy,orbiotherapyScreeningahealthypopulationorahighriskpopulationforthepresenceofcancerMakingadiagnosisofcancerorofaspecifictypeofcancerDeterminingtheprognosisinapatientMonitoringthecourseinapatientinremissionorwhilereceivingsurgery,radiation,chemotherapy,orbiotherapyApplicationofCancerBiomarkerIdentificationanddiagnosisIndividualsaffectedwithdiseasePeoplewhomaybeatriskbutdonotyetexhibitsymptomsMonitorprogressofdiseaseMonitoreffectsoftreatmentRemissionFollow-upCancersfoundinearlystage:lowmorbidityandrecurrenceratesCancersidentifiedinlatestage:highrecurrenceandmortalityratesCancerBiomarkerandTypesofCancer:statisticallysignificantassociationbetweenaparticularcancerandtheassociatedcancermarker(s)AFP=alphafetoproteinCEA=carcinogenicembryonicantigenCA15-3=carbohydrateantigen15-3CA19-9=carbohydrateantigen19-9CA125=carbohydrateantigen125PSA=freeprostatespecificantigen+prostatespecificantigen-alpha(1)antichymotrypsincomplexPSAF=freeprostatespecificantigenPSAC=prostatespecificantigen-alpha(1)antichymotrypsincomplexPAP=prostaticacidphosphatasehTG=humanthyroglobulinhCGb=humanchorionicgonadotropinbetaFerr=FerritinNSE=neuronspecificenolaseIL-2=interleukin2IL-6=interleukin6A2M=alpha2macroglobulinB2M=beta2microglobulinProblemsofCancerBiomarkerNocancerbiomarkerisabsolutelyspecificNocancerbiomarkertestisfreeoffalsenegativesNocancerbiomarkertestisfreeoffalsepositivesNocancerbiomarkerisabsolutelyspecificNocancerbiomarkertestisfreeoffalsenegativesNocancerbiomarkertestisfreeoffalsepositivesAntibodyBasedCancerBiomarkersCancerMarkerAntibodyCancerCA125OC125ovariancancerCA15-3DF3,115D8breastcancer,ovariancancerCA549BC4E549,BC4N154breastcancer,ovariancancerCA27-29B27.29breastcancerMCAb-12breastcancer,ovariancancerDU-PAN-2Du-PAN-2pancreaticcancer,ovariancancer,gastrointestinalcancers,lungcancerCA19-919-9pancreaticcancer,livercancer,gastrointestinalcancersCA19-519-5pancreaticcancer,ovariancancer,gastrointestinalcancersCA50C50pancreaticcancer,rectalcancer,gastrointestinalcancersCA72-4B27.3,cc49gastriccarcinoma,pancreaticcancer,ovariancancerCA242C242gastrointestinalcancerspancreaticcancerHer-2/neuHerceptinbreastcancerHAb18G/CD147HAb18hepatocellular

carcinoma,ectAntibodyBasedCancerBiomarkersHumangenomeis2.91-billionbasepairsinlength(1990)Thereareabout25,000genesexistinthehumangenome(42%haveanunknownfunction)Approximately12,000genesthatappeartohavethecapacitytomakesecretedproteins,allthegeneshavebeendeterminedtheentirenucleotidesequences(3141geneslocusonthefirstchromosome)AtMay2006,thefirstchromosomesequenceswerecompleted,atleast1,000newgeneswerefound.Thisistheendof16year’sHumanGenomicPlan.Thishasempoweredmoredirectmeansoftargetidentification,e.g.byexpandingproteindatabasesandenablingthemappingofnovelcancer-associatedgenes(Venteretal.2001).AntibodyBasedCancerBiomarkersCompletegenomesequenceshaveprovidedaplethoraofpotentialdrugtargets.Butthehardtaskoffindingtheirweakspotsisjustbeginning(about5000genescanbeusedasdrugtarget)Achallengingnewdevelopmentinthefieldofdrug-targetdiscoveryissystemsbiology,ortherecognitionthatgenes,orbetterthegeneproducts,arepartof,andfunction,inlargecomplexnetworks.Nature,428,225-231,2004.转化医学与公众健康EA.Zerhouni，NIH路线图计划（NIHRoadmap），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（BenchtoBedside,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ScienceTranslationalMedicine、JournalofTranslationalMedicine和TranslationalResearch三本国际性专业杂志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%中国的转化医学转化医学与4P医学PredictiveMedicine–预测医学PreventiveMedicine–预防医学PersonalizedMedicine–个体化医学ParticipateMedicine–参与医学PersonalizedPrevention&EarlyDetectionLifestylechangesScreeningChemopreventionProphylacticSurgery

Average Moderate High VeryHighRISKPersonalizedMedicine–个体化治疗No“onesizefitsall”drugMostdrugsworkfor30%to70%ofpatientsMultiplefactorsdeterminedrugresponsesPhamacogeneticsisessentialforindividualizedtherapyHerceptin,thefirstmarketedpersonalizedmedicine,wasapprovedusingacoordinateddrug/diagnosticapprovalprocessthatwillbecomemorecommon.PersonalizedMedicine–个体化治疗OurStudyMolecularDockingHAb18G&ItsAntibodiesHAb18G/CD147

&HAb18,6H8,5A12AntibodiesHAb18G/CD147ISetOurStudyCrystalStructureofHAb18G/CD147C2IXiao-LingYu,Zhi-NanChen,JBiolChem,283(26),2008Nationalpatent:200710018514.XPCTpatent:PCT/CN2007/003034PDBID:3B5HHAb18G/CD147OurStudyTissueAtlas-HAb18G/CD147HAb18G/CD147OurStudyTissueAtlas-HAb18G/CD147OurStudyTissueAtlas-HAb18G/CD147Shao-HuiHu,Zhi-NanChen,etal.Proteomics,7(13),2007LungLiverFetalNormalCancerCasePositivePositive

Rate(%)PFetal6711