分子生物学进展肿瘤标志物

分子生物学进展肿瘤标志物StatisticsMorethan11millionpeoplearediagnosedwithcancereveryyear.Itisestimatedthattherewillbe16millionnewcaseseveryyearby2023.Fromatotalof58milliondeathsworldwidein2023,canceraccountsfor7.6million(or13%)oftheglobalmortality.Deathsfromcancerintheworldareprojectedtocontinuerising,withanestimated9millionpeopledyingfromcancerin2023and11.4milliondyingin2030.IntheUSin2023,over1.4millionnewcasesofcancerwerediagnosed.Overhalfamillionpeoplediedfromthisdisease,accountingforapproximately25%ofalldeathsintheUSeachyearHowtoImprovetheSituation?PreventionDetectionCancerisadiseaseofgeneticprogressionthatisoftenassociatedwithspecificmolecular,geneticandhistologicalchanges.Theabilitytodevelopbiomarkersthatcandetectthecriticalcomponentsofthesehallmarksofcancertogetherprovidesapowerfulbasisfordiagnosing,monitoringandpredictingoutcomeandresponsetotreatment.HowtoImprovetheSituation?ConceptofCancerBiomarkersDefinitionofbiologicalmarkersBiologicalmarkers(biomarkers)havebeendefinedbyHulkaandcolleagues(1990)as“cellular,biochemicalormolecularalterationsthataremeasurableinbiologicalmediasuchashumantissues,cells,orfluids.”

HulkaBS.Overviewofbiologicalmarkers.In:Biologicalmarkersinepidemiology(HulkaBS,GriffithJD,WilcoskyTC,eds),pp3–15.NewYork:OxfordUniversityPress,1990.Morerecently,thedefinitionhasbeenbroadenedtoinclude“biologicalcharacteristicsthatcanbeobjectivelymeasuredandevaluatedasanindicatorofnormalbiologicalprocesses,pathogenicprocesses,orpharmacologicalresponsestoatherapeuticintervention”

NaylorS.Biomarkers:currentperspectivesandfutureprospects.ExpertRevMolDiagn3:525–529,2023.ConceptofCancerBiomarkers2.FormsofcancermarkersHormones,metabolites，aswellasdifferentfunctionalsubgroupsofproteinssuchasenzymes,glycoproteins,oncofetalantigensandreceptors.Furthermore,otherchangesintumors,suchasgeneticmutations,amplificationsortranslocations,andchangesinmicroarray-generatedprofiles(geneticsignatures),arealsoformsoftumormarkers.Themarkersareproducedeitherbythetumoritselforbyothertissues,inresponsetothepresenceofcancerorotherassociatedconditions,suchasinflammation.Cancerbiomarkerscanalsobeprocessessuchasapoptosis,angiogenesisorproliferation.ConceptofCancerBiomarkers3.FactorsthatareidealforatumormarkerProducedbythetumorcellsandentersthecirculationPresentatlowlevelsintheserumofhealthyindividualsandthosewithbenigndiseasebutincreasessubstantiallyincancer(preferablyinonecancertypeonly)EasilyquantifiablewithaninexpensiveassayPresentindetectable(orhigherthannormal)quantitiesatearlyorpreclinicalstagesQuantitativelevelsofthetumormarkerreflectthetumorburdenHighdiagnosticsensitivity(fewfalsenegatives)andspecificity(fewfalsepositives)ConceptofCancerBiomarkers3.FactorsthatareidealforaserologicaltumormarkerConceptofCancerBiomarkers4.Typesofcancerbiomarkers4.1.Diagnostic(screening)biomarkerAmarkerthatisusedtodetectandidentifyagiventypeofcancerinanindividual.ThesemarkersareexpectedtohavehighspecificityandsensitivityForexample,thepresenceofBence–JonesproteininurineremainsoneofthestrongestdiagnosticindicatorsofmultiplemyelomaConceptofCancerBiomarkersConceptofCancerBiomarkers4.3.Stratification(predictive)biomarkerThistypeofmarkerservestopredicttheresponsetoadrugbeforetreatmentisstarted.Thismarkerclassifiesindividualsaslikelyrespondersornonresponderstoaparticulartreatment.Thesebiomarkersmainlyarisefromarray-typeexperimentsthatmakeitpossibletopredictclinicaloutcomefromthemolecularcharacteristicsofapatient’stumor.CurrentapplicationsoftumormarkersandtheirlimitationsCurrentapplicationsoftumormarkersandtheirlimitationsCancerbiomarkersthatarecurrentlyinclinicaluseCancerbiomarkersthatarecurrentlyinclinicalusePhase1Determinationsoftoxicity,pharmacokinetics,andoptimaldoselevelsphase2DeterminationsofbiologicefficacyPhase3Definitivecontrolledtrialsofeffectsonclinicalendpoints.Foreachphase,guidelinesexistforsubjectselection,outcomemeasures,relevantcomparisonsforevaluatingstudyresults,andsoforth.Phasesofbiomarkerdevelopment

－MargaretSP;etal.UniversityofWashington(2023)Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)1.PreclinicalexploratorystudiesSpecimenSelectionTumortissuefromcasesubjectsshouldbeobtainedatdiagnosisandbeforetreatmentbecausetreatmentmayinterferewiththebehaviorofthebiomarker.Noncancercontrolsubjectsshouldbeselectedsothatfactorspotentiallyinfluencingthebiomarker,otherthanthecanceritself,aretightlymatchedtothoseofthecancercasesubjects.Thesefactorsmightincludeage,sex,race,andpossiblylifestyle-relatedcharacteristics,suchassmokinghabits.Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)1.PreclinicalexploratorystudiesSpecimenSelectionFactorsshouldbeconsideredwhenselectingtumorSpecimen1，样品旳一致性：取材部位、肿瘤亚型、年龄、性别、种族、生活习惯等。2，样品处理方式旳一致性：预处理条件、保存条件（涉及时间）、处理条件、操作等。Factorsshouldbeconsideredwhenselectingnontumor(control)Specimen1，对照样品和肿瘤样品旳对等性2，对照样品和肿瘤样品旳处理方式旳对等性Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)Phasesofbiomarkerdevelopmentforearlydetetion

－MargaretSP;etal.UniversityofWashington(2023)Phasesofbiomarkerdevelopmentforearlydetection

－MargaretSP;etal.UniversityofWashington(2023)Strategiesandtechniquesfordiscovery

ofcancerbiomarkersGenomiclevelcancerbiomarkerdiscovery1.1.GenomicaberrationSequencing:TheCancerGenomeAtlas(TCGA)isapplyinglarge-scalegenomesequencingtechnologytoidentifynovelgenesinvolvedincancerpathogenesis.Comparativegenomichybridization(CGH)array-CGH(aCGH)Spectralkaryotyping(SKY)1.2.SNPSequencingSNParray1.3.EpigeneticalternationsStrategiesandtechniquesfordiscovery

ofcancerbiomarkers2.Transcriptionallevelcancerbiomarkerdiscovery2.1.mRNAexpressionprofilecDNA-micro-array,Oligo-micro-arrayDifferentialdisplay-PCR(DD-PCR)Serialanalysisofgeneexpression(SAGE),cDNALibrarySubtraction,etc.Strategiesandtechniquesfordiscovery

ofcancerbiomarkers2.Transcriptionallevelcancerbiomarkerdiscovery2.2.miRNAPotentialimportanceofmiRNAs

ascancerbiomarkersExpressionofmicroRNAs(miRNAs)invarioustissueshasbeenassociatedwithavarietyofdiseases,includingcancers.

SerummiRNAscontainfingerprintsforvariousdiseases.

RelatedtechniquesSequencingmiRNA-arrayStrategiesandtechniquesfordiscovery

ofcancerbiomarkers3.Translationallevelcancerbiomarkerdiscovery3.1.Protein(orsubtypes:enzymes,antibodies,secretedproteins,etc)2-dimensioalelectrophoresis/massspectrometry(2-DE/MS)Surface-enhancedlaserdesorptionionizationtime-of-flightmassspectrometrytechnology(SELDI-TOP-MS):proteomicpatterndiagnosticsMulti-dimensionalproteinidentificationtechnology(MudPIT)/MSStrategiesandtechniquesfordiscovery

ofcancerbiomarkersPrincipleofSELDI-TOF-MSOnemicrolitreofraw,unfractionatedserumisappliedtothesurfaceofaprotein-bindingchip.Thechipisrinsedtoremoveunboundproteins,treatedwithaMATRIXCOMPOUND,washedanddried.Alaserirradiatesanddesorbstheadherentproteins.Thetime-of-flight(TOF)oftheionisdetectedbyanelectrode.Aproteomicsignatureoftheserumiscreated.Strategiesandtechniquesfordiscovery

ofcancerbiomarkersProteomicpatterndiagnosticsWiththisapproach,theunderlyingidentityoftheindividualcomponentsofthepatternisnotnecessaryforitsuseasapotentialdiagnosticfordisease.Strategiesandtechniquesfordiscovery

ofcancerbiomarkers3.Translationallevelcancerbiomarkerdiscovery3.1.Protein(orsubtypes:enzymes,antibodies,secretedproteins,etc)2-dimensioalelectrophoresis/massspectrometry(2-DE/MS)Surface-enhancedlaserdesorptionionizationtime-of-flightmassspectrometrytechnology(SELDI-TOP-MS):proteomicpatterndiagnosticsMulti-dimensionalproteinidentificationtechnology(MudPIT)/MSStrategiesandtechniquesfordiscovery

ofcancerbiomarkersPrincipleofMudPITStrategiesandtechniquesfordiscovery

ofcancerbiomarkers4.Post-translationalmodificationsofproteins(cleavageproducts,alteredglycosylation,etc)

4.1.Cleavageproductsoftumour-derivedproteinshavebeenproposedaspotentialcancerbiomarkers.4.2.Alteredproteinglycosylationincancerisanothersourceofpotentialcancerbiomarkers.Identificationofglycosylatedproteinsreliesonvariousglyco-capturestrategies,ameansofglycosylated-proteinsub-selectionbyaffinitychromatography.Electron-transferdissociation(电子转移解离)allowslabilemodificationstoremainintactwhileobtainingpeptidesequenceinformation,enablingthestudyofmodificationssuchasglycosylationandphosphorylation.CurrenttumormarkersunderdevelopmentContributionofoncoproteomicstocancerbiomarkerdiscoveryPublished:2April2023MolecularCancer2023,6:25PotentialimportanceofmiRNAs

ascancerbiomarkers1.HistoryDiscoveredinCaenorhabditiselegansin1993andformallynamedin2023Havebeenidentifiedineveryplantandanimalspeciesexamined2.Features2.1.GeneralfeaturesLength:AclassofnoncodingRNAs,18–25nucleotidesSpeciecs:miRNAshavebeenidentified5withupto1,000predicted.Location:miRNAsareencodedbyDNAthatmaybesituatedintheexonsorintronsofgenesorscatteredamongintergenicDNAPotentialimportanceofmiRNAs

ascancerbiomarkers2.2.Transcriptionandmaturation(i)nuclearprocessingintoaprimarymiRNA(pri-miRNA)andthenaprecursor(pre-miRNA);(ii)exportintothecytoplasm;(iii)furtherprocessingintomaturemiRNA;(iv)incorporationintoanRNA-inducedsilencingcomplex(RISC)withanArgonauteproteincatalystPotentialimportanceofmiRNAs

ascancerbiomarkers2.3.FunctionandtargetsThemiRNA-RISCcomplexhybridizestonucleotidesequencesofvaryingcomplementarityinthe3’untranslatedregion(UTR)ofmRNAandinhibitsproteinsynthesisordegradesthetargetmRNAPlayskeyrolesintheregulationoffundamentalcellularprocessesPotentialimportanceofmiRNAsascancerbiomarkersDysregulatedexpressionofmicroRNAs(miRNAs)invarioustissueshasbeenassociatedwithavarietyofdiseases,includingcancers.miRNAsexpressedincancermayactlikeoncogenesortumor-suppressorgenesbyregulatingproliferationand/orapoptosis.NormalandmalignanttissueshavespecificmiRNAsignaturesandshowdifferentialexpressionacrosstumortypes.OverexpressionorlackofexpressionofspecificmiRNAsappearstocorrelatewithclinicallyaggressiveormetastaticphenotype.miRNAexpressionhastissuespecificityandhasbeenusedfortumorclassification.——CancerbiomarkerprofilingwithmicroRNAsApril2023,NatureBiotechnologyVol26/4.PotentialimportanceofmiRNAincancerOrganizations1.TheInternationalCancerBiomarkersConsortium(ICBC)()UndertheleadershipofDr.LeeHartwell,PresidentandDirectorofFredHutchinsonCancerResearchCentertheICBCispioneeringanewmodelforbiomarkerdiscoveryanddevelopingtechnologiesandmethodologiestomakeitpossible.Organizations1.TheInternationalCancerBiomarkersConsortium(ICBC)()ThegoaloftheInternationalCancerBiomarkerConsortium(ICBC)istoadvancemedicalresearchandimprovepatientoutcomesbydiscoveringbiomarkers(indicators)formultipletypesofcancer.Throughalarge-scaleeffortsimilartotheHumanGenomeProject,theconsortiumaimstofacilitate