产品培训ctdna文献册

ctDNA1.1分1.2种1.3ctDNACTCs1986年，来自意大利的奖得主RenatoDulbecco在谈到为什么一定要测定整个人类组序列的时候说：这是打赢对的唯一途径。对于“是基因病”这一说法，科学家们已达成共识。2015年，在国情咨文目前，食品药品监督管理局（FDA）已强制要求在应用某些药物前进行EGFR、KRAS等，国立综合网络（ ）也已将EGFRKRASHER2、ERCC1等纳入治疗指南中。患者的外周循环血中含有多种与相关的突变，主要由肿瘤组织释ctDNActDNA作为一种高度灵敏、特异性的肿瘤新型标志物，可通过对其进行，分析出肿瘤中发生突变的，从而为患者在早期诊断、中，将液体活检技术列为肿瘤治疗领域的下一个十年趋势之一。ctDNA作为液体活检技术的重要一员已逐渐从科研临床，欧洲药品管理局（EMA）批准将ctDNA检测用于的伴随诊断更是标志着ctDNA临床应用的大规模实现。CirculatingTumorDNAandDNAMayHelpGuideTreatmentMorerecently,theliquidbiopsyconcepthasextendedtodetecting,capturing,andyzingcirculatingtumorDNA(ctDNA)andtumormicroRNA,minisculepiecesofthecancercell’sgeneticmaterialthatfloatlyinthebloodstream.RecentstudiesshowthatctDNAtestingcouldbeappliedtoallstagesofpatientcare–fromdetectionanddiagnosistoselectionoftreatmentsofctDNAlevelsintheboldmayalsobeusedtoquicklyestimatehowadvancedthecancerisandthepatient’ssurvivalchances,becausehigherlevelsofctDNAarestronglyassociatedwithshortersurvival.ysisofctDNAcouldalsobeusedforreal-timemonitoringofnewmutationsthatariseduringtumorprogressionandtherapyandtoselectthemostsuitabletreatments.Approximayadecadeago,researchersdiscoveredthatabnormalitiesinmicroRNAs–moleculesthatpreventcellsfrommakingspecificproteins–areassociatedwithcancerdevelopmentandprogression.SubsequentresearchhasshownthatmicroRNAysisisapromisingandcost-effectivewaytoidentifywhattypeofcancerapatienthas,estimatehowfastacancerisgrowing,monitorresponsetotherapy,anddetectrelapse.Althoughalltheseavenuesareunderintensiveresearch,confirmationinlarge-scaleclinicaltrialsisneededbeforeliquidbiopsies eroutineandwidelyavailableinIRESSAreceivesPpositiveopiniontoincludebloodbaseddiagnostictestinginEuropeanlabelAstraZenecatodayannouncedthattheCommitteeforMedicinalProductsforHumanUse(P)oftheEuropeanMedicinesAgency(EMA)hasadoptedapositiveopiniononaType-IIvariationupdatetotheEuropeanlabelforIRESSA®(gefitinib).Thelabelupdatewillhelpdoctorstoidentifylungcancerpatients-basedonthespecificgeneticdriversoftheirtumour-whocouldbenefitfromtreatmentwithIRESSAbutareunabletoprovideasuitabletumoursample.IRESSAisanepidermalgrowthfactorreceptortyrosinekinaseinhibitor(EGFRTKI)indicatedforthefirstlinetreatmentofadultpatientswithlocallyadvancedormetastaticnon-smallcelllungcancer(NSCLC)withactivatingmutationsoftheEGFRtyrosinekinase.Therefore,onlypatientswhosetumoursareEGFRmutationpositiveareeligibletoreceivetreatment.Tumoursamplesgainedthroughbiopsyaretheprimarymethodfordeterminingapatient’sEGFRmutationstatus,withoutwhichpatientsarenoteligiblefortreatmentwithanEGFRTKIsuchasIRESSA,whichisthestandardofcareinEurope.FollowingPopinion,IRESSAwillbethefirstEGFRTKIinEuropetoalabelFollowingPopinion,IRESSAwillbethefirstEGFRTKIinEuropetoalabelallowingtheuseofcirculatingtumourDNA(ctDNA)obtainedfromasample,tobeusedfortheassessmentofEGFRmutationstatusinthosepatientswhereatumoursampleisnotanoption.Theupdatewilltakeeffectimmedia yandwillbeapplicableinall28EuropeanUnionmembercountries.BriggsMorrison,Executive,GlobalMedicinesDevelopmentandMedicalOfficeratAstraZenecasaid:“AtAstraZeneca,wearecommittedtodevelotargetedmedicinesthatimprovehealth esforpatients.Understandingthenatureofanindividual’stumourandthereforewhichmedicineismostlikelytobenefitthemisvitalifwearetotransformthewaycancerpatientsaretreated.Ifdoctorsareunabletoassessthemutationstatusofatumour,thenpatients’accesstopotentiallylife-changingmedicinessuchasIRESSA esrestricted.Today’sdecisionbythe PtoendorsealabelupdateforIRESSAisasignificantstepforward.”AstraZenecahaspioneeredtheuseofinnovativeblood-baseddiagnostictestingforsolidtumoursandrecentlyannouncedapartnershipwithQiagentodevelopactDNAtestasacompaniondiagnosticforIRESSA.ctDNActDNA在了解ctDNA之前，首先需明确另一个与之相关的概念—cfDNA（circulating DNA，血液游离DNA即血浆中游离存在的DNA，主要来源于正常细胞、血液循环肿瘤DNA则是cfDNA中来源于肿瘤细胞的那一部分DNA，由肿瘤细胞释放入血，占cfDNA的0.01%-93%不等（Ref:NatRevClinOncol.2013Aug;10(8):472-84.。1947年，MandelcfDNA；1977年，Leon发现肿瘤患cfDNA1989年，StrouncfDNA中携ctDNA的研究热度持续上升，并开始应用于临床。ctDNA失（Indel、拷贝数变异（CNV）和结构变异（SV。ctDNA并非随机释放至血液，而是搭载其载体—核小体，核小体再以单个、双ctDNA166bp15分钟至数小时不等（Ref：NatRevCancer.2011Jun;11(6):426-37。ARMSBEAMing,0.01%or通过各种技术对ctDNA进序，可分析出其中突变及其突变丰度，为肿瘤患者提供有效信息。目前应用于ctDNA的检测技术有Sanger（又称一代、Pyrosequecing（焦磷酸、qPCR（tativeReal-timePCR，实时荧光PC（AmliiatinsystemNG（NeARMSBEAMing,0.01%orctDNA2015年指南提出，非小细胞肺癌患者治疗前应进行，然而1/3的患者没有足够的肿瘤组织样本进行（Ref:2015ASCOAnnualMeeting：e19082.。组织样本可直接反应肿瘤的谱，但也的确存在着一定的局限性，当近年来，ctDNA作为一种新的肿瘤标志物，在用药指导、疗效监测和复发监测等诊疗过程中表现出很大的潜力。但ctDNA的突变能否真实反映肿瘤组织的基非小细胞KRAS,EGFR,HER2,ClinCancerRes,-Cancer-PlosOne,KRASPLoSOne.NRAS0BRAFI--KRAS,EGFR,--ClinCancerRes,VNRAS--MolOncol.2015Sep25.BRAF--BRAF,cKIT,--胃肠道间--2013ASCOAnnual4--胰--50---MolOncol,异质性，使得所检测的组织样本不能完全代表患者肿瘤变异的全貌，因此可能存在组织中未检到而血液中检到突变的情况；2）血液中ctDNA是由的细胞释放至血液中，若在血液中未检测到组织中检到的突变，可能是这部分变异未另外，ctDNA作为一个系统性的检测，还可克服肿瘤异质性的问题。有研究发现，同一位肾癌患者的灶和转移灶不同部位的肿瘤组织在DNA水平上是有差异而有研究表示，ctDNA更能反映肿瘤谱的全貌，可克服肿瘤异质性，从而Capturingintra-tumorgeneticheterogeneitybydenovomutationprofilingofcirculatingcell-tumorDNA:aproof-of-principleDeMattos-ArrudaL,WeigeltB,CortesJ,etal.AnnOncol.2014Sep;25(9):1729-BACKGROUND:sma-derivedcell-tumorDNA(ctDNA)constitutesapotentialsurrogatefortumorDNAobtainedfromtissuebiopsies.Wepositthatmassivelyparallelsequencing(MPS)ysisofctDNAmayhelpdefinetherepertoireofmutationsinbreastcancerandmonitortumorsomaticaltionsduringthecourseoftargetedtherapy.PATIENTANDMETHODS:A66-year-oldpatientpresentedwithsynchronousestrogenreceptor-positive/HER2-negative,highlyproliferative,grade2,mixedinvasiveductal-lobularcarcinomawithboneandlivermetastasesatdiagnosis.DNAextractedfromarchivaltumormaterial,smaandperipheralbloodleukocyteswassubjectedtotargetedMPSusingatformcomprising300cancergenesknowntoharboractionablemutations.MultiplesmasampleswereduringthelineoftreatmentwithanAKTCONCLUSIONS:Thisproof-of-principlestudyisoneofthefirsttodemonstratethathigh-targetedMPSof sma-derivedctDNAconstitutesCONCLUSIONS:Thisproof-of-principlestudyisoneofthefirsttodemonstratethathigh-targetedMPSof sma-derivedctDNAconstitutesapotentialtoolfordenovomutationidentificationandmonitoringofsomaticgenetic tionsduringthecourseoftargetedandmaybeemployed总的来说，ctDNA不仅与肿瘤组织的检测结果有较高的一致性，还可在无法或ctDNA在cfDNA中的浓度为0.01%-93%不等，化差异极大。ctDNA能否在能被精准应用于临床，使患者获益。1.1分研究表明，几乎在所有中，都可检测到ctDNA，但不同癌种和不同分期患者的检出率差异较大。I-IV47%、55%、69%、82%，肿瘤越晚期，ctDNA的检出率越高。1.2种

Ref:SciTranslMed.2014FebctDNA，而脑部肿瘤的检出率不到10%——这可能和不同组织的特性，如血脑屏障有关。Ref:SciTranslMed.2014Feb者，均能检测到ctDNA，检出率为100%，特异度高达96%。就目前技术的灵敏度而言，ctDNA适用于肺癌、结直肠癌、、黑色素瘤、胃癌、胃肠道间质瘤、肝癌和胰的晚期患者。1.3ctDNA水平在患者治疗前和治疗后疾病进展后达到9TCGA数据库中非小NatMed.9TCGA数据库中非小NatMed.AnnOncol.PIK3CA,NEnglJMed.7exon9,11,13,14,17,AnnOncol.4胰ctDNA是近年来科学研究的热点，科学家们围绕其展开的研究数不胜数，以期Ref:SciTranslMed.2014Feb性，而不会波及肿瘤周围的正常组织细胞。早在1997年，食品药品监督管（FDA突变的肺癌患者的生存期，是性的变化。但靶向药物能否获得疗效主要取决于患者有无相关突变，若在不知患者有无突变状态情况下使用此类药，疗效EGFRmutationsdetectedinsmaareassociatedwithpatient esinerlotinibplusdocetaxel-treatednon-smallcelllungcancerMackPC,HollandWS,BurichRA,etal.JThoracOncol.2009Dec;4(12):1466-PURPOSE:Activatingmutationsintheepidermalgrowthfactorreceptor(EGFR)areassociatedwithenhancedresponsetoEGFRtyrosinekinaseinhibitorsinnon-smallcelllungcancer(NSCLC),whereasKRASmutationstranslateintopoorpatient es.WehypothesizedthatysisofsmaforEGFRandKRASmutationsfromshedtumorDNAwouldhaveclinicalutility.METHODS:Anallele-specificpolymerasechainreactionassayusingScorpion-amplificationrefractorymutationsystem(DxS,)wasusedtodetectmutationsinsmaDNAfrompatientswithadvancedstageNSCLCtreatedassecond-orthird-linetherapyonaphaseI/IItrialofdocetaxelplusintercalatedRESULTS:EGFRmutationsweredetectedin10of49patients(20%).Six(12%)hadsingleactivatingmutationsinEGFR,associatedwithimprovedprogression-survival(median,18.3months),comparedwithallotherpatients(median,3.9months;p=0.008),orthosewithwild-typeEGFR(median,4.0months;p=0.012).Fourof49patientsharboredadenovoT790M (medianprogression-survival,3.9months).EGFRmutationalstatuswasassociatedwithclinicalresponse(45assessable,p=0.0001);inthesixpatientswithactivatingmutations,allachievedcomplete(33%)orpartial(67%)response.AllCRpatientshadE19deldetectableinbothtumorandsma.KRASmutationsweredetectedintwoof49(4%)patients,bothofwhomhadrapidprogressivedisease.CONCLUSIONS:ActivatingEGFRmutationsdetectedinshedDNAinsmaaresignificantlyassociatedwithfavorable esinpatientswithadvancedNSCLCreceivingdocetaxelplusintercalatederlotinib.TheadditionofdocetaxelinthisscheduledidnotdiminishtheefficacyoferlotinibagainstpatientswithEGFRactivatingmutations.（OSACBCA：ctDNA突变与患者病情进展的关系，EGFRE19/21（EGFR-19delorEGFR-L858REGFR酪氨酸酸激酶抑制剂的耐药突变、mtKRAS和EGFR野生型患者的疗效不尽理想。B、CctDNAPFS/OS，EGFRE19/21PFSOS结论：ctDNAEGFR-19del/L858R突变的非小细胞肺癌患者，对厄洛替尼和多西他赛交替治疗的疗效良好，PFS和OS均高于其他型。靶向药物对不同型患者的疗效不同，在选择靶向治疗之前需进行ctDNAEvaluationofcirculatingtumorcellsandcirculatingtumorDNAinnon-smallcelllungcancer:associationwithclinicalendpointsinaphaseIIclinicaltrialofpertuzumabandPunnooseEA,AtwalS,LiuW,RajaR,etal.ClinCancerRes.2012Apr15;18(8):2391-PURPOSE:Elevatedlevelsorincreasesincirculatingtumorcells(CTC)portendpoorprognosisinpatientswithepithelialcancers.LessisknownaboutCTCsassurrogateendpointsortheiruseforpredictivebiomarkerevaluation.ThisstudyinvestigatedtheutilityofCTCenumerationandcharacterizationusingtheCellSearchtform,aswellasmutationdetectionincirculatingtumorDNA(ctDNA),inpatientswithadvancednon-smallcelllungcancer(NSCLC).EXPERIMENTALDESIGN:Forty-onepatientswereenrolledinasingle-armphaseIIclinicaltrialoferlotinibandpertuzumab.PeripheralbloodwasyzedforCTCenumeration,EGFRexpressioninCTCs,anddetectionofoncogenicmutationsinCTCsandctDNA.ChangesinCTClevelswerecorrelatedwith2[18F]fluoro-2-deoxy-D-glucose-positronemissiontomographic(FDG-PET)andcomputedtomographic(CT)imagingandsurvivalendpoints.RESULTS:CTCsweredetected(≥1CTC)atbaselinein78%ofpatients.GreatersensitivityformutationdetectionwasobservedinctDNAthaninCTCsanddetectedmutationswerestronglyconcordantwithmutationstatusinmatchedtumor.HigherbaselineCTCcountswereassociatedwithresponsetotreatmentbyResponseEvaluationCriteriainSolidTumors(RECIST,P=0.009)anddecreasedCTCcountsupontreatmentwereassociatedwithFDG-PETandRECISTresponse(P=0.014andP=0.019)andlongerprogression-survival(P=0.050).CONCLUSION:ThesedataprovideevidenceofacorrelationbetweendecreasesinCTCcountsandradiographicresponsebyeitherFDG-PETorRECISTinpatientswithadvancedNSCLC.ThesefindingsrequireprospectivevalidationbutsuggestapotentialroleforusingCTCdecreasesasanearlyindicationofresponsetotherapyandctDNAforreal-timeassessmentofmutationstatusfromblood.该研究对24位受试于帕妥珠单抗和厄洛替尼IIB患者进行ctDNA，并记录其PFS，以评估靶向药物对不同型患者的疗BB：EGFR56FDG-PETCT结果，1EGFR突变型患者在服ctDNAEGFRIF=EGFRmutationdetectioninctDNAfromNSCLCpatientsma:Across-tformcomparisonofleadingtosupporttheclinicaldevelopmentofAZD9291ThressKS,BrantR,CarrTH,etal.LungCancer.2015Oct9.OBJECTIVES:Toassesstheabilityofdifferenttechnologytformstodetectepidermalgrowthfactorreceptor(EGFR)mutations,includingT790M,fromcirculatingtumorDNA(ctDNA)inadvancednon-smallcelllungcancer(NSCLC)patients.MATERIALSANDMETHODS:AcomparisonofmultipletformsfordetectingEGFRmutationsinsmactDNAwasundertaken.smasampleswerecollectedfrompatientsenteringtheongoingAURAtrial(NCT ),investigatingthesafety,tolerability,andefficacyofAZD9291inpatientswithEGFR-sensitizingmutation-positiveNSCLC.smawascollectedpriortoAZD9291dosingbutfollowingclinicalprogressiononapreviousEGFR-tyrosinekinaseinhibitor(TKI).ExtractedctDNAwasyzedusingtwonon-digitaltforms(cobas®EGFRMutationTestandtherascreen™EGFRamplificationrefractorymutationsystemassay)andtwodigitaltforms(DropletDigital™PCRandBEAMingdigitalPCR[dPCR]).RESULTS:Preliminaryassessment(38samples)wasconductedusingallfourtforms.ForEGFR-TKI-sensitizingmutations,highsensitivity(78-100%)andspecificity(93-100%)wereobservedusingtissueasanon-referencestandard.FortheT790Mmutation,thedigitaltformsoutperformedthenon-digitaltforms.Subsequentassessmentusing72additionalbaselinesmasampleswasconductedusingthecobas®EGFRMutationTestandBEAMingdPCR.Thetwotformsdemonstratedhighsensitivity(82-87%)andspecificity(97%)forEGFR-sensitizingmutations.FortheT790Mmutation,thesensitivityandspecificitywere73%and67%,respectively,withthecobas®EGFRMutationTest,and81%and58%,respectively,withBEAMingdPCR.Concordancebetweenthetformswas>90%,showingthatmultipletformsarecapableofsensitiveandspecificdetectionofEGFR-TKI-sensitizingmutationsfromNSCLCpatientsma.CONCLUSION:Thecobas®EGFRMutationTestandBEAMingdPCRdemonstrateahighsensitivityforT790Mmutationdetection.GenomicheterogeneityofT790M-mediatedmayexinthereducedspecificityobservedwithsma-baseddetectionofT790Mmutationsversustissue.ThesedatasupporttheuseofbothtformsintheAZD9291clinical该研究用不同检测技术对非小细胞肺癌患者的ctDNA进行，评估第三代EGFR-TKI药物—AZD9291对不同型患者的疗效。为59%，疾病控制率为90%，均显著高于血液EGFR-T790M野生型患者。EGFR-T790MEGFRTKI类药物发生耐药的主要原因，在获得性耐药人群中的发生频率高达~50-65%（Ref:NatRevClinOncol.2014Aug;11(8):IF=EmergenceofKRASmutationsandacquiredtoanti-EGFRtherapyincolorectalcancerMisaleS,YaegerR,HoborS,etal.Nature.2012Jun28;486(7404):532-Amainlimitationoftherapiesthatselectivelytargetkinasesignallingpathwaysistheemergenceofsecondarydrug .Cetuximab,amonoclonalantibodythatbindstheextracellularofepidermalgrowthfactorreceptor(EGFR),iseffectiveinasubsetofKRASwild-typemetastaticcolorectalcancers.Afteraninitialresponse,secondaryinvariablyensues,therebylimitingtheclinicalbenefitofthisdrug.Themolecularbasesofsecondary tocetuximabincolorectalcancerarepoorlyunderstood.Hereweshowthatmolecularaltions(inmostinstancespointmutations)ofKRASarecausallyassociatedwiththeonsetofacquired toanti-EGFRtreatmentincolorectalcancers.ExpressionofmutantKRASunderthecontrolofitsendogenousgenepromoterwassufficienttoconfercetuximab ,butresistantcellsremainedsensitivetocombinatorialinhibitionofEGFRandmitogen-activatedprotein-kinasekinase(MEK).ysisofmetastasesfrompatientswhodeveloped tocetuximaborpanitumumabshowedtheemergenceofKRASamplificationinonesampleandacquisitionofsecondaryKRASmutationsin60%(6outof10)ofthecases.KRASmutantallelesweredetectableinthebloodofcetuximab-treatedpatientsasearlyas10monthsbeforeradiographicationofdiseaseprogression.Insummary,theresultsidentifyKRASmutationsasfrequentdriversofacquired tocetuximabincolorectalcancers,indicatethattheemergenceofKRASmutantclonescanbedetectednon-invasivelymonthsbeforeradiographicprogressionandsuggestearlyinitiationofaMEKinhibitorasarationalstrategyfordelayingorreversingdrug 服用单抗后，ctDNA中KRAS突变型患者的PFS明显低于KRAS野生型患者，P<0.05；但对于在服用单抗后，仍维持KRAS野生型和获得性KRAS突变的患者，二者的PFS差异无统计学结论：在结直肠癌中，KRAS突变可导致患者对EGFR单抗类药物耐药。单抗可有效改善ctDNAKRAS野生型的结直肠癌患者的PFS。肿瘤的治疗绝不是一蹴而就的事情，即使手术完全切除了肿瘤，还有可能会复发；即使服用了靶向药物，还会发展出耐药性；原位肿瘤不再发展，癌细胞还有可。NoninvasivedetectionofresponseandinEGFR-mutantlungcancerusingtativenext-generationgenotyofcell-smaDNAOxnardGR,PaweletzCP,KuangY,etal.ClinCancerRes.2014Mar15;20(6):1698-PURPOSE:Tumorgenotyusingcell-smaDNA(cfDNA)hasthepotentialtoallownoninvasiveassessmentoftumorbiology,yetmanyexistingassaysarecumbersomeandvulnerabletofalse-positiveresults.WesoughttodeterminewhetherdropletdigitalPCR(ddPCR)ofcfDNAwouldallowhighlyspecificandtativeassessmentoftumorgenotype.EXPERIMENTALDESIGN:ddPCRassaysforEGFR,KRAS,andBRAFmutationsweredevelopedusingsmacollectedfrompatientswithadvancedlungcancerormelanomaofaknowntumorgenotype.Sensitivityandspecificityweredeterminedusingcancerswithnonoverlapgenotypesaspositiveandnegativecontrols.Serialassessmentofresponseand wasstudiedinpatientswithEGFR-mutantlungcanceronaprospectivetrialoferlotinib.RESULTS:WeidentifiedareferencerangeforEGFRL858Randexon19deletionsinspecimensfromKRAS-mutantlungcancer,allowingidentificationofcandidatethresholdswithhighsensitivityand100%specificity.Receivedoperativecharacteristiccurveysisoffourassaysdemonstratedanareaunderthecurveintherangeof0.80to0.94.SensitivityimprovedinspecimenswithoptimalcfDNAconcentrations.SerialsmagenotyofEGFR-mutantlungcanceronerlotinibdemonstratedpretreatmentdetectionofEGFRmutations,completesmaresponseinmostcases,andincreasinglevelsofEGFRT790Memergingbeforeobjectiveprogression.CONCLUSIONS:NoninvasivegenotyofcfDNAusingddPCRdemonstratesassayqualitiesthatcouldalloweffectivetranslationintoaclinicaldiagnostic.Serialficationofsmagenotypeallowsnoninvasiveassessmentofresponseand,includingdetectionofmutationsupto16weeksbeforeradiographicprogression.周，患者出现疾病进展（PD。结论：EGFR-T790MEGFRTKIctDNAIF=Liquidbiopsies:genotycirculatingtumorDiazLAJr,BardelliA.JClinOncol.2014Feb20;32(6):579-Genotytumortissueinsearchofsomaticgeneticaltionsforactionableinformationhaseroutinepracticeinclinicaloncology.Althoughthesesequenceal tionsarehighlyinformative,samplingtumortissuehassignificantinherentlimitations;tumortissueisasinglesnapshotintime,issubjecttoselectionbiasresultingfromtumorheterogeneity,andcanbedifficulttoobtain.Cell-fragmentsofDNAareshedintothebloodstreambycellsundergoingapoptosisornecrosis,andtheloadofcirculatingcell-DNA(cfDNA)correlateswithtumorstagingandprognosis.Moreover,recentadvancesinthesensitivityandaccuracyofDNAysishaveallowedforgenotyofcfDNAforsomaticgenomical tionsfoundintumors.Theabilitytodetectandfytumormutationshasproveneffectiveintrackingtumordynamicsinrealtimeaswellasservingasaliquidbiopsythatcanbeusedforavarietyofclinicalandinvestigationalapplicationsnotpreviouslypossible.转移性结直肠癌患者的动态监测图：黄色实线代表APCctDNA突变水平作为基线：APC突变型、KRAS野生型。患者和NRAS突变和/或MET扩增，表明多种类型耐药突变的产生。随后出现临床耐药症状。中KRAS、MET等突变有关，且突变在影像学检出前数月即可发现。MutationtrackingincirculatingtumorDNApredictsrelapseinearlybreastGarcia-MurillasI,SchiavonG,WeigeltB,etal.SciTranslMed.2015Aug26;7(302):Theidentificationofearly-stagebreastcancerpatientsathighriskofrelapsewouldallowtailoringofadjuvanttherapyapproaches.WeassessedwhetherysisofcirculatingtumorDNA(ctDNA)insmacanbeusedtomonitorforminimalresidualdisease(MRD)inbreastcancer.Inaprospectivecohortof55earlybreastcancerpatientsreceivingneoadjuvantchemotherapy,detectionofctDNAinsmaaftercompletionofapparentlycurativetreatment-eitheratasinglepostsurgicaltimepointorwithserialfollow-upsmasamples-predictedmetastaticrelapsewithhighaccuracy[hazardratio,25.1(confidenceinterval,4.08to130.5;log-rankP<0.0001)or12.0(confidenceinterval,3.36to43.07;log-rankP<0.0001),respectively].Mutationtrackinginserialsamplesincreasedsensitivityforthepredictionofrelapse,withamedianleadtimeof7.9monthsoverclinicalrelapse.WefurtherdemonstratedthattargetedcapturesequencingysisofctDNAcoulddefinethegeneticeventsofMRD,andthatMRDsequencingpredictedthegeneticeventsofthesubsequentmetastaticrelapsemoreaccuraythansequencingoftheprimarycancer.Mutationtrackingcanthereforeidentifyearlybreastcancerpatientsathighriskofrelapse.SubsequentadjuvanttherapeuticinterventionscouldbetailoredtothegeneticeventspresentintheMRD,atherapeuticapproachthatcouldinpartcombatthechallengeposedbyintratumorgeneticheterogeneity.A2~4ctDNA，并统计患者术后三年的无病生存期。ctDNA阳性患者的无病生存率低于ctDNA患者，即ctDNA阳性患者的复发率显著高于ctDNA患者，其复发率为86%，是ctDNA患者的12倍。BctDNA，并统计患者的无病生存期。相较于仅在术后单次80%；ctDNA阳性患者的中位无病生存期为13.6个月。CirculatingmutantDNAtoassesstumorDiehlF,SidtK,ChotiMA,etal.NatMed.2008Sep;14(9):985-ThemeasurementofcirculatingnucleicacidshastransformedthemanagementofchronicviralinfectionssuchasHIV.Thedevelopmentofogousmarkersforindividualswithcancercouldsimilarlyenhancethemanagementoftheirdisease.DNAcontainingsomaticmutationsishighlytumorspecificandthus,intheory,canprovideoptimummarkers.However,thenumberofcirculatingmutantgenefragmentsissmallcomparedtothenumberofnormalcirculatingDNAfragments,makingitdifficulttodetectandfythemwiththesensitivityrequiredformeaningfulclinicaluse.Inthisstudy,weappliedahighlysensitiveapproachtofycirculatingtumorDNA(ctDNA)in162smasamplesfrom18subjectsundergoingmultimodalitytherapyforcolorectalcancer.WefoundthatctDNAmeasurementscouldbeusedtoreliablymonitortumordynamicsinsubjectswithcancerwhowereundergoingsurgeryorchemotherapy.Wesuggestthatthisalizedgeneticapproachcouldbegenerallyappliedtoindividualswithothertypesofcancer.18ctDNACEA对复AABB无复发生存率均高于CEA患者，即ctDNA患者的复发率显著低于CEA患者，假率较CEA低。CEA的灵敏度仅为56%。患者的复发率显著高于CEA阳性患者，假阳性率较CEA低。ctDNACT10个月发现复发征兆（Ref:Gut.2015Feb4.pii:gutjnl-2014-308859.。发现时大多已是晚期，这很大一部分原因是源于在的早期诊断方面尚无准确的检测，目前主要应用于临床的是肿瘤标志物和影像学检查，但其目前还没有足够的证实ctDNA可作为早诊的金标准，一方面是因为不同癌ctDNA水平差异较大，另一方面则源于目前仅有检测技术的灵敏度限制。但ctDNA作为肿瘤细胞分泌的特异性分子，相较传统早期诊断有着明显的情况，而不是数周甚至数月；并且有大量研究表明，ctDNA的水平可以反映肿瘤负荷，与临床病程相关。ctDNA在早期诊断方面仍是一个颇具潜力的指标。Circulatingcell-DNAinserumasabiomarkerfordiagnosisandprognosticpredictionofcolorectalcancerHaoTB,ShiW,ShenXJ,etal.BrJCancer.2014Oct14;111(8):1482-BACKGROUND:Toverifywhethertheconcentrationsandintegrityindexofcirculatingcell-DNA(ccf-DNA)inserummaybeclinicallyusefulforthediagnosisandprogressionmonitoringofcolorectalcancer(CRC)patients.METHODS:Serumsampleswerecollectedfrom104withprimaryCRC,85withoperatedCRC,16withrecurrent/metastaticCRC,63patientswithintestinalpolypsand110normalcontrols.Long(247bp)andshort(115bp)DNAfragmentsinserumweredetectedbyreal-timetativePCRbyamplifyingtheALUrepeats(ALU-qPCR).Serumcarcinoembryonicantigen(CEA)levelwasdetectedbyARCHITECTassay.RESULTS:ThemedianabsoluteserumALU115andALU247/115inprimaryCRCgroupwassignificantlyhigherthanthoseinintestinalpolypandnormalcontrolgroups(bothP<0.0001),inrecurrent/metastaticCRCwassignificantlyhighercomparedwithprimaryCRC(P=0.0021,P=0.0018)oroperatedCRC(P<0.0001,respectively)andduringfollow-up,ALU115andALU247/115wereincreasedbeforesurgeryanddecreasedsignificantlyaftersurgery.CONCLUSIONS:CombineddetectionofALU115,ALU247/115andCEAcouldimprovethediagnosticefficiencyforCRC.SerumDNAconcentrationsandintegrityindexmaybevaluableinearlycomplementarydiagnosisandmonitoringofprogressionandprognosisofCRC.含量和ALU247/115均显著高于肠息肉患者和健康人群。Valueofcirculatingcell-DNAindiagnosisofhepatocelluarcarcinoChenK,ZhangH,ZhangLN,etal.WorldJGastroenterol.2013May28;19(20):3143-AIM:Toinvestigatethevalueofcombineddetectionofcirculatingc