Molecular Biomarker Analyses Using Circulating Tumor Cells 英文参考文献.docVIP

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Molecular Biomarker Analyses Using Circulating Tumor Cells 英文参考文献.doc

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Molecular Biomarker Analyses Using Circulating Tumor Cells 英文参考文献

MolecularBiomarkerAnalysesUsingCirculatingTumor Cells ElizabethA.Punnoose1*,SiminderK.Atwal1,JillM.Spoerke1,HeidiSavage1,AjayPandita2 ,Ru-Fang Yeh3,AndreaPirzkall4,BernardM.Fine4,LukasC.Amler1,DanielS.Chen4,MarkR.Lackner1 1DepartmentofOncologyBiomarkerDevelopment,Genentech,Inc.,SouthSanFrancisco,California,UnitedStatesofAmerica,2DepartmentofMolecularDiagnostics andCancerCellBiology,Genentech,Inc.,SouthSanFrancisco,California,UnitedStatesofAmerica,3DepartmentofBiostatistics,Genentech,Inc.,SouthSanFrancisco, California, United States of America, 4Department of Oncology Exploratory Clinical Development, Genentech, Inc., South San Francisco, California, United States of America Abstract Background:Evaluationofcancerbiomarkersfrombloodcouldsignificantlyenablebiomarkerassessmentbyprovidinga relatively non-invasive source of representative tumor material. Circulating Tumor Cells (CTCs) isolated from blood of metastaticcancerpatientsholdsignificantpromiseinthisregard. Methodology/Principal Findings: Using spiked tumor-cells we evaluated CTC capture on different CTC technology platforms,includingCellSearchHandtwobiochipplatforms,andusedtheisolatedCTCstodevelopandoptimizeassaysfor molecularcharacterizationofCTCs.WereportsimilarperformanceforthevariousplatformstestedincapturingCTCs,and find that capture efficiency is dependent on the level of EpCAM expression. We demonstrate that captured CTCs are amenabletobiomarkeranalysessuchasHER2status,qRT-PCRforbreastcancersubtypemarkers,KRASmutationdetection, and EGFR staining by immunofluorescence (IF). We quantify cell surface expression of EGFR in metastatic lung cancer patientsamples.Inaddition,wedeterminedHER2statusbyIFandFISHinCTCsfrommetastaticbreastcancerpatients.In themajorityofpatients(89%)wefoundconcordancewithHER2statusfrompatienttumortissue,thoughinasubsetof patients(11%),HER2statusinCTCsdifferedfromthatobservedintheprimarytumor.Surprisingly,wefoundCTCcountsto behigherinER+patientsincomparisontoHER2+andtriplenegativepatie