Development of a TCR beta repertoire assay for profiling liquid biopsies from NSCLC donors  

作　　者：Leisa P.Jackson Benjamin A.Tjoa Hestia Mellert Gary A.Pestano 

机构地区：[1]Development,Biodesix Inc.,Boulder,CO 80301,USA. [2]Cellero,LLC,Bothell,Washington,WA 98021,USA.

出　　处：《Cancer Drug Resistance》2020年第3期563-571,共9页癌症耐药（英文）

摘　　要：Aim:The aim of this study was to demonstrate the utility of T-Cell receptor beta(TCRβ)sequencing as a robust method for assessing T-cell repertoire changes in donors with non-small cell lung cancer(NSCLC).We further demonstrated the use of the assay by monitoring repertoire modulation in a defined model antigen system,cytomegalovirus(CMV).Methods:Peripheral blood mononuclear cells from four healthy donors were challenged with a 1-week exposure to whole-cell lysate from CMV-infected cells or CMVpp65495-503 peptide(NLVPMVATV).T-cell repertoire perturbations were assessed using the Oncomine TCR Beta-SR Assay and Ion GeneStudio S5 Plus Sequencer.A pp65 tetramer flow cytometry assay was used as an orthogonal method to assess clonal expansion of a subset of CMV-specific T-cells.For evaluation of the assay in peripheral blood lymphocytes from NSCLC donors,five whole blood specimens were evaluated using the same sequencing workflow.Results:The TCR beta assay identified 6,683-61,936 unique clones from 1-2 million reads per sample,and an average of 80%of the total reads were usable for TCR profiling.In the NSCLC donors,TCR convergence and clonality values were consistent with published results and ranged 0.016-0.033 for convergence and 0.09-0.48 for clonality.In the CMV study,TCR sequencing detected the expansion of a common family of clones in all 4 samples in response to antigen stimulation.This expansion corresponded to an increase in pp65 tetramer staining by flow cytometry.Baseline TCR convergence scores ranged 0.009-0.041 and increased 5-fold in one sample as a result of pp65 antigen stimulation.Conclusion:The results of this study demonstrated the utility of profiling of the TCRβrepertoire in a model system and in donors with NSCLC.Additionally,we demonstrated the correlation between RNA-seq methods and protein-tetramer analysis using flow cytometry.These techniques represent an emerging solution that could complement other liquid and tissue diagnostic assays in the clinic and will be of value in predicting host r

关 键 词：TCRΒ liquid biopsy Ion Torrent immunotherapy 

分 类 号：R73[医药卫生—肿瘤]