Effective capture and analysis of a single circulating tumor cell(CTC)is instrumental for early diagnosis and personalized therapy of tumors.However,due to their extremely low abundance and susceptibility to interfere...Effective capture and analysis of a single circulating tumor cell(CTC)is instrumental for early diagnosis and personalized therapy of tumors.However,due to their extremely low abundance and susceptibility to interference from other cells,high-throughput isolation,enrichment,and single-cell-level functional protein analysis of CTCs within one integrated system remains a major challenge.Herein,we present an integrated multifunctional microfluidic system for highly efficient and label-free CTC isolation,CTC enrichment,and single-cell immunoblotting(ieSCI).The ieSCI-chip is a multilayer microfluidic system that combines an inertia force-based cell sorter with a membrane filter for label-free CTC separation and enrichment and a thin layer of a photoactive polyacrylamide gel with microwell arrays at the bottom of the chamber for single-cell immunoblotting.The ieSCI-chip successfully identified a subgroup of apoptosis-negative(Bax-negative)cells,which traditional bulk analysis did not detect,from cisplatin-treated cells.Furthermore,we demonstrated the clinical application of the ieSCI-chip with blood samples from breast cancer patients for personalized CTC epithelial-to-mesenchymal transition(EMT)analysis.The expression level of a tumor cell marker(EpCAM)can be directly determined in isolated CTCs at the single-cell level,and the therapeutic response to anticancer drugs can be simultaneously monitored.Therefore,the ieSCI-chip provides a promising clinical translational tool for clinical drug response monitoring and personalized regimen development.展开更多
基金We are grateful for financial support from Projects 22077079,31971327,and 81871448 of the National Natural Science Foundation of China(NSFC)a Project of the National Innovation Special Zone+11 种基金Projects 2017SHZDZX01,17DZ2203400,and 18430760500 of Shanghai Municipal Science and TechnologyProject G20180101 of the Shanghai Agriculture Applied Technology Development ProgramProject ZXWF082101 of the Shanghai Municipal Education CommissionProject 2017ZX10203205-006-002 of the National Key Research and Development Program of ChinaProjects 19×190020154,ZH2018ZDA01,YG2016QN24,and YG2016MS60 of the Shanghai Jiao Tong University Biomedical Interdisciplinary ProgramProjects ZH2018QNA54 and ZH2018QNA49 of the Medical-Engineering Cross Foundation of Shanghai Jiao Tong UniversityProject 2019CXJQ03 of the Innovation Group Project of the Shanghai Municipal Health CommissionProject 19MC1910800 of the Shanghai Clinical Medical Research CenterProject SD0820016 of the third batch of industrialization projects of the Innovation Incubation Fund of Nantong and Shanghai Jiao Tong UniversityProject SL2020MS026 of the Oceanic Interdisciplinary Program of Shanghai Jiao Tong UniversityProject Agri-X20200101 of Shanghai Jiao Tong Universityand the SJTU Global Strategic Partnership Fund(2020 SJTU-HUJI).In addition,we thank AEMD SJTU for the support.
文摘Effective capture and analysis of a single circulating tumor cell(CTC)is instrumental for early diagnosis and personalized therapy of tumors.However,due to their extremely low abundance and susceptibility to interference from other cells,high-throughput isolation,enrichment,and single-cell-level functional protein analysis of CTCs within one integrated system remains a major challenge.Herein,we present an integrated multifunctional microfluidic system for highly efficient and label-free CTC isolation,CTC enrichment,and single-cell immunoblotting(ieSCI).The ieSCI-chip is a multilayer microfluidic system that combines an inertia force-based cell sorter with a membrane filter for label-free CTC separation and enrichment and a thin layer of a photoactive polyacrylamide gel with microwell arrays at the bottom of the chamber for single-cell immunoblotting.The ieSCI-chip successfully identified a subgroup of apoptosis-negative(Bax-negative)cells,which traditional bulk analysis did not detect,from cisplatin-treated cells.Furthermore,we demonstrated the clinical application of the ieSCI-chip with blood samples from breast cancer patients for personalized CTC epithelial-to-mesenchymal transition(EMT)analysis.The expression level of a tumor cell marker(EpCAM)can be directly determined in isolated CTCs at the single-cell level,and the therapeutic response to anticancer drugs can be simultaneously monitored.Therefore,the ieSCI-chip provides a promising clinical translational tool for clinical drug response monitoring and personalized regimen development.
