title: An integrated microfluidic chip system for single-cell secretion profiling of rare circulating tumor cells.
abstract: Genetic and transcriptional profiling, as well as surface marker identification of single circulating tumor cells (CTCs) have been demonstrated. However, quantitatively profiling of functional proteins at single CTC resolution has not yet been achieved, owing to the limited purity of the isolated CTC populations and a lack of single-cell proteomic approaches to handle and analyze rare CTCs. Here, we develop an integrated microfluidic system specifically designed for streamlining isolation, purification and single-cell secretomic profiling of CTCs from whole blood. Key to this platform is the use of photocleavable ssDNA-encoded antibody conjugates to enable a highly purified CTC population with <75 'contaminated' blood cells. An enhanced poly-L-lysine barcode pattern is created on the single-cell barcode chip for efficient capture rare CTC cells in microchambers for subsequent secreted protein profiling. This system was extensively evaluated and optimized with EpCAM-positive HCT116 cells seeded into whole blood. Patient blood samples were employed to assess the utility of the system for isolation, purification and single-cell secretion profiling of CTCs. The CTCs present in patient blood samples exhibit highly heterogeneous secretion profile of IL-8 and VEGF. The numbers of secreting CTCs are found not in accordance with CTC enumeration based on immunostaining in the parallel experiments.
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Sun
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Minjiao
authors.initials: M
authors.lastname: Wang
authors.affiliation: Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Beiqin
authors.initials: B
authors.lastname: Yu
authors.affiliation: 1] Center for Bio-Detection and Bio-Instrumentation, Shanghai Jiao Tong University, Shanghai, China [2] School of Biomedicial Engineering, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Czajkowsky
authors.affiliation: 1] Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China [2] Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Bingya
authors.initials: B
authors.lastname: Liu
authors.affiliation: Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: 1] Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China [2] State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong University, Shanghai, China [3] Center for Bio-Detection and Bio-Instrumentation, Shanghai Jiao Tong University, Shanghai, China [4] School of Biomedicial Engineering, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Qihui
authors.initials: Q
authors.lastname: Shi
mesh_terms: D000368:Aged; D000951:Antigens, Neoplasm / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000097:blood* / Q000556:secondary; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D004044:Dietary Proteins / Q000378:metabolism*; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D045325:HCT116 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood* / Q000473:pathology; D008297:Male; D044085:Microfluidics / Q000379:methods*; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D040901:Proteomics / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D004044:Dietary Proteins; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; C012551:single cell proteins