title: Single-cell codetection of metabolic activity, intracellular functional proteins, and genetic mutations from rare circulating tumor cells.
abstract: The high glucose uptake and activation of oncogenic signaling pathways in cancer cells has long made these features, together with the mutational spectrum, prime diagnostic targets of circulating tumor cells (CTCs). Further, an ability to characterize these properties at a single cell resolution is widely believed to be essential, as the known extensive heterogeneity in CTCs can obscure important correlations in data obtained from cell population-based methods. However, to date, it has not been possible to quantitatively measure metabolic, proteomic, and genetic data from a single CTC. Here we report a microchip-based approach that allows for the codetection of glucose uptake, intracellular functional proteins, and genetic mutations at the single-cell level from rare tumor cells. The microchip contains thousands of nanoliter grooves (nanowells) that isolate individual CTCs and allow for the assessment of their glucose uptake via imaging of a fluorescent glucose analog, quantification of a panel of intracellular signaling proteins using a miniaturized antibody barcode microarray, and retrieval of the individual cell nuclei for subsequent off-chip genome amplification and sequencing. This approach integrates molecular-scale information on the metabolic, proteomic, and genetic status of single cells and permits the inference of associations between genetic signatures, energy consumption, and phosphoproteins oncogenic signaling activities in CTCs isolated from blood samples of patients. Importantly, this microchip chip-based approach achieves this multidimensional molecular analysis with minimal cell loss (<20%), which is the bottleneck of the rare cell analysis.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yin
authors.initials: Y
authors.lastname: Tang
authors.forename: Shuai
authors.initials: S
authors.lastname: Sun
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Wang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wu
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhao
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Czajkowsky
authors.affiliation: Shanghai Municipal Hospital of Traditional Chinese Medicine , Shanghai 200071, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Longhua Hospital, Shanghai University of Traditional Chinese Medicine , Shanghai 200032, P.R. China.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Tian
authors.affiliation: Longhua Hospital, Shanghai University of Traditional Chinese Medicine , Shanghai 200032, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California , Los Angeles, California 90095, United States.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Deng
authors.forename: Qihui
authors.initials: Q
authors.lastname: Shi
mesh_terms: D004252:DNA Mutational Analysis / Q000295:instrumentation*; D004867:Equipment Design; D023281:Genomics / Q000295:instrumentation; D005947:Glucose / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D009154:Mutation; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology; D061848:Optical Imaging / Q000295:instrumentation; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism*; D040081:Protein Array Analysis / Q000295:instrumentation; D059010:Single-Cell Analysis / Q000295:instrumentation*
chemical_list: D010750:Phosphoproteins; D005947:Glucose