title: Microfluidic enrichment for the single cell analysis of circulating tumor cells.
abstract: Resistance to drug therapy is a major concern in cancer treatment. To probe clones resistant to chemotherapy, the current approach is to conduct pooled cell analysis. However, this can yield false negative outcomes, especially when we are analyzing a rare number of circulating tumor cells (CTCs) among an abundance of other cell types. Here, we develop a microfluidic device that is able to perform high throughput, selective picking and isolation of single CTC to 100% purity from a larger population of other cells. This microfluidic device can effectively separate the very rare CTCs from blood samples from as few as 1 in 20,000 white blood cells. We first demonstrate isolation of pure tumor cells from a mixed population and track variations of acquired T790M mutations before and after drug treatment using a model PC9 cell line. With clinical CTC samples, we then show that the isolated single CTCs are representative of dominant EGFR mutations such as T790M and L858R found in the primary tumor. With this single cell recovery device, we can potentially implement personalized treatment not only through detecting genetic aberrations at the single cell level, but also through tracking such changes during an anticancer therapy.
authors.affiliation: Clearbridge Accelerator Pte Ltd, 81 Science Park Drive, The Chadwick, #02-03, Singapore Science Park 1, Singapore 118257, Singapore.
authors.forename: Trifanny
authors.initials: T
authors.lastname: Yeo
authors.affiliation: Clearbridge Accelerator Pte Ltd, 81 Science Park Drive, The Chadwick, #02-03, Singapore Science Park 1, Singapore 118257, Singapore.
authors.forename: Swee Jin
authors.initials: SJ
authors.lastname: Tan
authors.affiliation: School of Biological Science, National Technological University, 60 Nanyang Drive, Singapore 637551, Singapore.
authors.forename: Chew Leng
authors.initials: CL
authors.lastname: Lim
authors.affiliation: Cancer Therapeutics Research Laboratory, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.
authors.forename: Dawn Ping Xi
authors.initials: DP
authors.lastname: Lau
authors.affiliation: Department of Pathology, Singapore General Hospital, Outram Road, Singapore 169608, Singapore.
authors.forename: Yong Wei
authors.initials: YW
authors.lastname: Chua
authors.affiliation: Cancer Therapeutics Research Laboratory, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.
authors.forename: Sai Sakktee
authors.initials: SS
authors.lastname: Krisna
authors.affiliation: Cancer Therapeutics Research Laboratory, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.
authors.forename: Gopal
authors.initials: G
authors.lastname: Iyer
authors.affiliation: Department of Pathology, Singapore General Hospital, Outram Road, Singapore 169608, Singapore.
authors.forename: Gek San
authors.initials: GS
authors.lastname: Tan
authors.affiliation: Department of Pathology, Singapore General Hospital, Outram Road, Singapore 169608, Singapore.
authors.forename: Tony Kiat Hon
authors.initials: TK
authors.lastname: Lim
authors.affiliation: Cancer Therapeutics Research Laboratory, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.|Cancer Stem Cell Biology, Genome Institute of Singapore, 60 Biopolis St, #02-01, 138672, Singapore.|Division of Medical Oncology, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.
authors.forename: Daniel S W
authors.initials: DS
authors.lastname: Tan
authors.affiliation: Division of Medical Oncology, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610, Singapore.|Duke-NUS Medical School, 8 College Road, 169857, Singapore.|Institute of Molecular and Cell Biology, A*Star, 61 Biopolis Drive Proteos, 138673, Singapore.
authors.forename: Wan-Teck
authors.initials: WT
authors.lastname: Lim
authors.affiliation: Department of Biomedical Engineering, National University of Singapore, 4 Engineering Drive 3, Block E4, #04-08, Singapore 117583, Singapore.|Mechanobiology Institute of Singapore, 5A Engineering Drive 1, Singapore 117411, Singapore.
authors.forename: Chwee Teck
authors.initials: CT
authors.lastname: Lim
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics; D006801:Humans; D056656:Lab-On-A-Chip Devices; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D061986:MCF-7 Cells; D046210:Microfluidic Analytical Techniques; D009154:Mutation; D009360:Neoplastic Cells, Circulating / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D059010:Single-Cell Analysis
chemical_list: C512478:EGFR protein, human; D066246:ErbB Receptors