title: Expansion of CTCs from early stage lung cancer patients using a microfluidic co-culture model.
abstract: The potential utility of circulating tumor cells (CTCs) to guide clinical care in oncology patients has gained momentum with emerging micro- and nanotechnologies. Establishing the role of CTCs in tumor progression and metastasis depends both on enumeration and on obtaining sufficient numbers of CTCs for downstream assays. The numbers of CTCs are few in early stages of cancer, limiting detailed molecular characterization. Recent attempts in the literature to culture CTCs isolated from metastatic patients using monoculture have had limited success rates of less than 20%. Herein, we have developed a novel in-situ capture and culture methodology for ex-vivo expansion of CTCs using a three dimensional co-culture model, simulating a tumor microenvironment to support tumor development. We have successfully expanded CTCs isolated from 14 of 19 early stage lung cancer patients. Expanded lung CTCs carried mutations of the TP53 gene identical to those observed in the matched primary tumors. Next-generation sequencing further revealed additional matched mutations between primary tumor and CTCs of cancer-related genes. This strategy sets the stage to further characterize the biology of CTCs derived from patients with early lung cancers, thereby leading to a better understanding of these putative drivers of metastasis.
authors.affiliation: Department of Chemical Engineering, Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Hiroe
authors.initials: H
authors.lastname: Shiratsuchi
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Jules
authors.initials: J
authors.lastname: Lin
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Guoan
authors.initials: G
authors.lastname: Chen
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Rishindra M
authors.initials: RM
authors.lastname: Reddy
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Ebrahim
authors.initials: E
authors.lastname: Azizi
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Shamileh
authors.initials: S
authors.lastname: Fouladdel
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Chang
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Biostatistics, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109. Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Meghna
authors.initials: M
authors.lastname: Waghray
authors.affiliation: Department of Radiology, Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Gary
authors.initials: G
authors.lastname: Luker
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109. Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Diane M
authors.initials: DM
authors.lastname: Simeone
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Max S
authors.initials: MS
authors.lastname: Wicha
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, MI 48109.
authors.forename: David G
authors.initials: DG
authors.lastname: Beer
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109. Veterans Administration Ann Arbor Healthcare System, Ann Arbor, MI 48105.
authors.forename: Nithya
authors.initials: N
authors.lastname: Ramnath
authors.affiliation: Department of Chemical Engineering, Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109. Translational Oncology Program, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Sunitha
authors.initials: S
authors.lastname: Nagrath
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018920:Coculture Techniques / Q000379:methods*; D005260:Female; D005455:Fluorescent Antibody Technique; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood* / Q000473:pathology; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome