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Single-Cell Analysis of Circulating Tumor Cells

  • Jana- A. Thiele
  • Pavel Pitule
  • James Hicks
  • Peter KuhnEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1908)

Abstract

Circulating tumor cells (CTCs) are rare cells that can be found in the peripheral blood of cancer patients. They have been demonstrated to be useful prognostic markers in many cancer types. Within the last decade various methods have been developed to detect rare cells within a liquid biopsy from a cancer patient. These methods have revealed the phenotypic diversity of CTCs and how they can represent the complement of cells that are found in a tumor. Single-cell proteogenomics has emerged as an all-encompassing next-generation technological approach for CTC research. This allows for the deconstruction of cellular heterogeneity, dynamics of metastatic initiation and progression, and response or resistance to therapeutics in the clinical settings. We take advantage of this opportunity to investigate CTC heterogeneity and understand their full potential in precision medicine.The high-definition single-cell analysis (HD-SCA) workflow combines detection of the entire population of CTCs and rare cancer related cells with single-cell genomic analysis and may therefore provide insight into their subpopulations based on molecular as well as morphological data. In this chapter we describe in detail the protocols from isolation of a candidate cell from a microscopy slide, through whole-genome amplification and library preparation, to CNV analysis of identified cells from the HD-SCA workflow. This process may also be applicable to any platform starting with a standard microscopy slide or isolated cell of interest.

Key words

Circulating tumor cells CTC Copy number variation CNV Liquid biopsy Precision medicine Single-cell analysis 

Notes

Acknowledgment

Disclosure

James Hicks: Epic Sciences, Inc.—Clinical Advisory Board

Peter Kuhn: Epic Sciences, Inc.—Stock and Ownership Interests, Advisory Role, Patents, and Royalties

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jana- A. Thiele
    • 1
  • Pavel Pitule
    • 1
  • James Hicks
    • 2
  • Peter Kuhn
    • 2
    • 3
    • 4
    • 5
    Email author
  1. 1.Biomedical Center, Faculty of Medicine in PilsenCharles UniversityPilsenCzech Republic
  2. 2.USC Michelson Center for Convergent Biosciences, CSI-Cancer, Department of Biological Sciences, Dornsife College of Letters, Arts and SciencesUniversity of Southern CaliforniaLos AngelesUSA
  3. 3.Department of Biomedical Engineering and Aerospace and Mechanical Engineering, Viterbi School of EngineeringUniversity of Southern CaliforniaLos AngelesUSA
  4. 4.Department of Medicine, Keck School of MedicineUniversity of Southern CaliforniaLos AngelesUSA
  5. 5.Department of Urology, Keck School of MedicineUniversity of Southern CaliforniaLos AngelesUSA

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