Detection of Gene Rearrangements in Circulating Tumor Cells: Examples of ALK-, ROS1-, RET-Rearrangements in Non-Small-Cell Lung Cancer and ERG-Rearrangements in Prostate Cancer

Adv Exp Med Biol. 2017:994:169-179. doi: 10.1007/978-3-319-55947-6_9.

Abstract

Circulating tumor cells (CTCs) hold promise as biomarkers to aid in patient treatment stratification and disease monitoring. Because the number of cells is a critical parameter for exploiting CTCs for predictive biomarker's detection, we developed a FISH (fluorescent in situ hybridization) method for CTCs enriched on filters (filter-adapted FISH [FA-FISH]) that was optimized for high cell recovery. To increase the feasibility and reliability of the analyses, we combined fluorescent staining and FA-FISH and developed a semi-automated microscopy method for optimal FISH signal identification in filtration-enriched CTCs . Here we present these methods and their use for the detection and characterization of ALK-, ROS1-, RET-rearrangement in CTCs from non-small-cell lung cancer and ERG-rearrangements in CTCs from prostate cancer patients.

Keywords: Circulating tumor cells; FA-FISH; Filtration-enrichment; Fluorescent staining; Predictive biomarkers.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Anaplastic Lymphoma Kinase
  • Carcinoma, Non-Small-Cell Lung* / blood
  • Carcinoma, Non-Small-Cell Lung* / genetics
  • Female
  • Gene Rearrangement*
  • Humans
  • In Situ Hybridization, Fluorescence / instrumentation
  • In Situ Hybridization, Fluorescence / methods
  • Lung Neoplasms* / blood
  • Lung Neoplasms* / genetics
  • Male
  • Neoplastic Cells, Circulating / metabolism*
  • Prostatic Neoplasms* / blood
  • Prostatic Neoplasms* / genetics
  • Protein-Tyrosine Kinases* / genetics
  • Protein-Tyrosine Kinases* / metabolism
  • Proto-Oncogene Proteins c-ret* / genetics
  • Proto-Oncogene Proteins c-ret* / metabolism
  • Proto-Oncogene Proteins* / genetics
  • Proto-Oncogene Proteins* / metabolism
  • Receptor Protein-Tyrosine Kinases* / genetics
  • Receptor Protein-Tyrosine Kinases* / metabolism
  • Transcriptional Regulator ERG / genetics
  • Transcriptional Regulator ERG / metabolism

Substances

  • ERG protein, human
  • Proto-Oncogene Proteins
  • Transcriptional Regulator ERG
  • ALK protein, human
  • Anaplastic Lymphoma Kinase
  • Protein-Tyrosine Kinases
  • Proto-Oncogene Proteins c-ret
  • RET protein, human
  • ROS1 protein, human
  • Receptor Protein-Tyrosine Kinases