Single-cell mutational profiling and clonal phylogeny in cancer

Genome Res. 2013 Dec;23(12):2115-25. doi: 10.1101/gr.159913.113. Epub 2013 Sep 20.

Abstract

The development of cancer is a dynamic evolutionary process in which intraclonal, genetic diversity provides a substrate for clonal selection and a source of therapeutic escape. The complexity and topography of intraclonal genetic architectures have major implications for biopsy-based prognosis and for targeted therapy. High-depth, next-generation sequencing (NGS) efficiently captures the mutational load of individual tumors or biopsies. But, being a snapshot portrait of total DNA, it disguises the fundamental features of subclonal variegation of genetic lesions and of clonal phylogeny. Single-cell genetic profiling provides a potential resolution to this problem, but methods developed to date all have limitations. We present a novel solution to this challenge using leukemic cells with known mutational spectra as a tractable model. DNA from flow-sorted single cells is screened using multiplex targeted Q-PCR within a microfluidic platform allowing unbiased single-cell selection, high-throughput, and comprehensive analysis for all main varieties of genetic abnormalities: chimeric gene fusions, copy number alterations, and single-nucleotide variants. We show, in this proof-of-principle study, that the method has a low error rate and can provide detailed subclonal genetic architectures and phylogenies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Clonal Evolution*
  • DNA Copy Number Variations
  • Genetic Variation
  • Genomics / methods*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Multiplex Polymerase Chain Reaction
  • Mutation*
  • Neoplasms / genetics*
  • Phylogeny*
  • Polymorphism, Single Nucleotide
  • Single-Cell Analysis*

Associated data

  • GEO/GSE49215