Single-cell Analyses of Transcriptional Heterogeneity During Drug Tolerance Transition in Cancer Cells by RNA Sequencing

Proc Natl Acad Sci U S A. 2014 Nov 4;111(44):E4726-35. doi: 10.1073/pnas.1404656111. Epub 2014 Oct 22.

Abstract

The acute cellular response to stress generates a subpopulation of reversibly stress-tolerant cells under conditions that are lethal to the majority of the population. Stress tolerance is attributed to heterogeneity of gene expression within the population to ensure survival of a minority. We performed whole transcriptome sequencing analyses of metastatic human breast cancer cells subjected to the chemotherapeutic agent paclitaxel at the single-cell and population levels. Here we show that specific transcriptional programs are enacted within untreated, stressed, and drug-tolerant cell groups while generating high heterogeneity between single cells within and between groups. We further demonstrate that drug-tolerant cells contain specific RNA variants residing in genes involved in microtubule organization and stabilization, as well as cell adhesion and cell surface signaling. In addition, the gene expression profile of drug-tolerant cells is similar to that of untreated cells within a few doublings. Thus, single-cell analyses reveal the dynamics of the stress response in terms of cell-specific RNA variants driving heterogeneity, the survival of a minority population through generation of specific RNA variants, and the efficient reconversion of stress-tolerant cells back to normalcy.

Keywords: RNA-Seq; drug resistance; paclitaxel; single cell; tumor heterogeniety.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Breast Neoplasms* / drug therapy
  • Breast Neoplasms* / genetics
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Cell Line, Tumor
  • Drug Resistance, Neoplasm* / drug effects
  • Drug Resistance, Neoplasm* / genetics
  • Female
  • Humans
  • Paclitaxel / pharmacology*
  • RNA, Neoplasm* / biosynthesis
  • RNA, Neoplasm* / genetics
  • Sequence Analysis, RNA*
  • Transcription, Genetic* / drug effects
  • Transcription, Genetic* / genetics

Substances

  • Antineoplastic Agents, Phytogenic
  • RNA, Neoplasm
  • Paclitaxel

Associated data

  • SRA/SRP040309