Variation in RARG increases susceptibility to doxorubicin-induced cardiotoxicity in patient specific induced pluripotent stem cell-derived cardiomyocytes

Sci Rep. 2020 Jun 25;10(1):10363. doi: 10.1038/s41598-020-65979-x.

Abstract

Doxorubicin is a potent anticancer drug used to treat a variety of cancer types. However, its use is limited by doxorubicin-induced cardiotoxicity (DIC). A missense variant in the RARG gene (S427L; rs2229774) has been implicated in susceptibility to DIC in a genome wide association study. The goal of this study was to investigate the functional role of this RARG variant in DIC. We used induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) from patients treated with doxorubicin. iPSC-CMs from individuals who experienced DIC (cases) showed significantly greater sensitivity to doxorubicin compared to iPSC-CMs from doxorubicin-treated individuals who did not develop DIC (controls) in cell viability and optical mapping experiments. Using CRISPR/Cas9, we generated isogenic cell lines that differed only at the RARG locus. Genetic correction of RARG-S427L to wild type resulted in reduced doxorubicin-induced double stranded DNA breaks, reactive oxygen species production, and cell death. Conversely, introduction of RARG-S427L increased susceptibility to doxorubicin. Finally, genetic disruption of the RARG gene resulted in protection from cell death due to doxorubicin treatment. Our findings suggest that the presence of RARG-S427L increases sensitivity to DIC, establishing a direct, causal role for this variant in DIC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antibiotics, Antineoplastic / adverse effects
  • CRISPR-Cas Systems
  • Cardiotoxicity / etiology
  • Cardiotoxicity / metabolism
  • Cardiotoxicity / pathology*
  • Case-Control Studies
  • Doxorubicin / adverse effects*
  • Female
  • Follow-Up Studies
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / pathology*
  • Male
  • Middle Aged
  • Mutation*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / pathology*
  • Neoplasms / drug therapy*
  • Neoplasms / pathology
  • Receptors, Retinoic Acid / antagonists & inhibitors
  • Receptors, Retinoic Acid / genetics*
  • Receptors, Retinoic Acid / metabolism
  • Retinoic Acid Receptor gamma
  • Tumor Cells, Cultured

Substances

  • Antibiotics, Antineoplastic
  • Receptors, Retinoic Acid
  • Doxorubicin