Functional Validation of Doxorubicin-Induced Cardiotoxicity-Related Genes

Hananeh Fonoudi, Mariam Jouni, Romina B. Cejas, Tarek Magdy, Malorie Blancard, Ning Ge, Disheet A. Shah, Davi M. Lyra-Leite, Achal Neupane, Mennat Gharib, Zhengxin Jiang, Yadav Sapkota, Paul W. Burridge*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Background: Genome-wide association studies and candidate gene association studies have identified more than 180 genetic variants statistically associated with anthracycline-induced cardiotoxicity (AIC). However, the lack of functional validation has hindered the clinical translation of these findings. Objectives: The aim of this study was to functionally validate all genes associated with AIC using human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs). Methods: Through a systemic literature search, 80 genes containing variants significantly associated with AIC were identified. Additionally, 3 more genes with potential roles in AIC (GSTM1, CBR1, and ERBB2) were included. Of these, 38 genes exhibited expression in human fetal heart, adult heart, and hiPSC-CMs. Using clustered regularly interspaced short palindromic repeats/Cas9–based genome editing, each of these 38 genes was systematically knocked out in control hiPSC-CMs, and the resulting doxorubicin-induced cardiotoxicity (DIC) phenotype was assessed using hiPSC-CMs. Subsequently, functional assays were conducted for each gene knockout on the basis of hypothesized mechanistic implications in DIC. Results: Knockout of 26 genes increased the susceptibility of hiPSC-CMs to DIC. Notable genes included efflux transporters (ABCC10, ABCC2, ABCB4, ABCC5, and ABCC9), well-established DIC-associated genes (CBR1, CBR3, and RAC2), and genome-wide association study–discovered genes (RARG and CELF4). Conversely, knockout of ATP2B1, HNMT, POR, CYBA, WDR4, and COL1A2 had no significant effect on the in vitro DIC phenotype of hiPSC-CMs. Furthermore, knockout of the uptake transporters (SLC28A3, SLC22A17, and SLC28A1) demonstrated a protective effect against DIC. Conclusions: The present findings establish a comprehensive platform for the functional validation of DIC-associated genes, providing insights for future studies in DIC variant associations and potential mechanistic targets for the development of cardioprotective drugs.

Original languageEnglish (US)
Pages (from-to)38-50
Number of pages13
JournalJACC: CardioOncology
Volume6
Issue number1
DOIs
StatePublished - Feb 2024

Keywords

  • GWAS
  • cardiomyocytes
  • doxorubicin
  • genomics
  • human induced pluripotent stem cells

ASJC Scopus subject areas

  • Oncology
  • Cardiology and Cardiovascular Medicine

Fingerprint

Dive into the research topics of 'Functional Validation of Doxorubicin-Induced Cardiotoxicity-Related Genes'. Together they form a unique fingerprint.

Cite this