Functional Validation of Doxorubicin-Induced Cardiotoxicity-Related Genes

Area of Science:

• Cardiovascular Research
• Genetics and Genomics
• Stem Cell Biology
• Pharmacology

Background:

• Over 180 genetic variants are statistically linked to anthracycline-induced cardiotoxicity (AIC).
• Lack of functional validation impedes clinical translation of these genetic findings.

Purpose of the Study:

• To functionally validate genes associated with AIC using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).
• To identify novel genetic targets for mitigating doxorubicin-induced cardiotoxicity (DIC).

Main Methods:

• Systematic literature search identified 80 AIC-associated genes, plus 3 additional genes of interest.
• 38 genes with expression in cardiac tissues and hiPSC-CMs were selected for functional studies.
• CRISPR/Cas9 gene editing was used to knock out each of the 38 genes in hiPSC-CMs to assess DIC phenotype.

Main Results:

• Knockout of 26 genes, including efflux transporters (e.g., ABCC10, ABCC2) and known DIC genes (e.g., CBR1, RAC2), increased hiPSC-CM susceptibility to DIC.
• Knockout of uptake transporters (e.g., SLC28A3, SLC22A17) conferred a protective effect against DIC.
• Knockout of specific genes (e.g., ATP2B1, HNMT) showed no significant effect on the in vitro DIC phenotype.

Conclusions:

• Established a platform for functional validation of DIC-associated genes.
• Findings provide insights into DIC variant associations and potential targets for cardioprotective drug development.