Elucidating Anthracycline-Induced Cell Type-Specific Cardiovascular Toxicity with CRISPRi/a Screens

Abstract public domain (NIH RePORTER)

PROJECT SUMMARY As anti-cancer treatments have become more effective, treatment-related cardiovascular toxicity has emerged as a significant clinical challenge. Doxorubicin, a widely used and cost-efficient anthracycline in first-line chemotherapy, is particularly limited by its substantial cardiovascular toxicity through poorly understood mechanisms. This proposal aims to elucidate these mechanisms and identify potential protective compounds using an integrated approach. In Aims 1 and 2, we will employ CRISPR interference and activation (CRISPRi/a) screens of druggable genes in human induced pluripotent stem cell (iPSC)-derived 2D cardiomyocytes (CMs) and 3D vascularized cardioids (vCOs) to identify cell type-specific causative genes in doxorubicin-induced cytotoxicity. We will validate these findings across multiple iPSC-derived cardiovascular cell types (i.e., CMs, endothelial cells, cardiac smooth muscle cells, cardiac pericytes, and cardiac fibroblasts) and in 3D engineered heart tissues, assessing various cell type-dependent functional parameters. In Aim 3, we will leverage gene-drug interaction databases and molecular docking to identify promising small molecules, particularly from the Drug Repurposing Hub, that can mitigate doxorubicin-induced cardiovascular toxicity. Next, we will evaluate candidate compounds in iPSC-derived cardiovascular cells and in a PANC1 pancreatic cancer xenograft mouse model, assessing their ability to rescue doxorubicin-induced cardiovascular dysfunction. Finally, by integrating gene perturbation programs, cell type-specific phenotypes, and altered pathological signaling pathways, we will elucidate the mechanisms underlying doxorubicin-induced cardiovascular toxicity. The successful completion of this study will not only facilitate the development of novel cardioprotective therapies for cancer patients receiving anthracycline-based treatments but also establish a powerful platform for evaluating drug-induced cardiovascular toxicity beyond cardio-oncology.

Public health relevance public domain (NIH RePORTER)

PROJECT NARRATIVE Doxorubicin, a highly effective anthracycline used in first-line chemotherapy for various malignant cancers like breast cancer, is hampered by serious cardiovascular side effects, including hypertension, arrhythmia, thrombosis, and heart failure. This proposal will utilize high-throughput CRISPR interference and activation (CRISPRi/a) screens of druggable genes in human induced pluripotent stem cell-derived cardiomyocytes (iPSC- CMs) and vascularized cardioids (iPSC-vCOs) to identify cell type-specific causative genes and candidate compounds that mitigate doxorubicin-induced cardiotoxicity. Successful completion will provide effective countermeasures against anthracycline-induced cardiovascular toxicity and establish a reliable drug toxicity assessment platform for broader research applications.