CRISPRi/a screens in human iPSC-cardiomyocytes identify glycolytic activation as a druggable target for
Chun Liu1, Mengcheng Shen2, Yanxia Liu3
1Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Department of Physiology and Cancer Center, Milwaukee, WI, USA; Department of Medicine and Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA.
Abstract:
Doxorubicin is limited in its therapeutic utility due to its life-threatening cardiovascular side effects. Here, we present an integrated drug discovery pipeline combining human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iCMs), CRISPR interference and activation (CRISPRi/a) bidirectional pooled screens, and a small-molecule screening to identify therapeutic targets mitigating doxorubicin-induced cardiotoxicity (DIC) without compromising its oncological effects. The screens revealed several previously unreported candidate genes contributing to DIC, including carbonic anhydrase 12 (CA12). Genetic inhibition of CA12 protected iCMs against DIC by improving cell survival, sarcomere structural integrity, contractile function, and calcium handling. Indisulam, a CA12 antagonist, can effectively attenuate DIC in iCMs, engineered heart tissue, and animal models. Mechanistically, doxorubicin-induced CA12 potentiated a glycolytic activation in cardiomyocytes, contributing to DIC by interfering with cellular metabolism and functions. Collectively, our study provides a roadmap for future drug discovery efforts, potentially leading to more targeted therapies with minimal off-target toxicity.
Insights
Researchers identified carbonic anhydrase 12 (CA12) as a key factor in doxorubicin-induced cardiotoxicity. Inhibiting CA12 with indisulam protects heart cells from damage, offering a potential strategy to reduce chemotherapy side effects.
Area of Science:
- Cardiovascular Research
- Drug Discovery
- Stem Cell Biology
Background:
- Doxorubicin chemotherapy causes severe cardiotoxicity, limiting its clinical use.
- Identifying mechanisms of doxorubicin-induced cardiotoxicity (DIC) is crucial for developing safer cancer treatments.
Purpose of the Study:
- To discover novel therapeutic targets mitigating doxorubicin-induced cardiotoxicity (DIC).
- To develop strategies that reduce cardiotoxicity without affecting doxorubicin's anti-cancer efficacy.
Main Methods:
- Integrated drug discovery pipeline using human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iCMs).
- CRISPR interference and activation (CRISPRi/a) pooled screens and small-molecule screening.
- In vitro and in vivo validation using iCMs, engineered heart tissue, and animal models.
Main Results:
- Identified carbonic anhydrase 12 (CA12) as a novel contributor to DIC.
- Genetic inhibition of CA12 improved cardiomyocyte survival, structure, function, and calcium handling.
- CA12 antagonist indisulam attenuated DIC in multiple models, revealing a mechanism involving glycolytic activation.
Conclusions:
- Carbonic anhydrase 12 (CA12) plays a significant role in doxorubicin-induced cardiotoxicity.
- Targeting CA12 with indisulam presents a promising therapeutic approach to mitigate DIC.
- This study provides a framework for discovering targeted therapies with reduced cardiotoxicity.
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