Exploring Gene Editing Strategies for Heart Failure Protection

Samuel Y. Shin, Michael A. Fallon, Gabriel M. C. Longo, Benjamin P. Kleinstiver

Dilated cardiomyopathy (DCM) is a heart disease that is characterized by enlargement of one or both of the ventricles, which then leads to heart failure. DCM causes around 10,000 deaths and 46,000 hospitalizations annually in the U.S alone, and it is a progressive disease with no current cure.1 Around 30-50% DCM cases have an established genetic cause, however, alleles with increased risk for DCM have been mapped to over 20 loci within the genome.2 The genetic heterogeneity of DCM-promoting alleles makes it extremely difficult and inefficient to design patient-specific genetic therapies for each individual case of DCM. To overcome this allelic diversity, we sought to optimize genomic-editing methods to install a mutation in the BAG3 gene, which has been reported to have cardioprotective effects against heart failure. Inserting this protective allele could prevent DCM without having to correct each pathogenic mutation. In this experiment, we explored methods of installing the protective variant BAG3-C151R using CRISPR-Cas9-derived base editors (BEs).3 Base editors are composed of a Cas9 nickase fused to a deaminase domain, along with a guide RNA,4 allowing for precise nucleotide conversion either from C-to-T or A-to-G while also minimizing unwanted double-stranded DNA breaks that impact cell health or cause unwanted outcomes. By screening different combinations of BEs and guide RNAs, we identified editors that are most effective at installing the BAG3-C151R allele. These results suggest that precise genome editing may be used in future treatments for DCM and heart failure.

References

1. Mahmaljy H, Yelamanchili VS, Singhal M. Dilated Cardiomyopathy. [Updated 2023 Apr 7]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-.

2. Kärkkäinen, S., & Peuhkurinen, K. (2007). Genetics of dilated cardiomyopathy. Annals of Medicine, 39(2), 91–107. DOI: https://doi.org/10.1080/07853890601145821.

3. Perez-Bermejo, J.A., Judge, L.M., Jensen, C.L. et al. Functional analysis of a common BAG3 allele associated with protection from heart failure. Nat Cardiovasc Res 2, 615–628 (2023). https://doi.org/10.1038/s44161-023-00288-w.

4. Gaudelli, N., Komor, A., Rees, H. et al. Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551, 464–471 (2017). DOI: https://doi.org/10.1038/nature24644.

For more information: syshin@eckerd.edu

LinkedIn: www.linkedin.com/in/samuel-shin-204363298