Faculty, Staff and Student Publications
Language
English
Publication Date
1-2-2024
Journal
Journal of Clinical Investigation
DOI
10.1172/JCI175164
PMID
37856214
PMCID
PMC10760954
PubMedCentral® Posted Date
1-2-2024
PubMedCentral® Full Text Version
Post-print
Abstract
Cardiovascular diseases are the most common cause of worldwide morbidity and mortality, highlighting the necessity for advanced therapeutic strategies. Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ) is a prominent inducer of various cardiac disorders, which is mediated by 2 oxidation-sensitive methionine residues within the regulatory domain. We have previously shown that ablation of CaMKIIδ oxidation by CRISPR-Cas9 base editing enables the heart to recover function from otherwise severe damage following ischemia/reperfusion (IR) injury. Here, we extended this therapeutic concept toward potential clinical translation. We generated a humanized CAMK2D knockin mouse model in which the genomic sequence encoding the entire regulatory domain was replaced with the human sequence. This enabled comparison and optimization of two different editing strategies for the human genome in mice. To edit CAMK2D in vivo, we packaged the optimized editing components into an engineered myotropic adeno-associated virus (MyoAAV 2A), which enabled efficient delivery at a very low AAV dose into the humanized mice at the time of IR injury. CAMK2D-edited mice recovered cardiac function, showed improved exercise performance, and were protected from myocardial fibrosis, which was otherwise observed in injured control mice after IR. Our findings identify a potentially effective strategy for cardioprotection in response to oxidative damage.
Keywords
Mice, Animals, Humans, CRISPR-Cas Systems, Gene Editing, Heart, Cardiomyopathies, Cardiovascular Diseases, Cardiovascular disease, Gene therapy, Mouse models, Cardiology
Published Open-Access
yes
Recommended Citation
Lebek, Simon; Caravia, Xurde M; Straub, Leon G; et al., "CRISPR-Cas9 Base Editing of Pathogenic CaMKIIδ Improves Cardiac Function in a Humanized Mouse Model" (2024). Faculty, Staff and Student Publications. 764.
https://digitalcommons.library.tmc.edu/uthshis_docs/764
Graphical Abstract