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Updated: Jul 12, 2025
Adeno-Associated Virus-Mediated Delivery of CRISPR for Cardiac Gene Editing in Mice
Published on: August 2, 2018
CRISPR-Cas9 base editing of pathogenic CaMKIIδ improves cardiac function in a humanized mouse model
Simon Lebek1,2,3, Xurde M Caravia1,2, Leon G Straub4
1Department of Molecular Biology and.
Insights
Targeting calcium/calmodulin-dependent protein kinase IIδ (CaMKIIδ) oxidation via CRISPR-Cas9 base editing offers a promising cardioprotective strategy. Editing humanized CaMKIIδ in mice improved cardiac function and reduced fibrosis after injury.
Area of Science:
- Cardiovascular biology
- Molecular cardiology
- Gene editing technologies
Background:
- Cardiovascular diseases (CVDs) are a leading global cause of death, necessitating novel therapeutic approaches.
- Oxidation of Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ) contributes to cardiac dysfunction and disease progression.
- Previous studies demonstrated that ablating CaMKIIδ oxidation via CRISPR-Cas9 base editing promotes cardiac recovery from ischemia/reperfusion (IR) injury.
Purpose of the Study:
- To translate the therapeutic potential of CaMKIIδ oxidation ablation for clinical applications.
- To develop and optimize gene editing strategies for the human CAMK2D gene in a mouse model.
- To evaluate the efficacy of in vivo gene editing for cardioprotection against IR injury.
Main Methods:
- Generation of a humanized CAMK2D knockin mouse model with a human regulatory domain sequence.
- Development and packaging of optimized CRISPR-Cas9 base editing components into a myotropic adeno-associated virus (MyoAAV 2A).
- In vivo delivery of MyoAAV 2A to humanized mice at the time of IR injury to edit CAMK2D.
Main Results:
- CAMK2D-edited humanized mice exhibited significant recovery of cardiac function post-IR injury.
- Edited mice demonstrated improved exercise performance compared to control groups.
- Protection against myocardial fibrosis was observed in CAMK2D-edited mice following IR injury.
Conclusions:
- In vivo base editing of humanized CAMK2D provides effective cardioprotection against IR-induced oxidative damage.
- This strategy holds promise for clinical translation in treating cardiovascular diseases.
- Targeting CaMKIIδ oxidation represents a viable therapeutic avenue for heart protection.
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.
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