Long-Term Efficacy and Safety of Cardiac Genome Editing for Catecholaminergic Polymorphic Ventricular Tachycardia

Authors

Oliver M Moore
Yuriana Aguilar-Sanchez
Satadru K Lahiri
Mohit M Hulsurkar
J Alberto Navarro-Garcia
Tarah A Word
Joshua A Keefe
Dean Barazi
Elda M Munivez
Charles T Moore
Vaidya Parthasarathy
Jaysón Davidson
William R Lagor
So Hyun Park
Gang Bao
Christina Y Miyake
Xander H T Wehrens

Publication Date

1-1-2024

Journal

The Journal of Cardiovascular Aging

DOI

10.20517/jca.2023.42

PMID

38464671

PMCID

PMC10919902

PubMedCentral® Posted Date

1-4-2025

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

yes

Keywords

Catecholaminergic polymorphic ventricular tachycardia, ryanodine receptor, CRISPR/Cas9, RyR2, genome editing

Abstract

INTRODUCTION: Heterozygous autosomal-dominant single nucleotide variants in RYR2 account for 60% of cases of catecholaminergic polymorphic ventricular tachycardia (CPVT), an inherited arrhythmia disorder associated with high mortality rates. CRISPR/Cas9-mediated genome editing is a promising therapeutic approach that can permanently cure the disease by removing the mutant RYR2 allele. However, the safety and long-term efficacy of this strategy have not been established in a relevant disease model.

AIM: The purpose of this study was to assess whether adeno-associated virus type-9 (AAV9)-mediated somatic genome editing could prevent ventricular arrhythmias by removal of the mutant allele in mice that are heterozygous for

METHODS AND RESULTS: Guide RNA and SaCas9 were delivered using AAV9 vectors injected subcutaneously in 10-day-old mice. At 6 weeks after injection, R176Q/+ mice had a 100% reduction in ventricular arrhythmias compared to controls. When aged to 12 months, injected R176Q/+ mice maintained a 100% reduction in arrhythmia induction. Deep RNA sequencing revealed the formation of insertions/deletions at the target site with minimal off-target editing on the wild-type allele. Consequently, CRISPR/SaCas9 editing resulted in a 45% reduction of total

CONCLUSION: Taken together, AAV9-mediated CRISPR/Cas9 genome editing could efficiently disrupt the mutant Ryr2 allele, preventing lethal arrhythmias while preserving normal cardiac function in the R176Q/+ mouse model of CPVT.