biostudies-literatureMoore OMNIDDK NIH HHSNHLBI NIH HHSNHGRI NIH HHSNIH HHS8https://www.ebi.ac.uk/biostudies/studies/S-EPMC10919902biostudies-literatureUnknown4(1)<h4>Introduction</h4>Heterozygous autosomal-dominant single nucleotide variants in <i>RYR2</i> 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 <i>RYR2</i> allele. However, the safety and long-term efficacy of this strategy have not been established in a relevant disease model.<h4>Aim</h4>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 <i>Ryr2</i> variant p.Arg176Gln (R176Q/+).<h4>Methods and results</h4>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 <i>Ryr2</i> mRNA and a 38% reduction in RyR2 protein. Genome editing was well tolerated based on serial echocardiography, revealing unaltered cardiac function and structure up to 12 months after AAV9 injection.<h4>Conclusion</h4>Taken together, AAV9-mediated CRISPR/Cas9 genome editing could efficiently disrupt the mutant <i>Ryr2</i> allele, preventing lethal arrhythmias while preserving normal cardiac function in the R176Q/+ mouse model of CPVT.The journal of cardiovascular agingLong-term efficacy and safety of cardiac genome editing for catecholaminergic polymorphic ventricular tachycardia.PMC10919902R01 HL132840R01 HL153350R01 DK114356K23 HL136932R01 HL152314R01 DK124477R01 HL169761F30 HL156669UM1 HG006348R01 HL147108U42 OD026645R01 HL160992S10 OD032380T32 HL139430R01 HL089598Wehrens XHTBarazi DMiyake CYLahiri SKAlberto Navarro-Garcia JParthasarathy VLagor WRMunivez EMHulsurkar MMMoore OMBao GAguilar-Sanchez YPark SHMoore CTDavidson JWord TAKeefe JAfalseLong-term efficacy and safety of cardiac genome editing for catecholaminergic polymorphic ventricular tachycardia.<h4>Introduction</h4>Heterozygous autosomal-dominant single nucleotide variants in <i>RYR2</i> 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 <i>RYR2</i> allele. However, the safety and long-term efficacy of this strategy have not been established in a relevant disease model.<h4>Aim</h4>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 <i>Ryr2</i> variant p.Arg176Gln (R176Q/+).<h4>Methods and results</h4>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 <i>Ryr2</i> mRNA and a 38% reduction in RyR2 protein. Genome editing was well tolerated based on serial echocardiography, revealing unaltered cardiac function and structure up to 12 months after AAV9 injection.<h4>Conclusion</h4>Taken together, AAV9-mediated CRISPR/Cas9 genome editing could efficiently disrupt the mutant <i>Ryr2</i> allele, preventing lethal arrhythmias while preserving normal cardiac function in the R176Q/+ mouse model of CPVT.2024-01-01T00:00:00Z2024 Jan2026-06-01T11:07:34.408Z2025-04-04T12:02:46.443ZS-EPMC109199023846467110.20517/jca.2023.42