In Vivo Ryr2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia

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dc.citation.firstpage953
dc.citation.issueNumber8
dc.citation.journalTitleCirculation Research
dc.citation.lastpage963
dc.citation.volumeNumber123
dc.contributor.authorPan, Xiaolu
dc.contributor.authorPhilippen, Leonne
dc.contributor.authorLahiri, Satadru K.
dc.contributor.authorLee, Ciaran
dc.contributor.authorPark, So Hyun
dc.contributor.authorWord, Tarah A.
dc.contributor.authorLi, Na
dc.contributor.authorJarrett, Kelsey E.
dc.contributor.authorGupta, Rajat
dc.contributor.authorReynolds, Julia O.
dc.contributor.authorLin, Jean
dc.contributor.authorBao, Gang
dc.contributor.authorLagor, William R.
dc.contributor.authorWehrens, Xander H.T.
dc.date.accessioned2019-11-05T17:30:05Z
dc.date.available2019-11-05T17:30:05Z
dc.date.issued2018
dc.description.abstractRationale:Autosomal-dominant mutations in ryanodine receptor type 2 (RYR2) are responsible for ≈60% of all catecholaminergic polymorphic ventricular tachycardia. Dysfunctional RyR2 subunits trigger inappropriate calcium leak from the tetrameric channel resulting in potentially lethal ventricular tachycardia. In vivo CRISPR/Cas9-mediated gene editing is a promising strategy that could be used to eliminate the disease-causing Ryr2 allele and hence rescue catecholaminergic polymorphic ventricular tachycardia.Objective:To determine if somatic in vivo genome editing using the CRISPR/Cas9 system delivered by adeno-associated viral (AAV) vectors could correct catecholaminergic polymorphic ventricular tachycardia arrhythmias in mice heterozygous for RyR2 mutation R176Q (R176Q/+).Methods and Results:Guide RNAs were designed to specifically disrupt the R176Q allele in the R176Q/+ mice using the SaCas9 (Staphylococcus aureus Cas9) genome editing system. AAV serotype 9 was used to deliver Cas9 and guide RNA to neonatal mice by single subcutaneous injection at postnatal day 10. Strikingly, none of the R176Q/+ mice treated with AAV-CRISPR developed arrhythmias, compared with 71% of R176Q/+ mice receiving control AAV serotype 9. Total Ryr2 mRNA and protein levels were significantly reduced in R176Q/+ mice, but not in wild-type littermates. Targeted deep sequencing confirmed successful and highly specific editing of the disease-causing R176Q allele. No detectable off-target mutagenesis was observed in the wild-type Ryr2 allele or the predicted putative off-target site, confirming high specificity for SaCas9 in vivo. In addition, confocal imaging revealed that gene editing normalized the enhanced Ca2+ spark frequency observed in untreated R176Q/+ mice without affecting systolic Ca2+ transients.Conclusions:AAV serotype 9–based delivery of the SaCas9 system can efficiently disrupt a disease-causing allele in cardiomyocytes in vivo. This work highlights the potential of somatic genome editing approaches for the treatment of lethal autosomal-dominant inherited cardiac disorders, such as catecholaminergic polymorphic ventricular tachycardia.
dc.identifier.citationPan, Xiaolu, Philippen, Leonne, Lahiri, Satadru K., et al.. "In Vivo Ryr2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia." <i>Circulation Research,</i> 123, no. 8 (2018) American Heart Association: 953-963. https://doi.org/10.1161/CIRCRESAHA.118.313369.
dc.identifier.digitalnihms-1503253
dc.identifier.doihttps://doi.org/10.1161/CIRCRESAHA.118.313369
dc.identifier.urihttps://hdl.handle.net/1911/107600
dc.language.isoeng
dc.publisherAmerican Heart Association
dc.rightsThis is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by the American Heart Association
dc.titleIn Vivo Ryr2 Editing Corrects Catecholaminergic Polymorphic Ventricular Tachycardia
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpost-print
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