title: Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice.
abstract: Exon skipping mediated by antisense oligoribonucleotides (AON) is a promising therapeutic approach for genetic disorders, but has not yet been evaluated for cardiac diseases. We investigated the feasibility and efficacy of viral-mediated AON transfer in a Mybpc3-targeted knock-in (KI) mouse model of hypertrophic cardiomyopathy (HCM). KI mice carry a homozygous G>A transition in exon 6, which results in three different aberrant mRNAs. We identified an alternative variant (Var-4) deleted of exons 5-6 in wild-type and KI mice. To enhance its expression and suppress aberrant mRNAs we designed AON-5 and AON-6 that mask splicing enhancer motifs in exons 5 and 6. AONs were inserted into modified U7 small nuclear RNA and packaged in adeno-associated virus (AAV-U7-AON-5+6). Transduction of cardiac myocytes or systemic administration of AAV-U7-AON-5+6 increased Var-4 mRNA/protein levels and reduced aberrant mRNAs. Injection of newborn KI mice abolished cardiac dysfunction and prevented left ventricular hypertrophy. Although the therapeutic effect was transient and therefore requires optimization to be maintained over an extended period, this proof-of-concept study paves the way towards a causal therapy of HCM.
authors.affiliation: Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Gedicke-Hornung
authors.forename: Verena
authors.initials: V
authors.lastname: Behrens-Gawlik
authors.forename: Silke
authors.initials: S
authors.lastname: Reischmann
authors.forename: Birgit
authors.initials: B
authors.lastname: Geertz
authors.forename: Doreen
authors.initials: D
authors.lastname: Stimpel
authors.forename: Florian
authors.initials: F
authors.lastname: Weinberger
authors.forename: Saskia
authors.initials: S
authors.lastname: Schlossarek
authors.forename: Guillaume
authors.initials: G
authors.lastname: Précigout
authors.forename: Ingke
authors.initials: I
authors.lastname: Braren
authors.forename: Thomas
authors.initials: T
authors.lastname: Eschenhagen
authors.forename: Giulia
authors.initials: G
authors.lastname: Mearini
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lorain
authors.forename: Thomas
authors.initials: T
authors.lastname: Voit
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Dreyfus
authors.forename: Luis
authors.initials: L
authors.lastname: Garcia
authors.forename: Lucie
authors.initials: L
authors.lastname: Carrier
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017398:Alternative Splicing; D000818:Animals; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D002352:Carrier Proteins / Q000235:genetics*; D005091:Exons; D055879:Gene Knock-In Techniques; D015316:Genetic Therapy; D057809:HEK293 Cells; D006321:Heart / Q000503:physiopathology; D006801:Humans; D017379:Hypertrophy, Left Ventricular / Q000517:prevention & control; D051379:Mice; D009154:Mutation; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D020320:Oligoribonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012342:RNA, Small Nuclear / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D014161:Transduction, Genetic; D000099163:Myosin Binding Protein C
chemical_list: D002352:Carrier Proteins; D020320:Oligoribonucleotides, Antisense; D020033:Protein Isoforms; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D000099163:Myosin Binding Protein C; C405487:U7 small nuclear RNA
keywords: alternative splicing; antisense oligoribonucleotide; cardiac myosin-binding protein-C; exon skipping; hypertrophic cardiomyopathy