title: Reengineering a transmembrane protein to treat muscular dystrophy using exon skipping.
abstract: Exon skipping uses antisense oligonucleotides as a treatment for genetic diseases. The antisense oligonucleotides used for exon skipping are designed to bypass premature stop codons in the target RNA and restore reading frame disruption. Exon skipping is currently being tested in humans with dystrophin gene mutations who have Duchenne muscular dystrophy. For Duchenne muscular dystrophy, the rationale for exon skipping derived from observations in patients with naturally occurring dystrophin gene mutations that generated internally deleted but partially functional dystrophin proteins. We have now expanded the potential for exon skipping by testing whether an internal, in-frame truncation of a transmembrane protein γ-sarcoglycan is functional. We generated an internally truncated γ-sarcoglycan protein that we have termed Mini-Gamma by deleting a large portion of the extracellular domain. Mini-Gamma provided functional and pathological benefits to correct the loss of γ-sarcoglycan in a Drosophila model, in heterologous cell expression studies, and in transgenic mice lacking γ-sarcoglycan. We generated a cellular model of human muscle disease and showed that multiple exon skipping could be induced in RNA that encodes a mutant human γ-sarcoglycan. Since Mini-Gamma represents removal of 4 of the 7 coding exons in γ-sarcoglycan, this approach provides a viable strategy to treat the majority of patients with γ-sarcoglycan gene mutations.
authors.forename: Quan Q
authors.initials: QQ
authors.lastname: Gao
authors.forename: Eugene
authors.initials: E
authors.lastname: Wyatt
authors.forename: Jeff A
authors.initials: JA
authors.lastname: Goldstein
authors.forename: Peter
authors.initials: P
authors.lastname: LoPresti
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Castillo
authors.forename: Alec
authors.initials: A
authors.lastname: Gazda
authors.forename: Natalie
authors.initials: N
authors.lastname: Petrossian
authors.forename: Judy U
authors.initials: JU
authors.lastname: Earley
authors.forename: Michele
authors.initials: M
authors.lastname: Hadhazy
authors.forename: David Y
authors.initials: DY
authors.lastname: Barefield
authors.forename: Alexis R
authors.initials: AR
authors.lastname: Demonbreun
authors.forename: Carsten
authors.initials: C
authors.lastname: Bönnemann
authors.forename: Matthew
authors.initials: M
authors.lastname: Wolf
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: McNally
mesh_terms: D000818:Animals; D018389:Codon, Nonsense / Q000235:genetics; D003964:Diaphragm / Q000378:metabolism / Q000473:pathology; D029721:Drosophila Proteins / Q000172:deficiency / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D049028:Dystrophin-Associated Protein Complex / Q000737:chemistry*; D005091:Exons; D005355:Fibrosis; D015316:Genetic Therapy; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics / Q000628:therapy*; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000473:pathology / Q000628:therapy; D009154:Mutation; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology / Q000627:therapeutic use*; D015202:Protein Engineering; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D049031:Sarcoglycans / Q000096:biosynthesis / Q000737:chemistry / Q000172:deficiency / Q000235:genetics*; D012508:Sarcolemma / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D018389:Codon, Nonsense; D029721:Drosophila Proteins; D049028:Dystrophin-Associated Protein Complex; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049031:Sarcoglycans; C000605049:Scgdelta protein, Drosophila