OBJECTIVE: Duchenne muscular dystrophy (DMD) is caused by the inability to produce dystrophin protein at the myofiber membrane. A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for the mdx mouse and in four DMD patients by local intramuscular injection. We sought to test efficacy and toxicity of intravenous oligonucleotide (morpholino)-induced exon skipping in the DMD dog model. METHODS: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery methods (intramuscular injection and systemic intravenous injection). The efficiency and efficacy of multiexon skipping (exons 6-9) were tested at the messenger RNA, protein, histological, and clinical levels. RESULTS: Weekly or biweekly systemic intravenous injections with a three-morpholino cocktail over the course of 5 to 22 weeks induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels. This was accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized timed running tests, and clinical symptoms. Blood tests indicated no evidence of toxicity. INTERPRETATION: This is the first report of widespread rescue of dystrophin expression to therapeutic levels in the dog model of DMD. This study also provides a proof of concept for systemic multiexon-skipping therapy. Use of cocktails of morpholino, as shown here, allows broader application of this approach to a greater proportion of DMD patients (90%) and also offers the prospect of selecting deletions that optimize the functionality of the dystrophin protein.
OBJECTIVE:Duchenne muscular dystrophy (DMD) is caused by the inability to produce dystrophin protein at the myofiber membrane. A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for the mdx mouse and in four DMDpatients by local intramuscular injection. We sought to test efficacy and toxicity of intravenous oligonucleotide (morpholino)-induced exon skipping in the DMDdog model. METHODS: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery methods (intramuscular injection and systemic intravenous injection). The efficiency and efficacy of multiexon skipping (exons 6-9) were tested at the messenger RNA, protein, histological, and clinical levels. RESULTS: Weekly or biweekly systemic intravenous injections with a three-morpholino cocktail over the course of 5 to 22 weeks induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels. This was accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized timed running tests, and clinical symptoms. Blood tests indicated no evidence of toxicity. INTERPRETATION: This is the first report of widespread rescue of dystrophin expression to therapeutic levels in the dog model of DMD. This study also provides a proof of concept for systemic multiexon-skipping therapy. Use of cocktails of morpholino, as shown here, allows broader application of this approach to a greater proportion of DMDpatients (90%) and also offers the prospect of selecting deletions that optimize the functionality of the dystrophin protein.
Authors: Annemieke Aartsma-Rus; Wendy E Kaman; Rudie Weij; Johan T den Dunnen; Gert-Jan B van Ommen; Judith C T van Deutekom Journal: Mol Ther Date: 2006-06-06 Impact factor: 11.454
Authors: K Araishi; T Sasaoka; M Imamura; S Noguchi; H Hama; E Wakabayashi; M Yoshida; T Hori; E Ozawa Journal: Hum Mol Genet Date: 1999-09 Impact factor: 6.150
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Authors: M Koenig; A H Beggs; M Moyer; S Scherpf; K Heindrich; T Bettecken; G Meng; C R Müller; M Lindlöf; H Kaariainen; A de la Chapellet; A Kiuru; M L Savontaus; H Gilgenkrantz; D Récan; J Chelly; J C Kaplan; A E Covone; N Archidiacono; G Romeo; S Liechti-Gailati; V Schneider; S Braga; H Moser; B T Darras; P Murphy; U Francke; J D Chen; G Morgan; M Denton; C R Greenberg; K Wrogemann; L A Blonden; M B van Paassen; G J van Ommen; L M Kunkel Journal: Am J Hum Genet Date: 1989-10 Impact factor: 11.025
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Authors: Lawrence T Bish; Meg M Sleeper; Sean C Forbes; Bingjing Wang; Caryn Reynolds; Gretchen E Singletary; Dennis Trafny; Kevin J Morine; Julio Sanmiguel; Sylvain Cecchini; Tamas Virag; Adeline Vulin; Cyriaque Beley; Janet Bogan; James M Wilson; Krista Vandenborne; Joe N Kornegay; Glenn A Walter; Robert M Kotin; Luis Garcia; H Lee Sweeney Journal: Mol Ther Date: 2011-12-06 Impact factor: 11.454
Authors: Bo Wu; Peijuan Lu; Caryn Cloer; Mona Shaban; Snimar Grewal; Stephanie Milazi; Sapana N Shah; Hong M Moulton; Qi Long Lu Journal: Am J Pathol Date: 2012-06-07 Impact factor: 4.307
Authors: Sue Fletcher; Abbie M Adams; Russell D Johnsen; Kane Greer; Hong M Moulton; Steve D Wilton Journal: Mol Ther Date: 2010-03-23 Impact factor: 11.454
Authors: Eugene J Wyatt; Alexis R Demonbreun; Ellis Y Kim; Megan J Puckelwartz; Andy H Vo; Lisa M Dellefave-Castillo; Quan Q Gao; Mariz Vainzof; Rita C M Pavanello; Mayana Zatz; Elizabeth M McNally Journal: JCI Insight Date: 2018-05-03