title: Autologous skeletal muscle derived cells expressing a novel functional dystrophin provide a potential therapy for Duchenne Muscular Dystrophy.
abstract: Autologous stem cells that have been genetically modified to express dystrophin are a possible means of treating Duchenne Muscular Dystrophy (DMD). To maximize the therapeutic effect, dystrophin construct needs to contain as many functional motifs as possible, within the packaging capacity of the viral vector. Existing dystrophin constructs used for transduction of muscle stem cells do not contain the nNOS binding site, an important functional motif within the dystrophin gene. In this proof-of-concept study, using stem cells derived from skeletal muscle of a DMD patient (mdcs) transplanted into an immunodeficient mouse model of DMD, we report that two novel dystrophin constructs, C1 (ΔR3-R13) and C2 (ΔH2-R23), can be lentivirally transduced into mdcs and produce dystrophin. These dystrophin proteins were functional in vivo, as members of the dystrophin glycoprotein complex were restored in muscle fibres containing donor-derived dystrophin. In muscle fibres derived from cells that had been transduced with construct C1, the largest dystrophin construct packaged into a lentiviral system, nNOS was restored. The combination of autologous stem cells and a lentivirus expressing a novel dystrophin construct which optimally restores proteins of the dystrophin glycoprotein complex may have therapeutic application for all DMD patients, regardless of their dystrophin mutation.
authors.affiliation: The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Institute of Child Health, 30 Guilford Street, London, UK, WC1N 1EH.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Meng
authors.affiliation: The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Institute of Child Health, 30 Guilford Street, London, UK, WC1N 1EH.|UCL Cancer Institute, Paul O'Gorman Building, University College London, 72 Huntley Street, London, UK, WC1E 6BT.|Molecular and Cellular Immunology, Institute of Child Health, University College London, 30 Guilford Street, London, UK, WC1N 1EH.
authors.forename: John R
authors.initials: JR
authors.lastname: Counsell
authors.affiliation: John Walton Centre for Muscular Dystrophy Research, MRC Centre for Neuromuscular Diseases, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK, NE1 3BZ.
authors.forename: Mojgan
authors.initials: M
authors.lastname: Reza
authors.affiliation: John Walton Centre for Muscular Dystrophy Research, MRC Centre for Neuromuscular Diseases, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK, NE1 3BZ.
authors.forename: Steven H
authors.initials: SH
authors.lastname: Laval
authors.affiliation: UCL Cancer Institute, Paul O'Gorman Building, University College London, 72 Huntley Street, London, UK, WC1E 6BT.
authors.forename: Olivier
authors.initials: O
authors.lastname: Danos
authors.affiliation: Molecular and Cellular Immunology, Institute of Child Health, University College London, 30 Guilford Street, London, UK, WC1N 1EH.
authors.forename: Adrian
authors.initials: A
authors.lastname: Thrasher
authors.affiliation: John Walton Centre for Muscular Dystrophy Research, MRC Centre for Neuromuscular Diseases, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK, NE1 3BZ.
authors.forename: Hanns
authors.initials: H
authors.lastname: Lochmüller
authors.affiliation: The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Institute of Child Health, 30 Guilford Street, London, UK, WC1N 1EH.
authors.forename: Francesco
authors.initials: F
authors.lastname: Muntoni
authors.affiliation: The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Institute of Child Health, 30 Guilford Street, London, UK, WC1N 1EH.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Morgan
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D004195:Disease Models, Animal; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D018101:Mice, Inbred mdx; D008819:Mice, Nude; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism*; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000628:therapy; D032446:Myoblasts / Q000166:cytology* / Q000378:metabolism*; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism; D020286:Pericytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D011485:Protein Binding; D011993:Recombinant Fusion Proteins; D014161:Transduction, Genetic
chemical_list: D016189:Dystrophin; D011993:Recombinant Fusion Proteins; D052248:Nitric Oxide Synthase Type I
references.citation: Am J Pathol. 2008 May;172(5):1332-41
references.pmid: 18385524
references.citation: Nat Med. 2015 Dec;21(12):1455-63
references.pmid: 26569381
references.citation: Nat Cell Biol. 2007 Mar;9(3):255-67
references.pmid: 17293855
references.citation: Gene Ther. 2013 Jun;20(6):581-8
references.pmid: 23171920
references.citation: J Cell Biol. 1991 Dec;115(6):1685-94
references.pmid: 1757468
references.citation: Mol Ther. 2007 Nov;15(11):1947-54
references.pmid: 17700544
references.citation: Cell. 2008 Jul 11;134(1):37-47
references.pmid: 18614009
references.citation: Hum Gene Ther. 2012 Jan;23(1):98-103
references.pmid: 21933029
references.citation: Ann Neurol. 2012 Mar;71(3):304-13
references.pmid: 22451200
references.citation: J Gene Med. 2007 Jul;9(7):579-84
references.pmid: 17533614
references.citation: Neuromuscul Disord. 2003 Jan;13(1):21-31
references.pmid: 12467729
references.citation: Curr Gene Ther. 2015;15(4):395-415
references.pmid: 26159373
references.citation: Hum Gene Ther. 2004 Nov;15(11):1109-24
references.pmid: 15610611
references.citation: PLoS One. 2013;8(8):e72147
references.pmid: 24015212
references.citation: Mol Ther. 2010 Jan;18(1):206-13
references.pmid: 19888194
references.citation: J Clin Invest. 2009 Mar;119(3):624-35
references.pmid: 19229108
references.citation: PLoS One. 2010;5(12):e15286
references.pmid: 21187970
references.citation: Curr Gene Ther. 2015;15(1):64-81
references.pmid: 25429463
references.citation: Blood. 2011 May 19;117(20):5332-9
references.pmid: 21403130
references.citation: PLoS One. 2013;8(8):e73224
references.pmid: 23951345
references.citation: Gene Ther. 1998 Jan;5(1):59-64
references.pmid: 9536265
references.citation: Mol Ther. 2014 Apr;22(4):774-85
references.pmid: 24441399
references.citation: Hum Mol Genet. 2015 Aug 1;24(15):4225-37
references.pmid: 25935000
references.citation: Cell Stem Cell. 2007 Dec 13;1(6):646-57
references.pmid: 18371406
references.citation: Hum Mol Genet. 2015 Oct 15;24(20):5880-90
references.pmid: 26264580
references.citation: J Cell Biol. 2000 Sep 18;150(6):1399-410
references.pmid: 10995444
references.citation: Stem Cells. 2013 Aug;31(8):1611-20
references.pmid: 23606600
references.citation: Mol Ther Methods Clin Dev. 2014;1:null
references.pmid: 25580445
references.citation: Hum Gene Ther. 2001 Oct 10;12(15):1893-905
references.pmid: 11589831
references.citation: Mol Ther. 2012 Feb;20(2):462-7
references.pmid: 22086232
references.citation: Mol Ther. 2010 Mar;18(3):601-8
references.pmid: 19935780
references.citation: Lancet. 2011 Aug 13;378(9791):595-605
references.pmid: 21784508
references.citation: Mol Ther. 2014 May;22(5):1008-17
references.pmid: 24569833
references.citation: Hum Mol Genet. 2009 Apr 1;18(7):1209-20
references.pmid: 19131360
references.citation: Neuromuscul Disord. 2010 Jan;20(1):6-15
references.pmid: 20034794
references.citation: Cell. 1987 Dec 24;51(6):919-28
references.pmid: 3319190
references.citation: J Virol. 1998 Nov;72(11):8463-71
references.pmid: 9765382
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 8;110(2):525-30
references.pmid: 23185009
references.citation: Neuromuscul Disord. 1997 Jul;7(5):329-35
references.pmid: 9267847
references.citation: PLoS One. 2012;7(2):e31937
references.pmid: 22359642
references.citation: Exp Cell Res. 2014 Jan 1;320(1):33-45
references.pmid: 23899627
references.citation: Mol Ther. 2011 Jan;19(1):165-71
references.pmid: 20924363
references.citation: J Neurol Sci. 1990 Oct;99(1):9-25
references.pmid: 2250176
references.citation: PLoS One. 2011;6(3):e17454
references.pmid: 21408080
references.citation: Cell. 1995 Sep 8;82(5):743-52
references.pmid: 7545544
references.citation: N Engl J Med. 1988 May 26;318(21):1363-8
references.pmid: 3285207
references.citation: Neuromuscul Disord. 2007 Aug;17(8):631-8
references.pmid: 17588754
references.citation: Hum Mol Genet. 2004 Apr 1;13(7):693-702
references.pmid: 14962982
references.citation: Curr Gene Ther. 2014;14(4):276-88
references.pmid: 25039614
references.citation: Neuromuscul Disord. 2007 Apr;17(4):276-84
references.pmid: 17303423
references.citation: PLoS One. 2006;1:e54
references.pmid: 17183684
references.citation: Neuropathol Appl Neurobiol. 2004 Oct;30(5):540-5
references.pmid: 15488030
references.citation: Mol Ther. 2009 Oct;17(10):1771-8
references.pmid: 19623164