title: Genetic complementation of human muscle cells via directed stem cell fusion.
abstract: Duchenne muscular dystrophy (DMD) is caused by mutations in the X chromosome-linked DMD gene, which encodes the sarcolemma-stabilizing protein-dystrophin. Initial attempts at DMD therapy deployed muscle progenitor cells from healthy donors. The utilization of these cells is, however, hampered by their immunogenicity, while those from DMD patients are scarce and display limited ex vivo replication. Nonmuscle cells with myogenic capacity may offer valuable alternatives especially if, to allow autologous transplantation, they are amenable to genetic intervention. As a paradigm for therapeutic gene transfer by heterotypic cell fusion we are investigating whether human mesenchymal stem cells (hMSCs) can serve as donors of recombinant DMD genes for recipient human muscle cells. Here, we show that forced MyoD expression in hMSCs greatly increases their tendency to participate in human myotube formation turning them into improved DNA delivery vehicles. Efficient loading of hMSCs with recombinant DMD was achieved through a new tropism-modified high-capacity adenoviral (hcAd) vector directing striated muscle-specific synthesis of full-length dystrophin. This study introduces the principle of genetic complementation of gene-defective cells via directed cell fusion and provides an initial framework to test whether transient MyoD synthesis in autologous, gene-corrected hMSCs increases their potential for treating DMD and, possibly, other muscular dystrophies.
authors.affiliation: Virus and Stem Cell Biology Laboratory, Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. m.goncalves@lumc.nl
authors.forename: Manuel A F V
authors.initials: MA
authors.lastname: Gonçalves
authors.forename: Jim
authors.initials: J
authors.lastname: Swildens
authors.forename: Maarten
authors.initials: M
authors.lastname: Holkers
authors.forename: Anjali
authors.initials: A
authors.lastname: Narain
authors.forename: Gijsbert P
authors.initials: GP
authors.lastname: van Nierop
authors.forename: Marloes J M
authors.initials: MJ
authors.lastname: van de Watering
authors.forename: Shoshan
authors.initials: S
authors.lastname: Knaän-Shanzer
authors.forename: Antoine A F
authors.initials: AA
authors.lastname: de Vries
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002459:Cell Fusion / Q000379:methods; D002478:Cells, Cultured; D016189:Dystrophin / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D017570:MyoD Protein / Q000096:biosynthesis / Q000235:genetics; D032448:Myoblasts, Skeletal / Q000166:cytology / Q000378:metabolism
chemical_list: C484258:DMD protein, human; D016189:Dystrophin; D009124:Muscle Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein