Dystrophin expression in myofibers of Duchenne muscular dystrophy patients following intramuscular injections of normal myogenic cells.
Three Duchenne muscular dystrophy (DMD) patients received injections of myogenic cells obtained from skeletal muscle biopsies of normal donors. The cells (30 x 10 (6)) were injected in 1 cm3 of the tibialis anterior by 25 parallel injections. We performed similar patterns of saline injections in the contralateral muscles as controls. The patients received tacrolimus for immunosuppression. Muscle biopsies were performed at the injected sites 4 weeks later. We observed dystrophin-positive myofibers in the cell-grafted sites amounting to 9 (patient 1), 6.8 (patient 2), and 11% (patient 3). Since patients 1 and 2 had identified dystrophin-gene deletions these results were obtained using monoclonal antibodies specific to epitopes coded by the deleted exons. Donor dystrophin was absent in the control sites. Patient 3 had exon duplication and thus specific donor-dystrophin detection was not possible. However, there were fourfold more dystrophin-positive myofibers in the cell-grafted than in the control site. Donor-dystrophin transcripts were detected by RT-PCR (using primers reacting with a sequence int eh deleted exons) only in the cell-grafted sites in patients 1 and 2. Dystrophin transcripts were more abundant in the cell-grafted than in the control site in patient 3. Therefore, significant dystrophin expression can be obtained in teh skeletal muscles of DMD patients following specific conditions of cell delivery and immunosuppression.
Skuk
Daniel
D
Human Genetic Research Unit, laval Unibersity Hospital Center, Quebec, Canada.
Roy
Brigitte
B
Goulet
Marlyne
M
Chapdelaine
Pierre
P
Bouchard
Jean-Pierre
JP
Roy
Raynald
R
Dugré
Francine J
FJ
Lachance
Jean-Guy
JG
Deschênes
Louise
L
Hélène
Senay
S
Sylvain
Michel
M
Tremblay
Jacques P
JP
D000293:Adolescent; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D001706:Biopsy; D017690:Cell Transplantation / Q000379:methods; D002648:Child; D003598:Cytoskeletal Proteins / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D016189:Dystrophin / Q000096:biosynthesis* / Q000378:metabolism; D000939:Epitopes; D015316:Genetic Therapy / Q000379:methods*; D006239:Haplotypes; D006650:Histocompatibility Testing; D006801:Humans; D007150:Immunohistochemistry; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000378:metabolism; D020388:Muscular Dystrophy, Duchenne / Q000378:metabolism* / Q000628:therapy*; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D049031:Sarcoglycans; D016559:Tacrolimus / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D016189:Dystrophin; D000939:Epitopes; D007166:Immunosuppressive Agents; D008562:Membrane Glycoproteins; D049031:Sarcoglycans; D016559:Tacrolimus