Desmin-regulated lentiviral vectors for skeletal muscle gene transfer.
Lentiviral vectors (LVs) are highly attractive as a gene therapy agent as they are able to stably integrate their genomes in both dividing and nondividing cells and, in principle, provide long-term therapeutic benefit. However, their performance in skeletal muscle in adult animals has, to date, been disappointing. In order to gain clearer insight into their utility in this tissue type, we have conducted an extensive quantitative comparison of constitutive and muscle-specific promoter activities in skeletal muscle and nonmuscle systems following LV delivery in cell lines and neonatal mice. Our data show that LV delivery to hind leg skeletal muscle of neonatal mouse results in long-term transgene expression in adulthood. We find that the human desmin (DES) promoter/enhancer is the first muscle-specific control region to match the activity of the highly active constitutive human cytomegalovirus (hCMV) promoter/enhancer in skeletal muscle within a LV context both in vitro and in vivo. Furthermore, the DES promoter/enhancer provides six- to eightfold greater expression per viral copy than the muscle-specific human muscle creatine kinase (CKM) promoter/enhancer. DES also confers a more reproducible and tissue-specific transgene expression profile compared to CKM and is therefore a highly attractive regulatory element for use in muscle gene therapy vectors.
Talbot
Gillian E
GE
King's College London School of Medicine, Nuclear Biology Group, Department of Medical and Molecular Genetics, Guy's Hospital, London, UK.
Waddington
Simon N
SN
Bales
Olivia
O
Tchen
Rose C
RC
Antoniou
Michael N
MN
D000818:Animals; D002460:Cell Line; D003402:Creatine Kinase / Q000235:genetics; D003893:Desmin / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008957:Models, Genetic; D018482:Muscle, Skeletal / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D019076:Transgenes
D003893:Desmin; D049452:Green Fluorescent Proteins; D003402:Creatine Kinase