Circulating human or canine factor IX from retrovirally transduced primary myoblasts and established myoblast cell lines grafted into murine skeletal muscle.
We have used retroviral vectors to introduce human or canine factor IX cDNAs into cultured primary murine and canine myoblasts and into the established murine myoblast cell line C2C12. In all cases, the stably infected cells produced biologically active factor IX in culture and secreted detectable amounts into the culture medium both before and after differentiation of the cells into myotubes. Myoblasts and differentiated myotubes are therefore capable of performing all the posttranslational modifications of the coagulation factor required for biological activity. We have grafted the genetically modified myoblasts into skeletal muscles of nude mice and have detected stable levels of circulating human factor IX for up to two months after grafting. We propose that grafting genetically modified primary myoblasts or established myoblast cell lines into skeletal muscle may represent a useful approach to gene therapy for a variety of genetic diseases, including intrinsic muscle disease and defects in circulating proteins as in the hemophilias.
Roman
M
M
Department of Pediatrics, University of California School of Medicine, San Diego, La Jolla 92093-0634.
Axelrod
J H
JH
Dai
Y
Y
Naviaux
R K
RK
Friedmann
T
T
Verma
I M
IM
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004285:Dogs; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009132:Muscles / Q000378:metabolism / Q000637:transplantation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae; D014162:Transfection
D011994:Recombinant Proteins; D005164:Factor IX