Abstract
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.
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Literature cited
McKee, P.A. (1983).The Metabolic Basis of Inherited Disease (McGraw-Hill, New York), pp. 1531–1560.
Chung, D.S., Goldstein, J.C., and Roberts, H.R. (1980).Handbook Series in Clinical Laboratory Science (CRC, Boca Raton, Florida), p. 85.
White, G.C. II, Roberts, H.R., Kingdon, H.S., and Lundblad, R.L. (1977).Blood 49159–164.
White, G.C. II, Blatt, P.M., McMillan, W.P., Lesesne, H.R., and Roberts, H.R. (1980).South Med. J. 73155–159.
Remis, R.S., O'Shaughnessy, M.V., Tsoukas, C., Growe, G.H., Schechter, M.T., Palmer, R.W., and Lawrence, D.N. (1990).Can. Med. Assoc. J. 1421247–1254.
Axelrod, J.H., Read, M.S., Brinkhous, K.M., and Verma, I.M. (1990).Proc. Natl. Acad. Sci. U.S.A. 875173–5177.
St. Louis, D., and Verma, I.M. (1988).Proc. Natl. Acad. Sci. U.S.A. 853150–3154.
Anson, D.S., Hock, R.A., Austen, D., Smith, K.J., Brownlee, G.G., Verma, I.M., and Miller, A.D. (1987).Mol. Biol. Med. 411–20.
Palmer, T.D., Thompson, A.R., and Miller, A.D. (1989).Blood 73438–445.
Armentano, D., Thompson, A.R., Darlington, G., and Woo, S.L. (1990).Proc. Natl. Acad. Sci. U.S.A. 876141–6145.
Yao, S.N., Wilson, J.M., Nabel, E.G., Kurachi, S., Hachiya, H.L., and Kurachi, K. (1991).Proc. Natl. Acad. Sci. U.S.A. 888101–8105.
Evans, J.P., Brinkhous, K.M., Brayer, G.D., Reisner, H.M., and High, K.A. (1989).Proc. Natl. Acad. Sci. U.S.A. 8610095–10099.
Palmer, T.D., Rosman, G.J., Osborne, W.R.A., and Miller, A.D. (1991).Proc. Natl. Acad. Sci. U.S.A. 881330–1334.
Law, P.K., Goodwin, T.G., and Wang, M.G. (1988).Muscle Nerve 11525–533.
Law, P.K., Goodwin, T.G., and Li, H.J. (1988).Transplant. Proc. 201114–1119.
Morgan, J.E., Coulton, G.R., and Partridge, T.A. (1987).J. Muscle Res. Cell Motil. 8386–396.
Morgan, J.E., Watt, D.J., Sloper, J.C., and Partridge, T.A. (1988).J. Neurol. Sci. 86137–147.
Karpati, G., Pouliot, Y., Zubrzycka-Gaarn, E., Carpenter, S., Ray, P.N., Worton, R.G., and Holland, P. (1989).Am. J. Pathol. 13527–32.
Partridge, T.A., Morgan, J.E., Coulton, G.R., Hoffman, E.P., and Kunkel, L.M. (1989).Nature 337176–179.
Salminen, A., Elson, H.F., Mickley, L.A., Fojo, A.T., and Gottesman, M.M. (1991).Hum. Gene Ther. 215–26.
Dhawan, J., Pan, L.C., Pavlath, G.K., Travis, M.A., Lanctot, A.M., and Blau, H.M. (1991).Science 2541509–1511.
Barr, E., and Leiden, J.M. (1991).Science 2541507–1509.
Yaffe, D., and Saxel, O. (1977).Nature 270725–727.
Yaffe, D. (1968).Proc. Natl. Acad. Sci. U.S.A. 61477–483.
Southern, P.J., and Berg, P. (1982).J. Mol. Appl. Genet. 1327–341.
Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York).
Danos, O., and Mulligan, R.C. (1988).Proc. Natl. Acad. Sci. U.S.A. 856460–6464.
Price, J., Turner, D., and Cepko, C. (1987).Proc. Natl. Acad. Sci. U.S.A. 84156–160.
Chomczynski, P., and Sacchi, N. (1986).Anal. Biochem. 162156–159.
Bajaj, S.P., Rapaport, S.I., and Maki, S.L. (1985).J. Biol. Chem. 26011574–11580.
Bos, E.S., van der Doelen, A.A., van Rooy, N., and Shuurs, A.H.W.M. (1981).J. Immunoassay 2187–204.
Goldsmith, J.C., Chung, K.S., and Roberts, H.R. (1978).Thromb. Res. 12497–502.
Fujikawa, K., and Davies, E.W. (1976).Methods Enzymol. 4574–83.
Hughes, S.M., and Blau, H.M. (1990).Nature 345350–353.
Smith, B.F., Hoffman, R.K., Giger, U., and Wolfe, J.H. (1990).Mol. Cell. Biol. 103268–3271.
Wolff, J.A., Fisher, L.J., Xu, L., Jinnah, H.A., Langlais, P.J., Iuvone, P.M., O'Malley, K.L., Rosenberg, M.B., Shimohama, S., and Friedmann, T. (1989).Proc. Natl. Acad. Sci. U.S.A. 869011–9014.
Alameddine, H.S., Dehaupas, M., and Fardeau, M. (1989).Muscle Nerve 12544–555.
Karpati, G. (1990).Adv. Exp. Med. Biol. 28069–74.
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Roman, M., Axelrod, J.H., Dai, Y. et al. Circulating human or canine factor IX from retrovirally transduced primary myoblasts and established myoblast cell lines grafted into murine skeletal muscle. Somat Cell Mol Genet 18, 247–258 (1992). https://doi.org/10.1007/BF01233861
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DOI: https://doi.org/10.1007/BF01233861