This project aims to study muscle development and complementation mechanisms after normal myoblasts are transplanted into a growing and regenerating dystrophic muscle. Applicability of this new transplant technique to alleviate muscle weakness in hereditary myopathies will be tested in dystrophic mice. We have shown that surgically transplanted mesenchymal cells from normal mouse embryos improve the structure and function of dystrophic mouse muscles. The present myoblast transplantation is an extension designed to mimic future development in humans. Dystrophic cells degenerate because of the lack of the normal genome. The normal genome may become incorporated when normal donor myoblasts fuse with satellite cells of dystrophic hosts to form mosaic myofibers. In myopathies of recessive inheritance, gene products from the normal nuclei may restore the phenotype of these heterokaryotes to normal. Furthermore, surviving donor cells will develop and replace the myopathic tissue. They may further donate a diffusible gene product to supplement neighboring dystrophic fibers. These three mechanisms of complementation will be investigated. Cultured myoblasts from genetically normal mouse embryos will be transplanted into the right soleus of 20-day-old normal or dystrophic mice. Hosts and donors are of C57BL/6J background, and are immunocompatible. Survival and development of donor myoblasts in host muscles will be analyzed with electrophoresis of muscle isozymes of glucose phosphate isomerase (GPI). Donor cells carry the gpi-1b allele and produce GPI-1B. Host cells carry the gpi-1c allele and produce GPI-1C. Presence of the heterodimer, GPI-1BC, will substantiate intracellular mosaicism. Different groups of test and control solei will be compared before transplantation, and at 2, 4 and 6 months after transplantation. Muscle mechanophysiology, cellular resting potential and histology will be examined to determine if test muscles exhibit improved structure and function. Genetic mosaicism quantified each time with GPI electrophoresis will be correlated to phenotype indicated by physiology and histology. The effects of variation on donor cell number will be compared. The optimal quantity of donor cells for obtaining normal muscle function will be determined. The effects of transplanting myoblasts vs. fibroblasts will be compared. Dystrophic myoblasts will be transplanted into normal or dystrophic hosts and the results compared. Sham control for surgery will also be studied.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS020251-06
Application #
3400528
Study Section
Orthopedics and Musculoskeletal Study Section (ORTH)
Project Start
1983-12-01
Project End
1989-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
6
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Tennessee Health Science Center
Department
Type
Schools of Medicine
DUNS #
941884009
City
Memphis
State
TN
Country
United States
Zip Code
38163
Law, P K; Goodwin, T G; Fang, Q et al. (1993) Cell transplantation as an experimental treatment for Duchenne muscular dystrophy. Cell Transplant 2:485-505
Chen, M; Li, H J; Fang, Q et al. (1992) Dystrophin cytochemistry in mdx mouse muscles injected with labeled normal myoblasts. Cell Transplant 1:17-22
Fang, Q W; Chen, M; Li, H J et al. (1991) Vital marker for muscle nuclei in myoblast transfer. Can J Physiol Pharmacol 69:49-52
Law, P K; Goodwin, T G; Li, H J et al. (1990) Plausible structural/functional/behavioral/biochemical transformations following myoblast transfer therapy. Adv Exp Med Biol 280:241-9;discussion 249-50
Law, P K; Goodwin, T G; Li, H J et al. (1990) Myoblast transfer improves muscle genetics/structure/function and normalizes the behavior and life-span of dystrophic mice. Adv Exp Med Biol 280:75-84;discussion 84-7
Law, P K; Goodwin, T G; Li, H J (1988) Histoincompatible myoblast injection improves muscle structure and function of dystrophic mice. Transplant Proc 20:1114-9
Law, P K; Goodwin, T G; Wang, M G (1988) Normal myoblast injections provide genetic treatment for murine dystrophy. Muscle Nerve 11:525-33