Muscle SP cells are a heterogeneous population of primitive cells that demonstrated hematopoietic and myogenic differentiation potential in vivo. First discovered in mice, these cells hold promise for optimization of cell-based therapy for muscular dystrophy. To date, little is known about human-derived muscle SP cells and their presumed clinical potential. The goals of this proposal are to study the differentiation potential of human fetal and adult-derived muscle SP cells, and to optimize their use in pre-clinical experiments using newly generated muscular dystrophy mouse models. These goals will be achieved via the following specific aims:
Aim 1. 1. Study the potential of unfractionated human muscle-derived SP and MP cells to differentiate into myogenic or hematopoietic cells in vitro. Human samples will be obtained from autopsy fetal and adult tissues and from adult-derived discarded surgical tissue.
Aim 1. 2. Fractionate human fetal and adult muscle SP and MP cells based on the expression of the cell surface antigens CD34, CD90 and CD133. Compare the ability of fractionated versus unfractionated cells to differentiate into myogenic or hematopoietic cells in vitro.
Aim 2. 1. Compare the ability of unfractionated and fractionated muscle-derived SP and MP cells to fuse into dystrophic myofibers in vivo. Perform intramuscular injections into NOD/RAG1/null-DMD/mdx5cv mice using the fetal and adult-derived human muscle SP and MP cell fractions enriched for myogenic precursors (Aim 1.2).
Aim 2. 2. Assess the clinical potential of human fetal and adult-derived muscle SP or MP cells with myogenic activity to engraft dystrophic skeletal muscles in vivo. Perform intra-arterial and intravenous injections of human muscle SP or MP cells in non-irradiated NOD/RAG1/null-DMD/mdx5cv.
Aim 2. 3. Assess the ability of fractionated human fetal and adult-derived muscle SP or MP cells to differentiate into hematopoietic cells in vivo. Inject unfractionated and fractionated cells with hematopoietic activity (Aims 1.1 and 1.2) into the circulation of lethally irradiated NOD/RAG1/null/mdx5cv or NOD/RAG1/null-Pfp/null DMD/mdx5cv mice. These studies will enhance our basic knowledge on human muscle SP cells, their differentiation potential in fetal and adult tissue, and will evaluate their therapeutic promise for muscular dystrophy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS047727-02
Application #
6943910
Study Section
Special Emphasis Panel (ZRG1-MOSS-H (03))
Program Officer
Porter, John D
Project Start
2004-09-01
Project End
2008-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
2
Fiscal Year
2005
Total Cost
$257,497
Indirect Cost
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Robin, Jerome D; Wright, Woody E; Zou, Yaqun et al. (2015) Isolation and immortalization of patient-derived cell lines from muscle biopsy for disease modeling. J Vis Exp :52307
Huang, Ping; Schulz, Tim J; Beauvais, Ariane et al. (2014) Intramuscular adipogenesis is inhibited by myo-endothelial progenitors with functioning Bmpr1a signalling. Nat Commun 5:4063
Rozkalne, Anete; Adkin, Carl; Meng, Jinhong et al. (2014) Mouse regenerating myofibers detected as false-positive donor myofibers with anti-human spectrin. Hum Gene Ther 25:73-81
Wu, Melissa P; Doyle, Jamie R; Barry, Brenda et al. (2013) G-protein coupled receptor 56 promotes myoblast fusion through serum response factor- and nuclear factor of activated T-cell-mediated signalling but is not essential for muscle development in vivo. FEBS J 280:6097-113
Schulz, Tim J; Huang, Ping; Huang, Tian Lian et al. (2013) Brown-fat paucity due to impaired BMP signalling induces compensatory browning of white fat. Nature 495:379-83
Liadaki, Kalliopi; Casar, Juan Carlos; Wessen, McKenzie et al. (2012) ?4 integrin marks interstitial myogenic progenitor cells in adult murine skeletal muscle. J Histochem Cytochem 60:31-44
Lapan, Ariya D; Gussoni, Emanuela (2012) Isolation and characterization of human fetal myoblasts. Methods Mol Biol 798:3-19
Lapan, Ariya D; Rozkalne, Anete; Gussoni, Emanuela (2012) Human fetal skeletal muscle contains a myogenic side population that expresses the melanoma cell-adhesion molecule. Hum Mol Genet 21:3668-80
Lawlor, Michael W; Alexander, Matthew S; Viola, Marissa G et al. (2012) Myotubularin-deficient myoblasts display increased apoptosis, delayed proliferation, and poor cell engraftment. Am J Pathol 181:961-8
Wu, Melissa P; Gussoni, Emanuela (2011) Carbamylated erythropoietin does not alleviate signs of dystrophy in mdx mice. Muscle Nerve 43:88-93

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