The long-term objective of this proposal is to define mononuclear cells in human muscle that hold promisefor optimization of cell-based therapy for muscular dystrophy and myopathy. Recent findings from ourlaboratory have indicated that specific cell populations in human skeletal muscle may interact with oneanother, regulating proliferation and differentiation of muscle precursor cells. Using various methods ofpurification and by working closely with Project 1, which will perform parallel studies on muscle progenitors inzebrafish, we will fractionate several subsets of human muscle precursor cells. These isolated fractions willbe tested in both in vitro and in vivo models of skeletal muscle repair using three separate paradigms ofmyopathy: dystrophin, dysferlin and myotubularin deficiency. The overall goal of this application is that bybetter defining the mononuclear cells in human skeletal muscle and their interactions with one another, it willbe possible to identify a highly proliferative, myogenic cell fraction that is amenable to therapeuticapplications. This goal will be achieved via the following specific Aims:
Aim 1. Fractionate mononuclear cell populations in human fetal and adult skeletal muscles, study their cellsurface similarities, proliferative capacity and myogenic potential in vitro.
Aim 2. Use mRNA expression arrays to characterize human myogenic progenitors with high proliferativeand repair capacity.
Aim 3. Assay the capacity of fractionated human muscle progenitor cells to repair mutant muscle cells invitro.
Aim 4. Determine the restorative and myogenic potential of distinct mononuclear cell populations withinhuman muscle in vivo.The hope is that by working in close synergy with Project 1, we will be able to understand the signals thatcontrol human muscle cell proliferation, myogenic commitment and ability to repair diseased muscle. Bypursuing each one of these efforts, we will define the most effective cell population(s) in human muscle anddemonstrate their effectiveness in pre-clinical mouse models of muscular dystrophy and myopathy. Ifsuccessful, these findings are likely to make a significant step forward in the field of cell-based therapy formuscular dystrophy.
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