Genetically modified muscle cells which express neurotrophic factors

Abstract

An isolated muscle progenitor cell being MyoD positive, CD34 negative and CD45 negative is disclosed. The muscle progenitor cell is genetically modified to express at least one neurotrophic factor. In addition, cell populations are disclosed, comprising at least four subpopulations of muscle cells each being genetically modified to express a different neurotrophic factor, wherein said neurotrophic factor is selected from the group consisting of glial derived neurotrophic factor (GDNF), insulin growth factor (IGF-1), vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF). Uses of the cell populations are also disclosed.

What is claimed is: 
     
         1 . An isolated cell population, comprising at least four subpopulations of muscle progenitor cells, each of the at least four subpopulation being distinct in that they are genetically modified to express a different neurotrophic factor, wherein the neurotrophic factor is selected from the group consisting of glial derived neurotrophic factor (GDNF), insulin growth factor (IGF-1), vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF). 
     
     
         2 . The isolated cell population of  claim 1 , wherein said muscle progenitor cells comprise skeletal muscle progenitor cells. 
     
     
         3 . The isolated cell population of  claim 1 , wherein each of said at least four subpopulations are present in substantially equal amounts. 
     
     
         4 . The isolated cell population of  claim 1 , wherein each of said at least four subpopulations are present in alternate ratios. 
     
     
         5 . The isolated cell population of  claim 1 , wherein each of said at least four subpopulations is genetically modified to express a single neurotrophic factor. 
     
     
         6 . An isolated muscle progenitor cell being MyoD positive, CD34 negative and CD45 negative, genetically modified to express at least one neurotrophic factor. 
     
     
         7 . The isolated muscle progenitor cell of  claim 6 , being isolated by at least two rounds of differential plating, wherein a first round of said differential plating is effected on plastic plates and a second round of said differential plating is effected on a plate coated with a substance selected from the group consisting of collagen, gelatin and poly-lysine. 
     
     
         8 . The isolated muscle progenitor cell of  claim 6 , wherein said at least one neurotrophic factor is selected from the group consisting of glial derived neurotrophic factor (GDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4/5, Neurturin (NTN), Persephin, brain derived neurotrophic factor (BDNF), artemin (ART), ciliary neurotrophic factor (CNTF), insulin growth factor-I (IGF-1) and Neublastin. 
     
     
         9 . The isolated muscle progenitor cell of  claim 6 , wherein said neurotrophic factor is selected from the group consisting of glial derived neurotrophic factor (GDNF), vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF) and insulin growth factor-I (IGF-1). 
     
     
         10 . The isolated muscle progenitor cell of  claim 6 , being an adult, muscle progenitor cell. 
     
     
         11 . The isolated muscle progenitor cell of  claim 6 , being a skeletal muscle progenitor cell. 
     
     
         12 . The isolated muscle progenitor cell of  claim 6 , expressing at least two of said GDNF, VEGF, BDNF and IGF-1. 
     
     
         13 . The isolated muscle progenitor cell of  claim 6 , expressing each of said GDNF, VEGF, BDNF and IGF-1. 
     
     
         14 . A method of treating a nerve disease or damage in a subject, comprising introducing into the muscle cells of the subject a first polynucleotide which encodes a first neurotrophic factor, a second polynucleotide which encodes a second neurotrophic factor and a third polynucleotide which encodes a third neurotrophic factor, each of said first, said second and said third neurotrophic factor being mutually distinct, thereby treating the nerve disease or damage. 
     
     
         15 . The method of  claim 14 , further comprising introducing into the muscle cells of the subject a fourth polynucleotide which encodes a fourth neurotrophic factor, each of said first, said second, said third and said fourth neurotrophic factor being mutually distinct. 
     
     
         16 . The method of  claim 14 , wherein said first polynucleotide, said second polynucleotide and said third polynucleotide are comprised in a single nucleic acid construct. 
     
     
         17 . The method of  claim 14 , wherein said first polynucleotide, said second polynucleotide and said third polynucleotide are each comprised in a different nucleic acid construct. 
     
     
         18 . The method of  claim 15 , wherein said first neurotrophic factor is GDNF, said second neurotrophic factor is VEGF, said third neurotrophic factor is BDNF and said fourth neurotrophic factor is IGF-1. 
     
     
         19 . A method of treating a nerve disease or damage, comprising administering to a subject in need thereof a therapeutically effective amount of an isolated population of muscle progenitor cells, said muscle progenitor cells being MyoD positive, CD34 negative and CD45 negative, genetically modified to express at least one neurotrophic factor, thereby treating the nerve disease or damage. 
     
     
         20 . The method of  claim 19 , wherein said administering is effected by transplanting said isolated population of muscle cells into the muscle of said subject. 
     
     
         21 . The method of  claim 14 , wherein the nerve disease is a neuromuscular disease. 
     
     
         22 . The method of  claim 14 , wherein the nerve disease is a motor neuron disease. 
     
     
         23 . The method of  claim 14 , wherein the nerve damage is selected from the group consisting of peripheral nerve injury, peripheral nerve inflammation, autonomic nerve injury, pelvic nerve damage, burn, blunt trauma, back injury, back pain and sciatica. 
     
     
         24 . The method of  claim 21 , wherein the neuromuscular disease is selected from the group consisting of a spinal muscular atrophy, a amyotrophic lateral sclerosis (ALS), a Werdnig Hoffman disease, a Charcot-Marie tooth disease, multiple sclerosis, myasthenia gravis, muscular dystrophy and a myositis. 
     
     
         25 . The method of  claim 22 , wherein said motor neuron disease is selected from the group consisting of amyotrophic lateral sclerosis, primary lateral sclerosis (PLS), pseudobulbar palsy and progressive bulbar palsy. 
     
     
         26 . The method of  claim 24 , wherein the neuromuscular disorder is ALS.