Myostatin inhibition for enhancing muscle and/or improving muscle function

Abstract

The present invention relates to methods for inhibiting myostatin, a regulator of muscle mass, for muscle enhancement (including inducing hypertrophy and/or hyperplasia) as well as improving muscle function (including decreasing atrophy and/or increasing endurance, force and/or strength). Some of the methods involve delivering genes to cells using gene delivery or other delivery techniques known in the art in order to inhibit myostatin. Examples of genes to be delivered are genes encoding proteins such as Follistatin, Follistatin-related gene-1 (FLRG-1), growth differentiation factor associated protein-1 (GASP-1) and myostatin precursor propeptide. The genes can be delivered using, for example, a recombinant Adeno-associated virus (rAAV), lentivirus or equine-associated virus capable of infecting the cells. Following introduction, the genes are expressed in the cell body of the infected cell and the encoded proteins are secreted systemically. In other methods of the invention, expression of proteins such as activin IIb and myostatin is inhibited by oligonucleotide techniques to effect muscle enhancement. All the methods have applications in the treatment of musculoskeletal and neurodegenerative disorders among others, as well as enhancing muscle in livestock.

We claim: 
     
         1 . A method of delivering a myostatin inhibitor to an animal in need thereof, comprising the step of administering to the animal a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         2 . The method of  claim 1  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         3 . The method of  claim 1  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         4 . A method of enhancing muscle in an animal in need thereof comprising the step of administering to the animal a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         5 . The method of  claim 4  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         6 . The method of  claim 4  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         7 . A method of improving muscle function in an animal in need thereof comprising the step of administering to the animal a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         8 . The method of  claim 7  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         9 . The method of  claim 7  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         10 . A method of treating a musculoskeletal disease in an animal in need thereof comprising administering to the animal one or more effective doses of a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         11 . The method of  claim 10  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         12 . The method of  claim 10  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         13 . The method of  claim 10  wherein the musculoskeletal disease is a muscular dystrophy or inclusion body myositis (IBM). 
     
     
         14 . The method of  claim 13  wherein the muscular dystrophy is Duchenne muscular dystrophy. 
     
     
         15 . A method of treating a neurodegenerative disease in which muscle is adversely affected in an animal in need thereof comprising administering to the animal one or more effective doses of a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         16 . The method of  claim 15  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         17 . The method of  claim 15  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         18 . The method of  claim 15  wherein the neurodegenerative disease is Amyotrophic Lateral Sclerosis, multiple sclerosis or spinal muscular atrophy. 
     
     
         19 . A method of treating sarcopenia, cachexia, obesity, Type II diabetes, Pompe disease or lysosomal storage disorders in an animal in need thereof comprising administering to the animal one or more effective doses of a composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising AAV inverted terminal repeats flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         20 . The method of  claim 19  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         21 . The method of  claim 20  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         22 . The method of  claim 2 ,  5 ,  7 ,  11 ,  16  or  20  wherein the myostatin inhibitor protein is follistatin, follistatin-related gene (FLRG), myostatin precursor propeptide or growth differentiation factor associated protein-1 (GASP-1). 
     
     
         26 . The method of  claim 22  wherein the myostatin inhibitor protein is follistatin. 
     
     
         27 . The method of  claim 22  wherein the myostatin inhibitor protein is FLRG. 
     
     
         28 . The method of  claim 22  wherein the myostatin inhibitor protein is myostatin precursor propeptide. 
     
     
         29 . The method of  claim 22  wherein the myostatin inhibitor protein is GASP-1. 
     
     
         30 . The method of  claim 22  wherein the myostatin inhibitor protein binds to the myostatin receptor activin IIb. 
     
     
         31 . A recombinant adeno-associated virus (AAV) genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         32 . The recombinant AAV of  claim 31  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         33 . The recombinant AAV of  claim 31  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         34 . An infectious encapsidated rAAV particle (rAAV) comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         35 . A packaging cell producing an infectious encapsidated rAAV comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         36 . A composition comprising one or more infectious encapsidated rAAVs, each comprising a recombinant AAV genome comprising at least one AAV inverted terminal repeat flanking a polynucleotide encoding a myostatin inhibitor, wherein the genome lacks AAV rep and cap DNA. 
     
     
         37 . The composition of  claim 36  wherein the polynucleotide encodes a myostatin inhibitor protein and is operatively linked to transcriptional control DNA. 
     
     
         38 . The composition of  claim 36  wherein the myostatin inhibitor protein is follistatin, follistatin-related gene (FLRG), myostatin precursor propeptide or growth differentiation factor associated protein-1 (GASP-1). 
     
     
         39 . The composition of  claim 38  wherein the myostatin inhibitor protein is follistatin. 
     
     
         40 . The composition of  claim 38  wherein the myostatin inhibitor protein is FLRG. 
     
     
         41 . The composition of  claim 38  wherein the myostatin inhibitor protein is myostatin precursor propeptide. 
     
     
         42 . The composition of  claim 38  wherein the myostatin inhibitor protein is GASP-1. 
     
     
         43 . The composition of  claim 38  wherein the myostatin inhibitor protein binds to the myostatin receptor activin IIb. 
     
     
         44 . The composition of  claim 36  wherein the polynucleotide encodes a myostatin inhibitor RNA. 
     
     
         45 . The rAAV follistatin-344 comprising the DNA sequence of SEQ ID NO: 11. 
     
     
         46 . The rAAV MCK follistatin-344 comprising the DNA sequence of SEQ ID NO: 11.