Growth differentiation factor-8

Abstract

A transgenic non-human animal of the species selected from the group consisting of avian, bovine, ovine and porcine having a transgene which results in disrupting the production of and/or activity of growth differentiation factor-8 (GDF-8) chromosomally integrated into the germ cells of the animal is disclosed. Also disclosed are methods for making such animals, and methods of treating animals, including humans, with antibodies or antisense directed to GDF-8. The animals so treated are characterized by increased muscle tissue.

1 . A transgenic non-human animal having a transgene disrupting or interfering with expression of growth differentiation factor-8 (GDF-8) chromosomally integrated into the germ cells of the animal.  
     
     
         2 . The transgenic animal of  claim 1 , wherein the animal is selected from the group of species consisting of avian, bovine, ovine, piscine, murine, and porcine.  
     
     
         3 . The transgenic animal of  claim 1  where the species is avian.  
     
     
         4 . The transgenic animal of  claim 1  where the species is bovine.  
     
     
         5 . The transgenic animal of  claim 1  where the species is porcine.  
     
     
         6 . The transgenic animal of  claim 1  where the species is ovine.  
     
     
         7 . The transgenic animal of  claim 1  where the species is piscine.  
     
     
         8 . The transgenic animal of  claim 1 , wherein the transgene comprises GDF-8 antisense polynucleotide(s).  
     
     
         9 . The transgenic animal of  claim 1 , wherein the transgene comprises a gene encoding a dominant negative GDF-8 polypeptide.  
     
     
         10 . The transgenic animal of  claim 1 , wherein the animal is homozygous or heterozygous for GDF-8 polynucleotide.  
     
     
         11 . A chicken or turkey egg produced by the transgenic animal of  claim 3 .  
     
     
         12 . Beef obtained from the transgenic animal of  claim 4 .  
     
     
         13 . Milk obtained from the transgenic animal of  claim 4 .  
     
     
         14 . Pork obtained from the transgenic animal of  claim 5 .  
     
     
         15 . Lamb obtained from the transgenic animal of  claim 6 .  
     
     
         16 . Chicken or turkey meat produced by the transgenic animal of  claim 3 .  
     
     
         17 . A method of producing animal food products having increased muscle mass comprising: 
 a) introducing a transgene disrupting or interfering with expression of growth differentiation factor-8 (GDF-8) into an embryo into germ cells of a pronuclear embryo of the animal;    b) implanting the embryo into the oviduct of a pseudopregnant female thereby allowing the embryo to mature to full term progeny;    c) testing the progeny for presence of the transgene to identify transgene-positive progeny;    d) cross-breeding transgene-positive progeny to obtain further transgene-positive progeny; and    e) processing the progeny to obtain foodstuff.    
     
     
         18 . The method of  claim 17 , wherein the transgene comprises GDF-8 antisense polynucleotides.  
     
     
         19 . The method of  claim 17 , wherein the transgene comprises a gene encoding a dominant negative GDF-8 polypeptide.  
     
     
         20 . A method of producing avian food products having reduced cholesterol levels comprising: 
 a) introducing a transgene disrupting or interfering with expression of growth differentiation factor-8 (GDF-8) into an embryo of an avian animal;    b) culturing the embryo under conditions whereby progeny are hatched;    c) testing the progeny for presence of the transgene to identify transgene-positive progeny;    d) cross-breeding transgene-positive progeny; and    e) processing the progeny to obtain foodstuff.    
     
     
         21 . The method of  claim 20 , wherein the transgene comprises GDF-8 antisense polynucleotides.  
     
     
         22 . The method of  claim 20 , wherein the transgene comprises a gene encoding a dominant negative GDF-8 polypeptide.  
     
     
         23 . The transgenic animal of  claim 20 , wherein the transgene comprises a polynucleotide encoding a truncated GDF-8 polypeptide.  
     
     
         24 . A method for increasing the muscle mass in an animal comprising administering to the animal an antibody, or fragment thereof, which binds to GDF-8 polypeptide.  
     
     
         25 . The method of  claim 24 , wherein anti-GDF-8 antibody is administered to a domesticated animal.  
     
     
         26 . The method of  claim 24 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.  
     
     
         27 . The method of  claim 24 , wherein the anti-GDF-8 antibody is administered by intravenous, intramuscular, multiple bolus, or subcutaneous injections.  
     
     
         28 . The method of  claim 27 , wherein the anti-GDF-8 antibody is administered within a dose range of 0.1 ug/kg to 100 mg/kg.  
     
     
         29 . The method of  claim 27 , wherein the antibody is formulated in a formulation suitable for administration by injection into an animal.  
     
     
         30 . A method of inhibiting the growth regulating actions of GDF-8 comprising contacting a GDF-8 agent with fetal or adult muscle cells or progenitor cells.  
     
     
         31 . The method of  claim 30 , wherein the agent is selected from the group consisting of a monoclonal antibody, an antisense nucleic acid and a dominant negative encoding nucleic acid sequence or polypeptide.  
     
     
         32 . The method of  claim 31 , wherein the antibody is a humanized monoclonal antibody or a chimeric monoclonal antibody or fragment thereof.  
     
     
         33 . The method of  claim 30 , wherein the agent is administered to a patient suffering from a disorder selected from the group consisting of muscle wasting disease, neuromuscular disorder, muscle atrophy and aging.  
     
     
         34 . The method of  claim 30 , wherein the agent is administered to a patient suffering from a disorder selected from the group consisting of muscular dystrophy, spinal cord injury, traumatic injury, congestive obstructive pulmonary disease (COPD), AIDS and cachechia.  
     
     
         35 . The method of  claim 30 , wherein the agent is administered to a patient with muscle wasting disease or disorder by intravenous, intramuscular or subcutaneous injection.  
     
     
         36 . The method of  claim 31 , wherein the monoclonal antibody is administered within a dose range between about 0.1/kg to about 100 mg/kg.  
     
     
         37 . The method of  claim 31 , wherein the monoclonal antibody is formulated in a formulation suitable for administration to a patient.  
     
     
         38 . A method for treating a muscle or adipose tissue disorder in a subject, comprising administering a therapeutically effective amount of a GDF-8 agent to the subject, thereby inhibiting abnormal growth of muscle or adipose tissue.  
     
     
         39 . The method as in  claim 38 , wherein the GDF-8 agent is selected from the group consisting of an antisense polynucleotide, a polynucleotide encoding a dominant negative GDF-8 polypeptide, a GDF-8 antibody and a polynucleotide encoding a truncated GDF-8 polypeptide.  
     
     
         40 . The method of  claim 38 , wherein the disorder is a cancer selected from the group consisting of muscle, connective tissue, or bone.  
     
     
         41 . The method of  claim 38 , wherein the subject has an obesity disorder.  
     
     
         42 . A method of inhibiting the growth regulating actions of GDF-8 in a subject comprising administering to the subject, a GDF-8 agent that inhibits the action of GDF-8 in the subject.  
     
     
         43 . The method of  claim 42 , wherein the GDF-8 agent is selected from the group consisting of an antisense polynucleotide, a polynucleotide encoding a dominant negative GDF-8 polypeptide, a GDF-8 antibody and a polynucleotide encoding a truncated GDF-8 polypeptide.  
     
     
         44 . A method for identifying a compound that affects GDF-8 activity or gene expression comprising: 
 a) incubating the compound with GDF-8 polypeptide, or with a recombinant cell expressing GDF-8 under conditions sufficient to allow the components to interact; and    b) determining the effect of the compound on GDF-8 activity or expression.    
     
     
         45 . The method of  claim 44 , wherein the effect is inhibition of GDF-8 activity or expression.  
     
     
         46 . The method of  claim 44 , wherein the effect is stimulation of GDF-8 activity or expression.  
     
     
         47 . An isolated polynucleotide encoding a truncated GDF-8 polypeptide wherein the truncation is a loss of the C-terminal active fragment of GDF-8.  
     
     
         48 . The isolated polynucleotide of  claim 47 , wherein the polynucleotide is as shown in FIG. 12 a .