Disease prevention and alleviation by human myoblast transplantation

Abstract

Methods and materials are described for human genome prophylaxis and therapy of diseases using myoblast transfer. These methods result in gene transcript changes in multiple pathways. Linking the myoblast transfer technology development from DMD, cardiomyopathy, and Type-II diabetes, the myoblast transfer demonstrably mediates its effect through transfer of the normal myoblast nuclei that supply the complete human genome, in addition to just replenishing the missing gene(s) or the aberrant gene(s). The replacement genes then transcribe to produce the necessary proteins or factors for genetic repair. A variety of uses of this technology are described, including that for disease treatment, disease prevention, drug discovery, and selection of superior cells and clones for therapy

I claim: 
     
         1 . A method of treating Type II diabetes in a patient in need thereof, comprising:
 determining or verifying Type II diabetes status of the patient;   supplying a complete human genome to the patient through transfer of myoblast nuclei, comprising the steps of:
 a. culturing a supply of myoblasts from a muscle biopsy of a donor that is genetically normal with respect to insulin sensitivity; 
 b. administering a therapeutically effective dosage of an immunosuppressant to the patient; and 
 c. thereafter selecting and administering from the supply a therapeutically effective dose of myoblasts to at least one muscle of the patient, 
 d. allowing donor myoblasts to insert their nuclei, each of which contain a full complement of normal genes into the patient's genetically abnormal muscle cells to effect genetic complementation repair; and 
 e. allowing donor myoblasts to fuse among themselves to form genetically normal muscle fibers; 
 whereby muscle mediated glucose homeostasis is improved, and is detected by measuring an improvement in the Type II diabetes status of the patient. 
   
     
     
         2 . The method of  claim 1 , wherein said cultured myoblasts are histoincompatible with the patient, and derived from a genetically normal donor. 
     
     
         3 . The method of  claim 2 , wherein the donor is selected from a genetically normal group, including but not limited to the father or male siblings of the host. 
     
     
         4 . The method of  claim 1  wherein said cultured myoblasts are obtained from muscle biopsy of the patient before the patient has developed Type II diabetes. 
     
     
         5 . The method of  claim 1 , wherein the measurement of improvement in the Type II diabetes status of the patient is carried out by detecting changes in quantity or quality of direct transcripts of multiple genes that provide relief of disease symptoms. 
     
     
         6 . The method of  claim 1 , wherein said selected therapeutically effective dose of the cultured myoblasts is approximately 50 billion cells cells for Type II diabetes, and wherein the dose is administered at approximately 100 million cells per mL of patient serum. 
     
     
         7 . The method of  claim 1 , wherein said immunosuppressant comprises cyclosporine administered orally at 5 to 7 mg/Kg body weight per day, beginning at five days before myoblast implantation, and completely weaned at three weeks post-operatively. 
     
     
         8 . The method of  claim 7 , wherein cyclosporine whole blood trough level is monitored weekly and the cyclosporine dose is re-adjusted as needed to maintain a blood level of approximately 250 ng/mL until the last week when weaning begins. 
     
     
         9 . The method of  claim 1 , wherein the patient is identified as at risk of acquiring Type II diabetes, and whereby muscle mediated glucose homeostasis is improved, and is detected by measuring an improvement in the glucose response status of the patient. 
     
     
         10 . A method of treating genetic disease in a host by supplying a complete genome of the host species through transfer of myoblast nuclei, comprising the steps of:
 culturing a supply of genetically normal myoblasts from muscle biopsy of genetically normal donor;   administering a therapeutically effective dosage of an immunosuppressant to the host; and   thereafter selecting and administering from the said supply a therapeutically effective dose of myoblasts to at least one muscle of the host,   allowing donor myoblasts to insert their nuclei, each of which contain a full complement of normal genes into the patient's abnormal muscle cells to effect genetic complementation repair; and   allowing donor myoblasts to fuse among themselves to form genetically normal muscle fibers;   whereby detection of improvement in the host is carried out, including at least one of measurement of muscle function improvement, increase in locomotive capacity, blood ejection/vascularization, and transfer of biochemicals or ions across the muscle cell membrane.   
     
     
         11 . The method of  claim 10 , wherein said cultured myoblasts are histoincompatible with the host, being derived from donor selected from a genetically normal group, including but not limited to the father or male siblings of the host. 
     
     
         12 . The method of  claim 10 , wherein said cultured myoblasts are histocompatible with the host, with donor muscle biopsy having been obtained prior to being myopathic from the host. 
     
     
         13 . The method of  claim 10 , wherein detection of improvement in the host is carried out by detecting changes in the quantity or quality of direct transcripts of multiple genes that provide relief of disease symptoms. 
     
     
         14 . The method of  claim 10 , wherein said selected therapeutically effective dose of the cultured myoblasts is approximately 50 billion cells for muscular dystrophy, approximately one billion cells for cardiomyopathy, and approximately 50 billion cells for Type II diabetes, with said dose being administered at dosage of approximately 100 million cells per mL of host serum. 
     
     
         15 . The method of  claim 10 , wherein said immunosuppressant comprises cyclosporine administered orally at 5 to 7 mg/Kg body weight per day, beginning at five days before myoblast implantation, and completely weaned off at three weeks post-operatively. 
     
     
         16 . The method of  claim 15 , wherein cyclosporine whole blood trough level is monitored weekly and cyclosporine dose is re-adjusted if necessary to maintain at approximately 250 ng/mL until the last week when weaning begins. 
     
     
         17 . A composition for treating or preventing symptom onset of a genetic disease in a host by supplying a complete genome of the host species through transfer of myoblast nuclei, comprising: a supply quantum of cultured genetically normal myoblasts being administered at a therapeutically effective dose to at least one muscle of the host. 
     
     
         18 . The composition of  claim 17 , wherein said selected therapeutically effective dose of the cultured myoblasts is approximately 50 billion cells for muscular dystrophy, approximately one billion cells for cardiomyopathy, and approximately 50 billion cells for Type II diabetes, with said dose being administered at dosage of approximately 100 million cells per mL of host serum. 
     
     
         19 . The composition of  claim 17 , wherein said cultured myoblasts are histoincompatible with the host, being derived from donor selected from a genetically normal group, including but not limited to the father or male siblings of the host. 
     
     
         20 . The composition of  claim 17 , wherein said cultured myoblasts are histocompatible with the host, with donor muscle biopsy having been obtained prior to being myopathic from the host.