Cells genetically modified to comprise pancreatic islet glucokinase and uses thereof

Abstract

The present invention relates generally to a population of cells genetically modified to produce insulin in a glucose responsive manner and uses thereof. More particularly, the present invention relates to a population of cells genetically modified to produce insulin in response to physiologically relevant levels of glucose and uses thereof. The cells of the present invention are useful in a wide variety of applications, in particular in the context of therapeutic and prophylactic regimes directed to the treatment of diabetes and/or the amelioration of symptoms associated with diabetes, based on the transplantation of the cells of the present invention into mammals requiring treatment. Also facilitated is the design of in vitro based screening systems for testing the therapeutic effectiveness and/or toxicity of potential adjunctive treatment regimes.

1 . (canceled) 
     
     
         2 . A method of modulating glucose levels in a mammal said method comprising administering to said mammal an effective number of isolated genetically modified hepatocytes recombinantly expressing insulin protein, human pancreatic islet glucokinase protein, and GLUT2 protein, and wherein the isolated genetically modified hepatocytes intracellularly store insulin and commence secretion of said insulin when exposed to an extracellular glucose concentration ranging in an amount from about 3 mM to about 8 mM. 
     
     
         3 . The method of  claim 2 , wherein the insulin protein is a human insulin protein, and the GLUT2 protein is a human GLUT2 protein. 
     
     
         4 . The method of  claim 2 , wherein the human pancreatic islet glucokinase protein has an amino acid sequence of SEQ ID NO: 2 or a sequence that is at least 90% identical to SEQ ID NO: 2. 
     
     
         5 . The method of  claim 2 , wherein the isolated genetically modified hepatocytes are human hepatocytes. 
     
     
         6 . The method of  claim 2 , wherein the isolated genetically modified hepatocytes are Huh7 cells. 
     
     
         7 . The method of  claim 2 , the isolated genetically modified hepatocytes are autologous cells, allogenic cells, or combination thereof. 
     
     
         8 . The method of  claim 2 , isolated genetically modified hepatocytes are encapsulated. 
     
     
         9 . The method of  claim 2 , wherein the mammal is a human. 
     
     
         10 . The method of  claim 2 , wherein the mammal is diagnosed with diabetes. 
     
     
         11 . The method of  claim 10 , wherein the diabetes is type 1 diabetes, IDDM, gestational diabetes, slowly progressive IDDM, latent autoimmune diabetes, type 2 diabetes, or combinations thereof. 
     
     
         12 . The method of  claim 2 , wherein the administration comprises injection or transplantation.