Differentiation of non-insulin producing cells into insulin producing cells by GLP-1 or exendin-4 and uses thereof

Abstract

The present invention relates to a population of insulin producing cells made by a process comprising contacting non-insulin producing cells with a growth factor selected from the group consisting of GLP-1 or Exendin-4, growth factors having amino acid sequences substantially homologous to GLP-1 or Exendin-4, and fragments thereof. The present invention also relates to methods of differentiating non-insulin producing cells into insulin producing cells and of enriching a population of cells for insulin-producing cells. The present invention also relates to methods of treating diabetes.

1 - 52 . (canceled)  
     
     
         53 . A method of differentiating non-insulin-producing cells into insulin-producing cells in a subject with beta-cell dysfunction, comprising contacting the non-insulin-producing cells for at least two days with an amount of a substance effective to induce differentiation of the non-insulin-producing cells by administering the substance to the subject to provide said contact, wherein the substance comprises a peptide selected from the group consisting of exendin-4, an exendin-4 peptide, an exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of exendin-4, and a fragment of any one of the preceding exendin-4 peptides, wherein the exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells, and wherein administration of the substance is discontinued after insulin-producing cells are produced.  
     
     
         54 . The method of  claim 53 , wherein the contacting is for at least five days.  
     
     
         55 . The method of  claim 54 , wherein the contacting is for at least seven days.  
     
     
         56 . The method of  claim 55 , wherein the substance is administered subcutaneously.  
     
     
         57 . The method of  claim 55 , wherein the subject has type I diabetes.  
     
     
         58 . The method of  claim 55 , wherein the subject has type II diabetes.  
     
     
         59 . The method of  claim 55 , wherein the substance is exendin-4.  
     
     
         60 . The method of  claim 55 , wherein the non-insulin-producing cells are human pancreatic cells.  
     
     
         61 . The method of  claim 55 , wherein the administration of the substance is discontinued for at least two weeks after the insulin-producing cells are produced.  
     
     
         62 . The method of  claim 61 , wherein the administration of the substance is discontinued for at least two weeks after the insulin-producing cells are produced and the subject's fasting blood glucose is improved compared to that prior to administration of the substance.  
     
     
         63 . The method of  claim 62 , wherein the subject has type I diabetes.  
     
     
         64 . The method of  claim 63 , wherein the substance is exendin-4.  
     
     
         65 . The method of  claim 62 , wherein the subject has type II diabetes.  
     
     
         66 . The method of  claim 65 , wherein the substance is exendin-4.  
     
     
         67 . The method of  claim 61 , wherein the administration of the substance is discontinued for at least two weeks after the insulin-producing cells are produced and the subject's fasting blood insulin level is improved compared to that prior to administration of the substance.  
     
     
         68 . The method of  claim 67 , wherein the subject has type I diabetes.  
     
     
         69 . The method of  claim 68 , wherein the substance is exendin-4.  
     
     
         70 . The method of  claim 67 , wherein the subject has type II diabetes.  
     
     
         71 . The method of  claim 70 , wherein the substance is exendin-4.  
     
     
         72 . The method of  claim 53 , wherein the substance is administered at a dose of from about 1 pmole/kg to about 400 pmoles/kg.  
     
     
         73 . The method of  claim 72 , wherein the substance is administered at a dose of from about 10 pmoles/kg to about 40 pmoles/kg.  
     
     
         74 . The method of  claim 53 , wherein the substance is administered subcutaneously, intramuscularly, or intraperitoneally.  
     
     
         75 . The method of  claim 74 , wherein the substance is administered subcutaneously.  
     
     
         76 . The method of  claim 53 , wherein the subject has type I diabetes.  
     
     
         77 . The method of  claim 53 , wherein the subject has type II diabetes.  
     
     
         78 . The method of  claim 53 , wherein the administration of the substance is discontinued for at least 2 weeks after the insulin-producing cells are produced.  
     
     
         79 . The method of  claim 53 , wherein the substance is exendin-4.  
     
     
         80 . The method of  claim 53 , wherein the substance is an exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of exendin-4.  
     
     
         81 . The method of  claim 53 , wherein the substance is the fragment of exendin-4, an exendin-4 peptide, or an exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of exendin-4.  
     
     
         82 . The method of  claim 53 , wherein the non-insulin-producing cells are human pancreatic cells.  
     
     
         83 . The method of  claim 82 , wherein the human pancreatic cells are acinar cells.  
     
     
         84 . The method of  claim 53 , wherein the non-insulin-producing cells are stem cells.  
     
     
         85 . The method of  claim 84 , wherein the stem cells are human pancreatic stem cells.  
     
     
         86 . A method of differentiating non-insulin-producing cells into insulin-producing cells in a subject with beta-cell dysfunction, comprising contacting the non-insulin-producing cells for at least two days with an amount of a substance effective to induce differentiation of the non-insulin-producing cells by administering the substance to the subject to provide said contact, wherein the substance comprises a peptide selected from the group consisting of exendin-4, an exendin-4 peptide, an exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of exendin-4, and a fragment of any one of the preceding exendin-4 peptides, wherein the exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells, and wherein administration of the substance is discontinued after the subject's fasting blood glucose is improved compared to that prior to the administration of the substance.  
     
     
         87 . The method of  claim 86 , wherein the contacting is for at least seven days.  
     
     
         88 . The method of  claim 87 , wherein the administration of the substance is discontinued for at least two weeks after the subject's fasting blood glucose is improved compared to that prior to the administration of the substance.  
     
     
         89 . The method of  claim 88 , wherein the substance is administered subcutaneously.  
     
     
         90 . The method of  claim 88 , wherein the subject has type I diabetes.  
     
     
         91 . The method of  claim 90 , wherein the substance is exendin-4.  
     
     
         92 . The method of  claim 88 , wherein the subject has type II diabetes.  
     
     
         93 . The method of  claim 92 , wherein the substance is exendin-4.  
     
     
         94 . The method of  claim 88 , wherein the non-insulin-producing cells are human pancreatic cells.  
     
     
         95 . The method of  claim 88 , wherein the substance is exendin-4.  
     
     
         96 . The method of  claim 88 , wherein the substance is administered at a dose of from about 10 pmoles/kg to about 40 pmoles/kg.  
     
     
         97 . A method of differentiating non-insulin-producing cells into insulin-producing cells in a subject with beta-cell dysfunction, comprising contacting the non-insulin-producing cells for at least two days with an amount of a substance effective to induce differentiation of the non-insulin-producing cells by administering the substance to the subject to provide said contact, wherein the substance comprises a peptide selected from the group consisting of exendin-4, an exendin-4 peptide, an exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of exendin-4, and a fragment of any one of the preceding exendin-4 peptides, wherein the exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells, and wherein administration of the substance is discontinued after the subject's fasting blood insulin level is improved compared to that prior to the administration of the substance.  
     
     
         98 . The method of  claim 97 , wherein the contacting is for at least seven days.  
     
     
         99 . The method of  claim 98 , wherein the administration of the substance is discontinued for at least two weeks after the subject's fasting blood insulin level is improved compared to that prior to the administration of the substance.  
     
     
         100 . The method of  claim 99 , wherein the substance is administered subcutaneously.  
     
     
         101 . The method of  claim 99 , wherein the subject has type I diabetes.  
     
     
         102 . The method of  claim 101 , wherein the substance is exendin-4.  
     
     
         103 . The method of  claim 99 , wherein the subject has type II diabetes.  
     
     
         104 . The method of  claim 103 , wherein the substance is exendin-4.  
     
     
         105 . The method of  claim 99 , wherein the non-insulin-producing cells are human pancreatic cells.  
     
     
         106 . The method of  claim 99 , wherein the substance is exendin-4.  
     
     
         107 . The method of  claim 99 , wherein the substance is administered at a dose of from about 10 pmoles/kg to about 40 pmoles/kg.  
     
     
         108 . A method of differentiating non-insulin-producing cells into insulin-producing cells in a subject with beta-cell dysfunction, comprising contacting the non-insulin-producing cells for at least three days with an amount of a substance effective to induce differentiation of the non-insulin-producing cells by administering the substance to the subject to provide said contact, wherein the substance comprises a peptide selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding GLP-1 peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells, and wherein administration of the substance is discontinued after insulin-producing cells are produced.  
     
     
         109 . The method of  claim 108 , wherein the contacting is for at least seven days.  
     
     
         110 . The method of  claim 109 , wherein the substance is administered subcutaneously.  
     
     
         111 . The method of  claim 109 , wherein the subject has type I diabetes.  
     
     
         112 . The method of  claim 109 , wherein the subject has type II diabetes.  
     
     
         113 . The method of  claim 109 , wherein the non-insulin-producing cells are human pancreatic cells.  
     
     
         114 . The method of  claim 109 , wherein the administration of the substance is discontinued for at least 2 weeks after the insulin-producing cells are produced.  
     
     
         115 . The method of  claim 114 , wherein the administration of the substance is discontinued for at least 2 weeks after the insulin-producing cells are produced and the subject's fasting blood glucose or the subject's fasting blood insulin level is improved compared to that prior to administration of the substance.  
     
     
         116 . The method of  claim 115 , wherein the subject has type I diabetes.  
     
     
         117 . The method of  claim 115 , wherein the subject has type II diabetes.  
     
     
         118 . The method of  claim 109 , wherein the substance is a GLP-1 peptide.  
     
     
         119 . The method of  claim 109 , wherein the substance is a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.  
     
     
         120 . The method of  claim 109 , wherein the substance is the fragment of a GLP-1 peptide, or a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.