Induction of a Beta Cell Differentiation in Human Cells

Abstract

The present invention provides methods for inducing insulin gene expression in cultured pancreas cells, the method comprising contacting a culture of endocrine pancreas cells expressing a PDX-1 gene and a NeuroD/BETA2 gene with a GLP-1 receptor agonist, wherein the cells have been cultured under conditions such that the cells are in contact with other cells in the culture, thereby inducing insulin gene expression in the cells. The invention also provides high throughput screening methods for modulators of β-cell function, stable cultures of cells made by the methods of the invention, and methods of treating a human subject using the methods of the invention.

1 . A method for inducing β-cell function in a cultured cell, the method comprising the steps of: 
 (i) expressing a recombinant PDX-1 polynucleotide in a cells that have been cultured under conditions such that the cells are in contact with other cells in the culture; and    (ii) contacting the cell with a GLP-1 receptor agonist, thereby inducing β-cell function in the cells.    
   
   
       2 . The method of  claim 1 , further comprising expressing a recombinant NeuroD/BETA2 polynucleotide in the cells.  
   
   
       3 . The method of  claim 1 , wherein the β-cell function comprises insulin gene expression.  
   
   
       4 . The method of  claim 1 , wherein the β-cell function comprises insulin production.  
   
   
       5 . The method of  claim 1 , wherein the β-cell function comprises glucose-responsive insulin production.  
   
   
       6 . The method of  claim 1 , wherein the GLP-1 receptor agonist is a GLP-1 analog.  
   
   
       7 . The method of  claim 1 , wherein the GLP-1 receptor agonist has an amino acid sequence of a naturally occurring peptide.  
   
   
       8 . The method of  claim 7 , wherein the GLP-1 receptor agonist is GLP-1, exendin- 3 , or exendin-4.  
   
   
       9 . The method of  claim 1 , wherein the cells are cultured as aggregates in suspension.  
   
   
       10 . The method of  claim 1 , wherein the cells are endocrine pancreas β-cells.  
   
   
       11 . The method of  claim 1 , wherein the cells are βlox5 cells.  
   
   
       12 . The method of  claim 1 , wherein the cells are human cells.  
   
   
       13 . The method of  claim 1 , wherein the cells express a recombinant oncogene.  
   
   
       14 . The method of  claim 13 , wherein the cells expresses more than one recombinant oncogene.  
   
   
       15 . The method of  claim 1 , wherein the cell expresses a recombinant telomerase gene.