Induction of beta cell differentiation in human cells

• US 20050214269A1
• Filed: 05/09/2005
• Published: 09/29/2005
• Est. Priority Date: 10/18/2001
• Status: Abandoned Application

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Itkin-Ansari, Pamela, Levine, Fred, Dufayet, Dominique
• Application Number:
  US11/125,716
• Publication Number:
  US 20050214269A1
• Time in Patent Office: Days
• US Class Current: 424/93.210
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 5/0676 Pancreatic 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.

Claims

1-35. -35. (canceled)

36. A method of identifying a compound that modulates pancreatic cell function, the method comprising:

(i) expressing a recombinant NeuroD/BETA2 polynucleotide and a recombinant PDX-1 polynucleotide in pancreatic cells that have been cultured under conditions such that the cells are in contact with other cells in the culture; and

(ii) contacting the cells with a GLP-1 receptor agonist, thereby inducing insulin gene expression in the cells;

(iii) contacting the cells with a test compound; and

(iv) determining the effect of the test compound on pancreatic cell function to identify a compound that modulates pancreatic cell function.

37. The method of claim 36, wherein the pancreatic cell function comprises insulin production.

38. The method of claim 37, wherein the pancreatic cell function comprises glucose-responsive insulin production.

39. The method of claim 36, wherein the GLP-1 receptor agonist is a GLP-1 analog.

40. The method of claim 36, wherein the GLP-1 receptor agonist has an amino acid sequence of a naturally occurring peptide.

41. The method of claim 36, wherein the GLP-1 receptor agonist is GLP-1, exendin-3, or exendin-4.

42. The method of claim 36, wherein the cells are cultured as aggregates in suspension.

43. The method of claim 36, wherein the cells are human cells.

44. The method of claim 36, wherein the cells further express a recombinant oncogene.

45. The method of claim 44, wherein the cells express more than one recombinant oncogene.

46. The method of claim 36, wherein the cells further express a recombinant polynucleotide encoding telomerase.

47. The method of claim 36, wherein the cells are β

-cells.

48. The method of claim 47, wherein the cells are β

lox5 cells.

49. The method of claim 36, wherein the cells are a stable culture of pancreatic cells.