Endogenous neogenesis of beta cells

• US 10,071,172 B2
• Filed: 04/17/2015
• Issued: 09/11/2018
• Est. Priority Date: 04/23/2014
• Status: Active Grant

• Current Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Inventors: Gittes, George, Guo, Ping, Xiao, Xiangwei
• Primary Examiner(s): HILL, KEVIN KAI
• Application Number:
  US15/305,083
• Publication Number:
  US 20170087254A1
• Time in Patent Office: 1,243 Days
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 48/0058 Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
• CPC Class Codes: A61K 48/0075 characterised by an aspect of the delivery route, e.g. oral, subcutaneous
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Methods are disclosed for producing pancreatic beta cells in a subject. The methods include administering to the subject a vector encoding heterologous Pancreas duodenal homeobox protein (Pdx) 1 and MafA, wherein the vector does not encode Neurogenin 3 (Ngn3) and wherein the subject is not administered any other nucleic acid encoding Ngn3. The vector is administered intraductally into a pancreatic duct of the subject. Compositions are disclosed that include a) a viral vector comprising a promoter operably linked to a nucleic acids encoding Pdx1 and a nucleic acid encoding MafA, wherein the vector does not encode Ngn3; b) a buffer; and c) a contrast dye for endoscopic retrograde cholangiopancreatography. These compositions are of use in any of the methods disclosed herein.

Claims

1. A method of producing pancreatic beta cells in a subject, comprising

administering to the subject a vector comprising a glucagon promoter operably linked to a nucleic acid sequence encoding heterologous Pancreas duodenal homeobox protein (Pdx) 1 and a nucleic acid sequence encoding Musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), wherein the vector does not encode Neurogenin 3 (Ngn3), wherein the subject is not administered any other nucleic acid encoding Ngn3, wherein the glucagon promoter a) consists of the nucleic acid sequence of SEQ ID NO:

1, or b) comprises the nucleic acid sequence of SEQ ID NO;

3, and wherein the vector is administered intraductally into a pancreatic duct of the subject,

thereby inducing alpha cells to transform into pancreatic beta cells in the subject.

2. The method of claim 1, wherein the vector is an adenovirus vector or an adeno-associated virus vector.

3. The method of claim 2, wherein the vector is an adeno-associated virus vector, and wherein the adeno-associated virus vector is an adeno-associated virus 8 vector.

4. The method of claim 2, wherein the vector is an adeno-associated virus vector.

5. The method of claim 1, wherein the glucagon promoter consists of the nucleic acid sequence of SEQ ID NO:

1.

6. The method of claim 1, wherein the glucagon promoter comprises the nucleic acid sequence of SEQ ID NO:

3.

7. The method of claim 4, wherein the nucleic acid sequence encoding Pdx1 and the nucleic acid sequence encoding MafA are linked using a connector.

8. The method of claim 7, wherein the connector is a 2A connector.

9. The method of claim 2, wherein intraductally administering comprises the use of endoscopic retrograde cholangiopancreatography (ERCP).

10. The method of claim 2, wherein the subject is not administered an immunosuppressive agent.

11. The method of claim 1, wherein the subject is a human.

12. The method of claim 1, wherein the subject has type I diabetes.

13. A composition comprising:

a) an adeno-associated virus vector comprising a glucagon promoter operably linked to a nucleic acid sequence encoding Pdx1 and a nucleic acid sequence encoding MafA, wherein the vector does not encode Neurogenin 3 (Ngn3), and wherein the glucagon promoter a) consists of the nucleic acid sequence of SEQ ID NO;

1, or b) comprises the nucleic acid sequence of SEQ ID NO;

3;

b) a buffer; and

c) a contrast dye for endoscopic retrograde cholangiopancreatography.

14. The composition of claim 13, wherein the composition does not comprise a nucleic acid encoding Ngn3 or Ngn3 polypeptide.

15. The composition of claim 13, wherein the glucagon promoter comprises the nucleic acid sequence set forth as SEQ ID NO:

3.

16. The composition of claim 13, wherein the contrast dye is a low-osmolar low-viscosity non-ionic dye, a low-viscosity high-osmolar dye, or a dissociable high-viscosity dye.

17. The composition of claim 16, wherein the contrast dye is Iopromid, Ioglicinate, or Ioxaglinate.

18. The composition of claim 13, wherein the adeno-associated virus vector is an adeno-associated virus 8 vector.

19. The composition of claim 13, wherein the nucleic acid sequence encoding Pdx1 and the nucleic acid sequence encoding MafA are linked using a connector.

20. The composition of claim 19, wherein the connector is a 2A connector.

21. The composition of claim 13, wherein the adeno-associated virus vector comprises a nucleic acid sequence encoding a label.

22. The composition of claim 13, wherein the glucagon promoter consists of the nucleic acid sequence set forth as SEQ ID NO:

1.