Endogenous neogenesis of beta cells
View Patent ↗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.
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.