Methods for producing enteroendocrine cells that make and secrete insulin

• US 9,457,079 B2
• Filed: 05/12/2011
• Issued: 10/04/2016
• Est. Priority Date: 05/12/2010
• Status: Active Grant

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Talchai, Chutima, Accili, Domenico
• Primary Examiner(s): Gangle, Brian J
• Assistant Examiner(s): MCCOLLUM, ANDREA K
• Application Number:
  US13/697,792
• Publication Number:
  US 20130216554A1
• Time in Patent Office: 1,972 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12N 5/068 Stem cells; Progenitors
• CPC Class Codes: G01N 2333/62 Insulins
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5073 Stem cells

Abstract

Methods are described for producing enteroendocrine cells that make and secrete insulin in a mammal by blocking the expression or biological activity of one or more Foxo proteins or biologically active fragments or variants thereof.

Claims

1. A method for treating diabetes in a mammal, comprising administering to the mammal a therapeutically effective amount of an inhibitory oligonucleotide sufficiently complementary to an mRNA encoding Foxo1 to reduce its expression, wherein administering comprises delivering the inhibitory oligonucleotide to enteroendocrine progenitor cells in the mammal to produce glucose-responsive enteroendocrine cells that make and secrete insulin, and wherein the inhibitory oligonucleotide is orally administered in an enteric form so as to release the therapeutically effective amount in a gut region comprising enteroendocrine progenitor cells or is locally administered directly into or onto the gut region.

2. The method of claim 1, wherein the inhibitory oligonucleotide is selected from the group consisting of an antisense RNA, an antisense DNA and a chimeric antisense DNA/RNA.

3. The method of claim 1, wherein the inhibitory oligonucleotide is selected from the group consisting of small interfering RNA (siRNA) and small hairpin RNA (shRNA).

4. The method of claim 1, wherein the diabetes is diabetes type 1.

5. The method of claim 1, wherein the therapeutically effective amount is an amount that produces an effect selected from the group consisting of an increase in glucose responsiveness, an increase in serum insulin and the generation of enteroendocrine cells in the gastrointestinal tract that produce and secrete insulin.

6. The method of claim 1, wherein the gut region is the duodenum, ileum or colon.

7. A method for producing enteroendocrine cells that make and secrete insulin in a mammal, comprising administering to the mammal an inhibitory oligonucleotide sufficiently complementary to mRNA encoding Foxo1 to reduce its expression, wherein administering comprises delivering the inhibitory oligonucleotide to enteroendocrine progenitor cells in the mammal in an amount to produce glucose-responsive enteroendocrine cells that make and secrete insulin, and wherein the inhibitory oligonucleotide is orally administered in an enteric form so as to release said therapeutically effective amount in a gut region of the mammal that comprises enteroendocrine progenitor cells or is locally administered directly into or onto the gut region.

8. The method of claim 7, wherein the inhibitory oligonucleotide is selected from the group consisting of an antisense RNA, an antisense DNA and a chimeric antisense DNA/RNA.

9. The method of claim 7, wherein the inhibitory oligonucleotide is selected from the group consisting of small interfering RNA (siRNA) and small hairpin RNA (shRNA).

10. The method of claim 7, wherein the insulin-producing enteroendocrine cells further produce one or more pancreatic hormones selected from the group consisting of glucagon, pancreatic polypeptide, glucokinase, and glucose transport 2 (glut2), in response to administration of the inhibitory oligonucleotide.

11. The method of claim 7, wherein the insulin-producing enteroendocrine cells also produce one or more proteins selected from the group consisting of prohormone-convertase 2 (Pc2), pancreatic and duodenal homeobox1 (Pdx1), MafA, Nkx6.1, Nkx2.2, and paired box gene 4 (Pax4).

12. The method of claim 7, wherein the gut region is the duodenum, ileum or colon.

13. A method for making insulin-producing enteroendocrine cells comprising

a) isolating a population of non-insulin producing enteroendocrine progenitor cells from a gut region of a mammal,

b) contacting the population with an oligonucleotide sufficiently complementary to mRNA encoding Foxo1 to reduce its expression in an amount and under conditions that permit a portion of the population to produce insulin in a glucose-responsive manner, and

c) collecting the insulin-producing enteroendocrine cells.

14. The method of claim 13, wherein the inhibitory oligonucleotide is selected from the group consisting of an antisense RNA, antisense DNA, and a chimeric antisense DNA/RNA.

15. The method of claim 13, wherein the inhibitory oligonucleotide is selected from the group consisting of small interfering RNA (siRNA) and small hairpin RNA (shRNA).

16. The method of claim 13, wherein the population is isolated from a segment of the intestine or colon.

17. The method of claim 16, wherein the segment is taken from the duodenum or ileum and wherein the segment comprises crypts.

18. The method of claim 13, further comprising reintroducing the insulin-producing enteroendocrine cells into the mammal in sufficient numbers to treat or prevent a disease or disorder.

19. The method of claim 13, wherein the gut region is the duodenum, ileum or colon.

20. A method for treating Type I diabetes, the method comprising administering to the mammal a therapeutically effective amount of an inhibitory oligonucleotide that is sufficiently complementary to Foxo1 to reduce its expression, wherein administering comprises delivering the inhibitory oligonucleotide to enteroendocrine progenitor cells in the mammalian gut.

21. The method of claim 20, wherein the inhibitory oligonucleotide is delivered by oral administration in an enteric form that releases the therapeutically effective amount onto a gut region comprising duodenum, ileum and/or colon or by local administration into or onto the duodenum, ileum and/or colon.

22. The method of claim 20, wherein the inhibitory oligonucleotide is selected from the group consisting of an isolated small hairpin RNA (shRNA), small interfering RNA (siRNA), antisense RNA, antisense DNA, and chimeric antisense DNA/RNA.

23. A method for treating diabetes, the method comprising administering to the mammal a therapeutically effective amount of an inhibitory oligonucleotide that is sufficiently complementary to Foxo1 to reduce its expression, wherein administering comprises delivering the inhibitory oligonucleotide to enteroendocrine progenitor cells in the mammalian gut by oral administration in an enteric form that releases the therapeutically effective amount onto a gut region comprising duodenum, ileum and/or colon or by local administration into or onto the duodenum, ileum and/or colon.