Inhibition of serotonin expression in gut enteroendocrine cells results in conversion to insulin-positive cells

• US 10,487,314 B2
• Filed: 06/26/2015
• Issued: 11/26/2019
• Est. Priority Date: 06/26/2014
• 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: Accili, Domenico, Bouchi, Ryotaro
• Primary Examiner(s): Whiteman, Brian
• Application Number:
  US15/321,504
• Publication Number:
  US 20170204375A1
• Time in Patent Office: 1,614 Days
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 35/38 Stomach; Intestine; Goblet cells; Oral mucosa; Saliva
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61L 2430/22 for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/3813 Epithelial cells, e.g. keratinocytes, urothelial cells
• CPC Class Codes: A61L 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3882 Hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 16/40 against enzymes
• 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 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/12 catalytic nucleic acids, e.g. ribozymes
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2501/65 : MicroRNA
• CPC Class Codes: C12N 2501/825 : Serotonine (5-HT); Melatonine
• CPC Class Codes: C12N 2506/23 : from cells of the gastro-intestinal tract
• CPC Class Codes: C12N 2506/45 : from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0679 : Cells of the gastro-intestinal tract
• CPC Class Codes: C12Y 114/16004 : Tryptophan 5-monooxygenase (1.14.16.4)
• CPC Class Codes: C12Y 401/01028 : Aromatic-L-amino-acid decarboxylase (4.1.1.28), i.e. tryptophane-decarboxylase

Abstract

Disclosed herein are methods involving the targeting of 5HT biosynthesis in gut insulin-negative cells to convert them into insulin-positive cells. Also disclosed are methods for treating a disease or disorder in a mammal, preferably a human, associated with impaired pancreatic endocrine function, by administering a therapeutically effective amount of an enumerated active agent that reduces the expression, biosynthesis, signaling or biological activity of serotonin or increases its degradation, wherein administering comprises delivering the agent to Gut Ins− cells in the mammal. Other embodiments of the method are directed to therapy wherein an agent that significantly reduces FOXO1 expression, biosynthesis, signaling or biological activity or increases its degradation is administered in addition to the agent that reduces serotonin, or alternatively an agent that reduces FOXO1 expression is targeted to serotonin-positive gut enteroendocrine cells.

Claims

1. A method comprising administering to a mammal having a disease or disorder associated with impaired pancreatic endocrine function, a therapeutically effective amount of an agent selected from the group consisting of isolated small hairpin RNA (shRNA), small interfering RNA (siRNA), antisense RNA, antisense DNA, chimeric antisense DNA/RNA, and ribozymes that is sufficiently complementary to specifically bind to a gene or mRNA encoding Foxo1 protein thereby reducing the expression, biosynthesis, signaling or biological activity of Foxo1, wherein administering comprises delivering the agent to insulin-negative, serotonin-positive enteroendocrine cells in the gut (Gut-Ins−

cells) thereby producing insulin-positive cells that make and secrete biologically active insulin (Gut Ins+ cells) thereby treating the disease or disorder, and wherein the agent is targeted to serotonin-positive enteroendocrine cells.

2. The method of claim 1, wherein the insulin-positive cells are glucose-responsive cells.

3. The method of claim 1, wherein the agent is orally administered in an enteric form that releases the therapeutically effective amount in a region of the gut comprising insulin-negative, serotonin-positive enteroendocrine cells or is locally administered directly into or onto the gut region.

4. The method of claim 2, wherein the gut region is the duodenum, ileum or colon.

5. The method of claim 1, wherein the disease or disorder is selected from the group consisting of diabetes type 1, diabetes type 2, metabolic syndrome, glucose intolerance, hyperglycemia, decreased insulin sensitivity, increased fasting glucose, increased post-prandial glucose and obesity.

6. 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 tolerance, an increase in serum insulin, an increase insulin sensitivity, a decrease in fasting glucose, a decrease in post-prandial glucose, a decrease in weight gain, a decrease in fat mass, an increase in weight loss and the generation enteroendocrine cells in the gastrointestinal tract that produce and secrete insulin.

7. The method of claim 1, wherein the agent is administered orally, or parenterally or as a suppository.

8. A method for producing cells in the gut of a mammal that make and secrete biologically active insulin (Gut Ins+ cells), comprising administering to the mammal an agent selected from the group consisting of isolated small hairpin ins RNA (shRNA), small interfering RNA (siRNA), antisense RNA, antisense DNA, chimeric antisense DNA/RNA, and ribozymes that is sufficiently complementary to specifically bind to a gene or mRNA encoding Foxo1 protein thereby reducing the expression, biosynthesis, signaling or biological activity of Foxo1, wherein administering comprises delivering the agent to insulin-negative cells in the gut (Gut-Ins−

cells) comprising serotonin-positive, insulin-negative enteroendocrine cells thereby producing gut cells that make and secrete biologically active insulin (Gut Ins+ cells), wherein the agent is targeted to serotonin-positive enteroendocrine cells.

9. The method of claim 8, wherein the gut+cells are glucose responsive.