COMPOSITIONS AND METHODS FOR TREATING TYPE 1 AND TYPE 2 DIABETES AND RELATED DISORDERS

• US 20150368667A1
• Filed: 07/07/2015
• Published: 12/24/2015
• Est. Priority Date: 03/27/2014
• Status: Active Grant

• Current Assignee: Salk Institute for Biological Studies
• Original Assignee: Salk Institute for Biological Studies
• Inventors: EVANS, RONALD M., YOSHIHARA, EIJI, DOWNES, MICHAEL R., YU, RUTH T., ATKINS, ANNETTE R.
• Granted Patent: US 9,546,379 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/70567 Nuclear receptors, e.g. retinoic acid receptor [RAR], RXR, nuclear orphan receptors
• CPC Class Codes: C07K 14/721 Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/392 Sexual steroids
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2710/10032 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/10043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0676 : Pancreatic cells

Abstract

The invention features compositions comprising in vitro generated beta cells capable of glucose-stimulated insulin secretion, methods of inducing beta cell maturation from embryonic or induced pluripotent stem cell-derived beta-like cells, and methods of using in vitro generated beta cells for the treatment of type 1 diabetes, type 2 diabetes, or a related disorder.

Claims

1. A method for reprogramming a beta-like cell to a functional beta cell or generating a cell capable of glucose-stimulated insulin secretion, the method comprising expressing recombinant estrogen-related receptor (ERR) gamma in a beta-like cell, thereby reprogramming the beta-like cell to a functional beta cell or generating a cell capable of glucose-stimulated insulin secretion.

2. The method of claim 1, wherein the beta-like cell expresses one or more cell transcription factors selected from the group consisting of Nkx2.2, NeuroD1, Foxa2, Pax6 HNF4a, Pdx1, MafA and Nkx6-1 mRNA expression.

3. The method of claim 1, wherein the beta-like cell expresses one or more cell markers selected from the group consisting of Insulin 1, Insulin 2, glucagon and somatostatin.

4. The method of claim 1, wherein the beta-like cell is an embryonic stem cell, induced pluripotent stem cell, adipocyte-derived stem cell, human umbilical vein endothelial cell (Huvec), or a progenitor or stem cell thereof.

5. The method of claim 1, wherein the beta-like cell is modified to express ERRgamma in vitro or in vivo.

6. The method of claim 1, wherein the reprogrammed cell expresses insulin and/or secretes insulin in response to glucose stimulation.

7. The method of claim 1, wherein the beta-like cell is obtained by contacting an embryonic stem cell or induced pluripotent stem cell in culture with one or more of activin A, wingless-type MMTV integration site family member 3A (Wnt3a), insulin growth factor (IGF)-2, extendin (Ex)-4, fibroblast growth factor (FGF)-2, nicotinamide, and/or B27.

8. The method of claim 7, wherein the beta-like cell is treated with one or more of the following:

50-100 ng/ml Activin A each day for 3-4 days;

25 ng/ml Wnt3a on day 0; and

/or

50 ng/ml IGF-2, Ex-4 50 ng/ml, 10 ng/ml FGF-2, 10 mM nicotinamide, and 1% B27 on day 3.

9. The method of claim 1, wherein the obtained beta cell expresses one or more mRNA selected from the group consisting of Pdx1, insulin, Mafa, Pax6, NeuroD, GCK, CHGA, VAMP2, PC1/3, Glut2, Nkx6.1, GCG, SST, and U36B4.

10. The method of claim 1, wherein the beta-like cells is obtained by expressing in the embryonic stem cell or induced pluripotent stem cell one or more transcription factors selected from the group consisting of Oct4, Nanog, Sox17, FoxA2, Pdx1, Nkx6.1, and Ngn3.

11. A method of ameliorating hyperglycemia or treating type 1 or type 2 diabetes in a subject in need thereof, the method comprising administering to the subject a beta-like cell expressing recombinant estrogen related receptor (ERR) gamma, thereby ameliorating hyperglycemia or treating type 1 or type 2 diabetes in the subject.

12. The method of claim 11, wherein the method reduces or normalizes blood glucose level in the subject.

13. An expression vector comprising a beta cell-specific promoter operably linked to a nucleic acid sequence encoding an ERRgamma polypeptide.

14. The expression vector of claim 13, wherein the promoter is selected from the group consisting of insulin, insulin II, Pdx1, Mafa, Nkx6-1, pax4 and NeuroD1 promoters.

17. A host cell comprising the expression vector of claim 13, wherein the cell is an embryonic stem cell, neonate stem cell, adult stem cell, induced pluripotent stem cell, adipocyte-derived stem cell, human umbilical vein endothelial cell (Huvec), or a progenitor or stem cell thereof.

18. A tissue, organ or matrix comprising the host cell of claim 17.

19. A cellular composition comprising an effective amount of a host cell of claim 17 in a pharmaceutically acceptable excipient.

20. A packaged pharmaceutical comprising:

a) a host cell of claim 17; and

b) instructions for using said cell to ameliorate hyperglycemia in a subject.

21. A kit for treating hyperglycemia, the kit comprising an effective amount of a host cell of claim 17, and instructions for use thereof.

15. A method of ameliorating hyperglycemia in a subject in need thereof, the method comprising contacting a beta-like cell with an AAV vector encoding ERRgamma, thereby causing the cell to be capable of glucose-stimulated insulin secretion and administering the cell to the subject, thereby ameliorating hyperglycemia in the subject.

16. The method of claim 15, wherein the beta-like cell is derived from the subject or is not derived from the subject.

22. A method for generating a cell capable of glucose-stimulated insulin secretion, the method comprising expressing recombinant estrogen-related receptor (ERR)gamma and Pdx1 or ERRgama, Pdx1, and PGC-1alpha in an human adipose-derived stem cell (hADSC), thereby generating a cell capable of glucose-stimulated insulin secretion.

23. The method of claim 22, further comprising expressing Pax4 in the hADSC.