Methods for producing hormone secreting cells in a subject

• US 10,376,545 B2
• Filed: 04/03/2017
• Issued: 08/13/2019
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Agulnick, Alan D., D'Amour, Kevin Allen
• Primary Examiner(s): Fox, Allison M
• Application Number:
  US15/478,094
• Publication Number:
  US 20170202885A1
• Time in Patent Office: 862 Days
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/195 Heregulin, neu differentiation factor
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0012 : Cell encapsulation
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells

Abstract

A human immature endocrine cell population and methods for making an immature endocrine cell population are provided. Specifically, immature beta cells and methods for production of immature beta cells are described. Immature beta cells co-express INS and NKX6.1 and are uni-potent and thereby develop into mature beta cells when implanted in vivo. The mature beta cells in vivo are capable of producing insulin in response to glucose stimulation.

Claims

1. A method for producing hormone secreting cells, said method comprising:

maturing transplanted endocrine cells in vivo in a subject wherein the endocrine cells co-express INS+ and NKX6.1+, thereby producing hormone secreting cells.

2. The method of claim 1, wherein the endocrine cells are CHGA+.

3. The method of claim 1, wherein the hormone secreting cells secrete insulin and glucagon.

4. The method of claim 1, wherein the subject is a mammalian subject.

5. The method of claim 4, wherein the mammalian subject is a human subject.

6. The method of claim 5, wherein the human subject has been identified as having a condition which limits the ability of the subject to produce sufficient levels of insulin in response to physiologically high blood glucose concentrations.

7. The method of claim 1, further comprising detecting insulin secretion, wherein insulin secretion is detected by measuring C-peptide levels in a subject into whom the endocrine cells have been transplanted.

8. The method of claim 7, wherein C-peptide levels in the subject increase over time.

9. The method of claim 1, wherein the hormone secreting cells are insulin secreting cells that are responsive to blood glucose.

10. The method of claim 1, wherein the number of hormone-secreting cells increases over time.

11. The method of claim 1, wherein the hormone secreting cells are insulin secreting cells that form islet-like cell clusters.

12. The method of claim 1, wherein the endocrine cells are unipotent.

13. The method of claim 1, wherein the endocrine cells are aggregates.

14. The method of claim 13, wherein the aggregates are 50 to 300 microns in diameter.

15. The method of claim 1, wherein the endocrine cells are in a semipermeable encapsulation device.

16. The method of claim 1, wherein the endocrine cells do not substantially express NGN3.

17. A method for producing hormone secreting cells, said method comprising:

maturing transplanted human cells in vivo, wherein the transplanted human cells comprise unipotent human immature beta cell suspension aggregates and wherein at least 10% of cells in the beta cell suspension aggregates are human immature beta cells that co-express INS and NKX6.1,

thereby producing hormone secreting cells.

18. The method of claim 17, wherein the hormone secreting cells secrete insulin.