SC-beta cells and compositions and methods for generating the same

• US 10,655,106 B2
• Filed: 07/23/2018
• Issued: 05/19/2020
• Est. Priority Date: 06/11/2013
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Peterson, Quinn P., Pagliuca, Felicia J., Melton, Douglas A., Millman, Jeffrey R., Segel, Michael Saris, Gurtler, Mads
• Primary Examiner(s): Johnson, Kara D
• Application Number:
  US16/042,917
• Publication Number:
  US 20190040362A1
• Time in Patent Office: 666 Days
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• 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 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/375 Thyroid stimulating hormone [TSH]
• 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 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• 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 2506/03 from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2506/22 from pancreatic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12N 5/0696 : Artificially induced pluripotent stem cells, e.g. iPS

Abstract

Disclosed herein are methods, compositions, kits, and agents useful for inducing β cell maturation, and isolated populations of SC-β cells for use in various applications, such as cell therapy.

Claims

1. A device comprising a population of non-native pancreatic β

cells, wherein the non-native pancreatic β

cells exhibit an in vitro glucose-stimulated insulin secretion response to a glucose challenge, wherein the non-native pancreatic β

cells do not express at least one of somatostatin and glucagon, and wherein the device is configured to produce and release insulin when implanted into a subject.

2. The device of claim 1, wherein the non-native pancreatic β

cells exhibit an in vitro glucose-stimulated insulin secretion response to a first glucose challenge, a second glucose challenge, and a third glucose challenge, when the first glucose challenge, the second glucose challenge and the third glucose challenge are applied sequentially.

3. The device of claim 1, wherein the secretion of insulin by the non-native pancreatic β

cell in response to a glucose challenge is proportional to the concentration of glucose in the glucose challenge.

4. The device of claim 1, wherein the non-native pancreatic β

cells comprise one or more crystalline insulin granules.

5. The device of claim 1, wherein the non-native pancreatic β

cells exhibit cytosolic Ca²⁺ flux in response to sequential application of a first glucose challenge and a second glucose challenge.

6. The device of claim 1, wherein the non-native pancreatic β

cells are monohormonal.

7. The device of claim 1, wherein, upon incubation in the presence of at least 20 mM of glucose in vitro, at least 0.5 μ

IU insulin is secreted per 1000 cells per 30 minute incubation.

8. The device of claim 1, wherein the non-native pancreatic β

cells have a gene expression profile that is different from the gene expression profile of a native β

cell.

9. The device of claim 1, wherein the population of non-native pancreatic β

cells comprises between 340-748 million non-native pancreatic β

cells.

10. The device of claim 1, comprising non-native pancreatic β

cells encapsulated by a biomaterial selected from the group consisting of;

alginate, polyethylene glycol (PEG), chitosan, polyester (PES) hollow fibers, collagen, hyaluronic acid, dextran with arginine, glycine, aspartic acid (RGD), 5-ethyl-5-(hydroxymethyl)-13,13-dimethyl-1,3-dioxane-2-ethanol diacrylate (EHD), poly(ethylene glycol) diacrylate (PEGDA), poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate-co-p-vinylphenylboronic acid) (PMBV), poly(vinyl alcohol) (PVA), polyglycerol sialosides (PGSAS), agarose, agarose with gelatin, poly(lactic-co-glycolic acid) (PLGA), and any combinations thereof.

11. The device of claim 1, wherein the non-native pancreatic β

cells are genetically modified.

12. The device of claim 1, further comprising alpha cells and delta cells.

13. The device of claim 1, further comprising a semipermeable membrane, wherein the semipermeable membrane is configured to retain the non-native pancreatic β

cells in the device and permit passage of insulin secreted by the non-native pancreatic β

cells.

14. The device of claim 13, wherein the non-native pancreatic β

cells are separated by the semipermeable membrane in an islet chamber from one or more vascularizing chamber of the device.

15. The device of claim 13, wherein the semipermeable membrane is made of polysaccharide or polycation.

16. The device of claim 1, wherein at least one of the non-native pancreatic β

cells is modified with PEG.

17. The device of claim 1, comprising microcarriers or microcapsules.

18. The device of claim 1, further comprising an immune modulator.

19. The device of claim 1, further comprising an extracorporeal segment.

20. The device of claim 1, further comprising a resorbable or biodegradable matrix scaffold.

21. A method of treating a subject in need thereof, comprising implanting a device into the subject, wherein the device comprises a population of non-native pancreatic β

cells, wherein the non-native pancreatic β

cells exhibit an in vitro glucose-stimulated insulin secretion response to a glucose challenge, and wherein the non-native pancreatic β

cells do not express at least one of somatostatin and glucagon.

22. The method of claim 21, wherein the subject is diabetic.

23. The method of claim 21, wherein the subject has, or has an increased risk of developing a metabolic disorder.

24. The method of claim 21, wherein the subject has diabetes selected from the group consisting of:

Type I diabetes, Type II diabetes, and Type 1.5 diabetes.