Compositions and methods for producing and administering brown adipocytes

Patent metadata

Field	Value
Publication number	US-10150948-B2
Application number	US-201415036297-A
Country	US
Kind code	B2
Filing date	Dec 10, 2014
Priority date	Dec 11, 2013
Publication date	Dec 11, 2018
Grant date	Dec 11, 2018

Abstract

The present disclosure provides compositions comprising a hydrogel and a cell adhesion ligand that enhances the differentiation of mesenchymal stem cells into brown adipocytes, and/or enhances the maintenance of brown adipocytes. In some cases, a subject composition includes cells that are embedded within the hydrogel. The present disclosure also provides methods of making and using the subject compositions.

First claim

What is claimed is:

1. A method of increasing the number of brown adipocytes in an individual, the method comprising implanting into the individual a composition comprising:
a) a hydrogel;
b) a cell adhesion ligand embedded within the hydrogel, wherein the cell adhesion ligand enhances at least one of: (i) the differentiation of adipose derived mesenchymal stem cells (ADMSCs) into brown adipocytes; and (ii) the maintenance of brown adipocytes; and
c) ADMSCs embedded within the hydrogel, wherein at least 10% of the ADMSCs differentiate into brown adipocytes, thereby increasing the number of brown adipocytes in the individual.

2. The method of claim 1 , wherein said implanting provides for one or more of:
a) reduction of blood glucose to within a normal range;
b) increased insulin sensitivity;
c) increased metabolic rate;
d) weight loss; and
e) supplementation of endogenous heat production.

3. The method of claim 1 , wherein said implanting comprises injecting the composition into the individual prior to gelation of the hydrogel.

4. The method of claim 1 , wherein the ADMSCs are autologous ADMSCs obtained from the individual.

5. The method of claim 1 , wherein the cell adhesion ligand is a syndecan-1 ligand or an α 5 β 1/3 integrin ligand.

6. The method of claim 1 , wherein the cell adhesion ligand comprises the amino acid sequence RKRLQVQLSIRT.

7. The method of claim 1 , wherein the cell adhesion ligand comprises the amino acid sequence KAFDITYVRLKF.

8. The method of claim 1 , wherein the cell adhesion ligand has a length of from about 10 amino acids to about 100 amino acids.

9. The method of claim 1 , wherein the hydrogel comprises hyaluronic acid.

10. The method of claim 1 , further comprising enhancing the differentiation of ADMSCs to into brown adipocytes or enhancing the maintenance of brown adipocytes with an enhancement compound.

11. The method of claim 1 , wherein the hydrogel has a modulus in a range of from 5 pascals (Pa) to 500 kilopascals (kPa).

12. The method of claim 1 , wherein the number of ADMSCs embedded within the hydrogel is in a range of from 1×10 5 to 1×10 10 .

13. The method of claim 1 , wherein the hydrogel comprises a factor release molecule and/or a cleavable cross-linker.

14. A method of inducing adipose derived mesenchymal stem cells (ADMSCs) to differentiate into brown adipocytes, the method comprising culturing ADMSCs with a composition under conditions and for a time period that provide for differentiation of at least 30% of the ADMSCs into brown adipocytes, wherein the composition comprises:
a) a hydrogel; and
b) a cell adhesion ligand embedded within the hydrogel, wherein the cell adhesion ligand enhances at least one of: (i) the differentiation of adipose derived mesenchymal stem cells (ADMSCs) into brown adipocytes; and (ii) the maintenance of brown adipocytes.

15. The method of claim 14 , wherein said culturing occurs in vitro or in vivo.

16. The method of claim 14 , wherein said method comprises, prior to culturing the ADMSCs with said composition, culturing the ADMSCs in the absence of the hydrogel in a first differentiation medium for a period of time in a range of from 6 hours to 7 days, in the presence of the cell adhesion ligand,
wherein the first differentiation medium comprises at least one enhancement compound that enhances at least one of: (i) the differentiation of ADMSCs to into brown adipocytes; and (ii) the maintenance of brown adipocytes.

17. The method of claim 14 , wherein the cell adhesion ligand is a syndecan-1 ligand or an α 5 β 1/3 integrin ligand.

18. The method of claim 14 , wherein the cell adhesion ligand comprises the amino acid sequence RKRLQVQLSIRT.

19. The method of claim 14 , wherein the cell adhesion ligand comprises the amino acid sequence KAFDITYVRLKF.

20. The method of claim 14 , wherein the cell adhesion ligand has a length of from about 10 amino acids to about 100 amino acids.

21. The method of claim 14 , wherein the hydrogel comprises hyaluronic acid.

22. The method of claim 14 , wherein the hydrogel has a modulus in a range of from 5 pascals (Pa) to 500 kilopascals (kPa).

23. The method of claim 14 , wherein the number of ADMSCs embedded within the hydrogel is in a range of from 1×10 5 to 1×10 10 .

24. The method of claim 14 , wherein the hydrogel comprises a factor release molecule and/or a cleavable cross-linker.

25. The method of claim 16 , wherein the enhancement compound is selected from the group consisting of: vascular endothelial growth factor (VEGF), bone morphogetnic protein 7 (BMP7), sonic hedgehog (SHH), brain natriuretic peptide (BNP), brain-derived neurotrophic factor (BDNF), fibroblast growth factor 21 (FGF21), transforming growth factor, beta (TGFβ), wingless-type MMTV integration site family, member 10B (Wnt10b), 1-methyl-3-isobutylxanthine (IBMX), insulin, 3,3′,5 Triiodothyronine (T3), dexamethasone, rosiglitazone, and a combination thereof.

26. A method of increasing the number of brown adipocytes in an individual, the method comprising:
(a) culturing adipose derived mesenchymal stem cells (ADMSCs) in the absence of a hydrogel in a first differentiation medium for a period of time in a range of from 6 hours to 7 days, in the presence of a cell adhesion ligand, wherein the first differentiation medium comprises at least one enhancement compound that enhances at least one of: (i) the differentiation of ADMSCs to into brown adipocytes; and (ii) the maintenance of brown adipocytes, thereby producing a population of primed ADMSCs;
(b) mixing cells of the population of primed ADMSCs with a composition that includes a hydrogel and a cell adhesion ligand that enhances at least one of: (i) the differentiation of adipose derived mesenchymal stem cells (ADMSCs) into brown adipocytes; and (ii) the maintenance of brown adipocytes, thereby producing a cell-loaded loaded hydrogel; and
(c) implanting the cell-loaded hydrogel into the individual.

Assignees

• Univ California

Inventors

• Stahl Andreas
• Tharp Kevin
• Healy Kevin E
• Jha Amit Kumar
• Kraiczy Judith

Classifications

• C12N5/0653 Primary

  Adipocytes; Adipose tissue · CPC title

• C12N2506/1384

  from adipose-derived stem cells [ADSC], from adipose stromal stem cells · CPC title

• C12N2501/734

  Proteases (EC 3.4.) · CPC title

• C12N2533/80

  Hyaluronan · CPC title

• A61K35/35

  Fat tissue; Adipocytes; Stromal cells; Connective tissues (adipose-derived stem cells A61K35/28; collagen A61K38/39) · CPC title