Therapeutic immune modulation by stem cell secreted exosomes

• US 9,427,450 B2
• Filed: 01/31/2013
• Issued: 08/30/2016
• Est. Priority Date: 01/31/2012
• Status: Active Grant

• Current Assignee: Xon Cells, Inc (Precigen, Inc.)
• Original Assignee: Xon Cells, Inc (Precigen, Inc.)
• Inventors: Ichim, Thomas, Bogin, Vladimir
• Primary Examiner(s): BELYAVSKYI, MICHAIL A
• Application Number:
  US13/756,310
• Publication Number:
  US 20130195899A1
• Time in Patent Office: 1,307 Days
• Field of Search: None
• 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 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 39/0008 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• 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

Abstract

Disclosed are methods, compositions of matter, and protocols useful for the induction of a therapeutic immune modulatory response through administration of exosomes derived from a stem cell source. In one embodiment, said stem cell source is endometrial regenerative cells. Specifically, in one embodiment stem cell derived exosomes are used as a method of treating an autoimmune condition such as rheumatoid arthritis, multiple sclerosis, or systemic lupus erythromatosis.

Claims

1. A method of enhancing stem cell engraftment in a mammal comprising the steps of:

a) extracting a stem cell population;

b) culturing said stem cell population for a sufficient time point, under conditions permissible for exosome production;

c) collecting exosomes generated by said stem cell population;

d) concentrating said exosomes; and

e) administering said exosomes and said stem cells to said mammal.

2. The method of claim 1, wherein said stem cells are endometrial regenerative cells (ERC).

3. The method of claim 1, wherein said stem cells are induced to produce an increased amount of exosomes.

4. The method of claim 3, wherein said induction of exosomes from said stern cells is achieved by treatment of said stem cells with cytokines.

5. The method of claim 1, wherein said exosomes are concentrated by a means selected from the group consisting of a) ultracentrifugation;

b) column chromatography;

c) size exclusion; and

d) filtration through a device containing an affinity matrix selective towards exosomes.

6. The method of claim 1, wherein said exosomes are transfected with a genetic material so as to allow delivery of said genetic material to an immune cell.

7. The method of claim 1, wherein said cells producing said exosomes are transferred with a genetic material so as to allow delivery of said genetic material to an immune cell.

8. A method of enhancing stem cell engraftment in a mammal comprising

a) extracting a stem cell population;

b) culturing said stem cell population for a sufficient time point, under conditions permissible for exosome production;

c) collecting exosomes generated by said stem cell population;

d) concentrating said exosomes to at least 1 μ

g/mL; and

e) administering 1-2 μ

g of said exosomes and at least 100,000 of said stem cells to the mammal.

9. The method of claim 8, wherein said stem cells are endometrial regenerative cells (ERC).

10. The method of claim 8, further comprising incorporating one or more autoantigens into said exosomes generated from said stem cells.

11. The method of claim 8, further comprising transfecting the stem cell population with one or more autoantigens representing said autoimmune condition.

12. A method of treating a cardiac enhancing stem cell engraftment in a human patient in need of therapy comprising:

a) obtaining a mesenchymal stem cell;

b) introducing into said mesenchymal stem cell an antigen, wherein said antigen is present in one or more exosomes released by said mesenchymal stem cell;

c) purifying said exosomes and concentrating said exosomes to at least 1 μ

g/mL;

d) administering an amount of said exosomes and an amount of said stem cells into the human patient in need of therapy effective to inhibit T cell proliferation.

13. The method of claim 12, wherein said mesenchymal stem cell is an endometrial regenerative cell (ERC).

14. The method of claim 13 wherein ERC are cultured in a DMEM F12 medium comprising approximately 10% fetal calf serum for a period of 1-100 hours.

15. The method of claim 12, wherein said exosomes are concentrated by a means selected from:

a) ultracentrifugation;

b) column chromatography;

c) size exclusion; and

d) filtration through a device containing an affinity matrix selective towards exosomes.

16. The method of claim 12, wherein said antigen is incorporated into said exosome-producing mesenchymal stem cell by one or more of a) transfection with the gene encoding said antigen;

b) transfection with said antigenic protein; and

c) transfection with said antigenic peptide.

17. The method of claim 12, wherein said antigen is loaded onto said exosomes.

18. The method of claim 17, wherein said loading is accomplished through chemical means.