Universal donor stem cells and related methods

Patent metadata

Field	Value
Publication number	US-12031154-B2
Application number	US-202217993674-A
Country	US
Kind code	B2
Filing date	Nov 23, 2022
Priority date	May 8, 2015
Publication date	Jul 9, 2024
Grant date	Jul 9, 2024

Abstract

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and/or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

First claim

What is claimed is:

1. A genetically modified beta cell comprising:
reduced cell surface expression of one or more MHC-I human leukocyte antigen molecules and/or one or more MHC-II human leukocyte antigen molecules relative to an unmodified beta cell; and
increased cell surface expression of one or more tolerogenic factors relative to an unmodified beta cell, wherein a nucleic acid encoding the one or more tolerogenic factors is inserted into at least one allele of a safe harbor locus of the genetically modified beta cell.

2. The genetically modified beta cell of claim 1 , wherein the safe harbor locus comprises an AAVS1 locus.

3. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors inhibit an immune response when the genetically modified beta cell is administered to a subject.

4. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise CD47.

5. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise HLA-C.

6. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise HLA-E.

7. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise HLA-G.

8. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise PD-L1.

9. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise CTLA-4-Ig.

10. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise C1-inhibitor.

11. The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors comprise IL-35.

12. The genetically modified beta cell of claim 1 , comprising one or more indels in one or more genes encoding an MHC-I human leukocyte antigen molecule, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

13. The genetically modified beta cell of claim 12 , comprising one or more indels in an HLA-A gene, an HLA-B gene, an HLA-C gene, or a combination thereof in the genome of the genetically modified beta cell, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

14. The genetically modified beta cell of claim 13 , further comprising one or more indels in a class II major histocompatibility complex transactivator (CIITA) gene in the genome of the genetically modified beta cell, thereby resulting in the reduced cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

15. The genetically modified beta cell of claim 1 , comprising one or more indels in one or more genes encoding a transcriptional regulator of an MHC-I human leukocyte antigen molecule, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

16. The genetically modified beta cell of claim 1 , comprising one or more indels in a CIITA gene, a β2M gene, a TAPI gene, an NLRC5 gene, an RFX5 gene, an RFXAP gene, an RFXANK gene, an NFY-A gene, an NFY-B gene, an NFY-C gene, an IRF-1 gene, or a combination thereof, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules and/or the one or more MHC-II human leukocyte antigen molecules.

17. The genetically modified beta cell of claim 16 , comprising one or more indels in a class II major histocompatibility complex transactivator (CIITA) gene in the genome of the genetically modified beta cell, thereby resulting in the reduced cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

18. The genetically modified beta cell of claim 17 , wherein the one or more indels comprises a CIITA gene knock out.

19. The genetically modified beta cell of claim 16 , comprising one or more indels in a β2M gene in the genome of the genetically modified beta cell, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

20. The genetically modified beta cell of claim 19 , wherein the one or more indels comprises a β2M gene knock out.

21. The genetically modified beta cell of claim 1 , wherein the genetically modified beta cell is a CIITA −/− genetically modified beta cell.

22. The genetically modified beta cell of claim 1 , wherein the genetically modified beta cell is a β2M −/− genetically modified beta cell.

23. The genetically modified beta cell of claim 1 , wherein the genetically modified beta cell is a β2 −/− CIITA −/− genetically modified beta cell.

24. The genetically modified beta cell of claim 1 , wherein the genetically modified beta cell is derived from a stem cell.

25. The genetically modified beta cell of claim 24 , wherein the stem cell is an embryonic stem cell.

26. The genetically modified beta cell of claim 24 , wherein the stem cell is an induced pluripotent stem cell.

27. A composition comprising the genetically modified beta cell of claim 1 .

28. The genetically modified beta cell of claim 1 , wherein cell surface expression of the one or more MHC-I human leukocyte antigen molecules and/or the one or more MHC-II human leukocyte antigen molecules is eliminated.

29. The composition of claim 27 , wherein cell surface expression of the one or more MHC-I human leukocyte antigen molecules and/or the one or more MHC-II human leukocyte antigen molecules is eliminated.

30. The genetically modified beta cell of claim 1 , comprising one or more indels in one or more genes encoding an MHC-II human leukocyte antigen molecule in the genome of the genetically modified beta cell, thereby resulting in the reduced cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

31. The genetically modified beta cell of claim 1 , comprising one or more indels in one or more genes encoding a transcriptional regulator of an MHC-II human leukocyte antigen molecule, thereby resulting in the reduced cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

32. The composition of claim 27 , wherein the genetically modified beta cell is a β2M −/− genetically modified beta cell.

Assignees

• Harvard College

Inventors

• Meissner Torsten B
• Ferreira Leonardo M R
• Strominger Jack L
• Cowan Chad A

Classifications

• C12N2501/50

  Cell markers; Cell surface determinants · CPC title

• A61K39/001 Primary

  Preparations to induce tolerance to non-self, e.g. prior to transplantation · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C12N2501/998

  Proteins not provided for elsewhere · CPC title

• C12N2501/599

  with CD designations not provided for elsewhere · CPC title