Universal donor stem cells and related methods

Patent metadata

Field	Value
Publication number	US-12421493-B2
Application number	US-202217993659-A
Country	US
Kind code	B2
Filing date	Nov 23, 2022
Priority date	May 8, 2015
Publication date	Sep 23, 2025
Grant date	Sep 23, 2025

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 cell comprising:
reduced or eliminated cell surface expression of one or more MHC-I human leukocyte antigen molecules relative to an unmodified cell of the same type; and
increased cell surface expression of one or more tolerogenic factors relative to an unmodified cell of the same type, 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 cell.

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

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

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

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

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

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

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

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

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

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

12. The genetically modified 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 or eliminated cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

13. The genetically modified 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 cell, thereby resulting in the reduced or eliminated cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

14. The genetically modified 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 or eliminated cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

15. The genetically modified cell of claim 1 , comprising one or more indels in a B2M gene in the genome of the genetically modified cell, thereby resulting in the reduced or eliminated cell surface expression of the one or more MHC-I human leukocyte antigen molecules.

16. The genetically modified cell of claim 15 , wherein the one or more indels comprises a β2M knock out.

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

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

19. The genetically modified cell of claim 1 , wherein the genetically modified cell is a cardiomyocyte, an endothelial cell, a hepatocyte, a hepatocyte-like cell, a beta cell, a mesenchymal progenitor cell, a neural progenitor cell, a macrophage or a T cell.

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

21. The genetically modified cell of claim 20 , wherein the stem cell is an embryonic stem cell.

22. The genetically modified cell of claim 20 , wherein the stem cell is an induced pluripotent stem cell.

23. A composition comprising the genetically modified cell of claim 1 .

24. The genetically modified cell of claim 1 , further comprising reduced or eliminated cell surface expression of one or more MHC-II human leukocyte antigen molecules relative to an unmodified cell of the same type.

25. The genetically modified cell of claim 24 , 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 cell, thereby resulting in the reduced or eliminated cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

26. The genetically modified cell of claim 24 , 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 or eliminated cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

27. The genetically modified cell of claim 26 , comprising one or more indels in a class II major histocompatibility complex transactivator (CIITA) gene in the genome of the genetically modified cell, thereby resulting in the reduced or eliminated cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

28. The genetically modified cell of claim 27 , wherein the one or more indels comprises a CIITA knock out.

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

30. The composition of claim 23 , wherein the genetically modified cell further comprises reduced or eliminated cell surface expression of one or more MHC-II human leukocyte antigen molecules relative to an unmodified cell of the same type.

31. The composition of claim 23 , wherein the genetically modified cell is a CIITA −/− genetically modified cell.

32. The genetically modified cell of claim 24 , comprising one or more indels in a CIITA gene, a β2M gene, a TAP I 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 or eliminated 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.

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