Universal donor stem cells and related methods

Patent metadata

Field	Value
Publication number	US-2023303969-A1
Application number	US-202217993680-A
Country	US
Kind code	A1
Filing date	Nov 23, 2022
Priority date	May 8, 2015
Publication date	Sep 28, 2023
Grant date	—

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

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 the one or more tolerogenic factors comprise CD47.

2 . The genetically modified beta cell of claim 1 , 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.

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

4 . 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.

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

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

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

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

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

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

11 . The genetically modified beta cell of claim 1 , wherein the one or more tolerogenic factors further 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 and/or 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-I human leukocyte antigen molecules and/or the one or more MHC-II 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 and/or 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-I human leukocyte antigen molecules and/or the one or more MHC-II 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 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 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 β2M −/− 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 . (canceled)

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

29 . A method of administering a cell therapy to a subject in need thereof, the method comprising administering the composition of claim 28 to the subject.

30 . The method of claim 29 , wherein the genetically modified beta cell comprises:
one or more indels in a β2M gene, thereby resulting in the reduced cell surface expression of the one or more MHC-I human leukocyte antigen molecules, and one or more indels in a CIITA gene, thereby resulting in the reduced cell surface expression of the one or more MHC-II human leukocyte antigen molecules.

31 . (canceled)

32 . The method of claim 30 , wherein the genetically modified beta cell is a β2M −/− CIITA −/− genetically modified beta cell.

33 . 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.

34 . The composition of claim 28 , 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.

35 . The method of claim 29 , 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.

Assignees

• Harvard College

Inventors

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

Classifications

• C12N5/0606 Primary

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N5/0696

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title

• C12N15/86

  Viral vectors · CPC title

• C12N15/907

  in mammalian cells · CPC title

• A61K39/001 Primary

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