Universal donor stem cells and related methods

Patent metadata

Field	Value
Publication number	US-2019309259-A1
Application number	US-201615572776-A
Country	US
Kind code	A1
Filing date	May 9, 2016
Priority date	May 8, 2015
Publication date	Oct 10, 2019
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 stem cell comprising modulated expression of one or more MHC-I and MHC-II human leukocyte antigens relative to a wild-type stem cell, wherein the modulated expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises one or more genes encoding one or more transcriptional regulators of MHC-I or MHC-II, wherein the one or more genes are selected from the group consisting of HLA-A, HLA-B, HLA-C, and CIITA.

2 . (canceled)

3 . The stem cell of claim 1 , wherein the modulated expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises reduced expression of the one or more MHC-I and MHC-II human leukocyte antigens.

4 - 5 . (canceled)

6 . The stem cell of claim 1 , wherein the modulated expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises the one or more genes encoding one or more transcriptional regulators of MHC-I or MHC-II being deleted from at least one allele of the cell.

7 - 10 . (canceled)

11 . The stem cell of claim 1 , further comprising modulated expression of one or more tolerogenic factors relative to a wild-type stem cell.

12 . The stem cell of claim 11 , wherein the modulated expression of the tolerogenic factors comprises increased expression of the tolerogenic factors.

13 . The stem cell of claim 11 , wherein the tolerogenic factors are inserted into a safe harbor locus of at least one allele of the cell, wherein the safe harbor locus comprises an AAVS1 locus.

14 - 15 . (canceled)

16 . The stem cell of claim 11 , wherein the tolerogenic factors are selected from the group consisting of HLA-G, PD-L1, and CD47.

17 . The stem cell of claim 1 , wherein the stem cell is an embryonic stem cell.

18 - 20 . (canceled)

21 . The stem cell of claim 1 , wherein the cell does not express one or more of CIITA, HLA-A, HLA-B, and HLA-C.

22 . (canceled)

23 . The stem cell of claim 1 , wherein the cell has reduced expression of one or more of CIITA, HLA-A, HLA-B and HLA-C relative to a wild-type stem cell.

24 .- 42 . (canceled)

43 . A human stem cell that does not express one or more of CIITA, HLA-A, HLA-B, and HLA-C.

44 . The cell of claim 43 , wherein the cell is a CIITA −/− , HLA-A −/− , HLA-B −/− , and HLA-C −/− knockout stem cell.

45 - 48 . (canceled)

49 . The cell of claim 43 , wherein the cell expresses one or more tolerogenic factors.

50 - 52 . (canceled)

53 . The cell of claim 49 , wherein the tolerogenic factors are selected from the group consisting of HLA-G, PD-L1, CD47.

54 .- 60 . (canceled)

61 . A human stem cell comprising one or more tolerogenic factors inserted into a safe harbor locus of at least one allele of the cell, wherein the tolerogenic factors are selected from the group consisting of HLA-G, PD-L1, and CD47.

62 .- 67 . (canceled)

68 . A method of preparing a hypoimmunogenic stem cell, the method comprising modulating expression of one or more MHC-I and MHC-II human leukocyte antigens by the stem cell and thereby preparing the hypoimmunogenic stem cell, wherein the modulated expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises one or more genes encoding one or more transcriptional regulators of MHC-I or MHC-II, wherein the one or more genes are selected from the group consisting of HLA-A, HLA-B, HLA-C, and CIITA.

69 . (canceled)

70 . The method of claim 68 , wherein the modulating expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises reducing expression of the one or more MHC-I and MHC-II human leukocyte antigens.

71 - 72 . (canceled)

73 . The method of claim 68 , wherein the modulating expression of the one or more MHC-I and MHC-II human leukocyte antigens comprises deleting the one or more genes encoding one or more transcriptional regulators of MHC-I or MHC-II from at least one allele of the cell.

74 - 77 . (canceled)

78 . The method of claim 68 , further comprising modulating expression of one or more tolerogenic factors by the stem cell.

79 . The method of claim 78 , wherein the modulating expression of the tolerogenic factors comprises increasing the expression of the tolerogenic factors.

80 . The method of claim 78 , wherein the tolerogenic factors are inserted into a safe harbor locus of at least one allele of the cell, wherein the safe harbor locus comprises an AAVS1 locus.

81 - 82 . (canceled)

83 . The method of claim 78 , wherein the tolerogenic factors are selected from the group consisting of HLA-G, PD-L1, and CD47.

84 .- 112 . (canceled)

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

• C12N5/0696

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

• 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

• C12N2501/599

  with CD designations not provided for elsewhere · CPC title

• A61K39/001 Primary

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