Cells lacking b2m surface expression and methods for allogeneic administration of such cells

Patent metadata

Field	Value
Publication number	US-2025333473-A1
Application number	US-202418949619-A
Country	US
Kind code	A1
Filing date	Nov 15, 2024
Priority date	Nov 6, 2014
Publication date	Oct 30, 2025
Grant date	—

Abstract

Disclosed herein are cells and populations of cells comprising a genome in which the B2M gene has been edited to eliminate surface expression of MHC Class I protein in the cells or population of cells, and methods for allogeneic administration of such cells to reduce the likelihood that the cells will trigger a host immune response when the cells are administered to a subject in need of such cells.

First claim

1 .- 49 . (canceled)

50 . A method for treating or preventing a disorder associated with expression of a polynucleotide sequence in a subject, the method comprising:
(a) altering a target polynucleotide sequence associated with the disorder in a cell or a population of cells ex vivo by contacting the polynucleotide sequence with a clustered regularly interspaced short palindromic repeats-associated (Cas) protein and at least one ribonucleic acid, wherein the ribonucleic acid directs Cas protein to and hybridizes to a target motif of the target polynucleotide sequence associated with the disorder, wherein the target polynucleotide sequence associated with the disorder is cleaved; (b) altering a target B2M polynucleotide sequence in the cell or population of cells ex vivo by contacting the target B2M polynucleotide sequence with a clustered regularly interspaced short palindromic repeats-associated (Cas) protein and at least one ribonucleic acid selected from the group consisting of SEQ ID NOs: 9-23 and 419-2609; and (c) introducing the cell or population of cells into the subject, thereby treating or preventing a disorder associated with expression of the polynucleotide sequence.

51 . The method of claim 50 , wherein the disorder is selected from the group consisting of a genetic disorder, an infection, and cancer.

52 . The method of claim 50 , wherein the disorder comprises HIV or AIDs.

53 . The method of claim 50 , further comprising altering one or more additional polynucleotide sequences in the cell ex vivo.

54 . The method of claim 50 , wherein the Cas protein comprises a Cas9 protein or a functional portion thereof.

55 . The method of claim 50 , wherein the Cas protein comprises a Cpf1 protein or a functional portion thereof.

56 . The method of claim 50 , wherein the cell or population of cells are selected from the group consisting of a stem cell, a pluripotent cell, a progenitor cells, a hematopoietic stem and/or progenitor cell, a CD34 + mobilized peripheral blood cell, a CD34+ cord blood cell, a CD34+ bone marrow cell, a CD34 + CD38-Lineage-CD90 + CD45RA − cell, and a CD34+ hematopoietic stem and/or progenitor cell; a CD4+ T cell, a hepatocyte, a somatic cell, and a non-transformed cell.

57 . The method of claim 50 , wherein the at least one ribonucleic acid is selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23.

58 . The method of claim 50 , wherein the alteration of the target B2M polynucleotide sequence in the cell or population of cells results in the deletion of a contiguous stretch of genomic DNA, thereby eliminating surface expression of MHC Class I molecules in the cell or population of cells.

59 . The method of claim 50 , wherein the cell or population of cells are CD4+ T cells.

60 . The method of claim 50 , wherein the cell or population of cells are CD34+ cells.

Assignees

• Harvard College

Inventors

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

Classifications

• C12N2510/00

  Genetically modified cells · CPC title

• C12N15/11

  DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} · CPC title

• C12N5/0647

  Haematopoietic stem cells; Uncommitted or multipotent progenitors · CPC title

• A61K35/28

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• A61P31/18

  for HIV · CPC title