Cd33 exon 2 deficient donor stem cells for use with cd33 targeting agents

Patent metadata

Field	Value
Publication number	US-2024166721-A1
Application number	US-202318206269-A
Country	US
Kind code	A1
Filing date	Jun 6, 2023
Priority date	Aug 28, 2017
Publication date	May 23, 2024
Grant date	—

Abstract

Disclosed herein are compositions and methods relating to agents that target a lineage-specific cell-surface antigen and a population of hematopoietic cells that are deficient in the lineage-specific cell-surface antigen for immunotherapy of hematological malignancies.

First claim

1 - 28 . (canceled)

29 . A hematopoietic cell, or a descendant thereof, that is genetically engineered to comprise a mature transcript of CD33 having a mutated exon 2, wherein the hematopoietic cells are engineered by a CRISPR-Cas system comprising one or more guide nucleic acids comprising a sequence according to any one of SEQ ID NO: 52-61.

30 . The hematopoietic cell, or a descendant thereof, of claim 29 , wherein the hematopoietic cell comprises a CD33 with a mutated IgV domain.

31 . The hematopoietic cell, or a descendant thereof, of claim 29 , wherein the hematopoietic cell is not recognized by an agent targeting CD33, wherein the agent comprises an antigen-binding fragment that binds CD33.

32 . The hematopoietic cell, or a descendant thereof, of claim 31 , wherein the agent is an immune cell expressing a chimeric receptor that comprises the antigen-binding fragment, or wherein the agent is an antibody or fragment thereof comprising the antigen-binding fragment.

33 . The hematopoietic cell, or a descendant thereof, of claim 31 , wherein the agent is an antibody-drug conjugate that comprises the antigen-binding fragment.

34 . The hematopoietic cell, or a descendant thereof, of claim 29 , wherein the hematopoietic cell is a CD34+ hematopoietic stem cell, progenitor cell, myeloid progenitor cell, lymphoid progenitor cell, myeloid cell, or lymphoid cell.

35 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the hematopoietic cell is obtained from bone marrow cells or peripheral blood mononuclear cells (PBMCs).

36 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the hematopoietic cell is obtained from a healthy human donor or a subject that has a hematopoietic malignancy.

37 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the CRISPR-Cas system comprises a Cas9 nuclease or a Cpf1 nuclease.

38 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the CRISPR-Cas system comprises a nickase.

39 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the CRISPR-Cas system comprises a base editor.

40 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the hematopoietic cell has the capacity to proliferate and/or differentiate normally compared to a hematopoietic cell that has not been genetically engineered with the CRISPR-Cas system.

41 . The hematopoietic cell, or descendant thereof, of claim 29 , wherein the hematopoietic cell has the capacity to form colonies normally compared to a hematopoietic cell that has not been genetically engineered with the CRISPR-Cas system.

42 . A cell population comprising the hematopoietic cell, or descendant thereof, of claim 29 or a plurality thereof.

43 . A pharmaceutical composition comprising the cell population of claim 42 and a pharmaceutically acceptable carrier or excipient.

44 . A guide RNA (gRNA) comprising a sequence according to any one of SEQ ID NOs: 52-61.

45 . A method of preparing a population of genetically engineered hematopoietic cells, comprising
(i) providing a population of hematopoietic cells from a subject; (ii) introducing into cells of the population (a) a guide (RNA) comprising a sequence according to any one of SEQ ID NOs: 52-61; and (b) a Cas enzyme; thereby genetically engineering hematopoietic cells of the population to produce a population of genetically engineered hematopoietic cells.

46 . The hematopoietic cell, or descendant thereof, of claim 45 , wherein the Cas enzyme comprises a Cas9 nuclease, a Cpf1 nuclease, a nickase, or a base editor.

47 . The method of claim 45 , wherein the population of hematopoietic cells is obtained from a healthy donor subject or a subject that has a hematopoietic malignancy.

Assignees

• Univ Columbia

Inventors

• Mukherjee Siddhartha
• Ali Abdullah Mahmood
• Borot Florence

Classifications

• A61K40/421

  Immunoglobulin superfamily · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/15

  Natural-killer [NK] cells; Natural-killer T [NKT] cells · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/22

  Intracellular domain · CPC title