Demethylating therapy increases anti-CD123 CAR T cell cytotoxicity against acute myeloid leukemia

Nat Commun. 2021 Nov 8;12(1):6436. doi: 10.1038/s41467-021-26683-0.

Abstract

Successful treatment of acute myeloid leukemia (AML) with chimeric antigen receptor (CAR) T cells is hampered by toxicity on normal hematopoietic progenitor cells and low CAR T cell persistence. Here, we develop third-generation anti-CD123 CAR T cells with a humanized CSL362-based ScFv and a CD28-OX40-CD3ζ intracellular signaling domain. This CAR demonstrates anti-AML activity without affecting the healthy hematopoietic system, or causing epithelial tissue damage in a xenograft model. CD123 expression on leukemia cells increases upon 5'-Azacitidine (AZA) treatment. AZA treatment of leukemia-bearing mice causes an increase in CTLA-4negative anti-CD123 CAR T cell numbers following infusion. Functionally, the CTLA-4negative anti-CD123 CAR T cells exhibit superior cytotoxicity against AML cells, accompanied by higher TNFα production and enhanced downstream phosphorylation of key T cell activation molecules. Our findings indicate that AZA increases the immunogenicity of AML cells, enhancing recognition and elimination of malignant cells by highly efficient CTLA-4negative anti-CD123 CAR T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Disease
  • Animals
  • Azacitidine / administration & dosage*
  • Cell Line, Tumor
  • Cells, Cultured
  • Cytotoxicity, Immunologic
  • DNA Methylation / drug effects
  • Enzyme Inhibitors / administration & dosage
  • HEK293 Cells
  • HL-60 Cells
  • Humans
  • Immunotherapy, Adoptive / methods*
  • Interleukin-3 Receptor alpha Subunit / immunology*
  • Interleukin-3 Receptor alpha Subunit / metabolism
  • Kaplan-Meier Estimate
  • Leukemia, Myeloid / immunology
  • Leukemia, Myeloid / pathology
  • Leukemia, Myeloid / therapy*
  • Mice, Knockout
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Antigen, T-Cell / metabolism
  • Receptors, Chimeric Antigen / immunology
  • Receptors, Chimeric Antigen / metabolism
  • Single-Chain Antibodies / immunology*
  • Xenograft Model Antitumor Assays / methods*

Substances

  • Enzyme Inhibitors
  • Interleukin-3 Receptor alpha Subunit
  • Receptors, Antigen, T-Cell
  • Receptors, Chimeric Antigen
  • Single-Chain Antibodies
  • Azacitidine