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

• Published in: Nature communications Vol. 12; no. 1; pp. 6436 - 20
• Main Authors: El Khawanky, Nadia, Hughes, Amy, Yu, Wenbo, Myburgh, Renier, Matschulla, Tony, Taromi, Sanaz, Aumann, Konrad, Clarson, Jade, Vinnakota, Janaki Manoja, Shoumariyeh, Khalid, Miething, Cornelius, Lopez, Angel F., Brown, Michael P., Duyster, Justus, Hein, Lutz, Manz, Markus G., Hughes, Timothy P., White, Deborah L., Yong, Agnes S. M., Zeiser, Robert
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 08.11.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 59; 59/5; 631/250; 631/67/1990/283/1897; 64; 64/60; 692/4028/67/1059/2325; 96/31; Acute Disease; Acute myeloid leukemia; Animals; Antigens; Azacitidine - administration & dosage; CD123 antigen; CD28 antigen; Cell activation; Cell Line, Tumor; Cells, Cultured; Chimeric antigen receptors; Cytotoxicity; Cytotoxicity, Immunologic; DNA Methylation - drug effects; Enzyme Inhibitors - administration & dosage; HEK293 Cells; Hematopoietic stem cells; Hematopoietic system; HL-60 Cells; Humanities and Social Sciences; Humans; Immunogenicity; Immunotherapy, Adoptive - methods; Interleukin-3 Receptor alpha Subunit - immunology; Interleukin-3 Receptor alpha Subunit - metabolism; Intracellular signalling; Kaplan-Meier Estimate; Leukemia; Leukemia, Myeloid - immunology; Leukemia, Myeloid - pathology; Leukemia, Myeloid - therapy; Lymphocytes; Lymphocytes T; Mice; Mice, Knockout; multidisciplinary; Myeloid leukemia; Phosphorylation; Progenitor cells; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - metabolism; Receptors, Chimeric Antigen - immunology; Receptors, Chimeric Antigen - metabolism; Science; Science (multidisciplinary); Single-Chain Antibodies - immunology; Toxicity; Tumor necrosis factor-α; Xenograft Model Antitumor Assays - methods; Xenografts; Xenotransplantation
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-26683-0

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-4 negative anti-CD123 CAR T cell numbers following infusion. Functionally, the CTLA-4 negative 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-4 negative anti-CD123 CAR T cells. The success of CAR-T cells for treating acute myeloid leukaemia (AML) is hampered by toxicity to normal cells and low CAR-T cell persistence. Here, the authors show that the demethylating compound 5′-Azacitdine increases anti-CD123 CAR-T cell cytotoxicity against AML.