Preclinical evaluation of CD70-specific CAR T cells targeting acute myeloid leukemia

Front Immunol. 2023 Feb 10:14:1093750. doi: 10.3389/fimmu.2023.1093750. eCollection 2023.

Abstract

Backgrounds: Chimeric antigen receptor (CAR)-T cell therapy has achieved unprecedented success in treating hematopoietic malignancies. However, this cell therapy is hampered in treating acute myeloid leukemia (AML) due to lack of ideal cell surface targets that only express on AML blasts and leukemia stem cells (LSCs) but not on normal hematopoietic stem cells (HSCs).

Methods: We detected the CD70 expression on the surfaces of AML cell lines, primary AML cells, HSC, and peripheral blood cells and generated a second-generation CD70-specific CAR-T cells using a construct containing a humanized 41D12-based scFv and a 41BB-CD3ζ intracellular signaling domain. Cytotoxicity, cytokine release, and proliferation in antigen stimulation, CD107a assay, and CFSE assays were used to demonstrate the potent anti-leukemia activity in vitro. A Molm-13 xenograft mouse model was established to evaluate the anti-leukemic activity of CD70 CAR-T in vivo. CFU assay was explored to assess the safety of CD70 CAR-T on HSC.

Results: CD70 heterogeneously expressed on AML primary cells, including leukemia blasts, leukemic progenitor, and stem cells, but not expressed on normal HSCs and majority of blood cells. Anti-CD70 CAR-T cells exhibited potent cytotoxicity, cytokines production, and proliferation when incubated with CD70+ AML cell lines. It also displayed robust anti-leukemia activity and prolonged survival in Molm-13 xenograft mouse model. However, such CAR-T cell therapy did not completely eliminate leukemia in vivo.

Discussion: Our study reveals that anti-CD70 CAR-T cells are a new potential treatment for AML. However, such CAR-T cell therapy did not completely eliminate leukemia in vivo, suggesting that future studies aiming to generate innovative combinatorial CAR constructs or to increase CD70 expression density on leukemia cell surface to prolong the life-span of CAR-T cells in the circulation will be needed in order to optimize CAR-T cell responses for AML.

Keywords: CAR-T; CD70; acute myeloid leukemia; immunotherapy; leukemia stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD27 Ligand / metabolism
  • Cell Line, Tumor
  • Humans
  • Immunotherapy, Adoptive
  • Leukemia, Myeloid, Acute*
  • Mice
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / metabolism
  • T-Lymphocytes

Substances

  • Receptors, Chimeric Antigen
  • CD70 protein, human
  • CD27 Ligand

Grants and funding

This work was supported by the funds from Binjiang Institute of Zhejiang University (ZY202205SMKY004), the National Natural Science Foundation of China (No. 81830006, 82170219), and the Science Technology Department of Zhejiang Province (No. 2021C03117), and the Science Technology Department of Zhejiang Province (No. LGF19H080001).