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ORIGINAL RESEARCH article

Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
Volume 15 - 2024 | doi: 10.3389/fimmu.2024.1521778
This article is part of the Research Topic Precision Immunotherapy and Novel Target Discovery in Hematological Malignancy View all 4 articles

In vitro functional validation of anti-CD19 chimeric antigen receptor T cells expressing lysine-specific demethylase 1 short hairpin RNA for the treatment of diffuse large B cell lymphoma

Provisionally accepted
ZHI GUO ZHI GUO 1*Mingxin He Mingxin He 2Ning Liu Ning Liu 2*Yiqing Yang Yiqing Yang 3*Rui Sun Rui Sun 3*Jianxun Wang Jianxun Wang 3*Qiang Wang Qiang Wang 1*
  • 1 Wuhan University of Science and Technology, Wuhan, China
  • 2 Department of Hematology, The sixth Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, China
  • 3 Shenzhen Cell Valley Biomedical Co.,Ltd, Shenzhen, China

The final, formatted version of the article will be published soon.

    Background: Chimeric antigen receptor- T (CAR-T) cell therapy is more effective in relapsed or refractory diffuse large B cell lymphoma (DLBCL) than other therapies, but a high proportion of patients relapse after CAR-T cell therapy owing to antigen escape, limited persistence of CAR-T cells, and immunosuppression in the tumor microenvironment. CAR-T cell exhaustion is a major cause of relapse. Epigenetic modifications can regulate T cell activation, maturation and depletion; they can be applied to reduce T cell depletion, improve infiltration, and promote memory phenotype formation to reduce relapse after CAR-T cell therapy. Purpose: We propose to develop and validate in vitro the function of novel CAR-T cells for the treatment of DLBCL, which simultaneously express an anti-CD19 CAR with lysine-specific demethylase 1 (LSD1) short hairpin (sh)RNA to prevent depletion and prolong the survival of CAR-T cells. Results: We detected the expression of the CAR in the CAR-T cells by flow cytometry, and observed transduction rates of 31.5% for RNAU6 anti-CD19 CAR-T cells and 60.7% for LSD1 shRNA anti-CD19 CAR-T cells. The killing efficiency of LSD1 shRNA anti-CD19 CAR-T cells was significantly higher than that of RNAU6 anti-CD19 CAR-T cells at the low effector target ratio. We further found that LSD1 shRNA anti-CD19 CAR-T cells secreted more IFN-γ and granzyme B than RNAU6 anti-CD19 CAR-T cells. CAR-T cells proliferated after U-2932 cell stimulation and were able to sustain proliferation. After stimulation via U-2932 cell co-culture, both RNAU6 anti-CD19 CAR-T and LSD1 shRNA anti-CD19 CAR-T populations had increased proportions of cells with the TCM phenotype, with a higher percentage among LSD1 shRNA anti-CD19 CAR-T cells. Conclusion: We developed a novel, feasible CD19-LSD1 shRNA CAR-T cell strategy for the treatment of DLBCL. Our in vitro assay results showed that LSD1 shRNA anti-CD19 CAR-T cells more effectively killed target cells than RNAU6 anti-CD19 CAR-T cells, and developed a higher proportion of TCM phenotype cells. LSD1 shRNA anti-CD19 CAR-T cells may represent a potential treatment for DLBCL.

    Keywords: LSD1 shRNA 1, diffuse large B cell lymphomaDLBCL 2, anti-CD19 CAR-T cells 3, In vitro function 4, cell therapy 5

    Received: 02 Nov 2024; Accepted: 24 Dec 2024.

    Copyright: © 2024 GUO, He, Liu, Yang, Sun, Wang and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    ZHI GUO, Wuhan University of Science and Technology, Wuhan, China
    Ning Liu, Department of Hematology, The sixth Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, China
    Yiqing Yang, Shenzhen Cell Valley Biomedical Co.,Ltd, Shenzhen, China
    Rui Sun, Shenzhen Cell Valley Biomedical Co.,Ltd, Shenzhen, China
    Jianxun Wang, Shenzhen Cell Valley Biomedical Co.,Ltd, Shenzhen, China
    Qiang Wang, Wuhan University of Science and Technology, Wuhan, China

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