Spermidine Promotes Nb CAR-T Mediated Cytotoxicity to Lymphoma Cells Through Elevating Proliferation and Memory

Hongxia Wang; Dan Jiang; Liyuan Liu; Yanting Zhang; Miao Qin; Yuliang Qu; Liyan Wang; Shan Wu; Haijin Zhou; Tao Xu; Guangxian Xu

doi:10.2147/OTT.S382540

Spermidine Promotes Nb CAR-T Mediated Cytotoxicity to Lymphoma Cells Through Elevating Proliferation and Memory

Onco Targets Ther. 2022 Oct 18:15:1229-1243. doi: 10.2147/OTT.S382540. eCollection 2022.

Authors

Hongxia Wang^#^{1

2}, Dan Jiang^#^{1

2}, Liyuan Liu², Yanting Zhang², Miao Qin², Yuliang Qu², Liyan Wang², Shan Wu², Haijin Zhou¹, Tao Xu¹, Guangxian Xu^{1

2}

Affiliations

¹ Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, School of Medical Technology, The First Dongguan Affiliated Hospital, Guangdong Medical University, Dongguan, People's Republic of China.
² School of Clinical Medicine, Ningxia Medical University, Yinchuan, People's Republic of China.

^# Contributed equally.

Abstract

Purpose: Due to the natural advantages of spermidine in immunity, we investigated the effects of spermidine pretreatment on nanobody-based CAR-T cells (Nb CAR-T) mediated cytotoxicity and potential mechanism.

Patients and methods: The optimal concentration of spermidine was determined by detecting its impact on viability and proliferation of T cells. The phenotypic characteristic of CAR-T cells, which were treated with spermidine for 4 days, was examined by flow cytometry. The expansion ability of CAR-T cells was monitored in being cocultured with tumor cells. Additionally, CAR-T cells were stimulated by lymphoma cells to test its cytotoxicity in vitro, and the supernatant in co-culture models were collected to test the cytokine production. Furthermore, xenograft models were constructed to detect the anti-tumor activity of CAR-T cells in vivo.

Results: The optimal concentration of spermidine acting on T cells was 5μM. The antigen-dependent proliferation of spermidine pretreatment CD19 CAR-T cells or Nb CAR-T cells was increased compared to control. Central memory T cells(TCM) dominated the CAR-T cell population in the presence of spermidine. When spermidine pretreatment CAR-T cells were stimulated with Daudi cells, the secretion of IL-2 and IFN-γ has been significantly enhanced. The ability of CAR-T cells to lysis Daudi cells was enhanced with the help of spermidine, even at higher tumor loads. Pre-treated Nb CAR-T cells with spermidine were able to control tumor cells in vivo, and therefore prolong mice survival.

Conclusion: Our results revealed that spermidine could promote Nb CAR-T mediated cytotoxicity to lymphomas cells through enhancing memory and proliferation, and provided a meaningful approach to strengthen the anti-tumor effect of CAR-T cells.

Keywords: CAR-T; exhaustion; nanobody; spermidine.

Grants and funding

This study was supported by the Discipline Construction Project of Guangdong Medical University (No: 4SG21278P) and Scientific Research Start-up Fund for High-end Talents of Guangdong Medical University (4SG21240G).