Asparaginase enhances CAR-T cell antitumor immunity by asparagine metabolic reprogramming and central memory induction in ALL

  • Mol Ther. 2025 Aug 12:S1525-0016(25)00650-1. doi: 10.1016/j.ymthe.2025.08.019.
Xinting Zhu  1 ,  Leng Han  1 ,  Dingyuan Bai  1 ,  Lei Yi  2 ,  Yonghong Zhao  3 ,  Shuaibing Liu  4 ,  Run Gan  1 ,  Bo Xin  1 ,  Yixing Tu  1 ,  Jianping Zhang  1 ,  Yonglong Han  1 ,  Juan Hao  5 ,  Zixue Xuan  6 ,  Cheng Guo  7 ,  Quanjun Yang  8
Affiliations
  • 1. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China.
  • 2. Department of Burn, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
  • 3. Department of Surgery, Shanghai Sixth People's Hospital Affiliated Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China.
  • 4. Department of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
  • 5. Department of Endocrinology, Shanghai TCM-Integrated Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Baoding 230, Hongkou 200086, Shanghai, China.
  • 6. Center for Clinical Pharmacy, Cancer Center, Department of Pharmacy, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou 310014, Zhejiang, China. Electronic address: [email protected].
  • 7. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China. Electronic address: [email protected].
  • 8. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China. Electronic address: [email protected].
PMID: 40808257 DOI: 10.1016/j.ymthe.2025.08.019
Abstract

High levels of asparagine synthetase (ASNS) in acute lymphoblastic leukemia (ALL) lead to immunotherapy resistance. Our study showed that ASNS overexpression (OE) in NALM6-GL Cancer cells attenuated chimeric antigen receptor (CAR)-T cell-mediated cancer Cell Lysis. Asparaginase (ASPG) is an approved drug that breaks down circulating asparagine in leukemia cells, thereby depriving Cancer cells of asparagine and inhibiting Cancer growth. We proposed a hypothesis that ASPG-engineered CAR-T cells undergo phenotype switching to overcome immunotherapy resistance in ALL. Coculture killing assay showed ASPG-OE CAR-T cells exhibited increased killing efficacy against ASNS-OE Cancer cells by enhancing the expression of granzyme B, interferon gamma, and tumor necrosis factor alpha, whereas ASPG-knockout (KO) CAR-T cells showed decreased cancer Cell Lysis efficiency. Phenotypic analysis revealed that ASPG-OE CAR-T cells exhibited distinct phenotypes, including increasing central memory T cells percentage, while decreasing effector memory T cells and effector memory cells that re-expressed CD45RA cells proportions. This distinct phenotype switch of ASPG-OE CAR-T cells toward central memory T cells exerted the increased killing efficacy against NALM6-GL cells even without ASNS-OE. The in vivo xenograft mouse model confirmed that ASPG-OE CAR-T cells exhibited superior Anticancer activity against NALM6-GL Cancer cells, while ASPG-KO CAR-T cells exhibited inferior Anticancer activity. Taken together, ASPG orchestrates CAR-T cell distinct phenotype toward central memory T cells and reprogramming of asparagine metabolism for enhancing antitumor immunity.

Keywords
acute lymphoblastic leukemia; antitumor immunity; asparaginase; asparagine; asparagine synthetase; cancer metabolism; chimeric antigen receptor; immunotherapy; leukemia; metabolic reprogramming.
Products