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Sub-lethal doses of chemotherapeutic agents induce senescence in T cells and upregulation of PD-1 expression

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Abstract

Cellular senescence is a stable cell cycle arrest, usually in response to internal and/or external stress, including telomere dysfunction, abnormal cellular growth, and DNA damage. Several chemotherapeutic drugs, such as melphalan (MEL) and doxorubicin (DXR), induce cellular senescence in cancer cells. However, it is not clear whether these drugs induce senescence in immune cells. We evaluated the induction of cellular senescence in T cells were derived from human peripheral blood mononuclear cells (PBMNCs) in healthy donors using sub-lethal doses of chemotherapeutic agents. The PBMNCs were kept overnight in RPMI 1640 medium with 2% phytohemagglutinin and 10% fetal bovine serum and then cultured in RPMI 1640 with 20 ng/mL IL-2 and sub-lethal doses of chemotherapeutic drugs (2 μM MEL and 50 nM DXR) for 48 h. Sub-lethal doses of chemotherapeutic agents induced phenotypes associated with senescence, such as the formation of γH2AX nuclear foci, cell proliferation arrest, and induction of senescence-associated beta-galactosidase (SA-β-Gal) activity, (control vs. MEL, DXR; median mean fluorescence intensity (MFI) 1883 (1130–2163) vs. 2233 (1385–2254), 2406.5 (1377–3119), respectively) in T cells. IL6 and SPP1 mRNA, which are senescence-associated secretory phenotype (SASP) factors, were significantly upregulated by sublethal doses of MEL and DXR compared to the control (P = 0.043 and 0.018, respectively). Moreover, sub-lethal doses of chemotherapeutic agents significantly enhanced the expression of programmed death 1 (PD-1) on CD3 + CD4 + and CD3 + CD8 + T cells compared to the control (CD4 + T cells; P = 0.043, 0.043, and 0.043, respectively, CD8 + T cells; P = 0.043, 0.043, and 0.043, respectively). Our results suggest that sub-lethal doses of chemotherapeutic agents induce senescence in T cells and tumor immunosuppression by upregulating PD-1 expression on T cells.

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Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.”

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Authors and Affiliations

  1. Department of Laboratory Sciences, Gunma University Graduate School of Health Sciences, 3-39-22 Showa-Machi, Maebashi, Gunma, 371-8514, Japan

    Tetsuhiro Kasamatsu, Maaya Awata-Shiraiwa, Rei Ishihara, Yuki Murakami, Yuta Masuda, Nanami Gotoh, Hirokazu Murakami & Takayuki Saitoh

  2. Gunma University of Health and Welfare, 191-1 Kawamagari-Cho, Maebashi, Gunma, 371-0823, Japan

    Maaya Awata-Shiraiwa, Rei Ishihara, Yuki Murakami, Yuta Masuda & Hirokazu Murakami

  3. Institute of Molecular and Cellular Regulation, Gunma University, 3-39-22 Showa-Machi, Maebashi, Gunma, 371-8511, Japan

    Tsukasa Oda

  4. Blood Transfusion Service, Gunma University Hospital, 3-39-15 Showa-Machi, Maebashi, Gunma, 371-8511, Japan

    Akihiko Yokohama

  5. Department of Hematology, Gunma University, 3-39-22 Showa-Machi, Maebashi, Gunma, 371-0034, Japan

    Ikuko Matsumura & Hiroshi Handa

  6. Oncology Center, Gunma University Hospital, 3-39-15 Showa-Machi, Maebashi, Gunma, 371-8511, Japan

    Norifumi Tsukamoto

Authors
  1. Tetsuhiro Kasamatsu
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Contributions

All authors contributed to the study conception and design. TK designed the research, performed most of the experiments, performed analysis and wrote the manuscript; MA-S, RI, YM, YM, NG and TO performed some of the experiments and data collection; AY, IM, HH and NT collected the samples; HM and TS supervised all of the research work.

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Correspondence to Tetsuhiro Kasamatsu.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institutional Review Board of Gunma University Hospital (Approval # HS2019-247) and with the 1964 Declaration of Helsinki.

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Informed consent was obtained from all individual participants included in the study.

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Kasamatsu, T., Awata-Shiraiwa, M., Ishihara, R. et al. Sub-lethal doses of chemotherapeutic agents induce senescence in T cells and upregulation of PD-1 expression. Clin Exp Med 23, 2695–2703 (2023). https://doi.org/10.1007/s10238-023-01034-z

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