Translational Medical Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China; Department of Hematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Translational Medical Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Cytokine. 2024 Dec;184:156758. doi: 10.1016/j.cyto.2024.156758. Epub 2024 Sep 17.
Chimeric antigen receptor (CAR)-T therapy has demonstrated sustained clinical remission in numerous hematologic malignancies and has expanded to encompass solid tumors and autoimmune diseases. While progress is being made in establishing optimal culture conditions for CAR-T cells, the identification of the most effective cytokine for promoting their persistence in vitro remains elusive.
Here, we employed scRNA-seq (single-cell RNA sequencing) analysis to investigate the potential alterations in biological processes within CAR-T cells following exposure to cytokines (IL-2, IL-12, and IL-21) and antigens. Transcriptomic changes in diverse CAR-T groups were compared following various treatments, with a focus on epigenetic modifications, metabolic shifts, cellular senescence, and exhaustion.
Our study reveals that CAR-T cells treated with antigen, IL-2, and IL-12 exhibit signs of exhaustion and senescence, whereas those treated with IL-21 do not display these characteristics. The activities of glycolysis and epigenetic changes were significantly increased by treatments with antigens, IL-2, and IL-12, while IL-21 treatment maintained the oxidative phosphorylation (OXPHOS) of CAR-T cells.
Our findings suggest that IL-21 may play a role in preventing senescence and could be utilized in combination with other strategies, such as IL-2 and IL-12, for CAR-T culture.
嵌合抗原受体（CAR）-T 疗法在许多血液恶性肿瘤中表现出持续的临床缓解，并已扩展到涵盖实体瘤和自身免疫性疾病。虽然在建立 CAR-T 细胞的最佳培养条件方面取得了进展，但确定最有效的细胞因子来促进其在体外的持久性仍然难以捉摸。
在这里，我们采用 scRNA-seq（单细胞 RNA 测序）分析来研究 CAR-T 细胞在暴露于细胞因子（IL-2、IL-12 和 IL-21）和抗原后，其生物学过程中可能发生的潜在变化。比较不同 CAR-T 组在不同治疗后的转录组变化，重点关注表观遗传修饰、代谢变化、细胞衰老和衰竭。
我们的研究表明，用抗原、IL-2 和 IL-12 处理的 CAR-T 细胞表现出衰竭和衰老的迹象，而用 IL-21 处理的细胞则没有这些特征。用抗原、IL-2 和 IL-12 处理显著增加了糖酵解和表观遗传变化的活性，而 IL-21 处理维持了 CAR-T 细胞的氧化磷酸化（OXPHOS）。
我们的研究结果表明，IL-21 可能在预防衰老中发挥作用，并可与 IL-2 和 IL-12 等其他策略联合使用，用于 CAR-T 培养。
