CAR-T（CAR-T）细胞疗法在非肿瘤性疾病中的应用
Chimeric antigen receptor T (CAR-T) cell therapy in non-oncological diseases.
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
靶向 CCR7-KMT2D 通过抑制治疗诱导的衰老增强 B 细胞非霍奇金淋巴瘤中 CAR-T 细胞疗效
英文原题：Targeting CCR7-KMT2D enhances CAR-T cell efficacy by suppressing therapy-induced senescence in B-cell non-Hodgkin lymphoma.
查看英文原题
Targeting CCR7-KMT2D enhances CAR-T cell efficacy by suppressing therapy-induced senescence in B-cell non-Hodgkin lymphoma.
分数与星级只用于站内排序 —— 不代表疗效、安全性或个人适用性。
研究概要
我们的发现表明，阻断 KMT2D-CCR7 介导的衰老可增强 CD19 CAR-T 细胞在 B-NHL 中的抗肿瘤活性。
中文摘要
背景
抗CD19嵌合抗原受体（CAR）T细胞疗法已在复发/难治性B细胞非霍奇金淋巴瘤（B-NHL）患者中显示疗效。然而，治疗诱导衰老（TIS）已被认为是一种新型耐药机制，可能加重免疫抑制并损害CAR-T 细胞功能。
方法
采用40 nM多柔比星（Dox）处理Raji和SU-DHL-2细胞72小时以诱导TIS，并使用10 μg/mL Cap-100抑制CCR7。通过SA-β-Gal染色和流式细胞术评估细胞衰老，通过CCK-8和Annexin V/PI分别评估增殖和凋亡，并以流式细胞术检测CAR-T 细胞毒性。测量B-NHL患者及健康对照T细胞上的CCR7表达。采用慢病毒转导调节CCR7表达，通过蛋白质印迹和免疫共沉淀（Co-IP）分析蛋白表达和相互作用。分别使用GS143和Y27632抑制NF-κB及ARHGAP/RhoA通路。
结果
CCR7过表达增强衰老相关分泌表型（SASP），而敲低CCR7则减弱该表型。在Dox诱导的TIS模型中，KMT2D表达降低，而H3K9me3、CCR7和LGALS9表达增加；抑制CCR7可逆转这些变化。Cap-100通过缓解共培养中的耗竭，提高CD19 CAR-T 细胞杀伤能力。从机制上看，Cap-100稳定IκB以抑制NF-κB，抑制NF-κB或ROCK均可逆转促衰老效应。Cap-100降低T细胞增殖，但不影响凋亡或耗竭。临床上，B-NHL患者T细胞上的CCR7低于健康对照，较高CCR7与更好的T细胞质量相关。转录组分析显示，Dox激活TIM3/Galectin 9和PD-1/PD-L1通路，而Dox联合Cap-100可抑制这些通路。Co-IP证实CCR7、KMT2D和LGALS9之间存在相互作用。总体而言，Dox诱导TIS并上调CCR7，进而增加SASP和LGALS9；后者与TIM-3相互作用，引发免疫细胞耗竭并降低杀伤效率。阻断CCR7可逆转这一状态并增强细胞毒性。
结论
我们的研究表明，阻断KMT2D-CCR7介导的衰老可增强CD19 CAR-T 细胞对B-NHL的抗肿瘤活性。
展开英文摘要原文
BACKGROUND
The efficacy of anti-CD19 Chimeric Antigen Receptor (CAR) T-cell therapy has been demonstrated in patients with relapsed or refractory B-cell non-Hodgkin lymphoma (B-NHL). However, therapy-induced senescence (TIS) has been identified as a novel resistance mechanism, potentially exacerbating immunosuppression and impairing CAR-T cell function.
METHODS
Raji and SU-DHL-2 cells were treated with 40 nM doxorubicin (Dox) for 72 h to induce TIS, and CCR7 was inhibited with 10 g/mL Cap-100. Cellular senescence was assessed by SA- -Gal staining and flow cytometry; proliferation and apoptosis by CCK-8 and Annexin V/PI, respectively. CAR-T cytotoxicity was evaluated via flow cytometry. CCR7 expression on T cells from B-NHL patients and healthy controls was measured. Lentiviral transduction regulated CCR7 expression; protein expression and interactions were analyzed by Western blot and Co-IP. NF- B and ARHGAP/RhoA pathways were inhibited using GS143 and Y27632, respectively.
RESULTS
CCR7 overexpression enhanced SASP, while knockdown attenuated it. In the Dox-induced TIS model, KMT2D decreased, whereas H3K9me3, CCR7, and LGALS9 increased; CCR7 inhibition reversed these changes. Cap-100 improved CD19 CAR T killing by alleviating exhaustion in co-culture. Mechanistically, Cap-100 stabilized I B to inhibit NF B, and inhibition of NF B or ROCK reversed pro-senescence. Cap-100 reduced T cell proliferation but did not affect apoptosis or exhaustion. Clinically, CCR7 was lower on T cells from B NHL patients than healthy controls, and higher CCR7 correlated with better T cell quality. Transcriptomics showed Dox activated TIM3/Galectin 9 and PD- 1/PD-L1 pathways, while Dox + Cap-100 suppressed them. Co-IP confirmed interactions among CCR7, KMT2D, and LGALS9. Collectively, Dox induced TIS to upregulate CCR7, increasing SASP and LGALS9, which interacted with TIM-3, causing immune cell exhaustion and reducing killing efficiency; CCR7 blockade reversed this state and enhanced cytotoxicity.
CONCLUSIONS
Our findings demonstrate that blockade of KMT2D-CCR7-mediated senescence enhances CD19 CAR-T cell anti-tumor activity in B-NHL.
论文信息
- 作者
- Li J、Huang K、Wang H、Chen W、Zhang Y、Jiang S、Cao N、Zhang W
- 第一作者单位
- Department of Hematology, The Second Hospital of Anhui Medical University, Hefei, 230601, Anhui, China.China
- 通讯作者单位
- Department of Hematology, The Second Hospital of Anhui Medical University, Hefei, 230601, Anhui, China. wangjiyu1992@126.com.China
- 期刊
- BMC medicine2026 Jun 26
- 原文标识
- PubMed 42363110 · DOI 10.1186/s12916-026-05030-4
在 PubMed 查看 → 出版商原文（DOI） 全文 PDF（PMC）· 可下载 治疗专题与资料阅读指南 资料来源与翻译说明 报告译文或资料问题 →