Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Guangdong Provincial Key Laboratory of Single Cell Technology and Application, Southern Medical University, Guangzhou, Guangdong, China.
Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
J Immunother Cancer. 2024 Nov 9;12(11):e009212. doi: 10.1136/jitc-2024-009212.
Bendamustine-rituximab (BR) therapy stands out as a promising alternative for elderly patients with diffuse large B-cell lymphoma (DLBCL), demonstrating notable efficacy when conventional regimens pose challenges. Despite its clinical success, the intricate mechanisms underlying BR therapy have remained elusive.
DLBCL cell lines were used to investigate the mechanism of BR therapy in vitro. RNA-seq and Western blot were used to explore the target pathways of BR therapy. STING was knocked out using Crispr-cas9 and inhibited using H-151 to investigate its role in BR therapy. Bulk RNA-seq and single-cell RNA-seq data from patients were analyzed to investigate the association between STING and pyroptosis pathways, validated using STING downregulated cells. Flow cytometry, transwell experiments and co-culture experiments were performed to investigate the inflammatory phenotype of DLBCL cells after BR treatment and its effect on T-cell recruitment and activation.
This study elucidates that BR elicits direct tumoricidal effects by promoting apoptosis and inducing cell cycle arrest. The synergistic impact with rituximab is further potentiated by complement addition, demonstrating the pivotal role of in vivo antibody-dependent cellular cytotoxicity. Moreover, our investigation reveals that, through a cGAS-STING-dependent pathway, prolonged exposure to BR induces pyroptosis in DLBCL cells. Activation of the cGAS-STING pathway by BR therapy triggers the release of inflammatory factors and upregulates major histocompatibility complex molecules, shaping an immunologically hot tumor microenvironment.
This unique dual influence not only directly targets DLBCL cells but also engages the patient's immune system, paving the way for innovative combination therapies. The study provides comprehensive insights into the multifaceted actions of BR in DLBCL, offering a foundation for refined and personalized treatment strategies in elderly patients.
苯达莫司汀-利妥昔单抗（BR）治疗方案在治疗老年弥漫性大 B 细胞淋巴瘤（DLBCL）患者中表现出良好的疗效，为常规方案治疗困难的患者提供了新的选择。尽管 BR 治疗取得了显著的临床疗效，但其中的复杂机制仍不清楚。
采用 DLBCL 细胞系在体外研究 BR 治疗的机制。RNA-seq 和 Western blot 用于研究 BR 治疗的靶通路。使用 Crispr-cas9 敲除 STING 并使用 H-151 抑制 STING，以研究其在 BR 治疗中的作用。分析患者的批量 RNA-seq 和单细胞 RNA-seq 数据，以研究 STING 与细胞焦亡通路之间的关联，并用 STING 下调细胞进行验证。流式细胞术、Transwell 实验和共培养实验用于研究 BR 治疗后 DLBCL 细胞的炎症表型及其对 T 细胞募集和激活的影响。
本研究阐明了 BR 通过促进细胞凋亡和诱导细胞周期停滞来发挥直接的肿瘤杀伤作用。与利妥昔单抗的协同作用进一步增强了补体的加入，表明体内抗体依赖性细胞毒性的关键作用。此外，我们的研究还揭示了 BR 通过 cGAS-STING 依赖性途径在 DLBCL 细胞中诱导细胞焦亡。BR 治疗激活 cGAS-STING 通路触发炎症因子的释放，并上调主要组织相容性复合体分子，塑造免疫活跃的肿瘤微环境。
这种独特的双重影响不仅直接靶向 DLBCL 细胞，还激活患者的免疫系统，为创新的联合治疗开辟了道路。该研究全面深入地探讨了 BR 在 DLBCL 中的多方面作用，为老年患者的精细化和个体化治疗策略提供了基础。
