套细胞淋巴瘤 (MCL) 患者经常对依鲁替尼产生耐药性。淋巴瘤相关巨噬细胞 (LAM) 可能在这种耐药性中发挥因果作用，但在当前文献中仍未得到充分探索。为了阐明 LAM 在介导 MCL 依鲁替尼耐药中的作用。我们使用 CD206 抗体通过多参数流式细胞术 (MPFC) 研究了巨噬细胞极化以及对依鲁替尼耐药和敏感的 MCL 患者血液和组织样本中的 CD86。随后，我们开发了一种利用 MCL 细胞系的体外共培养模型来识别与依鲁替尼耐药和巨噬细胞 M2 极化相关的细胞因子。使用 MPFC、RNA 测序和蛋白质印迹分析检查了耐药性的机制。此外，我们还评估了 CXCR2 抑制剂 SB225002 是否可以通过 CCK-8 和 caspase-3 检测逆转依鲁替尼耐药性，以及在涉及依鲁替尼耐药性 MCL 细胞系的小鼠异种移植模型中逆转依鲁替尼耐药性。与敏感患者相比，M2 至 M1 LAM 的数量显着更高。在 LAM 和 MCL 细胞的共培养中，M2 巨噬细胞的百分比、依鲁替尼的 IC50 值以及 IL-8 和 CXCL5 的浓度显着升高。从机制上讲，LAM 分泌的 CXCL5 与 MCL 细胞上的 CXCR2 相互作用，导致在依鲁替尼存在的情况下激活 Akt、p38 和 STAT3 信号通路；阻断 CXCL5/CXCR2 轴后，该活性减弱。与单独使用依鲁替尼治疗相比，SB225002 和依鲁替尼的组合显着增强了 MCL 细胞凋亡，抑制了异种移植模型中的淋巴瘤生长，并重新编程了巨噬细胞表型。我们的数据表明，M2 极化 LAM 与 MCL 模型中的依鲁替尼耐药相关。 CXCR2 抑制剂可以逆转这种耐药性。这些发现提出了一种潜在的新治疗策略。版权所有 © 2024。由 Elsevier B.V. 制作和托管。

Patients with mantle cell lymphoma (MCL) frequently develop resistance to ibrutinib. Lymphoma-associated macrophages (LAMs) may play a causal role in this resistance but remain underexplored in current literature.To elucidate the role of LAMs in mediating ibrutinib resistance in MCL.We investigated macrophage polarization through multiparameter flow cytometry (MPFC) using antibodies against CD206 and CD86 in blood and tissue samples from patients with MCL, both resistant and sensitive to ibrutinib. Subsequently, we developed an in vitro co-culture model utilizing MCL cell lines to identify cytokines associated with ibrutinib resistance and macrophage M2 polarization. The mechanisms underlying resistance were examined using MPFC, RNA sequencing, and Western blot analysis. Additionally, we assessed whether SB225002, a CXCR2 inhibitor, could reverse ibrutinib resistance through CCK-8 and caspase-3 assays, as well as in a mouse xenograft model involving an ibrutinib-resistant MCL cell line.In patients exhibiting ibrutinib resistance, the ratio of M2 to M1 LAMs was significantly higher compared to sensitive patients. In co-cultures of LAMs and MCL cells, the percentage of M2 macrophages, the IC50 value for ibrutinib, and the concentrations of IL-8 and CXCL5 were significantly elevated. Mechanistically, CXCL5 secreted by LAMs interacted with the CXCR2 on MCL cells, leading to the activation of the Akt, p38, and STAT3 signaling pathways in the presence of ibrutinib; this activity was diminished upon blockade of the CXCL5/CXCR2 axis. The combination of SB225002 and ibrutinib significantly enhanced MCL cell apoptosis, suppressed lymphoma growth in the xenograft model, and reprogrammed macrophage phenotype compared to treatment with ibrutinib alone.Our data indicate that M2-polarized LAMs are associated with ibrutinib resistance in a model of MCL, and that a CXCR2 inhibitor can reverse this resistance. These findings suggest a potential new therapeutic strategy.Copyright © 2024. Production and hosting by Elsevier B.V.