PO.CL01.06 · 临床研究
硬脂酸-巨噬细胞串扰将MIF-CD44信号鉴定为肺癌起始中的治疗靶点
Stearic acid-macrophage crosstalk identifies MIF-CD44 signaling as a therapeutic target in lung cancer initiation
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：肺癌（LC）是癌症相关死亡的主要原因，但肿瘤起始的过程仍知之甚少。我们课题组的既往工作表明，硬脂酸（SA）等生物分子以及MIP1alpha和MIP1beta等促炎细胞因子在早期LC患者的荷瘤肺叶中选择性升高。因此，我们假设SA-巨噬细胞（MΦ）炎症轴促进了肺上皮的肿瘤性转化。
方法：比较了早期LC患者（n=5）和健康对照（n=6）之间的血浆细胞因子谱。在PBMC来源的MΦ和人单核细胞系（U937）中评估了SA暴露后在转录和翻译水平上的细胞因子产生，以确立两个系统之间的一致性。将来自SA处理的MΦ的条件培养基（CM）（SA-CM）应用于BEAS2B上皮细胞（非转化的肺上皮细胞），通过软琼脂和球体形成实验测量转化。通过mRNA/蛋白分析定量了MIP1alpha（CCR5）和MIF（CD44）的受体表达。使用小分子拮抗剂（分别为Maraviroc和Verbascoside）对CCR5和CD44进行功能性阻断，以确定驱动转化的关键细胞因子。
结果：与健康个体相比，在早期LC患者的血浆样本中观察到MIP1alpha升高。SA处理增强了PBMC和U937来源MΦ的MIP1alpha和MIF表达。同时，来自PBMC和U937来源MΦ的SA-CM在软琼脂基质中诱导了显著更多的集落形成，并在低黏附条件下诱导了强劲的球体生长，提示肿瘤性转化。受体分析显示BEAS2B细胞上CD44丰富而CCR5表达有限。用Verbascoside阻断CD44可消除SA-CM诱导的球体形成，而抑制CCR5则无效果。
结论：基于这些观察，我们得出结论：SA重编程MΦ的转录状态，促进MΦ驱动的促炎环境，进而诱导肺上皮细胞发生MIF-CD44依赖性的肿瘤性转化。这些发现提示了一个参与LC起始早期致癌事件的新型免疫代谢轴，并支持将靶向SA-MIF-CD44信号作为潜在预防策略。
查看英文原文 English abstract
Background: Lung cancer (LC) is the leading cause of cancer-related mortality, yet the process of tumor initiation is poorly understood. Prior work from our group indicated that biomolecules like stearic acid (SA) and proinflammatory cytokines like MIP1alpha and MIP1beta were selectively elevated in the tumor bearing lung lobes of early-stage LC patients. Hence, we hypothesized that a SA-macrophage (MΦ) inflammatory axis contributes to neoplastic transformation in lung epithelium.
Method: Plasma cytokine profiles were compared between early-stage LC patients (n=5) and healthy controls (n=6). Cytokine production following SA exposure at transcriptional and translational level were assessed in PBMC-derived MΦs and human monocytic cell lines (U937) to establish synchrony between two systems. Conditioned media (CM) from SA-treated MΦs (SA-CM) was applied to BEAS2B epithelial cells (non-transformed lung epithelial cells), with transformation measured by soft agar and spheroid assays. Receptor expression for MIP1alpha (CCR5) and MIF (CD44) was quantified by mRNA/protein analyses. Functional blockades of CCR5 and CD44 were performed using small-molecule antagonists (Maraviroc and Verbascoside respectively) to identify critical cytokine driving transformation.
Results: Elevated MIP1alpha were observed in plasma samples from early-stage LC patients compared to healthy individuals. SA treatment augmented MIP1alpha and MIF expression from both PBMC and U937 derived MΦs. Simultaneously, SA-CM from both PBMC and U937 derived MΦs induced significantly greater colony formation in soft agar matrix and robust spheroid growth under low-attachment conditions, indicating neoplastic transformation. Receptor analysis revealed abundant CD44 but limited CCR5 expression on BEAS2B cells. Blockade of CD44 with Verbascoside abrogated SA-CM-induced spheroid formation, whereas CCR5 inhibition had no effect.
Conclusion: Based on these observations we conclude that SA reprograms MΦ transcriptional states promoting a MΦ driven proinflammatory environment that induces MIF-CD44-dependent neoplastic transformation of lung epithelial cells. These findings implicate a novel immunometabolic axis involved in the early oncogenic events of LC initiation and support therapeutic targeting of SA-MIF-CD44 signaling as a potential preventive strategy.
利益披露 Disclosure
A. Roy, None..
G. Forbes, None..
D. Prabhakara, None.