DataSheet_1_Arachidonate 15-lipoxygenase-mediated production of Resolvin D5n-3 DPA abrogates pancreatic stellate cell-induced cancer cell invasion.pdf

Activation of pancreatic stellate cells (PSCs) to cancer-associated fibroblasts (CAFs) is responsible for the extensive desmoplastic reaction observed in PDAC stroma: a key driver of pancreatic ductal adenocarcinoma (PDAC) chemoresistance leading to poor prognosis. Specialized pro-resolving mediators (SPMs) are prime modulators of inflammation and its resolution, traditionally thought to be produced by immune cells. Using liquid chromatography–tandem mass spectrometry (LC-MS/MS)-based lipid mediator profiling PSCs as well as primary human CAFs express enzymes and receptors to produce and respond to SPMs. Human PSC/CAF SPM secretion profile can be modulated by rendering these cells activated [transforming growth factor beta (TGF-β)] or quiescent [all-trans retinoic acid (ATRA)]. ATRA-induced nuclear translocation of arachidonate-15-lipoxygenase (ALOX15) was linked to increased production of n-3 docosapentaenoic acid-derived Resolvin D5 (RvD5n-3 DPA), among other SPMs. Inhibition of RvD5n-3 DPA formation increases cancer cell invasion, whereas addback of this molecule reduced activated PSC-mediated cancer cell invasion. We also observed that circulating concentrations of RvD5n-3 DPA levels were decreased in peripheral blood of metastatic PDAC patients when compared with those measured in plasma of non-metastatic PDAC patients. Together, these findings indicate that RvD5n-3 DPA may regulate cancer–stroma cross-talk and invasion.

胰腺星状细胞（pancreatic stellate cells, PSCs）激活为癌相关成纤维细胞（cancer-associated fibroblasts, CAFs）是胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）间质中广泛促纤维增生反应的核心诱因，而该反应是驱动PDAC化疗耐药、导致不良预后的关键因素。特异性促炎症消退介质（specialized pro-resolving mediators, SPMs）是炎症及其消退的主要调控因子，传统观点认为其仅由免疫细胞产生。本研究通过基于液相色谱-串联质谱（liquid chromatography–tandem mass spectrometry, LC-MS/MS）的脂质介质谱分析，发现原代人PSCs及CAFs均表达可合成并响应SPMs的酶类与受体。人PSCs/CAFs的SPMs分泌谱可因细胞被诱导为激活状态（经转化生长因子β（transforming growth factor beta, TGF-β）处理）或静息状态（经全反式维甲酸（all-trans retinoic acid, ATRA）处理）而发生改变。全反式维甲酸诱导的花生四烯酸15-脂氧合酶（arachidonate-15-lipoxygenase, ALOX15）核转位，与包括n-3二十二碳五烯酸衍生的消退素D5（n-3 docosapentaenoic acid-derived Resolvin D5, RvD5n-3 DPA）在内的多种SPMs的生成增加密切相关。抑制RvD5n-3 DPA的生成会增强癌细胞侵袭能力，而外源性补充该分子则可降低激活的PSCs介导的癌细胞侵袭水平。本研究还观察到，与非转移性PDAC患者的血浆样本相比，转移性PDAC患者外周血中RvD5n-3 DPA的循环浓度显著降低。综上，上述研究结果表明RvD5n-3 DPA或可调控癌-间质串扰与癌细胞侵袭过程。