Lorazepam stimulates IL-6 production and is associated with poor survival outcomes in pancreatic cancer

LOR is associated with worse progression-free survival (PFS) while alprazolam (ALP) is associated with improved PFS, in pancreatic cancer patients receiving chemotherapy. LOR promotes desmoplasia (fibrosis and extracellular matrix protein deposition), inflammatory signaling, and ischemic necrosis. GPR68 is preferentially expressed on human PDAC CAFs, and n-unsubstituted BZDs, such as LOR, significantly increase IL-6 expression and secretion in CAFs in a pH and GPR68-dependent manner. Conversely, ALP, and other GPR68 n-substituted BZDs decrease IL-6 in human CAFs in a pH and GPR68-independent manner. Across many cancer types, LOR is associated with worse survival outcomes relative to ALP and patients not receiving BZDs. We demonstrate that LOR stimulates fibrosis and inflammatory signaling, promotes desmoplasia and ischemic necrosis, and is associated with decreased pancreatic cancer patient survival. This research investigates the association between benzodiazepines (BZDs) and cancer patient survival outcomes, the pancreatic cancer tumor microenvironment, and cancer-associated fibroblast (CAF) signaling. Multivariate Cox regression modeling was used to retrospectively measure associations between Roswell Park cancer patient survival outcomes and BZD prescription records. Immunohistochemistry, H&E, Masson’s trichrome, RNAscope, and RNA sequencing were used to evaluate the impact of lorazepam (LOR) on the murine PDAC tumor microenvironment. ELISA and qPCR were used to determine the impact of BZDs on IL-6 expression or secretion by human immortalized pancreatic CAFs. PRESTO-Tango assays, reanalysis of PDAC single cell sequencing/TCGA datasets, and GPR68 CRISPRi knockdown CAFs were used to determine the impact of BZDs on GPR68 signaling. 8 samples. Four from DMSO and LOR conditions

本研究以接受化疗的胰腺癌患者为研究对象，发现劳拉西泮（LOR，lorazepam）与更差的无进展生存期（progression-free survival, PFS）显著相关，而阿普唑仑（ALP，alprazolam）则与更佳的无进展生存期相关。劳拉西泮可促进结缔组织增生（即纤维化与细胞外基质蛋白沉积）、激活炎症信号通路并诱发缺血性坏死。G蛋白偶联受体68（GPR68）在人胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）的癌症相关成纤维细胞（cancer-associated fibroblasts, CAF）中呈偏好性表达；未经N取代的苯二氮䓬类药物（benzodiazepines, BZDs，如LOR）可通过酸碱度（pH）及GPR68依赖的途径，显著上调CAFs中白细胞介素6（interleukin-6, IL-6）的表达与分泌水平。反之，ALP及其他经N取代的苯二氮䓬类药物，则可通过非酸碱度及非GPR68依赖的途径，降低人CAFs中的IL-6水平。在多种癌症类型中，相较于ALP治疗组及未接受BZDs治疗的患者，LOR治疗均与更差的生存结局相关。本研究证实，劳拉西泮可刺激纤维化进程与炎症信号通路激活，促进结缔组织增生及缺血性坏死，并与胰腺癌患者生存期缩短显著相关。本研究旨在探究苯二氮䓬类药物与癌症患者生存结局、胰腺癌肿瘤微环境以及癌症相关成纤维细胞信号通路之间的关联。本研究采用多因素Cox回归模型，对罗斯韦尔帕克癌症患者的生存结局与苯二氮䓬类药物处方记录之间的关联进行回顾性分析。采用免疫组化、苏木精-伊红（H&E）染色、马松三色（Masson’s trichrome）染色、RNAscope原位杂交技术以及RNA测序，评估劳拉西泮对小鼠胰腺导管腺癌肿瘤微环境的影响。采用酶联免疫吸附测定（ELISA）与实时定量聚合酶链反应（qPCR），探究苯二氮䓬类药物对永生化人胰腺CAFs分泌或表达IL-6的影响。采用PRESTO-Tango检测法、胰腺导管腺癌单细胞测序/癌症基因组图谱（The Cancer Genome Atlas, TCGA）数据集的重新分析以及GPR68 CRISPR干扰（CRISPRi）敲低CAFs模型，探究苯二氮䓬类药物对GPR68信号通路的影响。本研究共纳入8份样本，其中4份来自二甲基亚砜（Dimethyl Sulfoxide, DMSO）对照组与LOR处理组。