Impact of benzodiazepines on the pancreatic tumor microenvironment

苯二氮卓类药物对胰腺肿瘤微环境的影响

• 批准号: 10386004
• 负责人: Abigail Carissa Cornwell
• 金额: $ 3.13万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386004
• 关键词: Address; Adverse effects; Allografting; Alprazolam; Anti-Anxiety Agents; Anxiety; Automobile Driving; Benzodiazepines; Blood Vessels; Chemoresistance; Clinical; Collagen; Deposition; Desmoplastic; Development; Disease; Drug Delivery Systems; Fibroblasts; Fibrosis; G-Protein-Coupled Receptors; GPR68 gene; Genes; Genetically Engineered Mouse; Goals; Growth; Histologic; Human; Image; Impairment; In Vitro; Inflammation; Inflammatory; Ischemia; KRASG12D; Lorazepam; Malignant Neoplasms; Malignant neoplasm of pancreas; Methods; Modeling; Mus; Nausea; Necrosis; Organoids; Outcome; Pain; Pancreatic Ductal Adenocarcinoma; Patient-Focused Outcomes; Patients; Perfusion; Pharmaceutical Preparations; Phenotype; Progression-Free Survivals; Proteins; Protons; Recommendation; Research; Role; Signal Transduction; Signaling Molecule; Sleeplessness; Survival Rate; Testing; Treatment Protocols; Work; base; cancer risk; chemotherapy; clinically relevant; clinically significant; cytokine; epidemiology study; improved; in vivo; mouse model; overexpression; pancreatic cancer patients; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; response; subcutaneous; treatment response; tumor; tumor growth; tumor microenvironment

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDA), is a lethal malignancy, due in part to a dense, desmoplastic (fibrotic) tumor microenvironment which aids tumor growth and inhibits drug delivery. Poor survival rates, harsh treatments, and tumor intrinsic aspects of the disease such as pain and altered cytokine pools, promote high levels of anxiety in PDA patients. Nearly 25% of pancreatic cancer patients are prescribed benzodiazepines (BZDs) to treat anxiety, insomnia, or to relieve nausea. Epidemiological studies indicate that BZDs may increase the risk of cancer development but the role of BZDs in modifying the tumor microenvironment is unknown. Recent studies indicate that n-unsubstituted BZDs promote the signaling of the proton-sensing G protein-coupled receptor, GPR68, under acidic conditions, such as those present in the PDA tumor microenvironment. GPR68 is overexpressed by PDA cancer-associated fibroblasts (CAFs), and its expression drives desmoplasia, fibrosis, and inflammation, microenvironmental features associated with impaired drug delivery and chemoresistance. To establish clinical relevance, covariate adjusted analyses was conducted of pancreatic cancer patients who received chemotherapy at Roswell Park from 2004 to 2020. Patients receiving lorazepam (LOR), a strong GPR68 activator, had significantly decreased progression-free survival (PFS) relative to non-users (HR 3.83 (1.53,9.57)), while patients receiving alprazolam (ALP), a GPR68 non-activator, had significantly improved PFS (HR 0.38 (0.16-0.92)). Preliminary studies using subcutaneous allografts derived from KPC mice, a genetically engineered mouse model of PDA, indicated that LOR promoted collagen deposition, desmoplasia, and ischemic necrosis in vivo, supporting that this BZD may promote an unfavorable tumor microenvironment that could negatively impact PDA patient survival. Additionally, LOR-treated CAFs had increased expression of pro- inflammatory and pro-fibrotic genes, suggesting that CAFs may be driving the observed phenotype. These findings led to the central hypothesis that GPR68-activating BZDs modulate CAF signaling, which will increase desmoplasia, subsequently constricting the tumor vasculature. The long-term goal is to determine the impact of BZDs on the PDA tumor microenvironment and chemotherapeutic efficacy. This hypothesis will be tested with two aims. Aim 1 will determine the impact of LOR/ALP on the PDA vasculature and desmoplasia in tumor-bearing KPC mice, using imaging and histological methods. Aim 2 will evaluate the role of BZDs in modifying CAF signaling and activity. Changes in the expression/secretion of pro-fibrotic proteins, inflammatory cytokines, and GPR68 downstream signaling molecules by BZD-treated immortalized CAFs will be quantified. Alterations in PDA organoid proliferation and collagen contraction by BZD-treated CAFs will also be quantified. This research is clinically significant because it will indicate if BZDs commonly prescribed to PDA patients modify the PDA tumor microenvironment in a manner which may decrease chemotherapeutic efficacy.

项目总结/摘要 胰腺导管腺癌（PDA）是一种致命的恶性肿瘤，部分原因是由于致密的促结缔组织增生（纤维化） 肿瘤微环境，其有助于肿瘤生长并抑制药物递送。存活率低， 治疗和疾病的肿瘤内在方面，如疼痛和改变的细胞因子库，促进高表达。 PDA患者的焦虑水平。近25%的胰腺癌患者服用苯二氮卓类药物 （BZD）治疗焦虑、失眠或缓解恶心。流行病学研究表明，BZD可能会增加 癌症发展的风险，但BZD在改变肿瘤微环境中的作用尚不清楚。最近 研究表明，n-未取代的BZD促进质子敏感G蛋白偶联的信号传导， 受体，GPR 68，在酸性条件下，如那些存在于PDA肿瘤微环境。GPR68 由PDA癌相关成纤维细胞（CAF）过表达，其表达驱动结缔组织增生，纤维化， 以及炎症、与受损的药物递送和化学抗性相关的微环境特征。 为了建立临床相关性，对胰腺癌患者进行了协变量校正分析， 从2004年到2020年在罗斯威尔公园接受化疗。接受劳拉西泮（LOR）的患者， GPR 68激活剂，相对于非使用者，具有显著降低的无进展生存期（PFS）（HR 3.83 (1.53，9.57）），而接受阿普唑仑（ALP）（一种GPR 68非激活剂）的患者PFS显著改善 (HR 0.38（0.16-0.92））。使用来自KPC小鼠的皮下同种异体移植物的初步研究， PDA的工程小鼠模型，表明LOR促进胶原沉积，结缔组织增生和缺血性血管生成， 体内坏死，支持这种BZD可能促进不利的肿瘤微环境， 对PDA患者的生存率有负面影响。此外，LOR处理的CAFs表达增加， 炎症和促纤维化基因，表明CAF可能驱动观察到的表型。 这些发现导致了核心假设，即GPR 68激活BZD调节CAF信号传导， 将增加结缔组织增生，随后收缩肿瘤血管。长期目标是 确定BZD对PDA肿瘤微环境和化疗功效的影响。这 我们将以两个目标来检验这个假设。目的1将确定LOR/ALP对PDA血管系统的影响 和结缔组织增生在荷瘤KPC小鼠，使用成像和组织学方法。目标2将评估 BZD在修饰CAF信号传导和活性中的作用。促纤维化蛋白表达/分泌的变化， 炎症细胞因子和GPR 68下游信号分子的BZD处理的永生化CAF将被 量化。BZD处理的CAFs引起的PDA类器官增殖和胶原收缩的改变也将被观察到。 量化。这项研究具有临床意义，因为它将表明BZD是否常用于PDA 患者以可能降低化疗功效的方式改变PDA肿瘤微环境。