Characterizing the fibrogenic role of NADPH oxidase 1 in the transition from chronic pancreatitis to pancreatic cancer

表征 NADPH 氧化酶 1 在慢性胰腺炎向胰腺癌转变中的纤维形成作用

基本信息

批准号：
10515175
负责人：
Maria Eugenia Sabbatini
金额：
$ 44.4万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10515175
关键词：
Address Attenuated Basal Cell Basal lamina Biological Assay Caerulein Cancer cell line Carcinoma in Situ Cells Clinical Collagen Type IV Coomassie blue Detection Digestion Down-Regulation E-Cadherin Early treatment Enzymes Event Exposure to Extracellular Matrix Proteins Fluorescence Fluorescent in Situ Hybridization Free Radical Formation Future Gelatinase B General Population Generations Genetic Goals High Pressure Liquid Chromatography Hydrogen Peroxide Immunohistochemistry In Vitro Inflammation Mediators Inflammatory Intervention KPC model Knockout Mice Knowledge Laminin Lead Lesion Location Luciferases Malignant neoplasm of pancreas Mass Spectrum Analysis Matrix Metalloproteinases Mentors Molecular Morphology Mus NADP NADPH Oxidase NADPH Oxidase 1 Organ Outcome Oxides Pancreas Pancreatic Ductal Adenocarcinoma Pancreatic Intraepithelial Neoplasia Patients Process Production Proteins RNA Interference Reactive Oxygen Species Regulation Reporter Research Retroviridae Role Saline Signal Transduction Small Interfering RNA Stains Stimulus Students Supervision Tamoxifen Testing Thinness Tissues Translational Research Universities Up-Regulation Vertebral column Western Blotting base chronic pancreatitis design experience experimental study gel electrophoresis high risk hydroethidine in vitro testing in vivo in vivo evaluation innovation overexpression oxidation pancreatic cancer cells pancreatic stellate cell prevent programs promoter pyrroline student participation targeted treatment transcription factor translational impact undergraduate research undergraduate research experience undergraduate student

项目摘要

PROJECT SUMMARY/ABSTRACT The problem: Patients suffering from chronic pancreatitis (CP) have a higher risk of pancreatic ductal adenocarcinoma (PDAC). CP is characterized by an activated pancreatic stellate cell (PaSC)-rich stroma, which has facilitated the progression of non-invasive PanIN lesions to invasive PDAC. A critical barrier to progress in preventing the CP-to-PDAC transition is the gap of knowledge regarding the mechanism by which quiescent PaSCs become activated by inflammatory mediators, expand, and synthesize stroma and matrix metalloproteinases (MMPs), facilitating the progression of non-invasive PanIN lesions to invasive PDAC. Among the inflammatory mediators of CP are reactive oxygen species (ROS), which activate PaSCs. ROS generation can occur as a primary product of NADPH oxidase (Nox) enzymes. Previously, we showed that Nox1 signaling in CP-activated PaSCs: i) forms fibrotic tissue, ii) up-regulates both the transcription factor E-cadherin repressor Twist1 and MMP-9, and iii) facilitates the invasion of pancreatic cancer cell lines both in vitro and in vivo. The objective: To address the gaps in our knowledge regarding the mechanisms by which Nox1/Twist1/MMP-9 signaling in CP-activated PaSCs facilitates the progression of non-invasive PanIN lesions to invasive PDAC. The central hypothesis: The induction of CP generates ROS by Nox1 in PaSCs, which lead to a sustained expression of Twist1. Twist1, in turn, induces the expression of MMP-9, which promotes the progression of non- invasive PanIN lesions to invasive PDAC by degrading the basal lamina. The hypothesis will be tested by pursuing two specific aims: 1) Under the first aim, we will test in vitro the prediction that inhibiting Nox1/Twist1/MMP-9 signaling in CP-activated PaSCs attenuates the degradation of collagen IV and laminin. Under the second aim, we will test in vivo the prediction that inhibiting Nox1 signaling in CP-activated PaSCs prevents the progression of non-invasive PanIN lesions to invasive PDAC by attenuating the degradation of basal lamina. Our outcomes will include 1) new knowledge of CP-related mechanisms of progression of PDAC, 2) new knowledge of Nox1-related mechanisms of progression of PDAC, 3) high impact research experiences for undergraduate students. The approach is innovate because it will assess the extent to which the generation of Nox1-derived ROS from CP-activated PaSCs can facilitate the progression of non-invasive PanIN lesions to invasive PDAC. The proposed research is significant because a finding that the lack of Nox1 in CP-activated PaSCs prevents the progression of PanIN lesions to invasive PDAC by attenuating the degradation of basal lamina will establish the feasibility and premise for future translational research directed at developing rational, stroma-targeted therapies of early-stages of PDAC (e.g., carcinoma in situ) that, in combination with other approaches might lead to clinical strategies to increase the survival of patients. Moreover, these studies are designed to promote opportunities for a diverse undergraduate student participation providing them with exposure to hands-on experiments and close mentoring.

项目概要/摘要问题：慢性胰腺炎 (CP) 患者发生胰管癌的风险较高腺癌（PDAC）。 CP 的特征是富含活化的胰腺星状细胞 (PaSC) 的基质，促进了非侵袭性 PanIN 病变进展为侵袭性 PDAC。进步的关键障碍阻止 CP 向 PDAC 转变的原因是关于静态 PaSC 被炎症介质激活、扩张并合成基质和基质金属蛋白酶 (MMP)，促进非侵袭性 PanIN 病变进展为侵袭性 PDAC。之中 CP 的炎症介质是活性氧 (ROS)，它可以激活 PaSC。 ROS生成可以作为 NADPH 氧化酶 (Nox) 的初级产物出现。之前，我们展示了 Nox1 信号传导在 CP 激活的 PaSC 中：i) 形成纤维化组织，ii) 上调转录因子 E-钙粘蛋白阻遏蛋白 Twist1 和 MMP-9，以及 iii) 在体外和体内促进胰腺癌细胞系的侵袭。这目标：弥补我们在 Nox1/Twist1/MMP-9 机制方面的知识空白 CP 激活的 PaSC 中的信号传导促进非侵袭性 PanIN 病变进展为侵袭性 PDAC。中心假设：CP 的诱导通过 PaSC 中的 Nox1 产生 ROS，从而导致持续的 Twist1 的表达式。 Twist1 反过来诱导 MMP-9 的表达，从而促进非通过降解基底层，将侵袭性 PanIN 病变转化为侵袭性 PDAC。该假设将通过以下方式进行检验追求两个具体目标：1）在第一个目标下，我们将在体外测试抑制的预测 CP 激活的 PaSC 中的 Nox1/Twist1/MMP-9 信号传导可减弱 IV 型胶原蛋白和层粘连蛋白的降解。在第二个目标下，我们将在体内测试抑制 CP 激活的 PaSC 中的 Nox1 信号传导的预测通过减弱 PDAC 的降解来防止非侵袭性 PanIN 病变进展为侵袭性 PDAC 基底层。我们的成果将包括 1) 对 PDAC 进展的 CP 相关机制的新认识， 2) Nox1 相关 PDAC 进展机制的新知识，3) 高影响力的研究经验对于本科生。这种方法是创新的，因为它将评估一代人的影响程度 CP 激活的 PaSC 中 Nox1 衍生的 ROS 可以促进非侵袭性 PanIN 病变进展为侵入性 PDAC。拟议的研究意义重大，因为发现 CP 激活中缺乏 Nox1 PaSCs 通过减弱基底细胞的降解来防止 PanIN 病变进展为侵袭性 PDAC lamina 将为未来的转化研究建立可行性和前提，旨在开发合理的、基质靶向治疗的早期阶段 PDAC（例如，原位癌）与其他相结合方法可能会导致提高患者生存率的临床策略。此外，这些研究是旨在促进多元化本科生参与的机会，为他们提供接触实践实验和密切指导。