Autophagy in Alcoholic Pancreatitis

酒精性胰腺炎中的自噬

• 批准号: 9925046
• 负责人: Wen-Xing Ding
• 金额: $ 34.43万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925046
• 关键词: Acetaldehyde; Acetates; Acinar Cell; Acute; Address; Adenoviruses; Agonist; Alcohol abuse; Alcohol consumption; Alcoholic Pancreatitis; Alcohols; Amylases; Antiinflammatory Effect; Autophagocytosis; Binding; Biogenesis; Blood; CNR2 gene; Cells; Chronic; Consumption; Dependovirus; Development; Drug Kinetics; Edema; Enzyme Precursors; Enzymes; Esters; Ethanol Metabolism; Excision; Exocrine pancreas; Extracellular Signal Regulated Kinases; FRAP1 gene; Fatty Acids; Goals; Human; Impairment; In Vitro; Infiltration; Inflammatory; Injury; Intervention; Intramural Research Program; Kansas; Knockout Mice; Light; Link; Lipase; Lysosomal Function Inhibition; Lysosomes; Mediating; Medical center; Microtubules; Modeling; Molecular; Morbidity - disease rate; Mus; National Institute on Alcohol Abuse and Alcoholism; Normal Cell; Oral; Outcome; Pancreas; Pancreatic enzyme; Pancreatitis; Pathogenesis; Pathway interactions; Pattern; Pharmacology; Pharmacology and Toxicology; Process; Proteins; Protocols documentation; Research; Risk Factors; Rodent; Role; Serum; Testing; Therapeutic Intervention; Time; Tissues; Toxic effect; Translations; Trypsin; Universities; Zymogen Granules; alcohol measurement; cell injury; chronic alcohol ingestion; chronic pancreatitis; clinical application; cytokine; feeding; improved; in vivo; mortality; mouse model; novel; novel therapeutics; overexpression; prevent; problem drinker; receptor; repaired; transcription factor

Project Summary  Alcohol abuse is a major risk factor in initiating pancreatitis in humans and in rodents. Experimental evidence indicates that alcohol consumption leads to the destabilization of lysosomes and increases zymogen fragility in acinar cells. Cells can adapt and protect themselves by removing these damaged lysosomes and zymogen granules (ZGs) by autophagy and lysosomal biogenesis. However, these protective mechanisms are impaired after chronic alcohol consumption. Cannabinoid receptor 2 (CB2) agonists have potent anti-inflammatory effects and protect against experimental pancreatitis in mice. However, the underlying molecular mechanisms by which CB2 agonists protect against pancreatitis are not known. Moreover, what is also missing is the availability of CB2 agonists with optimal pharmacokinetics and absence of major toxicity as tested in relevant alcoholic pancreatitis models. In our preliminary studies, we established a novel alcohol pancreatitis mouse model using the recently established chronic plus acute alcohol binge (Gao-binge) protocol. Gao-binge induces significant pathogenesis in mouse pancreas, including edema, increased ZGs, elevated expression of inflammatory cytokines, increased infiltration of inflammatory cells and release of pancreatic enzymes (amylase and lipase) into the blood. We also demonstrated that Gao-binge decreases expression of pancreatic cannabinoid receptor 2 (CB2) and inactivates TFEB, a master regulator of lysosomal biogenesis. A selective CB2 agonist activates TFEB in mouse pancreas and increases autophagic flux in cultured acinar cells. Here we propose a novel hypothesis that ethanol metabolism leads to decreased CB2 expression and impaired TFEB-mediated lysosomal biogenesis, insufficient autophagy, accumulation of fragile ZGs, damaged lysosomes, and subsequent pancreatitis. Our long-term goal is to understand the molecular mechanisms for alcohol impairment of lysosomal biogenesis in pancreatic acinar cells, in order to identify steps in the repair pathway that are points for intervention in alcoholic pancreatitis. The objective of this proposal is to understand how CB2 agonists activate TFEB-mediated lysosomal biogenesis and selective zymophagy and lysophagy to protect against alcohol-induced pancreatitis. Using complimentary expertise in CB2 receptor pharmacology/toxicology (NIAAA intramural research program) and modeling of lysosomal biogenesis and alcoholic pancreatitis in vitro and in vivo (University of Kansas Medical Center) we propose to test a set of novel, highly selective and orally available CB2 agonists assuring quick translation toward clinical application for the treatment of alcoholic pancreatitis. We will determine how the selective CB2 agonists activate TFEB- mediated lysosomal biogenesis and their ability to improve the insufficient autophagy to remove damaged lysosomes and ZGs induced by chronic alcohol consumption. This proposal brings together research expertise that will address key research questions on alcoholic pancreatitis and the potential intervention by CB2 agonists, which would not otherwise be possible.

项目摘要 酒精滥用是引发人类和啮齿动物胰腺炎的主要危险因素。实验证据 表明酒精消耗导致溶酶体不稳定，并增加酶原脆性， 腺泡细胞细胞可以通过清除这些受损的溶酶体和酶原来适应和保护自己 颗粒（ZGs）通过自噬和溶酶体生物合成。然而，这些保护机制受损 慢性酒精消耗后。大麻素受体2（CB 2）激动剂具有有效的抗炎作用， 对小鼠实验性胰腺炎的影响。然而，潜在的分子机制 CB 2激动剂通过何种途径预防胰腺炎尚不清楚。此外，还缺少 CB 2激动剂的可用性，具有最佳的药代动力学和无主要毒性，如在相关研究中所测试的， 酒精性胰腺炎模型。在我们的初步研究中，我们建立了一种新的酒精性胰腺炎小鼠， 模型使用最近建立的慢性加急性酒精狂欢（高-狂欢）协议。高宾格诱导 在小鼠胰腺中的显著发病机制，包括水肿、ZGs增加、 炎性细胞因子、炎性细胞浸润增加和胰腺酶释放 （淀粉酶和脂肪酶）进入血液。我们还证明，高斌格减少胰腺癌细胞的表达， 大麻素受体2（CB 2）和失活TFEB，溶酶体生物发生的主要调节剂。选择性 CB 2激动剂激活小鼠胰腺中的TFEB并增加培养的腺泡细胞中的自噬通量。这里 我们提出了一个新的假设，即乙醇代谢导致CB 2表达减少， TFEB介导的溶酶体生物发生，自噬不足，脆性ZG积累，受损 溶酶体和随后的胰腺炎。我们的长期目标是了解 酒精损伤胰腺腺泡细胞中溶酶体生物发生，以确定修复步骤 这些途径是酒精性胰腺炎的干预点。本提案的目的是了解 CB 2激动剂如何激活TFEB介导的溶酶体生物发生和选择性酶噬和溶噬， 防止酒精引起的胰腺炎。利用CB 2受体的补充专业知识 药理学/毒理学（NIAAA内部研究计划）和溶酶体生物发生建模， 酒精性胰腺炎在体外和体内（堪萨斯大学医学中心），我们建议测试一组 新型、高选择性和口服CB 2激动剂，确保快速转化为临床应用 酒精性胰腺炎的治疗方法我们将确定选择性CB 2激动剂如何激活TFEB-1。 介导的溶酶体生物发生和它们改善不充分的自噬以去除受损的 溶酶体和ZGs诱导的慢性酒精消耗。这项提案汇集了研究专家 这将解决酒精性胰腺炎的关键研究问题和CB 2的潜在干预作用， 激动剂，否则是不可能的。