Exocrine pancreatic zymogen activation

外分泌胰腺酶原激活

基本信息

批准号：
8694878
负责人：
Fred Sanford Gorelick
金额：
$ 27.13万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-07-15 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8694878
关键词：
5&apos -AMP-activated protein kinase Acinar Cell Acinus organ component Affect Apical Blood flow Calcium Catalytic Domain Cause of Death Cell Death Co-Immunoprecipitations Data Development Disease Drug Targeting Drug usage Effectiveness Enzyme Activation Enzyme Precursors Enzymes Exocrine pancreas Exposure to Genetic Hand Human Inflammation Inflammation Mediators Inflammatory Injury Islets of Langerhans Transplantation Knockout Mice Knowledge Lead Link Mediator of activation protein Metformin Modeling Molecular Target Mus Pancreas Pancreatitis Pathologic Pathway interactions Perfusion Pharmaceutical Preparations Phase Phosphorylation Preparation Process Protein Isoforms Protein Kinase Protein Kinase C Publications Publishing Rattus Reporting Rodent Role Severities Severity of illness Signal Transduction Signaling Molecule Small Interfering RNA Therapeutic Time Trypsinogen Wild Type Mouse acute pancreatitis base cellular targeting in vivo in vivo Model prevent protective effect public health relevance response salicylate therapeutic target trafficking vacuolar H+-ATPase

5&apos -AMP-activated protein kinase Acinar Cell Acinus organ component Affect Apical Blood flow Calcium Catalytic Domain Cause of Death Cell Death Co-Immunoprecipitations Data Development Disease Drug Targeting Drug usage Effectiveness Enzyme Activation Enzyme Precursors Enzymes Exocrine pancreas Exposure to Genetic Hand Human Inflammation Inflammation Mediators Inflammatory Injury Islets of Langerhans Transplantation Knockout Mice Knowledge Lead Link Mediator of activation protein Metformin Modeling Molecular Target Mus Pancreas Pancreatitis Pathologic Pathway interactions Perfusion Pharmaceutical Preparations Phase Phosphorylation Preparation Process Protein Isoforms Protein Kinase Protein Kinase C Publications Publishing Rattus Reporting Rodent Role Severities Severity of illness Signal Transduction Signaling Molecule Small Interfering RNA Therapeutic Time Trypsinogen Wild Type Mouse acute pancreatitis base cellular targeting in vivo in vivo Model prevent protective effect public health relevance response salicylate therapeutic target trafficking vacuolar H+-ATPase

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项目摘要

Acute pancreatitis, an inflammatory disease of the pancreas, causes death in over 30% of those with severe disease. Specific therapies to prevent pancreatitis or reduce injury are lacking. The disease is thought to begin in the pancreatic acinar cell after exposure to insults that initiate pathologic changes in this cell's signaling. Subsequent pathologic acinar cell responses are followed by changes in blood flow, inflammation, and cell death. We propose to examine a previously undescribed mediator of intracellular signaling that modulates pancreatitis responses, AMP-regulated protein kinase (AMPK). The enzyme may be an attractive therapeutic target. Based on our preliminary findings and recent publication, we hypothesize that AMPK serves a protective role in pancreatitis within the acinar cell and does so by inhibiting key mediators of pancreatitis, including the vacuolar ATPase (vATPase). Further, its protective role appears to be reduced in the acinar cell during the early phases of pancreatitis when AMPK activity rapidly decreases in selective cellular compartments. In preliminary studies, the proposed protective role for AMPK was supported by findings in cellular and in vivo pancreatitis models which showed that: a) clinically used drugs that can activate AMPK reduce disease severity, b) genetically reducing AMPK using either siRNA in pancreatic acini (groups of 20-100 acinar cells) or a knockout mouse lacking the essential AMPK ¿1 subunit, increases the severity of pancreatitis. Finally, preliminary studies indicate that human acinar cells contain AMPK and have an essential subunit isoform (¿1) that is required for drug targeting. To examine the role of AMPK in pancreatitis, the following Specific Aims are planned: 1) Examine agents that activate AMPK in isolated pancreatic acini, 2) Examine agents that activate AMPK using in vivo models of pancreatitis, and 3) Determine the molecular and cellular targets of AMPK. Studies will be carried out with isolated groups of pancreatic acinar cells (acini) and in vivo pancreatitis models that produce mild and severe disease. Effects observed in isolated rodent acinar cells will be confirmed in a similar preparation of human acinar cells. Genetic knockdown of AMPK in acini has been performed and intact mice with deletions of AMPK's catalytic subunits are on hand and will be used to confirm AMPK's role in pancreatitis and to show that drugs found to reduce disease severity are acting through AMPK. The effects of AMPK activating drugs on AMPK translocation and downstream processes such as vATPase activation will be examined. We have identified AMPK activators that are currently available medications with safe profiles that also decrease the severity of acute pancreatitis in our models. Thus, the proposed studies have strong translational and therapeutic potential.

急性胰腺炎是胰腺的炎症性疾病，导致30％以上的严重患者死亡疾病。缺乏预防胰腺炎或减少损伤的特定疗法。人们认为这种疾病在暴露于引发该细胞病理变化的损伤后，在胰腺腺泡细胞中开始信号。随后的病理性腺泡细胞反应之后是血流，炎症的变化，和细胞死亡。我们建议检查先前未描述的细胞内信号传导中介人调节胰腺炎反应，AMP调节的蛋白激酶（AMPK）。酶可能是一个吸引人的治疗靶标。根据我们的初步发现和最新出版物，我们假设AMPK 在腺泡细胞内发挥了受保护的作用胰腺炎，包括真空ATPase（VATPase）。此外，其保护作用似乎减少了当AMPK活性在选择性中迅速降低时，胰腺炎早期的腺泡细胞细胞室。在初步研究中，AMPK的拟议保护作用得到了支持细胞和体内胰腺炎模型中的发现表明：a）临床使用的药物可以激活AMPK降低疾病的严重程度，b）在胰腺中使用任何一个siRNA通常会降低AMPK （一组20-100个腺泡细胞）或缺乏必需AMPK的基因敲除小鼠增加，增加了胰腺炎的严重程度。最后，初步研究表明人腺泡细胞包含AMPK，并且具有药物靶向必需的必需亚基同工型（1）。检查AMPK在胰腺炎，计划以下特定目的：1）检查激活AMPK的药物孤立的胰腺acini，2）检查使用体内模型激活AMPK的剂胰腺炎和3）确定AMPK的分子和细胞靶标。研究将是用孤立的胰腺腺泡细胞（acini）和体内胰腺炎模型进行产生轻度和重度疾病。在孤立的啮齿动物细胞中观察到的效果将在A中得到确认人类腺泡细胞的类似制备。 ACINI中AMPK的遗传敲低已经进行了，并且具有AMPK催化亚基的缺失的完整小鼠在手上，将用于确认AMPK的角色在胰腺炎中，以表明发现降低疾病严重程度的药物正在通过AMPK作用。效果在AMPK易位和下游过程（例如VATPase激活）上激活药物的AMPK激活药物将被检查。我们已经确定了AMPK激活剂，这些AMPK激活剂当前可用的药物安全在我们的模型中，还会减少急性胰腺炎的严重程度。那就是拟议的研究强大的翻译和治疗潜力。