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

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.

急性胰腺炎是一种胰腺炎症性疾病，导致 30% 以上的重症患者死亡人们认为缺乏预防胰腺炎或减少损伤的具体疗法。在暴露于导致该细胞发生病理变化的损伤后，在胰腺腺泡细胞中开始随后的病理性腺泡细胞反应伴随着血流、炎症、我们建议检查一种以前未描述过的细胞内信号传导介质。调节胰腺炎反应，AMP 调节蛋白激酶 (AMPK) 可能是一种有吸引力的酶。根据我们的初步研究结果和最近发表的文章，我们采用了 AMPK。在腺泡细胞内的胰腺炎中发挥保护作用，并通过抑制关键介质来实现这一作用胰腺炎，包括空泡 ATP 酶（vATP 酶），此外，其保护作用似乎有所减弱。胰腺炎早期阶段，当 AMPK 活性选择性迅速降低时，腺泡细胞在初步研究中，提出的 AMPK 保护作用得到了支持。在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 降低疾病严重程度，b) 在胰腺腺泡中使用任一 siRNA 从基因上降低 AMPK （20-100 个腺泡细胞组）或缺乏必需 AMPK 的基因敲除小鼠 ¿ 1 个亚基，增加最后，初步研究表明，人类腺泡细胞含有 AMPK 和具有药物靶向所需的重要亚基亚型 (¿1) 以检查 AMPK 在其中的作用。胰腺炎，计划了以下具体目标：1）检查激活 AMPK 的药物分离的胰腺腺泡，2) 使用体内模型检查激活 AMPK 的药物胰腺炎，以及 3) 确定 AMPK 研究的分子和细胞靶点。用分离的胰腺腺泡细胞（acini）组和体内胰腺炎模型进行在分离的啮齿动物腺泡细胞中观察到的影响将在实验中得到证实。已对腺泡细胞中的 AMPK 进行了类似的制备，并进行了类似的制备。现有 AMPK 催化亚基缺失的完整小鼠，将用于确认 AMPK 的作用并证明可降低疾病严重程度的药物是通过 AMPK 发挥作用的。 AMPK 激活药物对 AMPK 易位和 vATP 酶激活等下游过程的影响我们已确定 AMPK 激活剂是目前可用的安全药物。因此，拟议的研究也降低了我们模型中急性胰腺炎的严重程度。强大的转化和治疗潜力。