Exocrine pancreatic zymogen activation

外分泌胰腺酶原激活

• 批准号: 8820251
• 负责人: Fred Sanford Gorelick
• 金额: $ 22.78万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/8820251
• 关键词: 5' -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; Health; 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; Salicylic Acids; 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; response; therapeutic target; trafficking; vacuolar H+-ATPase

DESCRIPTION (provided by applicant): 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. Subsequen 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 pancreatiti 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 alpha-1 subunit, increases the severity of pancreatitis. Finally, preliminary studies indicate that human acinar cells contain AMPK and have an essential subunit isoform (beta-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.

描述（由申请人提供）：胰腺炎性疾病急性胰腺炎，导致30％的严重疾病患者死亡。缺乏预防胰腺炎或减少损伤的特定疗法。人们认为该疾病是在胰腺腺泡中开始的 暴露于侮辱后，会引发该细胞信号传导中的病理变化。随后的病理腺泡细胞反应随后是血流，炎症和细胞死亡的变化。我们建议检查一个先前未描述的细胞内信号传导介质，该介体调节胰腺炎反应，AMP调节的蛋白激酶（AMPK）。该酶可能是一个有吸引力的治疗靶标。根据我们的初步发现和最近的出版物，我们假设AMPK在腺泡细胞内的胰腺炎中发挥了保护作用，并通过抑制包括液泡ATPase（VATPase）在内的胰腺炎的关键介质来做到这一点。此外，当AMPK活性在选择性细胞室中迅速降低时，在胰腺早期的早期阶段，其保护作用似乎会降低。在初步研究中，在细胞和体内胰腺炎模型中的发现支持了AMPK提出的保护作用，该模型表明：a）可以激活AMPK的临床使用的药物，降低了疾病的严重性，b）遗传上使用任何一个sirna在pancreatic acini中使用sirna的遗传性AMPK，或者缺乏20-100个基础的nigner acnun acnun acnun acnun nockout and nockout小鼠），这是一个基础，是一个良好的and and and and and nockout小鼠。胰腺炎的严重程度。最后，初步研究表明，人类腺泡细胞包含AMPK，并具有药物靶向所需的必需亚基同工型（beta-1）。为了检查AMPK在胰腺炎中的作用，计划了以下特定目的：1）检查激活孤立胰腺acini中AMPK的剂，2）检查使用体内胰腺炎模型激活AMPK的药物，以及3）确定AMPK的分子和细胞靶标。研究将使用胰腺腺泡细胞（ACINI）的孤立组和产生轻度和严重疾病的体内胰腺炎模型进行。在人类腺泡细胞的类似制备中，将确认在分离的啮齿动物细胞中观察到的效果。 AMPK在ACINI中的遗传敲低已经进行了，并且完整的小鼠在手头上删除了AMPK催化亚基的缺失，并将用于确认AMPK在胰腺炎中的作用，并表明发现降低疾病严重程度的药物通过AMPK作用。将检查AMPK激活药物对AMPK易位和下游过程（例如VATPase激活）的影响。我们已经确定了当前可用的具有安全特征的药物的AMPK激活剂，这些药物也会减少我们模型中急性胰腺炎的严重程度。因此，拟议的研究具有强大的转化和治疗潜力。