The Minor Secretory Pathway in Pancreatitis

胰腺炎的次要分泌途径

• 批准号: 8743018
• 负责人: Fred Sanford Gorelick
• 金额: $ 25.72万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8743018
• 关键词: Acinar Cell; Adaptor Signaling Protein; Affect; Animals; Apical; Autophagocytosis; Binding; Biochemical; Biogenesis; Biostatistics Core; Cause of Death; Cell Fractionation; Cell membrane; Characteristics; Clathrin Adaptors; Data; Disease; Electron Microscopy; Enzyme Precursors; Enzymes; Event; Exocytosis; Exposure to; Functional disorder; Genetic; Goals; Health; Human; Inflammation Mediators; Inflammatory; Injury; Late Effects; Link; Lysosomes; Mediating; Membrane Protein Traffic; Membrane Proteins; Minor; Modeling; Molecular; Mus; Mutate; Organelles; Pancreas; Pancreatitis; Pathologic; Pathology; Pathway interactions; Peptide Hydrolases; Physiological; Preparation; Process; Proteins; Research Personnel; Resources; Role; Secretory Vesicles; Signal Transduction; Site; Stress; TPD52 gene; Techniques; Vesicle; Zymogen Granules; abstracting; acute pancreatitis; cell injury; chronic pancreatitis; design; genetic regulatory protein; in vivo; in vivo Model; light microscopy; lysosomal proteins; prevent; programs; protein expression; protein function; protein transport; rab GTP-Binding Proteins; response; trafficking; tumor; vector

Project 2: Summary/Abstract 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 usually originates in the pancreatic acinar cell after exposure to specific insults that cause pathologic acinar cell signaling. These changes in acinar cell signaling change the function of specific target proteins and cause organelle dysfunction. Among the most characteristic acinar cell early pancreatitis responses are intracellular activation of digestive enzymes (especially proteases) and inhibited secretion. Recent studies from Program investigators have also shown that there is decreased lysosomal function in acute pancreatitis. This Project will examine two recently characterized proteins (AP3 and D52) that can modulate zymogen activation, secretion, and may also affect autophagy and lysosomal function. We hypothesize that these two proteins regulate the biogenesis and trafficking of the minor secretory compartment (MSC), a pathway needed to form zymogen granules (ZG) from immature secretory granules (ISG). We posit that this compartment also regulates apical secretion and contributes to the normal function of the endo-lysosomal pathway. Finally, we propose a new model for zymogen activation at the onset of pancreatitis that begins with zymogen activation in the MSC then proceeds to decreased degradation of active enzymes by lysosomes. Three Specific Aims relating to AP3 and D52 and its binding partner, Rab5, are planned: 1) Investigate their role in regulating physiologic secretion and membrane trafficking 2) Examine their expression and localization in pancreatitis models 3) Determine whether changing their expression levels affects pancreatitis responses. Finally, we will collaborate with other Projects to determine whether these molecules affect other pathways under study. Studies are planned using isolated mouse and human pancreatic acinar cells and in vivo mouse pancreatitis models. Our preliminary studies indicate that during pancreatitis, the levels of expression of these proteins change. Further, over-expression or deletion of these proteins by genetic or molecular means has dramatic effects on lysosomal protein trafficking, zymogen activation, and cell injury. Preliminary findings also suggest that the functions of AP3 and D52 may interact with pathways under study by other projects, such as Rab GTPases in Project 1, autophagic pathways in Project 3, and ER stress in Project 4. Extensive use of the Program's Animal and Pathology, Vector and Human Acinar, and Resource Management and Biostatistics Cores is planned.

项目2：摘要/摘要 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。该疾病通常起源于 暴露于导致病理腺泡细胞信号传导的特定损伤后，胰腺腺泡细胞。这些 腺泡细胞信号的变化会改变特定靶蛋白的功能，并引起细胞器功能障碍。 在最有特征性的腺泡细胞中，早期胰腺炎反应是消化的细胞内激活 酶（尤其是蛋白酶）并抑制分泌。计划调查人员的最新研究也 表明急性胰腺炎的溶酶体功能降低。 该项目将检查两个最近表征的蛋白质（AP3和D52），它们可以调节Zymogen 激活，分泌，也可能影响自噬和溶酶体功能。我们假设这两个 蛋白质调节次要分泌室（MSC）的生物发生和运输，这是所需的途径 从未成熟的分泌颗粒（ISG）中形成酶原颗粒（ZG）。我们认为这个隔间也 调节顶端分泌，并有助于内部溶酶体途径的正常功能。最后，我们 在胰腺炎发作时提出了一种新的模型，用于酶原激活，该模型始于酶原激活 然后，MSC通过溶酶体降低了活性酶的降解。 计划针对AP3和D52及其约束伙伴Rab5的三个具体目标： 1）研究它们在调节生理分泌和膜贩运方面的作用 2）检查它们在胰腺炎模型中的表达和定位 3）确定改变其表达水平是否影响胰腺炎反应。 最后，我们将与其他项目合作，以确定这些分子是否影响 其他正在研究的途径。使用孤立的小鼠和人类胰腺腺泡细胞计划进行研究 和体内小鼠胰腺炎模型。我们的初步研究表明，在胰腺炎期间， 这些蛋白质的表达发生了变化。此外，通过遗传或 分子平均值对溶酶体蛋白运输，酶原激活和细胞损伤具有巨大影响。 初步发现还表明，AP3和D52的功能可能与正在研究的途径相互作用 其他项目，例如项目1中的RAB GTPASE，项目3中的自噬途径以及项目4中的ER压力。 广泛使用该计划的动物和病理学，向量和人类腺泡以及资源管理 计划了生物统计学核心。