Protein Homeostasis in AL Amyloidosis

描述(由申请人提供)：这个项目的长期目标是描述蛋白质稳态的分子基础及其调节，并了解该系统的能力如何以及为什么随着年龄的增长而下降，从而导致功能性疾病的丧失和获得。本项目的具体目标1将描述控制淀粉样变轻链(LC)折叠和内质网(ER)相关降解(ERAD)途径之间的平衡的蛋白质动态平衡网络，以了解人类疾病淀粉样蛋白LC淀粉样变性(AL)的基础。对分泌淀粉样蛋白LC的细胞系进行高通量筛选(HTS)将使我们能够进行全基因组筛选，以发现新的靶点，将平衡转向降解，而不是容易错误折叠的LC的分泌，这一结果被预测将对这种疾病产生实质性的影响，甚至可能治愈这种疾病。特定目标2还将使用HTS格式来识别化学文库中的小分子，这些小分子减少淀粉样蛋白LCS的分泌，有利于ERAD，从而导致组织中细胞外淀粉样蛋白负荷的减少。在这两个特定的目标中，我们将探索一个普遍的假设，即蛋白质稳态网络在我们年轻时可以控制这些疾病，但这些组件在衰老过程中的失败可能会导致功能获得蛋白毒性和器官衰竭，原因是淀粉样蛋白负荷超过修复和降解途径的能力。要实现这两个具体目标，需要研究人员之间强有力的四方合作，他们拥有有效互动的记录，并拥有强大和互补的专业知识，应用细胞生物学、化学以及小鼠和蠕虫模型方法来了解AL疾病，以应对老龄化计划。项目简介：该项目描述了一种新的高通量筛选方法，以确定在衰老过程中影响人类骨髓瘤疾病轻链淀粉样变性(AL)的发病和病理的小分子化合物和途径。对这些途径和化合物的识别将导致对病理生理学的关键新见解，并可能治愈疾病。

DESCRIPTION (provided by applicant): The long-term goal of this project is to characterize the molecular basis of protein homeostasis and its regulation, and to understand how and why the capacity of this system declines with aging leading to both loss and gain of function diseases. Specific Aim 1 of this project will be to characterize the protein homeostasis network that controls the balance between amyloidogenic light chain (LC) folding and secretion versus degradation by endoplasmic reticulum (ER) associated degradation (ERAD) pathways to understand the basis for the human disease amyloid LC amyloidosis (AL). A high throughput screening (HTS) application of biologicals (siRNA and cDNA) to cell lines secreting amyloidogenic LCs will allow us to do a genome wide screen to uncover new targets that will shift the balance towards degradation and away from secretion of misfolding prone LCs, a result that is predicted to have a substantial impact on this disease, and possibly even cure the disease. Specific Aim 2 will also use a HTS format to identify small molecules in chemical libraries that diminish secretion of amyloidogenic LCs in favor of ERAD and therefore lead to reduction of extracellular amyloid load in tissue. In both specific aims we will explore the general hypothesis that protein homeostasis networks keep these diseases in check when we are young, but that a failure of these components during aging can contribute to gain-of-function proteotoxicity and organ failure due to amyloid burden that exceeds the capacity of repair and degradation pathways. Accomplishing these two specific aims involves a strong 4-way collaboration amongst investigators who have a track record of effective interaction, and have strong and complimentary expertise to apply cell biological, chemical and mouse and worm model approaches to understand AL disease in response to the aging program. Project Narrative: This project describes a new high-throughput screening approach to identify small molecule compounds and pathways that affect the onset and pathology of human myeloma disease light chain amyloidosis (AL) during aging. Identification of these pathways and compounds will lead to critical new insights into pathophysiology and a possible cure for disease.