Protein Homeostasis in AL Amyloidosis

描述（由申请人提供）：该项目的长期目标是表征蛋白质稳态及其调控的分子基础，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，从而导致功能疾病的丧失和获得。该项目的具体目标1将是表征控制淀粉样蛋白轻链（LC）折叠和分泌与内质网（ER）相关降解（ERAD）途径降解之间平衡的蛋白质稳态网络，以了解人类疾病淀粉样蛋白LC淀粉样变性（AL）的基础。将生物制剂（siRNA和cDNA）高通量筛选（HTS）应用于分泌淀粉样变性LCs的细胞系，将使我们能够进行全基因组筛选，以发现新的靶点，这些靶点将使平衡转向降解，而不是分泌容易错误折叠的LCs，预计这一结果将对该疾病产生重大影响，甚至可能治愈该疾病。Specific Aim 2还将使用HTS格式识别化学文库中的小分子，这些小分子减少淀粉样蛋白的分泌，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，我们将探索蛋白质稳态网络在我们年轻时控制这些疾病的一般假设，但随着年龄的增长，这些成分的失败可能导致功能获得性蛋白质毒性和器官衰竭，这是由于淀粉样蛋白负担超过了修复和降解途径的能力。实现这两个特定目标需要研究者之间强有力的4-way合作，这些研究者具有有效互动的记录，并且具有强大的互补专业知识，可以应用细胞生物学，化学和小鼠和蠕虫模型方法来了解AL疾病对衰老程序的反应。项目描述：本项目描述了一种新的高通量筛选方法，用于识别影响衰老过程中人类骨髓瘤轻链淀粉样变性（AL）发病和病理的小分子化合物和途径。这些途径和化合物的鉴定将导致对病理生理学和可能的治疗疾病的关键的新见解。

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.