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节的分子基础，并了解该系统的能力如何以及为什么随着衰老而下降，导致功能疾病的丧失和获得。本项目的具体目标1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，以了解人类淀粉样蛋白LC淀粉样变性（AL）疾病的基础。将生物制剂（siRNA和cDNA）高通量筛选（HTS）应用于分泌淀粉样蛋白生成LC的细胞系将允许我们进行基因组范围的筛选以发现新的靶点，所述新的靶点将使平衡转向降解并远离易错误折叠LC的分泌，预测该结果对该疾病具有实质性影响，甚至可能治愈该疾病。Specific Aim 2还将使用HTS格式来鉴定化学文库中的小分子，这些小分子减少了淀粉样蛋白生成LC的分泌，有利于ERAD，因此导致组织中细胞外淀粉样蛋白负荷的减少。在这两个具体的目标，我们将探讨的一般假设，蛋白质稳态网络保持这些疾病在检查时，我们年轻，但这些组件在衰老过程中的失败可能会导致获得的功能蛋白毒性和器官衰竭，由于淀粉样蛋白的负担，超过了修复和降解途径的能力。实现这两个具体目标涉及研究人员之间的强有力的4方合作，这些研究人员具有有效互动的跟踪记录，并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疾病对衰老计划的反应。 项目叙述：该项目描述了一种新的高通量筛选方法，用于识别影响衰老期间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。这些途径和化合物的鉴定将导致对病理生理学的重要新见解和疾病的可能治愈。

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.