Role of p62 in Protein Aggregation and Neurodegeneration in ALS

p62 在 ALS 蛋白质聚集和神经变性中的作用

基本信息

批准号：
7832202
负责人：
Haining Zhu
金额：
$ 23.39万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7832202
关键词：
Adaptor Signaling Protein Affect American Amyotrophic Lateral Sclerosis Autophagocytosis Cells Cessation of life Characteristics Co-Immunoprecipitations Data Degradation Pathway Disease Progression Equipment and Supplies Etiology Failure Familial Amyotrophic Lateral Sclerosis Funding Genes Grant Life Link Lysosomes Mediating Molecular Chaperones Motor Neurons Mus Mutation Names Nerve Degeneration Neurodegenerative Disorders Parents Pathway interactions Play Positioning Attribute Proteins Publishing Reagent Recovery Reporting Request for Applications Research Research Personnel Role Series Spinal System Testing Time Toxic effect Transgenic Mice Ubiquitin United States National Institutes of Health Work age related base biological adaptation to stress copper zinc superoxide dismutase design domain mapping heat-shock factor 1 histone deacetylase 6 in vivo inhibitor/antagonist insight multicatalytic endopeptidase complex mutant neurotoxicity new therapeutic target protein aggregate protein aggregation protein degradation protein misfolding protein transport public health relevance research study response

项目摘要

DESCRIPTION (provided by applicant): This Competitive Revision application is in response to NOT-OD-09-058 "NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications." We request competitive supplement to our NIH grant R21AG032567 entitled "Role of p62 in Protein Aggregation and Neurodegeneration in ALS." Protein aggregates containing mutant copper-zinc superoxide dismutase (SOD1) are a hallmark of familial amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease), an age-related neurodegenerative disease. P62/Sequestosome 1 (referred as p62 in this proposal) is a multifunctional protein involved in both of the two major protein degradation mechanisms: ubiquitin-proteasome system (UPS) and autophagy-lysosome pathway. The central hypothesis to be tested in the parent R21 project is that p62 can recognize misfolded mutant SOD1 and that p62 can ameliorate the mutant SOD1 induced toxicity by shuttling such misfolded proteins to UPS and/or autophagy. We previously reported that p62 was co-localized with mutant SOD1 in the protein aggregates in spinal motor neurons in G93A SOD1 transgenic mice. Co-immunoprecipitation experiments showed that p62 specifically recognized mutant SOD1, but not the WT protein. Three specific aims were designed to test the hypothesis at the time. Aim 1 was to map the domains of p62 essential for recognizing and interacting with mutant SOD1. Aim 2 was to determine whether and how p62 mediated the autophagy activation induced by mutant SOD1. Aim 3 was to study how p62 would influence protein aggregation and ALS disease progression in vivo using p62 KO mice. We have largely accomplished the Aims 1 and 2 and are currently performing experiments proposed in Aim 3. This Competitive Revision is built on the progress made in the R21 project and the new preliminary data suggesting that HDAC6 is likely an adaptor protein between p62 and mutant SOD1. In addition, HDAC6 has been shown to play a critical role in autophagy activation and stress response. We thus propose to expand the parent R21 project to test a new hypothesis that HDAC6 is an adaptor protein mediating mutant SOD1-p62 interaction, regulate the autophagic degradation of mutant SOD1, and mediate the stress response induced by mutant SOD1. Three new specific aims are proposed to test this hypothesis. Aim 1 is to dissect the detailed mechanism how HDAC6 functions as an adaptor between mutant SOD1 and p62. Aim 2 is to determine the role of HDAC6 in aggregation and autophagic degradation of mutant SOD1. Aim 3 is to study the significance of HDAC6 and p62 in mutant SOD1 induced stress response. Results from this study will provide invaluable insights into the role of HDAC6 and p62 in protein aggregation and neurodegeneration in ALS, which will result in better understanding of ALS etiology and potential discovery of new therapeutic target for ALS treatment. Relevance to the American Recovery & Reinvestment Act (ARRA): This application will create two new scientific researcher positions as soon as it is funded. Purchase of necessary equipment, supplies and reagents for the proposed research will also stimulate the economy as intended by the ARRA. PUBLIC HEALTH RELEVANCE: Amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease) is an age-related neurodegenerative disorder characteristic of the preferential motor neurons death. Mutations in the gene encoding copper-zinc superoxide dismutase (SOD1) have been linked to a subset of familial ALS cases. Protein aggregates containing mutant SOD1 are a hallmark of familial ALS. P62/Sequestome 1 (referred as p62 in this proposal) is a multifunctional protein involved in both of the two major protein degradation mechanisms: ubiquitin-proteasome system (UPS) and autophagy-lysosome pathway. The parent R21 was to test the hypothesis that p62 could recognize misfolded mutant SOD1 and ameliorate the mutant SOD1 induced toxicity by shuttling such misfolded proteins to the UPS and/or autophagy. We have largely accomplished the Aims 1 and 2 proposed in the parent R21 and are currently performing experiments proposed in Aim 3. Based on the progress and new preliminary results, we propose to test the significance of an adaptor protein named histone deacetylase 6 (HDAC6) in mutant SOD1 protein degradation, aggregation and neurotoxicity. The specific hypothesis to be tested in the expanded project is that HDAC6 is an adaptor protein mediating mutant SOD1-p62 interaction, regulates the autophagic degradation of mutant SOD1, and mediates the stress response induced by mutant SOD1. Results from this study will provide new insights into the role of HDAC6 and p62 in protein aggregation and neurodegeneration in ALS, which will result in better understanding of ALS etiology and potential discovery of new therapeutic target for ALS treatment. Moreover, this application will create two new scientific researcher positions as soon as it is funded. Purchase of necessary equipment, supplies and reagents for the proposed research will also stimulate the economy as intended by the American Recovery & Reinvestment Act.

描述（申请人提供）：此竞争性修订申请是对NOT-OD-09-058的回应，“ NIH宣布为竞争性修订申请提供恢复法案资金。”我们要求对NIH授予R21AG032567的竞争补充，题为“ p62在ALS中的蛋白质聚集和神经变性中的作用”。含有突变铜锌超氧化物歧化酶（SOD1）的蛋白质聚集体是家族性肌萎缩性侧面硬化症的标志（ALS，Lou Gehrig氏病），这是一种与年龄相关的神经退行性疾病。 p62/sequestosome 1（该提案中称为p62）是参与两种主要蛋白质降解机制的多功能蛋白：泛素蛋白 - 蛋白酶体系统（UPS）和自噬 - 溶酶体途径。在母体R21项目中要测试的中心假设是p62可以识别错误折叠的突变体SOD1，并且p62可以通过将这种错误折叠的蛋白穿过UPS和/或自噬来改善突变体SOD1引起的毒性。我们先前曾报道过，p62与G93A SOD1转基因小鼠的脊柱运动神经元中的蛋白质聚集体中的突变SOD1共定位。共免疫沉淀实验表明，p62特异性识别的突变体SOD1，而不是WT蛋白。当时设计了三个特定目标来检验该假设。目的1是映射p62的域对于识别和与突变体Sod1相互作用所必需的域。 AIM 2是确定P62是否以及如何介导突变SOD1诱导的自噬激活。 AIM 3是研究P62如何使用p62 KO小鼠在体内影响蛋白质聚集和ALS疾病进展。我们在很大程度上完成了目标1和2，目前正在进行AIM 3中提出的实验。这种竞争性修订构建基于R21项目中的进度，而新的初步数据表明HDAC6可能是p62和Mutant Sod1之间的适配器蛋白。此外，HDAC6已被证明在自噬激活和应力反应中起着至关重要的作用。因此，我们建议扩展父r21项目，以检验一个新的假设，即HDAC6是一种介导突变体SOD1-P62相互作用的衔接蛋白，调节突变体SOD1的自噬降解，并介导突变体SOD1诱导的应激响应。提出了三个新的特定目标来检验这一假设。目的1是剖析HDAC6在突变SOD1和p62之间的适配器的详细机制。目的2是确定HDAC6在突变SOD1的聚集和自噬降解中的作用。 AIM 3是研究HDAC6和P62在突变体SOD1诱导的应力反应中的重要性。这项研究的结果将为HDAC6和p62在ALS中的蛋白质聚集和神经变性中的作用提供宝贵的见解，这将更好地了解ALS病因以及潜在地发现ALS治疗的新治疗靶标。与《美国恢复与再投资法》（ARRA）相关：一旦资助，此应用程序将立即创建两个新的科学研究人员职位。购买必要的设备，供应和试剂为拟议的研究也将刺激ARRA预期的经济。公共卫生相关性：肌萎缩性侧索硬化症（ALS，Lou Gehrig病）是与年龄有关的神经退行性疾病的特征，是优先运动神经元死亡的特征。编码铜锌超氧化物歧化酶（SOD1）的基因中的突变已与家族性ALS病例的一部分相关。含有突变SOD1的蛋白质聚集体是家族性ALS的标志。 p62/sequestome 1（该提案中称为p62）是一种参与两种主要蛋白质降解机制的多功能蛋白：泛素蛋白 - 蛋白酶体系统（UPS）和自噬 - 溶酶体途径。母体R21是测试p62可以识别错误折叠的突变体SOD1并通过将这种错误折叠的蛋白穿过UPS和/或自噬来改善突变体SOD1引起的毒性的假设。我们已经在很大程度上完成了在母体R21中提出的目标1和2，目前正在进行AIM 3中提出的实验。根据进度和新的初步结果，我们建议测试称为组蛋白脱乙酰基酶6（HDAC6）的衔接蛋白的重要性（HDAC6）（HDAC6）（HDAC6）在突变SOD1蛋白降解中，聚集和神经毒性。在扩展项目中要测试的特定假设是HDAC6是一种介导突变体SOD1-P62相互作用的衔接蛋白，调节突变体SOD1的自噬降解，并介导突变体SOD1诱导的应激反应。这项研究的结果将为ALS中HDAC6和p62在蛋白质聚集和神经退行性中的作用提供新的见解，这将更好地了解ALS病因，并潜在地发现ALS治疗的新治疗靶点。此外，该应用程序将在资助后立即创建两个新的科学研究人员职位。购买必要的设备，供应和试剂为拟议的研究也将刺激《美国恢复与再投资法》的意图。