Determining the role of AR transcriptional function in SBMA

确定 AR 转录功能在 SBMA 中的作用

基本信息

批准号：
10210450
负责人：
DIANE E MERRY
金额：
$ 44.02万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-20 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10210450
关键词：
Acetylation Adult Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Androgen Receptor Androgen Response Element Androgens Animal Model Antibodies Behavioral Binding Biochemical CRISPR/Cas technology Cell Nucleus Cell model Characteristics Clinical Trials Cultured Cells DNA DNA Binding DNA Binding Domain Data Development Disease Disease model Drosophila genus Evaluation Event Excision Family Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Transcription Goals Hormones Huntington Disease In Vitro Inherited Investigation Knock-in Knock-in Mouse Laboratories Lead Leuprolide Ligands Mediating Metabolism Modeling Molecular Molecular Conformation Motor Motor Neurons Mus Muscle Cells Muscle Fibers Mutation Neurodegenerative Disorders Neuromuscular Diseases Neuronal Dysfunction Neurons Nuclear Outcome Parkinson Disease Pathogenesis Pathogenicity Pathology Pathway interactions Patients Pharmacologic Substance Pharmacology Phosphorylation Play Process Production Proteins Proteolysis Proteomics Publishing RIPK1 gene Receptor Aggregation Research Role Spinocerebellar Ataxias Structure Symptoms System Testis Testosterone Therapeutic Toxic effect Transgenic Mice Work androgenic design disease phenotype effective therapy functional restoration in vivo in vivo Model induced pluripotent stem cell insight mouse model muscle physiology mutant neuron loss neurotoxic novel novel therapeutics overexpression polyglutamine prevent protein misfolding receptor function spinal and bulbar muscular atrophy therapeutic development transcriptomics

项目摘要

Many neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease and the polyglutamine diseases, result from protein misfolding and accumulation due to a variety of genetic and/or environmental causes. Spinal and bulbar muscular atrophy (SBMA) is an adult-onset, inherited neuromuscular disease that is caused by polyglutamine expansion within the androgen receptor (AR); it is related to other neurodegenerative diseases caused by polyglutamine expansion, including Huntington's disease and several spinocerebellar ataxias. Although the precise pathway leading to neuronal dysfunction and death is unknown, the evaluation of transgenic mouse and cell models of these diseases has yielded mechanistic insights to disease pathogenesis. SBMA stands apart from other polyglutamine diseases in that its onset and progression are dependent on AR androgenic ligands. Our cell and mouse models of SBMA reproduce the androgen- and polyglutamine-dependent nuclear AR aggregation seen in patients, as well as its consequent toxicity, making these models highly useful for the analysis of the mechanistic basis for upstream events involved in AR toxicity. Previous studies from our group and others revealed that the nuclear localization, N/C interaction, phosphorylation and acetylation of the AR are all required for its aggregation and toxicity. Inhibiting each of these steps through genetic or pharmacological manipulation prevents mutant AR aggregation as well as its toxicity. Notably, inhibition of each of these steps also prevents AR transcriptional activity, highlighting a central question in the understanding of SBMA pathogenesis of the role of AR transcriptional activity in disease. While earlier studies in a Drosophila model of SBMA supported such a role, studies in mammalian systems suggest otherwise. Answering this important question is critical to the development of effective therapies. We propose with this application to answer this central question of the role of AR transcriptional activity in SBMA, through the use of novel cell and animal models, and to use these models to investigate the molecular pathways that mediate disease. We predict that our proposed studies will reveal further details about the role of AR transcriptional activity in disease pathogenesis and provide robust insights into ideal therapeutic directions in SBMA. To achieve these goals, we propose three specific aims: 1) To determine the role of AR DNA binding in disease, in vivo, using novel, genetic knock-in mouse models of SBMA; 2) To evaluate the role of DNA binding in cell models of SBMA; and 3) To use these models to identify common pathogenic pathways through transcriptional profiling and quantitative analysis of mutant AR-interacting proteins. We anticipate that results from these studies will lead us to a new understanding of the molecular pathogenesis of SBMA and enhance our development of new therapies for SBMA.

许多神经退行性疾病，包括阿尔茨海默病、帕金森病和多聚谷氨酰胺疾病，是由于多种遗传和/或原因导致的蛋白质错误折叠和积累引起的环境原因。脊髓和延髓肌萎缩症 (SBMA) 是一种成人发病的遗传性由雄激素受体 (AR) 内的聚谷氨酰胺扩张引起的神经肌肉疾病；这是与多聚谷氨酰胺扩张引起的其他神经退行性疾病有关，包括亨廷顿舞蹈病疾病和几种脊髓小脑性共济失调。尽管通往神经元的精确途径功能障碍和死亡未知，这些疾病的转基因小鼠和细胞模型的评估对疾病发病机制产生了机制见解。 SBMA 与其他聚谷氨酰胺不同疾病的发生和进展依赖于 AR 雄激素配体。我们的细胞和老鼠 SBMA 模型再现了雄激素和多谷氨酰胺依赖性核 AR 聚集患者及其随之而来的毒性，使得这些模型对于分析涉及 AR 毒性的上游事件的机制基础。我们小组和其他人之前的研究揭示了 AR 的核定位、N/C 相互作用、磷酸化和乙酰化都是其聚集和毒性所必需的。通过遗传或药理学抑制这些步骤中的每一步操作可防止突变体 AR 聚集及其毒性。值得注意的是，抑制每个步骤也阻止 AR 转录活动，突出了理解 SBMA 的一个核心问题 AR转录活性在疾病中的作用的发病机制。虽然早期对果蝇的研究 SBMA 模型支持这种作用，但哺乳动物系统的研究表明并非如此。回答这个重要的问题对于开发有效的疗法至关重要。我们建议通过此申请通过使用新型细胞回答 AR 转录活性在 SBMA 中的作用这一核心问题和动物模型，并使用这些模型来研究介导疾病的分子途径。我们预测我们提出的研究将进一步揭示 AR 转录活性在疾病发病机制，并为 SBMA 的理想治疗方向提供深入的见解。达到为了实现这些目标，我们提出了三个具体目标：1) 确定 AR DNA 结合在疾病中的作用，体内，使用新型 SBMA 基因敲入小鼠模型； 2) 评估DNA结合在细胞中的作用 SBMA 模型； 3) 使用这些模型来识别常见的致病途径突变 AR 相互作用蛋白的转录谱和定量分析。我们预计这些研究的结果将使我们对 SBMA 的分子发病机制有新的认识并加强我们对 SBMA 新疗法的开发。