Determining the role of AR transcriptional function in SBMA

确定 AR 转录功能在 SBMA 中的作用

• 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
• 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' 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

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，包括阿尔茨海默氏病，帕金森氏病和 聚谷氨酰胺疾病是由蛋白质折叠和积累引起的，原因是多种遗传和/或 环境原因。脊柱和鳞茎肌肉萎缩（SBMA）是一种成人发言权，遗传 神经肌肉疾病是由雄激素受体（AR）内聚谷氨酰胺膨胀引起的；这是 与多谷氨酰胺扩张引起的其他神经退行性疾病有关，包括亨廷顿的 疾病和几种脊椎动物共济失调。虽然导致神经元的精确途径 功能障碍和死亡尚不清楚，这些疾病的转基因小鼠和细胞模型的评估 已经对疾病发病机理产生了机械见解。 SBMA与其他聚谷氨酰胺不同 疾病的发作和进展取决于AR雄激素配体。我们的细胞和鼠标 SBMA的模型重现了在 患者以及随之而来的毒性，使这些模型可用于分析 与AR毒性有关的上游事件的机械基础。我们小组和其他人的先前研究 揭示了AR的核定位，N/C相互作用，磷酸化和乙酰化均为 其汇总和毒性所必需的。通过遗传或药理抑制这些步骤 操作可防止突变AR聚集及其毒性。值得注意的是，抑制每个步骤 还可以防止AR转录活动，突出了对SBMA的理解中的一个核心问题 AR转录活性在疾病中的作用的发病机制。虽然较早的果蝇研究 SBMA的模型支持了这种作用，哺乳动物系统中的研究表明。回答这个 重要的问题对于有效疗法的发展至关重要。我们将此申请提出建议 通过使用新细胞来回答AR转录活性在SBMA中的作用的中心问题 和动物模型，并使用这些模型研究介导疾病的分子途径。 我们预测，我们提出的研究将揭示有关AR转录活性作用的更多细节 疾病发病机理，并为SBMA中理想的治疗方向提供了强有力的见解。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，在 体内使用SBMA的新型遗传敲击小鼠模型； 2）评估DNA结合在细胞中的作用 SBMA的模型； 3）使用这些模型通过 突变体Arteracting蛋白的转录分析和定量分析。我们预料到这一点 这些研究的结果将使我们对SBMA分子发病机理有了新的了解 并增强我们针对SBMA的新疗法的开发。