Regulation of Polyglutamine Aggregation and Toxicity

描述（由申请人提供）：聚谷氨酰胺神经退行性疾病是一种毁灭性的遗传性疾病家族，包括亨廷顿病和脊髓球性肌萎缩症（SBMA）。该项目的长期目标是确定聚谷氨酰胺神经退行性疾病的分子机制，确定特定的治疗靶点，并开发基于机制的治疗方法。先前的研究表明，rho相关激酶p160ROCK的抑制剂Y-27632在细胞和果蝇模型中降低了聚谷氨酰胺的聚集和毒性。目标1：识别和表征新的调控途径和靶分子。我们将完成一个生物活性小分子库的筛选。我们还将分析两个先前筛选的生物活性化合物。结果将采用遗传和药理学方法相结合的系统方式进行分析，以确定潜在意义的新途径。目的2：确定p160ROCK信号通路影响聚谷氨酰胺聚集和毒性的分子机制。p160ROCK信号通路的特定组分的作用将在果蝇中进行测试。将测试聚谷氨酰胺蛋白与肌动蛋白结合的分子基础，并确定其在调节聚谷氨酰胺聚集中的作用。目的3：检测Y-27632在体内预防神经退行性变的活性。应确定Y-27632在脑中的生物活性和全身毒性，以便计划适当的给药方案。Y-27632在体内对多谷氨酰胺依赖病理的抑制作用应在R6/2亨廷顿病小鼠模型中采用多种行为、旋转体、病理和生化分析进行测试。

DESCRIPTION (provided by applicant): Polyglutamine neurodegenerative diseases are a devastating family of inherited disorders that include Huntington Disease and spinobulbar muscular atrophy (SBMA). Long-term goals of this project are to identify molecular mechanisms of polyglutamine neurodegenerative disease, to determine specific therapeutic targets, and to develop mechanism-based therapies. Previous work demonstrated that Y-27632, an inhibitor of the rho-associated kinase p160ROCK, reduced polyglutamine aggregation and toxicity in cell and Drosophila models. Aim 1: Identify and characterize novel regulatory pathways and target molecules. We will complete a screen of a library of biologically active small molecules. We will also analyze hits from two prior screens of biologically active compounds. Results will be analyzed in a systematic fashion using a combination of genetic and pharmacologic approaches to determine new pathways of potential significance. Aim 2: Determine the molecular mechanism by which p160ROCK signaling influences polyglutamine aggregation and toxicity. The role of specific components of the p160ROCK signaling pathway will be tested in Drosophila. The molecular basis of polyglutamine protein association with actin will be tested, and its role in modulating polyglutamine aggregation determined. Aim 3: Test the activity of Y-27632 in preventing neurodegeneration in vivo. Bioactivity of Y-27632 in brain, and systemic toxicity shall be determined in order to plan an appropriate dosing regimen. Y-27632 inhibition of polyglutamine-dependent pathology in vivo shall be tested using a variety of behavior, rotarod, pathological and biochemical analyses in the R6/2 mouse model of Huntington's disease.