Small Molecule Screen for Novel Regulators of Chaperone Expression

描述（由申请人提供）：在亨廷顿病、阿尔茨海默病、帕金森病和肌萎缩性侧索硬化症（ALS）的各种模型系统中，分子伴侣蛋白的升高表达已被证明可以抑制蛋白质错误折叠/聚集和毒性。与这些疾病相关的各自蛋白质的突变导致采用交替构象的错误折叠物种的出现。这些观察结果导致提出突变的亨廷顿蛋白、tau蛋白、α -突触核蛋白和超氧化物歧化酶（SOD1）的一个共同特征是自我结合并形成有毒物质和蛋白质聚集体的交替折叠状态的出现。分子伴侣具有识别和隔离受损和错误折叠物种的独特特性，因此为治疗提供了有趣的靶点。因此，伴侣可能在蛋白质稳态中发挥核心作用，以防止慢性错误折叠物种的有害后果，随着时间的推移，导致细胞发育不良和病理，如神经退行性疾病和其他与蛋白质错误折叠相关的疾病中发生的那样。然而，由于伴侣蛋白在体内以网络的形式发挥作用，因此越来越明显的是，单个伴侣蛋白的单独表达要么无效，要么远不如多个伴侣蛋白的协同表达来实现最大的网络功能。我们提出三个具体目标：目标1。建立热休克反应小分子调控因子的筛选机制；目标2。表征候选细胞毒性和诱导HS基因表达和伴侣蛋白水平的动力学；和Aim 3。在表达突变亨廷顿蛋白和SOD1的细胞的二次分析中检测热休克反应的小分子调节因子。虽然我们的实验将只检查新型小分子伴侣蛋白表达调节因子对神经退行性疾病模型系统的影响，但我们预计我们的结果可能会很好地扩展到与细胞质中错误折叠蛋白通量相关的其他病理。

DESCRIPTION (provided by applicant): Elevated expression of molecular chaperones has been shown to suppress protein misfolding/aggregation and toxicity in various model systems of Huntington's disease, Alzheimer's disease, Parkinson's disease, and Amyotrophic Lateral Sclerosis (ALS). Mutations in the respective proteins associated with these diseases results in the appearance of misfolded species that adopt alternate conformations. These observations have led to the proposal that a common feature of mutant huntingtin, tau, alpha-synuclein, and superoxide dismutase (SOD1) is the appearance of alternate folded states that self-associate and form toxic species and protein aggregates. Molecular chaperones offer intriguing targets for therapeutics because of their unique characteristic to recognize and sequester damaged and misfolded species. Consequently, chaperones may have a central role in protein homeostasis to prevent the deleterious consequences of chronic misfolded species, that, over time results in cell dysgenesis and pathologies as occurs in neurodegenerative diseases and other diseases associated with protein misfolding. However, because chaperones function in vivo as networks, it has also become increasingly evident that the expression of individual chaperones alone is either ineffective or much less effective than the coordinated expression of multiple chaperones to achieve maximal network functionality. We propose three Specific Aims: Aim 1. To establish a robust primary screen for small molecule regulators of the heat shock response; Aim 2. To characterize the candidates for cytotoxicity and kinetics of induction of HS gene expression and chaperone levels; and Aim 3. To test small molecule regulators of the heat shock response in secondary assays of cells expressing mutant huntingtin and SOD1. Although our experiments will only examine the consequences of novel small molecule regulators of chaperone expression on model systems of neurodegenerative disease, we anticipate that our results may well extend to other pathologies associated with the flux of misfolded proteins in the cytoplasm.