Small Molecule Screen for Novel Regulators of Chaperone Expression

描述（由申请人提供）：已显示分子伴侣的表达升高抑制亨廷顿氏病、阿尔茨海默氏病、帕金森氏病和肌萎缩性侧索硬化症（ALS）的各种模型系统中的蛋白质错误折叠/聚集和毒性。与这些疾病相关的相应蛋白质中的突变导致采用交替构象的错误折叠种类的出现。这些观察结果导致了这样的建议，即突变亨廷顿蛋白、tau蛋白、α-突触核蛋白和超氧化物歧化酶（SOD 1）的共同特征是交替折叠状态的出现，这些折叠状态自缔合并形成有毒物质和蛋白质聚集体。分子伴侣由于其识别和隔离受损和错误折叠的物种的独特特性，为治疗提供了有趣的靶标。因此，分子伴侣可能在蛋白质稳态中具有中心作用，以防止慢性错误折叠种类的有害后果，随着时间的推移，慢性错误折叠种类导致细胞发育不全和病理学，如在神经退行性疾病和与蛋白质错误折叠相关的其他疾病中发生的。然而，由于分子伴侣在体内作为网络起作用，因此也越来越明显的是，单独的分子伴侣的表达对于实现最大网络功能性是无效的或比多个分子伴侣的协调表达有效得多。我们提出三个具体目标：目标1。建立一个强大的初步筛选小分子调节剂的热休克反应;目的2。表征候选物的细胞毒性和诱导HS基因表达和伴侣蛋白水平的动力学;以及目的3。在表达突变型亨廷顿蛋白和SOD 1的细胞的二次试验中检测热休克反应的小分子调节剂。虽然我们的实验将只检查的结果，新的小分子伴侣蛋白表达调节剂的模型系统的神经退行性疾病，我们预计，我们的结果可能会很好地扩展到其他病理与错误折叠的蛋白质在细胞质中的流量。

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.