High-Content Cell-Based Screening for Modulators of Autophagy

描述（由申请人提供）：自噬是一种细胞过程，其中一部分细胞内容物被双膜结构隔离，然后通过溶酶体降解。这一基本过程对于细胞存活、细胞器稳态、能量代谢和防御细胞内病原体是重要的。自噬在神经退行性疾病、癌症和微生物感染的发病机制中具有重要意义。研究表明，在动物模型中，调节自噬可以改善癌症治疗，促进感染的解决并减轻神经变性。然而，关于自噬的分子机制以及在这些环境中的调控仍然有很多需要了解的。在本申请中，我们提出了一种高含量的基于细胞的筛选新的化学物质，可以调节自噬。我们已经进行了初步的研究，以证明这种方法是可行的，并可以优化测定条件。我们的短期目标是，通过这种筛选，我们将能够找到更有效，毒性更小，更特异的独特化合物。我们近期的目标是确定有前途的化合物的结构与其生物学效应的关系。我们还想确定具体的分子靶点，以了解该化合物的工作原理。我们的长期希望是，通过这项研究开发的知识和材料将对未来的临床应用有价值。

DESCRIPTION (provided by applicant): Autophagy is a cellular process in which a part of cellular contents are sequestered by a double membrane structure, which are then degraded through the lysosome. This fundamental process is important for cell survival, organelle homeostasis, energy metabolism, and defense against intracellular pathogens. Autophagy has significant implications in the pathogenesis of neurodegenerative diseases, cancer and microbial infections. Studies have shown that modulating autophagy could improve cancer therapy, promote the resolution of infection and lessen neurodegeneration in animal models. However, there are still very much to be learnt about the molecular mechanism of autophagy and the regulation in these settings. We propose in this application a high content cell based screening for novel chemicals that can modulate autophagy. We have conducted preliminary studies to demonstrate that this approach is feasible and the assay conditions can be optimized. Our short-term goal is that through this screening we will be able to find unique compounds that are more potent, less toxic and more specific. Our immediate future goal is to define the structure of promising compounds in relationship with their biological effects. We would also like to determine the specific molecular targets to understand how the compound works. Our long term hope is that the knowledge and the materials developed through this study will be valuable for future clinical applications.