A Chemical Genetics Approach to the Discovery and Synthesis of Bioactive Marine Natural Products

该提案的主要目标是使用化学遗传学方法来发现新的生物活性海洋天然产物（MNP），这些产物可用作自噬、MRSA中心蛋白、组蛋白甲基化和F508 del-CFTR校正的基础研究工具。第二个目标将是新的MNP的功能导向的合成，以确定其药效团的SAR，产生用于蛋白质靶点鉴定的探针，并为体内实验提供材料。化学遗传学使用小分子来改变细胞环境中单个基因产物的功能，以便将特定基因与表型联系起来。正向化学遗传学筛选在基于细胞的测定中引起感兴趣的表型的分子，然后鉴定蛋白质靶。反向化学遗传学筛选纯蛋白质的抑制剂，然后将细胞暴露于抑制剂以揭示表型。我们将专注于在人类生物学和疾病中很重要的生物过程，但在蛋白质或遗传水平上没有很好的表征。我们的海洋无脊椎动物和培养微生物的粗提物库将是新的小分子蛋白质配体的来源。每个项目都将涉及与一名生物过程专家的既定合作。研究的意义是多方面的。新MNP的发现为已知化学实体的库增加了独特的多样性，并揭示了未知的生物合成途径和药效团。复杂的生物活性MNP是合成有机化学家的有吸引力的目标，推动了新的合成策略和方法的发展。MNP调节自噬、MRSA生物学、组蛋白甲基化和F508 del-CFTR校正中涉及的蛋白质的发现将为在单细胞和动物中在遗传和蛋白质水平上研究这些重要的生物学过程提供工具，并有助于确定新的药物靶点。新的生物活性MNP将有可能为结核病、MRSA、癌症和囊性纤维化药物的开发提供灵感。我们的跨学科研究提供培训，使HQP能够获得高层次的工业，政府和学术工作。

The main objective of the proposal will be to use a chemical genetics approach to discover new bioactive marine natural products (MNPs) that can be used as tools in fundamental studies of autophagy, MRSA hub proteins, histone methylation, and F508del-CFTR correction. A secondary objective will be the function-oriented synthesis of the new MNPs in order to define the SAR for their pharmacophores, generate probes for protein target identification, and provide material for in vivo experiments. Chemical genetics uses small molecules to alter the function of a single gene product within a cellular context in order to link a specific gene to a phenotype. Forward chemical genetics screens for molecules that elicit a phenotype of interest in a cell-based assay and then identifies the protein target. Reverse chemical genetics screens for inhibitors of pure proteins and then exposes cells to the inhibitors to reveal the phenotype. We will focus on biological processes that are important in human biology and disease, but are not well characterized at the protein or genetic level. Our library of crude extracts of marine invertebrates and cultured microorganisms will be the source of novel small molecule protein ligands. Each project will involve an established collaboration with an expert on the biological process. The significance of the research will be multifold. Discovery of new MNPs adds unique diversity to the pool of known chemical entities and reveals unknown biosynthetic pathways and pharmacophores. Complex bioactive MNPs are attractive targets for synthetic organic chemists, driving the development of new synthetic strategies and methodology. Discovery of MNPs that modulate proteins involved in autophagy, MRSA biology, histone methylation, and F508del-CFTR correction will provide tools to study these important biological processes at the genetic and protein level in single cells and animals, and help identify new drug targets. The new bioactive MNPs will have the potential to provide the inspiraton for development of drugs for TB, MRSA, cancer, and cystic fibrosis. Our interdisciplinary research provides training that will enable HQP to obtain high-level industrial, government, and academic jobs.