Small Molecule Signaling in Caenorhabditis elegans

描述（由申请人提供）：线虫秀丽隐杆线虫是生物医学研究中最重要的模式生物之一，因为它具有生物学上的易处理性，而且它的许多生理途径与人类中的相应途径具有很强的相似性。该项目的目的是补充高度发达的C.线虫具有其代谢组的全面结构和功能特征，令人惊讶的是，其仅在非常有限的程度上进行了探索。这一努力的动机是由几条证据表明，在很大程度上不确定的结构的小分子在C。elegans内分泌和外分泌信号，特别是在调节寿命，发育和代谢的关键途径。拟议研究的核心是使用新的NMR光谱方法，该方法允许分析复杂的小分子混合物，并大大加快了检测到的化合物的结构解析过程和功能表征。该方法允许比较来自不同C. elegans突变株，并鉴定其生物合成由于特定突变而被强烈上调或下调的化合物的化学结构。对于该项目，选择了少量突变株，其表型表明影响寿命，发育或脂肪代谢的小分子信号传导缺陷。NMR光谱将用于检测和鉴定与这些表型相关的化合物。随后，将对已鉴定化合物的合成样品进行化学遗传筛选，以确定对寿命、发育调节和脂肪代谢调节的影响。在野生型C.线虫将接受代表与衰老和发育相关的关键遗传途径的突变体或RNAi菌株的额外测定。该项目的成功完成将为C. elegans代谢组，大大增加了我们对C.线虫生物学和哺乳动物中相应的疾病相关途径。产生的小分子知识不仅将使未来的努力，旨在更多样化的化学遗传筛选探索的生物学和生态学的其他方面的C。线虫，但也与农业或医学相关的线虫物种。此外，还建立了C. elegans信号分子将促进对来自其他模式生物的小分子代谢物的结构和功能表征的类似研究。

DESCRIPTION (provided by applicant): The nematode Caenorhabditis elegans is one of the most important model organisms for biomedical research, because of its biological tractability and because many of its physiological pathways show strong analogies to corresponding pathways in humans. The goal of this project is to complement the highly developed genomics and proteomics of C. elegans with a comprehensive structural and functional characterization of its metabolome, which, surprisingly, has been explored to only a very limited extent. This effort is motivated by several lines of evidence indicating that small molecules of largely undetermined structure play important roles in C. elegans endocrine and exocrine signaling, specifically in key pathways regulating lifespan, development, and metabolism. Central to the proposed research is the use of new NMR-spectroscopic methodology that permits the analysis of complex small molecule mixtures and greatly accelerates both the structure elucidation process and the functional characterization of the detected compounds. This methodology permits to compare complex metabolite samples derived from different C. elegans mutant strains, and to identify the chemical structures of compounds whose biosynthesis is strongly up- or downregulated as a result of a specific mutation. For this project, a small number of mutant strains were selected whose phenotypes suggest a defect in small-molecule signaling affecting lifespan, development, or fat metabolism. NMR-spectroscopy will be used to detect and identify compounds that correlate with these phenotypes. Subsequently, synthetic samples of the identified compounds will be subjected to chemical genetic screens to determine effects on lifespan, developmental regulation and fat metabolism regulation. Compounds that show activity in wild-type C. elegans will be subjected to additional assays with mutant or RNAi strains representative of key genetic pathways related to ageing and development. Successful conclusion of this project will provide a partial structural and functional annotation of the C. elegans metabolome, substantially increasing our understanding of fundamental pathways in C. elegans biology and corresponding disease-relevant pathways in mammals. The small-molecule knowledge generated will not only enable future efforts aimed at more varied chemical genetic screens exploring additional aspects of the biology and ecology of C. elegans, but also of nematode species relevant in agriculture or medicine. Furthermore, methodology developed for characterizing C. elegans signaling molecules will facilitate similar studies toward structural and functional characterization of small molecule metabolites from other model organisms.