Identifying New Aging and Longevity Genes in Caenorhabditis elegans

描述(由申请人提供)：应聘者是Dana-Farber癌症研究所的肿瘤学研究员，曾接受过哺乳动物细胞凋亡和免疫学博士研究生的研究培训。在中断了5年的临床培训后，他回到了实验室，包括内科的实习和住院医生，以及内科肿瘤学的专科临床培训。他对衰老和长寿产生了兴趣，并希望从事一项职业发展计划，该计划将在(1)衰老生物学、(2)功能基因组学和(3)线虫作为模式生物和工具方面提供培训。职业发展计划要求在达纳-法伯癌症研究所的马克·维达尔博士和马萨诸塞州总医院的加里·鲁夫昆博士的双重指导下进行培训。维达尔博士是线虫功能基因组方法的先驱，加里·鲁夫昆博士是线虫衰老和寿命方面的知名研究员。此外，附近还有许多其他机构，它们共同构成了一个非常丰富和多样化的培训环境。该研究计划建议使用组合功能基因组学方法在线虫中识别新的衰老和长寿基因。线虫中与长寿有关的已知蛋白质，包括daf-2胰岛素样信号通路，将被用作全基因组酵母双杂交筛选的诱饵(一种被称为蛋白质相互作用图谱的方法)。通过这些筛选确定的新的相互作用因子将受到功能分析的影响，该分析最初将包括RNA介导的干扰(RNAi)，以观察功能丧失的表型。最初要检查的表型是Dauer形成，因为在历史上，改变Dauer形成的daf-2途径中的基因突变也会产生长寿表型。达尔幼虫是一种休眠、不活跃、寿命长的幼虫，可以在恶劣的条件下发育，但当条件变得更有利时，可以恢复正常发育。初步鉴定了7个新的相互作用蛋白，其中2个通过RNAi给出了Dauer表型。还提出了双杂交确定的候选基因的寿命分析。预计蛋白质相互作用图谱和基于RNAi的分析相结合的方法将发现许多与Dauer形成、衰老和长寿有关的新基因。

DESCRIPTION (provided by applicant): The candidate is an oncology fellow at the Dana-Farber Cancer Institute with prior research training as an MD-PhD student in mammalian apoptosis and immunology. He has returned to the laboratory after a 5 year hiatus spent pursuing clinical training, including internship and residency in Internal Medicine and subspecialty clinical training in Medical Oncology. He has developed an interest in aging and longevity and wishes to pursue a program of career development that will provide training in the areas of (1) biology of aging (2) functional genomics and (3) C. elegans as a model organism and tool. The career development plan calls for training under dual mentors Dr. Marc Vidal at Dana-Farber Cancer Institute, who is a pioneer in functional genomic approaches in C. elegans, as well as Dr. Gary Ruvkun at Massachusetts General Hospital, who is an established investigator in aging and longevity in C. elegans. In addition, there are many other institutions in the immediate vicinity that form a collectively very rich and diverse training environment. The research plan proposes to identify new aging and longevity genes in C. elegans by using a combined functional genomic approach. The known proteins involved in longevity in C. elegans, which comprise the daf-2 insulin-like signaling pathway, will be used as baits in genome-wide yeast two-hybrid screens (an approach termed protein interaction mapping). New interactors identified by these screens will be subject to a functional analysis that will initially consist of RNA-mediated interference (RNAi) to look at loss-of-function phenotypes. The phenotype to be initially examined is dauer formation, since historically mutations in genes in the daf-2 pathway that alter dauer formation also give longevity phenotypes. The dauer form is a dormant, hypoactive and long-lived larval form that can develop under harsh conditions, but that can resume normal development when conditions become more favorable. Preliminary data is presented identifying 7 novel interacting proteins, 2 of which give dauer phenotypes by RNAi. Longevity assays of candidate genes identified by two-hybrid are also proposed. It is anticipated that the combined approach of protein interaction mapping and RNAi-based analysis will identify many new genes involved in dauer formation, aging and longevity.