Identifying New Aging and Longevity Genes in Caenorhabditis elegans

描述（由申请人提供）：候选人是丹娜法伯癌症研究所的肿瘤学研究员，之前接受过哺乳动物细胞凋亡和免疫学医学博士-博士生的研究培训。经过五年的临床培训（包括内科实习和住院医师培训以及肿瘤内科亚专科临床培训）后，他回到了实验室。他对衰老和长寿产生了兴趣，并希望从事职业发展计划，该计划将提供以下领域的培训：（1）衰老生物学（2）功能基因组学和（3）作为模式生物和工具的秀丽隐杆线虫。该职业发展计划要求在丹纳法伯癌症研究所的双重导师 Marc Vidal 博士和麻省总医院的 Gary Ruvkun 博士的指导下进行培训。Marc Vidal 博士是线虫功能基因组学方法的先驱，而马萨诸塞州总医院的 Gary Ruvkun 博士是线虫衰老和长寿领域的知名研究者。此外，附近还有许多其他机构，共同形成了非常丰富和多样化的培训环境。该研究计划建议通过使用组合的功能基因组方法来识别秀丽隐杆线虫中新的衰老和长寿基因。参与线虫寿命的已知蛋白质（包含 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（一种称为蛋白质相互作用图谱的方法）中的诱饵。通过这些筛选鉴定出的新相互作用因子将接受功能分析，最初包括 RNA 介导的干扰 (RNAi)，以观察功能丧失表型。最初要检查的表型是 dauer 形成，因为历史上改变 dauer 形成的 daf-2 途径基因突变也会产生长寿表型。 dauer 形式是一种休眠、活动减退且寿命长的幼虫形式，可以在恶劣条件下发育，但当条件变得更有利时可以恢复正常发育。初步数据鉴定了 7 种新型相互作用蛋白，其中 2 种通过 RNAi 给出了 dauer 表型。还提出了通过两种杂交鉴定的候选基因的寿命测定。预计蛋白质相互作用作图和基于 RNAi 的分析相结合的方法将鉴定出许多与 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.