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，曾在哺乳动物细胞凋亡和免疫学方面接受过医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，包括内科实习和住院医师培训，以及肿瘤医学的亚专科临床培训。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，该计划将在以下领域提供培训：(1)衰老生物学(2)功能基因组学和(3)秀丽隐杆线虫作为模式生物和工具。职业发展计划要求在达纳-法伯癌症研究所（Dana-Farber Cancer Institute）的马克·维达尔（Marc Vidal）博士和马萨诸塞州总医院（Massachusetts General Hospital）的加里·鲁夫昆（Gary Ruvkun）博士的双重导师下进行培训，前者是秀丽隐杆线虫功能基因组方法的先驱，后者是秀丽隐杆线虫衰老和长寿方面的知名研究者。此外，附近还有许多其他机构，共同形成了非常丰富多样的培训环境。该研究计划提出利用组合功能基因组方法在秀丽隐杆线虫中鉴定新的衰老和长寿基因。已知的与秀丽隐杆线虫长寿有关的蛋白质，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（一种称为蛋白质相互作用作图的方法）的诱饵。通过这些筛选确定的新相互作用物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），以观察功能丧失表型。最初要研究的表型是道尔形成，因为从历史上看，daf-2通路中改变道尔形成的基因突变也会产生长寿表型。水虫是一种休眠的、低活性的、长寿命的幼虫，它可以在恶劣的条件下发育，但当条件变得更有利时，它可以恢复正常的发育。初步数据鉴定了7种新的相互作用蛋白，其中2种通过RNAi提供了更强的表型。还提出了双杂交鉴定的候选基因的寿命测定方法。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识别出许多与细胞形成、衰老和寿命有关的新基因。

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.