Elucidating the Gene Regulatory Networks that Specify Invasive Behavior

项目7：项目概要/摘要 我的长期目标是在一家顶级研究机构中成为一名独立的科学家 研究基因调控网络（GRNs），调节细胞形态发生运动， 发展和癌症。为此，拟议的研究推进了我在以下方面的训练： 功能基因组学和系统生物学方法，补充了我以前的研究， 细胞和发育生物学技术。GRN包括物理和监管 转录因子（TF）之间的关系以及TF与其靶基因之间的关系， 特定的细胞生物学过程。尽管它们在调节细胞侵袭行为方面很重要， 转录因子及其指定侵袭性的下游靶标的身份在很大程度上是未知的。细胞 通过基底膜（BM）的侵入作为细胞分散的基础机制， 正常发育期间的器官形成，免疫监视，以及癌症期间的错误调节 转移杜克大学的舍伍德实验室建立了一个简单的体内模型， 独特地结合了锚细胞的强大遗传、功能基因组和单细胞视觉分析 (AC)在C.线虫幼虫发育我们 目前对AC入侵的理解包括三种TF的鉴定和一些假定的 下游靶点调节专门的侵入性膜的建立， AC在侵入期间去除BM的能力。在指导期间提出的实验 该奖项的开始阶段将通过以下方式来表征GRNs通过BM的潜在细胞侵袭：1） 鉴定bZIP癌基因TF的结合配偶体fos-1a和FOS-1A结合的身份 在C. elegans基因组;和2）鉴定调节AC侵入的另外的TF， 测试它们是否在体外调节癌细胞侵袭中显示保守功能。培训 在功能基因组技术和系统生物学方法将加快通过 高级课程的工作，参加国家会议，与玛丽安·瓦尔豪特博士的合作 实验室（马萨诸塞州大学医学院），并由菲利普·本尼迪克特博士， 杜克大学系统生物学小组和基因组科学研究所的主任， 政策（IGSP）。人类癌症体外分析的培训将在Dr. Stephen韦斯（密歇根大学）。这些培训经验和在培训期间获得的数据 指导阶段将为开展独立的研究生涯提供基础。研究 在该奖项的未指导阶段提出的包括1）表征基因组调控 含有TF结合位点的区域，所述TF结合位点在侵袭期间控制AC中的基因表达，以及2） 确定TF的核心补充，这是必要的，也是足够的，以概括一个 在正常发育和人类癌症侵袭中的侵袭程序。

Item 7: Project Summary / Abstract My long-term goal is to establish myself as an independent scientist at a top-tier research institution studying the gene regulatory networks (GRNs) that regulate cell morphogenetic movements during development and cancer. Towards this end, the proposed research advances my training in functional genomics and systems biology approaches that complement my previous research in cellular and developmental biology techniques. GRNs encompass both the physical and regulatory relationships amongst transcription factors (TFs) and between TFs and their target genes that drive specific cell biological processes. Despite their importance in regulating cell invasive behavior, the TFs and the identity of their downstream targets that specify invasiveness is largely unknown. Cell invasion through basement membrane (BM) serves as a mechanism underlying cell dispersal and organ formation during normal development, immune surveillance, and is mis-regulated during cancer metastasis. The Sherwood laboratory at Duke University has established a simple in vivo model that uniquely combines powerful genetic, functional genomic, and single cell visual analyses of anchor cell (AC) invasion through BM into the vulval epithelium during C. elegans larval development. Our current understanding of AC invasion includes the identification of three TFs and a handful of putative downstream targets that regulate both the establishment of a specialized invasive membrane and the ability of the AC to remove BM during invasion. The proposed experiments during the mentored phase of the award will begin to characterize the GRNs underlying cell invasion through BM by 1) identifying the binding partner of the bZIP oncogene TF, fos-1a, and the identity of FOS-1A binding sites within the C. elegans genome; and 2) identifying additional TFs that regulate AC invasion and testing whether they show conserved functions in regulating carcinoma cell invasion in vitro. Training in functional genomic techniques and systems biology approaches will be accelerated through advanced course work, attendance at national conferences, a collaboration with Dr. Marian Walhout's laboratory (University of Massachusetts Medical School), and co-mentorship by Dr. Philip Benfey, the director of Duke University's Systems Biology group and the Institute for Genome Sciences and Policy (IGSP). Training in human cancer in vitro assays will be carried out in the laboratory of Dr. Stephen Weiss (University of Michigan). These training experiences and the data acquired during the mentored phase will provide the basis to launch an independent research career. The research proposed in the unmentored phase of the award includes 1) characterizing the genomic regulatory regions that contain TF-binding sites that control gene expression in the AC during invasion and 2) determining the core complement of TFs that are both necessary and sufficient to recapitulate an invasion program in normal development and in human cancer invasion.