Elucidating the Gene Regulatory Networks that Specify Invasive Behavior

项目7：项目摘要/摘要 我的长期目标是让自己成为一家顶级研究机构的独立科学家 研究调控细胞形态发生运动的基因调控网络 发展和癌症。为此，拟议的研究推进了我在以下方面的培训 功能基因组学和系统生物学方法，补充了我之前在 细胞和发育生物学技术。GRN涵盖物理和监管两个方面 转录因子之间以及转录因子与驱动它们的靶基因之间的关系 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，但 TFS及其下游目标的身份指定侵入性在很大程度上是未知的。细胞 通过基底膜(BM)的侵袭是细胞弥散和 正常发育过程中的器官形成，免疫监视，以及癌症过程中的错误调节 转移。杜克大学舍伍德实验室建立了一个简单的活体模型 独一无二地结合了锚细胞的强大遗传、功能基因组和单细胞可视化分析 (Ac)在线虫幼虫发育期间，通过BM侵入外阴上皮。我们的 目前对AC侵袭的理解包括识别三个TF和几个假定的 下游靶标，调节专门侵袭性膜的建立和 AC在入侵过程中清除BM的能力。在指导期间提出的实验 颁奖阶段将开始通过1)确定通过BM进行细胞入侵的GRN的特征 鉴定bZIP癌基因Tf、FOS-1a的结合伙伴及FOS-1a结合的同源性 线虫基因组中的位置；以及2)识别更多调控AC入侵和 检测它们在体外是否具有调节癌细胞侵袭的保守功能。培训 在功能基因组技术和系统生物学方面，将通过 高级课程工作，参加国家会议，与玛丽安·沃尔胡特博士合作 实验室(马萨诸塞大学医学院)，菲利普·本菲博士共同指导 杜克大学系统生物学小组和基因组科学研究所所长 政策(IGSP)。人类癌症体外分析的培训将在Dr。 史蒂芬·韦斯(密歇根大学)。这些培训经验和在培训期间获得的数据 指导阶段将为开始独立的研究生涯提供基础。这项研究 在该奖项的非指导阶段提出的建议包括1)描述基因组调控 包含控制AC在入侵过程中基因表达的TF结合位点的区域和2) 确定既必要又充分的转换函数的核心补充，以概括 侵袭程序在正常发育和人类肿瘤的侵袭中。

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.