Elucidating the Gene Regulatory Networks that Specify Invasive Behavior

阐明指定入侵行为的基因调控网络

• 批准号: 8321493
• 负责人: David Matus
• 金额: $ 13.42万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-17 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8321493
• 关键词: Architecture; Automobile Driving; Award; Basement membrane; Behavior; Binding; Binding Sites; Biological; Biological Process; Blood Vessels; C. elegans genome; Caenorhabditis elegans; Cancer Model; Cells; Cellular biology; Collaborations; Complement; Complex; Data; Development; Developmental Biology; Dimerization; Dissection; Disseminated Malignant Neoplasm; Elements; Epithelium; Excision; Family member; Fingers; Gene Components; Gene Expression; Gene Targeting; Genes; Genetic; Genome; Genomics; Goals; Growth; Homologous Gene; Human; Human Genome; Hybrids; Immunologic Surveillance; In Vitro; Institutes; Institution; Intervention; Laboratories; Malignant Epithelial Cell; Malignant Neoplasms; Massachusetts; Membrane; Mentors; Mentorship; Michigan; Modeling; Movement; Neoplasm Metastasis; Nucleic Acid Regulatory Sequences; Organ; Orthologous Gene; Pathway Analysis; Phase; Phenotype; Phylogenetic Analysis; Pregnancy; Process; Property; Proteins; RNA Interference; Regulation; Regulator Genes; Regulatory Element; Research; Research Proposals; Resolution; Role; Science Policy; Scientist; Site; Specific qualifier value; Systems Biology; Techniques; Testing; Time; Training; Transcription Factor AP-1; Transcription Factor Oncogene; Universities; Vertebrates; Visual; Work; Yeasts; anti-cancer therapeutic; base; career; dimer; expectation; experience; functional genomics; in vitro Assay; in vivo; in vivo Model; innovation; insight; jun Oncogene; medical schools; new therapeutic target; novel strategies; programs; promoter; research study; symposium; therapeutic target; tool; transcription factor

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：我的长期目标是建立自己作为一个独立的科学家在一个顶级的研究机构研究基因调控网络（GRNs），在发展和癌症调节细胞形态发生运动。为此，拟议的研究推进了我在功能基因组学和系统生物学方法方面的培训，补充了我以前在细胞和发育生物学技术方面的研究。GRN包括转录因子（TF）之间以及TF与其驱动特定细胞生物学过程的靶基因之间的物理和调节关系。尽管它们在调节细胞侵袭行为中的重要性，但TF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。通过基底膜（BM）的细胞侵袭充当在正常发育、免疫监视期间的细胞分散和器官形成的基础机制，并且在癌症转移期间被错误调节。杜克大学的舍伍德实验室建立了一个简单的体内模型，该模型独特地结合了强大的遗传学、功能基因组学和单细胞视觉分析，用于分析锚细胞（AC）在C.线虫幼虫发育我们目前对AC侵袭的理解包括鉴定三种TF和少数推定的下游靶点，其调节专门的侵袭膜的建立和AC在侵袭期间去除BM的能力。在该奖项的指导阶段，拟议的实验将开始通过1）鉴定bZIP癌基因TF的结合伴侣fos-1a和C.线虫基因组;和2）鉴定调节AC侵袭的其它TF并测试它们在体外调节癌细胞侵袭中是否显示保守功能。将通过高级课程、参加国家会议、与Marian Walhout博士的实验室（马萨诸塞州大学医学院）合作以及由杜克大学系统生物学组和基因组科学与政策研究所主任Philip Benhout博士共同指导，加快功能基因组技术和系统生物学方法的培训。将在Stephen韦斯博士（密歇根大学）的实验室进行人类癌症体外测定的培训。这些培训经验和在指导阶段获得的数据将为启动独立的研究生涯提供基础。在该奖项的未指导阶段提出的研究包括1）表征包含TF结合位点的基因组调控区域，这些位点在侵袭期间控制AC中的基因表达，以及2）确定TF的核心补体，这些TF对于在正常发育和人类癌症侵袭中重现侵袭程序是必要且足够的。