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: 8028801
• 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 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8028801
• 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. PUBLIC HEALTH RELEVANCE: Cell invasion is not only a fundamental cell biological process during times of normal development (e.g., blood vessel growth and pregnancy), but is the underlying mechanism driving the spread of metastatic cancer in humans. Determining the core genes involved in promoting cell invasion in both C. elegans and human cancer will allow for the identification of new therapeutic targets to halt the lethality associated with cancer.

我的长期目标是成为一家顶级研究机构的独立科学家，研究在发育和癌症期间调节细胞形态发生运动的基因调控网络(GRN)。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，这些方法补充了我之前在细胞和发育生物学技术方面的研究。GRN包括转录因子之间的物理和调控关系，以及转录因子与其驱动特定细胞生物学过程的靶基因之间的关系。尽管它们在调节细胞侵袭行为方面很重要，但它们及其下游靶标的身份指定侵袭性在很大程度上是未知的。细胞通过基底膜(BM)侵袭是正常发育、免疫监视过程中细胞扩散和器官形成的一种机制，在肿瘤转移过程中受到错误调控。杜克大学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、功能基因组和单细胞视觉分析，研究了线虫幼虫发育过程中锚定细胞(AC)通过BM侵入外阴上皮的情况。我们目前对AC侵袭的理解包括识别三个TF和几个假定的下游靶点，这些靶点既调节特定侵袭膜的建立，也调节AC在侵袭过程中清除BM的能力。在该奖项的指导阶段提出的实验将开始通过1)确定bZIP癌基因Tf，FOS-1a的结合伙伴，以及线虫基因组中FOS-1A结合位点的身份；以及2)确定其他调节AC侵袭的因子，并测试它们是否在调节癌细胞体外侵袭中显示保守功能，从而表征GRN通过BM侵袭细胞的特性。功能基因组技术和系统生物学方法方面的培训将通过高级课程作业、出席国家会议、与玛丽安·沃尔胡特博士的实验室(马萨诸塞大学医学院)的合作以及杜克大学系统生物学小组和基因组科学与政策研究所(IGSP)主任菲利普·本菲博士的共同指导来加速。人类癌症体外分析方面的培训将在Stephen Weiss博士(密歇根大学)的实验室进行。这些培训经验和在指导阶段获得的数据将为开始独立的研究生涯提供基础。在该奖项的无指导阶段提出的研究包括1)表征基因组调节区，该基因组调节区包含控制AC在侵袭过程中基因表达的TF结合位点，以及2)确定对于概括正常发育和人类癌症侵袭过程中的侵袭计划来说是必要且充分的TF的核心补充。 公共卫生相关性：细胞侵袭不仅是正常发育时期(如血管生长和怀孕)的基本细胞生物学过程，而且是推动人类转移性癌症扩散的潜在机制。确定在线虫和人类癌症中促进细胞侵袭的核心基因将允许识别新的治疗靶点，以阻止与癌症相关的致命性。