Control of Proliferation-related Genes and miRs by Nkx2-1 in Lung Development

Nkx2-1 对肺发育中增殖相关基因和 miR 的控制

• 批准号: 9274335
• 负责人: Jean-Bosco Tagne
• 金额: $ 16.6万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9274335
• 关键词: Address; Adult; Binding; Biological Assay; Birth; Boston; Cell Culture Techniques; Cell Differentiation process; Cell Proliferation; Cell Proliferation Regulation; Cells; Cellular biology; ChIP-on-chip; Complex; Cyclin B; Development; Differentiation and Growth; Distal; Environment; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Faculty; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Goals; Grant; In Vitro; Knock-in Mouse; Link; Lung; Lung Neoplasms; Lung diseases; Mediating; Medicine; Mentors; Mesenchymal; Messenger RNA; MicroRNAs; Molecular Biology; Morphogenesis; Mus; Mutant Strains Mice; Mutation; Nucleic Acid Regulatory Sequences; Oncogenes; Pathogenesis; Pattern; Phenotype; Phosphorylation; Prognostic Factor; Program Development; Proteins; Regulation; Reporter; Research; Research Personnel; Research Training; Role; Structure; Structure of parenchyma of lung; System; Teacher Professional Development; Testing; Time; Training; Translating; Tumor Cell Line; Universities; Writing; base; career; cell behavior; chromatin immunoprecipitation; deletion analysis; design; fetal; in vivo; insight; lung development; medical schools; meetings; mutant; nano; novel; promoter; skills; transcription factor

DESCRIPTION (provided by applicant): The research and training plans included in this application are designed to increase my expertise in lung cell and molecular biology, and to develop skills to pursue an independent career in lung research. With this KO1 project I expect to advance my skills in project design, grant writing, project and lab management to succeed in a timely transition to independent investigator. My immediate goals are to acquire advanced training in the analysis of transcriptional mechanisms of lung gene regulation, and of complex lung phenotypes. With the support of my mentor and co-mentors, experts in these fields, I propose to analyze Nkx2-1 regulation of proliferation-related genes and their effect on lung epithelial cell proliferation in Nkx2-1 mutant mice. The Nkx2-1 transcription factor is essential fr lung branching and distal epithelial cell differentiation during development. In the adult, Nkx2-1 maintains distal epithelial phenotypes by cell specific gene regulation. In lung tumors, Nkx2-1 is a lineage survival oncogene and a prognostic factor. A role of Nkx2-1 in control of cell proliferation has been suggested, but the target genes that may mediate proliferation in vivo are unknown. The goal of this project is to evaluate the role of Nkx2-1 direct target genes in the regulation of cell proliferation attributed to Nkx2-1 in mouse lung development and disease. Our previous ChIP-on-chip analyses revealed an overrepresentation of proliferation-related genes and a number of miRNAs bound by Nkx2-1 in early lung development. Here I hypothesize that Nkx2-1 regulates cell proliferation in the lung through these genes and miRNAs, and that they contribute to the pathogenesis of the abnormal lung phenotypes when altered by disruption of Nkx2-1 function. This hypothesis will be tested by: 1) Analyzing the developmental expression of selected cell proliferation-related Nkx2-1 targets previously identified in our screen, and evaluating Nkx2-1 binding to these targets in lung tissues, 2) Characterizing expression of Nkx2-1-regulated miRNAs and their mRNAs linked to cell proliferation in vitro and correlate their expression to that of downstream genes to establish an Nkx2-1 driven miRNA regulatory network involved in lung cell proliferation; 3) Evaluating the contribution of these Nkx2-1 target genes and miRNAs to the phenotype of Nkx2-1 null mutant lungs in which branching morphogenesis and differentiation are disrupted, and in Nkx2-1 phosphorylation mutant lungs in which the lung is hypoplastic in spite of normal branching. These will provide insights into the differential role of phosphorylated, unphosphorylated or absent Nkx2-1 in control of cell proliferation genes in vivo. The Pulmonary Center at Boston University School of Medicine is widely recognized for its supporting environment, helping Junior Faculty to transition to an independent career. The weekly discussions with my mentor and co-mentors, the work-in-progress meetings and the Faculty Development Program offered by the Department of Medicine are key components of my training plan. (End of Abstract)

描述(申请人提供)：本申请中包括的研究和培训计划旨在增加我在肺细胞和分子生物学方面的专业知识，并培养在肺部研究中独立发展的技能。通过这个KO1项目，我希望提高我在项目设计、拨款撰写、项目和实验室管理方面的技能，以成功地及时过渡到独立研究人员。我目前的目标是接受高级培训，分析肺基因调控的转录机制，以及复杂的肺表型。在我的导师和其他导师、这些领域的专家的支持下，我提议分析Nkx2-1对增殖相关基因的调控及其对Nkx2-1突变小鼠肺上皮细胞增殖的影响。Nkx2-1转录因子是发育过程中肺分支和远端上皮细胞分化所必需的。在成人中，Nkx2-1通过细胞特异性基因调控维持远端上皮细胞表型。在肺肿瘤中，Nkx2-1是一种谱系生存癌基因，是一种预后因素。Nkx2-1在控制细胞增殖中的作用已被提出，但可能在体内介导增殖的靶基因尚不清楚。本项目的目的是评估Nkx2-1直接靶基因在调节Nkx2-1在小鼠肺发育和疾病中的细胞增殖中的作用。我们之前的芯片分析显示，在早期肺发育中，与增殖相关的基因和Nkx2-1结合的一些miRNAs过度表达。在这里，我假设Nkx2-1通过这些基因和miRNAs调节肺内细胞的增殖，当Nkx2-1功能被破坏时，它们参与了异常肺表型的发病机制。这一假设将通过以下几个方面得到验证：1)分析我们筛选出的与细胞增殖相关的Nkx2-1靶标的发育表达，并评估Nkx2-1在肺组织中与这些靶标的结合；2)鉴定Nkx2-1调控的miRNAs及其与细胞增殖相关的mRNAs的表达，并将其与下游基因的表达相关联，以建立Nkx2-1驱动的参与肺细胞增殖的miRNA调控网络；3)评价Nkx2-1靶基因和miRNAs在Nkx2-1缺失突变肺分支形态发生和分化受阻以及Nkx2-1磷酸化突变肺分支正常但发育不良的表型中的作用。这些结果将为研究Nkx2-1的磷酸化、非磷酸化或缺失在体内控制细胞增殖基因中的不同作用提供依据。波士顿大学医学院的肺中心因其支持环境而得到广泛认可，帮助初级教师过渡到独立的职业生涯。每周与我的导师和合作导师的讨论，工作进行中的会议和医学部提供的教师发展计划是我培训计划的关键组成部分。(摘要结束)