Characterizing Novel Regulators of Branching Morphogenesis within the Ureteric Bud Tip

输尿管芽尖内分支形态发生的新型调节因子的表征

• 批准号: 9325509
• 负责人: Elisabeth Rutledge
• 金额: $ 3.76万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-18 至 2018-08-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325509
• 关键词: Address; Alpha Cell; Biochemical; Biological Assay; Birth; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell Separation; Cells; Characteristics; Complex; Congenital Abnormality; Data; Defect; Development; Developmental Process; Diagnosis; Disintegrins; Dissection; Duct (organ) structure; Embryo; Employee Strikes; Epithelial; Epithelium; Etiology; Event; Gene Expression; Genes; Genitourinary system; Growth; Health; Human; Image; In Situ; In Situ Hybridization; In Vitro; Individual; Infection; Kidney; Kidney Diseases; Knowledge; Literature; Logic; Lung; Maps; Mediating; Mediator of activation protein; Mesenchyme; Metalloproteases; Metanephric Diverticulum; Microdissection; Modeling; Molecular; Molecular Analysis; Morphogenesis; Mus; Nephrons; Organ; Organogenesis; Pattern; Peptide Hydrolases; Phenotype; Platelet Factor 4; Play; Population; Prevention; Process; Regulator Genes; Renal function; Reporter; Research; Role; Signal Induction; Signal Pathway; Signal Transduction; Small Interfering RNA; Stem cells; System; Techniques; Thrombospondins; Transcript; Tretinoin; Tubular formation; Ureter; Urinary tract; Vascular System; base; body system; cell immortalization; cell type; comparative; congenital anomaly; developmental disease; extracellular; follow-up; genetic approach; human disease; immortalized cell; improved; insight; interest; kidney vascular structure; knock-down; mutant; nephrogenesis; novel; progenitor; programs; public health relevance; spatiotemporal; stem; transcriptome sequencing

 DESCRIPTION (provided by applicant): A process of branching morphogenesis underpins the formation of arborized epithelial networks in many organ systems including the lung and collecting duct system of the mammalian kidney. The collecting duct initiates on formation of the ureteric bud (UB) from the nephric duct. Cells at the bud tip, and at subsequent branch tips of the expanding ureteric epithelium, respond to inductive signals from the adjacent mesenchyme that promote and regulate branching morphogenesis, and maintain an uncommitted progenitor population at the branch tip. Progenitors exiting the tip to populate stalk regions differentiate ito the regionally distinct cell types of the collecting duct network. I aim to ascertain the unique molecular features of these tip cells to gain vital insights into stem cell dynamics and branching morphogenesis. I have utilized RNA sequencing data comparing gene expression in tip and stalk domains expression of the developing mouse kidney. These comparisons have identified a strong, tip-enriched set of genes that include all known tip regulatory genes from the literature. In Aim 1, I will perform a detailed in situ expression analysis of the tip- enriched gene set in th developing mouse kidney and lung to identify general and kidney specific regulators of epithelial morphogenesis. In Aim 2, I will develop a cell culture model of the ureteric tip population to systematically analyze the function of tip-enriched genes through genetic approaches. In Aim 3, I will explore the specific actions of Adamts18 (encoding a metalloproteinase with disintegrin-like and thrombospondin type 1 motif repeats) in the branching process. Adamts18 was identified as tip-enriched in both the kidney and lung. We have analyzed Adamts18 null mutants and shown that a subset of mutants display enlarged kidneys with double ureters. Thus, Adamts18 is normally required to restrict kidney branching and has emerged as a novel regulator of branching morphogenesis.

 描述（由申请人提供）：分支形态发生过程支持许多器官系统（包括哺乳动物肾脏的肺和集合管系统）中树枝状上皮网络的形成。集合管起始于输尿管芽（UB）从肾管形成。在芽尖的细胞，并在随后的扩张输尿管上皮分支尖端，响应于诱导信号从相邻的间充质，促进和调节分支形态发生，并保持一个未定型的祖细胞群体在分支尖端。祖细胞离开顶端进入茎区，分化成集合管网络的区域性不同的细胞类型。我的目标是确定这些尖端细胞的独特分子特征，以获得对干细胞动力学和分支形态发生的重要见解。我利用RNA测序数据比较了发育中小鼠肾脏的顶端和茎域表达的基因表达。这些比较已经确定了一个强大的，尖端丰富的基因集，包括所有已知的尖端调控基因的文献。在目的1中，我将对发育中的小鼠肾脏和肺中的尖端富集基因集进行详细的原位表达分析，以鉴定上皮形态发生的一般和肾脏特异性调节因子。目的二：建立输尿管尖端细胞培养模型，通过遗传学方法系统分析尖端富集基因的功能。在目标3中，我将探索Adamts 18（编码一种具有去整合素样功能的金属蛋白酶）的特异性作用。 和血小板反应蛋白1型基序重复）。Adamts18在肾脏和肺中均被鉴定为尖端富集。我们已经分析了Adamts18无效突变体，并显示突变体的一个子集显示双输尿管扩大的肾脏。因此，Adamts18通常需要限制肾脏分支，并已成为分支形态发生的新调节因子。