A novel epithelial cell migration drives nephron repatterning and convolution

一种新的上皮细胞迁移驱动肾单位重新模式和卷积

基本信息

批准号：
7918918
负责人：
Aleksandr Vasilyev
金额：
$ 13.88万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-21 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7918918
关键词：
Actins Anterior Architecture Biological Assay Cancer Biology Catalytic Domain Chemicals Competence Confocal Microscopy Congenital Abnormality Cues Defect Development Dominant-Negative Mutation Epithelial Epithelial Cells Epithelium Fluorescent Antibody Technique Focal Adhesions Future Genes Homeostasis Human Image Image Analysis Immigration In Situ Lesion Injury Investigation Kidney Knowledge Label Life Ligands Link Liquid substance Malignant Neoplasms Mediating Mediator of activation protein Molecular Molecular Profiling Morphogenesis Morphology Mutation Myosin ATPase Natural regeneration Nephrons Oligonucleotides Organ PIK3CG gene Pathology Pathway interactions Phenotype Play Positioning Attribute Principal Investigator Process Pronephric structure Recovery Relative (related person)Renal function Renal tubule structure Resolution Role Screening procedure Seminal Shapes Signal Pathway Signal Transduction Small Molecule Chemical Library System Technology Time Tissues Transgenic Model Transgenic Organisms Tubular formation Work Zebrafish cell motility fluid flow in vivo inhibitor/antagonist kidney cell migration mutant notch protein novel prevent public health relevance regenerative small molecule tool

Actins Anterior Architecture Biological Assay Cancer Biology Catalytic Domain Chemicals Competence Confocal Microscopy Congenital Abnormality Cues Defect Development Dominant-Negative Mutation Epithelial Epithelial Cells Epithelium Fluorescent Antibody Technique Focal Adhesions Future Genes Homeostasis Human Image Image Analysis Immigration In Situ Lesion Injury Investigation Kidney Knowledge Label Life Ligands Link Liquid substance Malignant Neoplasms Mediating Mediator of activation protein Molecular Molecular Profiling Morphogenesis Morphology Mutation Myosin ATPase Natural regeneration Nephrons Oligonucleotides Organ PIK3CG gene Pathology Pathway interactions Phenotype Play Positioning Attribute Principal Investigator Process Pronephric structure Recovery Relative (related person)Renal function Renal tubule structure Resolution Role Screening procedure Seminal Shapes Signal Pathway Signal Transduction Small Molecule Chemical Library System Technology Time Tissues Transgenic Model Transgenic Organisms Tubular formation Work Zebrafish cell motility fluid flow in vivo inhibitor/antagonist kidney cell migration mutant notch protein novel prevent public health relevance regenerative small molecule tool

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项目摘要

DESCRIPTION (provided by applicant): The human nephron is subdivided into segments, each playing a distinct role in overall kidney function. The segments differ in their position, architecture, and molecular profile. Little is known about factors determining the exact positioning of segment boundaries as well as the forces determining the morphology of various nephron segments, such as tubule convolution. We discovered that in zebrafish pronephros epithelial cells are engaged in a concerted migration towards the glomerulus that depends on the flow of luminal fluid. This migration results in establishing the position of nephron segment boundaries and in convolution of the proximal kidney. In the proposed study, we will explore the mechanism of this novel phenomenon and establish new tools for the study of cell migration. In Aim1, we will investigate the subcellular mechanisms of pronephric migration by combining live fluorescent imaging with fluorescent antibody assays and confocal microscopy, all within the framework of advanced image analysis. In Aim 2, we will expand on our finding that in zebrafish mindbomb mutant which interferes with Notch signaling, the pronephric migration is eliminated and the tubule fails to form the proximal convolution. We will use a combination of morpholino oligonucleotide, chemical inhibitor and inducible dominant negative transgenic approaches to define the exact molecular substrate and the mode of action of Notch signaling underlying this phenotype. In Aim 3, we will probe specific molecular mechanisms underlying the pronephric migration. We found that Pi3K inhibitor LY294002 interferes with kidney cell migration implicating Pi3K and possibly Cdc42. We will disrupt the function of PI3K and Cdc42 by expressing inducible dominant negative constructs and examine the role of these two molecules in pronephric migration. Lastly, we will use a targeted small molecule chemical library to screen for effects of known inhibitors on pronephric migration. In summary, we propose an in-depth investigation of a novel process of cell migration within polarized epithelium, which links organ function with organ morphogenesis. PUBLIC HEALTH RELEVANCE: This study will significantly advance our knowledge of nephron morphogenesis and help us develop future regenerative technologies. In addition, it will make a seminal contribution to the study of epithelial migration of which very little is currently known. This process also has potential implications in cancer biology, such as cancer invasion and spread of in-situ lesions.

描述（由申请人提供）：人的肾单位细分为细分市场，每个细分都在整个肾脏功能中起着独特的作用。这些段在其位置，结构和分子剖面上有所不同。关于确定片段边界的确切定位以及确定各种肾单位段形态（例如小管卷积）的力学的因素，知之甚少。我们发现，在斑马鱼中，pronephros中皮细胞与肾小球的一致迁移，取决于腔流体的流动。这种迁移导致建立肾单位段边界的位置和近端肾脏的卷积。在拟议的研究中，我们将探讨这种新现象的机制，并为细胞迁移研究建立新的工具。在AIM1中，我们将通过将活荧光成像与荧光抗体测定和共聚焦显微镜相结合，研究促脑迁移的亚细胞机制，均在高级图像分析的框架内。在AIM 2中，我们将扩展我们的发现，在斑马鱼思维炸弹突变体中会干扰Notch信号传导，消除了倾斜迁移，并且小管未形成近端卷积。我们将使用吗啡寡核苷酸，化学抑制剂和可诱导的显性负转基因方法的组合来定义确切的分子底物以及该表型的凹口信号传导的作用方式。在AIM 3中，我们将探测促脑迁移的基于的特定分子机制。我们发现PI3K抑制剂LY294002干扰了肾细胞迁移，这可能是PI3K和可能的CDC42。我们将通过表达可诱导的主要负面构建体来破坏PI3K和CDC42的功能，并检查这两个分子在俯卧迁移中的作用。最后，我们将使用靶向的小分子化学文库来筛选已知抑制剂对俯卧迁移的影响。总而言之，我们提出了对偏振上皮内细胞迁移过程的深入研究，该过程将器官功能与器官形态发生联系起来。公共卫生相关性：这项研究将大大提高我们对肾脏形态发生的了解，并帮助我们发展未来的再生技术。此外，它将为研究目前知之甚少的上皮迁移的研究做出开创性的贡献。该过程还对癌症生物学具有潜在的影响，例如癌症侵袭和原位病变的扩散。