Wnt5a and Planar Cell Polarity in Kidney Development and Cyst formation

Wnt5a 和平面细胞极性在肾脏发育和囊肿形成中的作用

基本信息

批准号：
8383256
负责人：
Liwei Huang
金额：
$ 15.51万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-15 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8383256
关键词：
Affect American Anatomy Binding Birth Cell Culture Techniques Cell Line Cell Polarity Cell division Cells Cellular Structures Child Childhood Collagen Congenital Abnormality Cyst Cystic kidney Cytoskeletal Proteins Defect Development Developmental Gene Developmental Process Disease Drosophila genus Duct (organ) structure Elements Embryo End stage renal failure Endoderm Engineering Epithelial Epithelium Family Gel Gene Family Genes Glycoproteins Goals Human Illinois In Vitro Intermediate Mesoderm Kidney Knock-out Knockout Mice MAPK8 gene MDCK cell Measurement Mesenchymal Mesenchyme Messenger RNA Metanephric Diverticulum Metanephric structure Modeling Molecular Profiling Morphogenesis Morphology Mus Newborn Infant Nuclear Organelles Partner in relationship Pathway interactions Pattern Phenotype Physiological Play Polycystic Kidney Diseases Process Prostate Prostatic Tissue Protein Family Proteins Renal tubule structure Reporting Role Signal Pathway Signal Transduction Structure System Testing Therapeutic Intervention Tissues Tubular formation Universities Urinary tract Wild Type Mouse Zebrafish cell motility design directional cell functional group in vivo kidney cell member neonate nephrogenesis novel overexpression prenatal receptor research study zebrafish development

项目摘要

DESCRIPTION (provided by applicant): Congenital anomalies of the kidney and urinary tract (CAKUT) constitutes 20-30% of anomalies identified in the prenatal period, and is estimated to play a causative role in up to 50% of cases of end-stage kidney disease in children. CAKUT, affecting approximately 600,000 Americans, encompases a large group of disorders that result from abnormal renal developmental processes. Planar cell polarity (PCP) is the coordinated orientation of cells and cellular structures along an axis within the plane of an epithelial surfac. PCP plays a central role during renal development when newly formed tubules undergo progressive lengthening by directed cell migration and cell division. Wnt family proteins have been shown to control cell polarity and directional cell movement in many developmental systems. The mechanisms by which Wnt signaling pathways integrate the organization of receptors, organelles, and cytoskeletal proteins to confer cell polarity and directional cell movement are not completely understood. The goal here is to understand the mechanism by which Wnt5a integrates downstream organelles and cytoskeletal proteins, as well as PCP proteins, to confer cell polarity and directional cell movement during kidney development. Wnt5a is a noncanonical Wnt that binds to its receptor Frizzled, acts through disheveled (Dsh), and activates both the Wnt/Ca2+ and PCP pathways. I previously showed that Wnt5a nul mice develop defects in prostate gland lumenization and planar cell polarity, and Wnt5a null mice, which die at birth, lack kidneys. Wnt5a appears to be expressed in the mesenchyme/stroma of the developing mouse kidney. The hypothesis, therefore, that we propose testing is that Wnt5a regulates kidney development via actions on the PCP pathway. Accordingly, in Aim 1, I will use in vitro cel culture to investigate the detailed mechanism by which Wnt5a regulates PCP. In Aim 2, I will determine the in vivo role of Wnt5a signaling for guiding pronephric kidney development, using antisense Wnt5a morpholinos in zebrafish. Use of zebrafish will allow me to rapidly confirm and expand on the mechanistic in vitro results from Aim 1. Finally, in Aim 3, I will determine the role of Wnt5a in controlling PCP and regulating metanephric kidney development in vivo by utilizing the Cre/loxP binary mouse system to generate kidney-specific Wnt5a knockout mice. Wnt5a floxed mice are available to us and I plan to do a whole expression profile of Wnt5a in the developing mouse kidney to confirm reports of Wnt5a expression in developing kidney mesenchyme/stroma. Mesenchymal kidney Cre drivers, such as FoxD1, are commercially available, and I can knockout Wnt5a in both a kidney-specific and temporal manner by mating Wnt5a floxed mice with FoxD1-CreERT2 mice. I will evaluate oriented cell division and other PCP parameters in the Wnt5a knockout model and then investigate downstream elements including PCP proteins. Successful completion of these experiments will result in the identification of novel candidate targets for therapeutic intervention in CAKUT, a disorder for which no approved treatments currently exist. PUBLIC HEALTH RELEVANCE: Congenital anomalies of the kidney and urinary tract (CAKUT) constitute 20-30% of all birth defects identified in newborns, and is the cause of up to 50% of childhood end-stage kidney disease. Wnt5a is a member of the Wnt gene family, a group of very important developmental genes, and mice without Wnt5a lack kidneys. The proposed experiments are designed to determine the mechanism by which Wnt5a regulates kidney development, with the successful completion of these experiments resulting in the identification of novel candidate targets for therapeutic intervention in CAKUT, a disorder for which no treatments currently exist.

描述（由申请人提供）：肾脏和尿路（Cakut）的先天性异常占产前时期鉴定的异常的20-30％，据估计，在儿童的最终末期肾脏病病例中最多可在50％的病例中起致病作用。卡库特（Cakut）影响了大约60万美国人，它包含了由于肾脏发育过程异常而导致的大量疾病。平面细胞极性（PCP）是沿上皮表面平面内轴的细胞和细胞结构的协调方向。当新形成的小管通过定向细胞迁移和细胞分裂进行逐步延长时，PCP在肾发育过程中起着核心作用。 Wnt家族蛋白已显示可控制许多发育系统中的细胞极性和定向细胞运动。 Wnt信号通路整合受体，细胞器和细胞骨架蛋白的组织以赋予细胞极性和定向细胞运动的机制。这里的目的是了解WNT5A整合下游细胞器和细胞骨架蛋白以及PCP蛋白的机制，以在肾脏发育过程中赋予细胞极性和方向性细胞运动。 Wnt5a是一种非规范的Wnt，与其受体卷曲结合，通过衣衫e（DSH）起作用，并激活Wnt/Ca2+和PCP途径。我先前表明，wnt5a小鼠在前列腺透明度和平面细胞极性中出现缺陷，而生于出生时死亡的Wnt5a无效小鼠缺乏肾脏。 Wnt5a似乎在发育中的小鼠肾脏的间充质/基质中表达。因此，我们提出的测试的假设是WNT5A通过在PCP途径上的作用来调节肾脏发育。因此，在AIM 1中，我将使用体外培养物来研究WNT5A调节PCP的详细机制。在AIM 2中，我将使用反义WNT5A吗磷脂在斑马鱼中确定Wnt5a信号传导在引导倾斜肾脏发育中的体内作用。斑马鱼的使用将使我能够迅速确认和扩展AIM 1的体外结果。最后，在AIM 3中，我将确定Wnt5a在控制PCP和调节体内跨肾上腺素肾脏发育中的作用，通过使用CRE/LOXP二进制型二进制型肾小管型肾脏型肾小管肾脏特异性Wntney Wntney Wntney Wnt5A敲除小鼠。我们可以使用Wnt5a Floxed小鼠，我计划在发育中的小鼠肾脏中进行WNT5A的全面表达曲线，以确认在发育中的肾脏间充质/基质中的Wnt5a表达的报道。诸如FOXD1之类的间充质肾脏CRE驱动器是可商购的，我可以通过将Wnt5a Floxed小鼠与FoxD1-Creert2小鼠交配，以肾脏特异性和时间的方式敲除Wnt5a。我将评估Wnt5a基因敲除模型中的定向细胞分裂和其他PCP参数，然后研究包括PCP蛋白在内的下游元素。这些实验的成功完成将导致确定在Cakut治疗干预措施的新型候选靶标，该疾病目前尚无批准的治疗方法。公共卫生相关性：肾脏和尿路的先天异常（Cakut）占新生儿中所有出生缺陷的20-30％，是童年末期肾脏疾病的50％的原因。 Wnt5a是Wnt Gene家族的成员，Wnt Gene家族是一组非常重要的发育基因，而没有Wnt5a的小鼠缺乏肾脏。提出的实验旨在确定WNT5A调节肾脏发育的机制，这些实验成功地完成了这些实验，从而确定了Cakut治疗干预的新型候选靶标，这是当前不存在治疗的疾病。