Calcineurin in Congenital Urinary Tract Obstruction

先天性尿路梗阻中的钙调神经磷酸酶

• 批准号: 7194966
• 负责人: FENG CHEN
• 金额: $ 30.68万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7194966
• 关键词: Alleles; BMP4; Behavior; Calcineurin; Calcineurin Pathway; Cell Line; Cells; Childhood; Complement; Development; Diagnostic Procedure; Epithelial; Epithelium; Etiology; Event; Genomics; Genotype; Human; Kidney Diseases; Kidney Failure; Knowledge; Lesion; Mesenchymal; Mesenchyme; Metanephric Diverticulum; Methods; Molecular; Molecular Genetics; Mus; Mutant Strains Mice; Obstruction; Pathogenesis; Pathway interactions; Proteomics; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Testing; Therapeutic; Tissues; Transducers; Transgenes; Ureteropelvic junction obstruction; Urinary tract; extracellular; immortalized cell; in vivo; insight; knowledge base; mutant; novel diagnostics; programs; response; tool; urinary; urinary tract obstruction

DESCRIPTION (provided by applicant): Congenital obstructive nephropathy is the principal cause of pediatric renal failure. The molecular and cellular lesions leading to congenital obstruction, however, are still largely undetermined. We have shown that the absence of calcineurin in the developing urinary tract mesenchyme, in the Pax3Cre Cnb1 mutants we generated, led to congenital obstructive nephropathy resembling Ureteropelvic Junction (UPJ) obstruction in humans. Our study has demonstrated a clear requirement for calcineurin in urinary tract development and provided us with materials and knowledge to further investigate the indispensable signaling events involved and the etiology of congenital obstructive nephropathy. We propose to first study the interactions between calcineurin and other signaling pathways known to be involved in urinary tract development. This will test the hypothesis that calcineurin is an indispensable signaling transducer for urinary tract development. This will also effectively connect our observations to the existing knowledge in this field and will provide a more comprehensive explanation, at the molecular and cellular level, for the pathogenesis of the congenital obstructive nephropathy. Then, we will develop primary and immortalized cell lines carrying the Cnb1 deletion to determine the effects of calcineurin inactivation in cellular behavior and signaling to test the hypothesis that calcineurin is required for interpreting various extracellular signals in cells of the urinary tract. We will also use genomic/proteomic approaches and molecular genetic methods to identify factors downstream of calcineurin in urinary tract development. Furthermore, we will complement the Pax3Cre-Cnb1 study with Cnb1 deletion in the ureteric bud (UB) derivatives to test the hypothesis that calcineurin has additional indispensable in vivo function in the UB and in the epithelial-mesenchymal interactions during urinary tract development. We will use these mutants to further analyze the interactions of the relevant signaling pathways in vivo. Results from the proposed studies will provide mechanistic insights into the function of calcineurin in normal and abnormal development of the urinary tract and the etiology of congenital obstructive nephropathy.

描述（由申请人提供）：先天性梗阻性肾病是小儿肾衰竭的主要原因。然而，导致先天性梗阻的分子和细胞病变仍然很大程度上尚未确定。我们已经证明，在我们产生的 Pax3Cre Cnb1 突变体中，发育中的尿路间充质中缺乏钙调神经磷酸酶，导致先天性梗阻性肾病，类似于人类肾盂输尿管连接处 (UPJ) 梗阻。我们的研究证明了尿路发育对钙调神经磷酸酶的明确需求，并为我们进一步研究先天性梗阻性肾病所涉及的不可或缺的信号事件和病因学提供了材料和知识。 我们建议首先研究钙调神经磷酸酶和已知参与泌尿道发育的其他信号通路之间的相互作用。这将检验钙调神经磷酸酶是泌尿道发育不可或缺的信号转导器的假设。这也将有效地将我们的观察结果与该领域的现有知识联系起来，并在分子和细胞水平上为先天性梗阻性肾病的发病机制提供更全面的解释。然后，我们将开发携带 Cnb1 缺失的原代细胞系和永生化细胞系，以确定钙调磷酸酶失活对细胞行为和信号传导的影响，以检验钙调磷酸酶是解释泌尿道细胞中各种细胞外信号所必需的假设。我们还将使用基因组/蛋白质组学方法和分子遗传学方法来识别泌尿道发育中钙调神经磷酸酶下游的因素。此外，我们将通过输尿管芽（UB）衍生物中的 Cnb1 缺失来补充 Pax3Cre-Cnb1 研究，以检验钙调神经磷酸酶在 UB 和泌尿道发育过程中上皮间质相互作用中具有额外不可或缺的体内功能的假设。我们将利用这些突变体进一步分析体内相关信号通路的相互作用。拟议研究的结果将为钙调神经磷酸酶在泌尿道正常和异常发育中的功能以及先天性梗阻性肾病的病因学提供机制见解。