NPHP2 in ciliary function, renal fibrosis and cyst formation

NPHP2 在纤毛功能、肾纤维化和囊肿形成中的作用

• 批准号: 10736919
• 负责人: ZHAOXIA SUN
• 金额: $ 58.7万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736919
• 关键词: Autosomal Dominant Polycystic Kidney; Biochemical; Biological Assay; Cells; Child; Cilia; Clustered Regularly Interspaced Short Palindromic Repeats; Coupled; Cyst; Cystic Kidney Diseases; Cystic kidney; Data; Development; Disease; Disease Progression; End stage renal failure; Epithelial Cells; Epithelial cyst; Epithelial-Stromal Communication; Epithelium; Etiology; Excision; Extracellular Matrix Proteins; Feedback; Fibroblasts; Fibrosis; Foundations; Future; Gene Silencing; Generations; Genes; Genetic; Genetic Transcription; Grant; Investigation; Kidney; Knock-out; Knockout Mice; Knowledge; Left; Ligands; Link; Mediating; Messenger RNA; Modeling; Molecular; Mus; Mutagenesis; Myofibroblast; Nature; Neonatal; Nephronophthisis; Outcome; PKD1 gene; PKD2 protein; Pathway Analysis; Pathway interactions; Patients; Phenotype; Physiological; Play; Post-Transcriptional Regulation; Proteins; Proteomics; Research; Role; Severities; Signal Transduction; Stromal Cells; Symptoms; Testing; Tissues; Work; Zebrafish; cell type; ciliopathy; cilium biogenesis; in vivo; infancy; insight; interstitial; interstitial cell; kidney cell; kidney fibrosis; mouse model; mutant; novel; polycystic kidney disease 1 protein; posttranscriptional; renal epithelium; response; smoothened signaling pathway; targeted treatment; transcriptomic profiling; translatome; young adult

This proposal focuses on both cell autonomous and non-autonomous function of the nephronophthisis (NPHP) gene, Nphp2, in renal fibrosis and cyst formation. NPHP is a recessive renal ciliopathy characterize by interstitial fibrosis and cysts. Although NPHP accounts for a significant portion of end stage renal disease in children and young adults, currently no targeted therapy is available for this disease and the underlying molecular etiology, particular for interstitial fibrosis, is not well understood, even though NPHP proteins have been put into different modules biochemically. The lack of knowledge hampers the effort to develop targeted therapy for this disease. Previous work showed that mouse whole-body knockout model of Nphp2 displays renal fibrosis and cysts at the neonatal stage. By generating and analyzing epithelial and complementary stromal specific knockout mouse models of Nphp2, we pinpointed defective epithelial cells as the driver for both interstitial fibrosis and epithelial cyst formation. In addition, myofibroblast activation occurred early during disease progression and preceded detectable cyst formation in the epithelial specific Nphp2 mutants. Moreover, abrogation of cilia genetically partially suppresses the phenotype of Nphp2 mutants, suggesting that cilia play a significant role in Nphp2 function. The central hypothesis of this project is that Nphp2 inhibits a cilia-dependent profibrotic and pro-cystic pathway in renal epithelial cells and that both cell autonomous and non-autonomous responses contribute to disease progression. Two specific aims were proposed to test this hypothesis. Aim 1 focuses on renal epithelial cells. A combination of approaches, from in vivo phenotype characterization, transcriptome profiling to pathway analysis to identify key pathways that are disrupted in Nphp2 mutant epithelial cells. Aim 2 focuses on epithelial-interstitial crosstalk and will determine how stromal cells respond to epithelial cells with defective NPHP2 and whether stromal cells modify phenotypes of Nphp2 mutants. Completion of this project will provide critical insight into the molecular and cellular etiology of NPHP and cilia-mediated signaling under diverse physiological and disease states.

该建议集中于肾单位缺损（NPHP）基因Nphp2在肾纤维化和囊肿形成中的细胞自主和非自主功能。NPHP是一种以间质纤维化和囊肿为特征的隐性肾纤毛病。虽然NPHP占儿童和年轻人终末期肾病的很大一部分，但目前尚无针对这种疾病的靶向治疗，并且即使NPHP蛋白已被生物化学地放入不同的模块中，但其潜在的分子病因学，特别是间质纤维化的分子病因学仍不清楚。知识的缺乏阻碍了为这种疾病开发靶向治疗的努力。先前的工作表明，Nphp2的小鼠全身敲除模型在新生儿阶段显示肾纤维化和囊肿。通过生成和分析上皮和互补基质特异性敲除Nphp2的小鼠模型，我们确定了有缺陷的上皮细胞作为间质纤维化和上皮囊肿形成的驱动因素。此外，肌成纤维细胞活化发生在疾病进展的早期，并在上皮特异性Nphp2突变体中可检测到囊肿形成之前。此外，废除纤毛遗传部分抑制Nphp2突变体的表型，这表明纤毛在Nphp2功能中发挥重要作用。该项目的中心假设是Nphp 2抑制肾上皮细胞中依赖于纤毛的促纤维化和促囊性途径，并且细胞自主和非自主反应都有助于疾病进展。提出了两个具体目标来检验这一假设。目的1：研究肾上皮细胞。从体内表型表征，转录组分析到途径分析的方法组合，以确定在Nphp2突变上皮细胞中被破坏的关键途径。目的2侧重于上皮间质串扰，并将确定基质细胞如何响应上皮细胞与缺陷NPHP2和基质细胞是否修改NPHP2突变体的表型。本项目的完成将提供关键的洞察NPHP和纤毛介导的信号在不同的生理和疾病状态下的分子和细胞病因学。