Molecular and cellular basis of the renal diseases associated with Alagille Syndrome

阿拉吉尔综合征相关肾脏疾病的分子和细胞基础

基本信息

批准号：
10209370
负责人：
Kameswaran Surendran
金额：
$ 36.52万
依托单位：
SANFORD RESEARCH/USD
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10209370
关键词：
Address Alagille Syndrome Alleles Cell Culture Techniques Cell division Cell physiology Cellular Assay Child Chronic Kidney Failure Cilia Code Congenital Abnormality Cyst Cystic kidney DNA Sequence Alteration Defect End stage renal failure Enhancers Ensure Epithelial Epithelial Cells Exons Genes Genetic Genetic Models Genetic Variation Genome Head Cancer Health Human Kidney Kidney Diseases Knowledge Length Life Ligands Link Maintenance Malignant neoplasm of lung Mediating Mediator of activation protein Methods Molecular Multicystic Dysplastic Kidney Mus Mutation Neck Cancer Notch Signaling Pathway Pathogenicity Pathway interactions Patients Penetrance Peptides Phenotype Plants Process Proteins Renal carcinoma Reporting Repression Retrospective Studies Scaffolding Protein Severities Signal Transduction Signaling Protein Skin Cancer Structure Syndrome Testing Tumor Suppressor Proteins Urinary tract Variant autosomal dominant mutation cell type congenital anomaly exome sequencing improved kidney cell kidney epithelial cell mouse model mutant nephrogenesis nephron progenitor notch protein notch-2 protein novel prevent rare variant receptor renal epithelium small molecule stem

项目摘要

Project Summary The leading cause of end stage renal disease among children under 5 years is congenital abnormalities of the kidney and urinary tract (CAKUT). Among the several genetic mutations that have been linked to CAKUT are mutations in Notch signaling pathway genes, JAG1 and NOTCH2. Mutations in these Notch pathway genes are predicted to reduce the level of Notch signaling activity, and are associated with Alagille Syndrome (ALGS). One component of ALGS is the variable occurrence of kidney disease including that of small multicystic, dysplastic kidneys. For instance, a retrospective study determined that 40% of ALGS patients with JAG1 mutations had some form of kidney disease with dysplastic kidneys with or without cysts occurring in 60% of ALGS patients with kidney disease. The underlying cellular and molecular mechanisms by which Notch signaling ensures normal kidney development and maintenance are poorly understood. We know that JAG1 can function as a ligand to activate NOTCH2, and assume the canonical Notch signaling pathway prevents kidney disease associated with ALGS. However, mutations in the other canonical Notch signaling pathway components have not been associated with ALGS. Additionally, the high degree of variability in the manifestation of kidney disease among ALGS patients is puzzling. In our mouse models the severity of multi-cystic kidney disease increases with increasing number of Notch1 and Notch2 alleles inactivated in the developing renal epithelium. Here we propose to determine the Renal Epithelial Notch Signaling Network (RENSN) and perform whole exome sequencing of ALGS patients to determine if additional variations in RENSN genes in addition to JAG1 and/or NOTCH2 determine the occurrence of multicystic/dysplastic kidneys in ALGS. Additionally, we will test if NOTCH2 mutations associated with ALGS alter the orientation of renal epithelial cell division, primary cilia structure and expression of renal epithelial Notch target genes in different renal epithelial cell cultures. We will also determine the proximal interacting proteins of wild type versus ALGS associated NOTCH2 variants using BioID. Additionally, we will apply three strategies to molecularly intervene and prevent the Notch-signaling- deficient kidney defects. These studies will establish the underlying cellular and molecular mechanisms of kidney disease associated with ALGS and identify methods to increase Notch signaling specifically in kidney cell types by determining the molecular interface between Notch signaling and renal epithelial functions.

项目概要 5岁以下儿童终末期肾病的主要原因是先天性肾功能异常肾脏和泌尿道（CAKUT）。与 CAKUT 相关的几种基因突变包括 Notch 信号通路基因 JAG1 和 NOTCH2 发生突变。这些Notch途径基因的突变是预计会降低 Notch 信号活性水平，并与 Alagille 综合征 (ALGS) 相关。一 ALGS 的组成部分是肾脏疾病的可变发生，包括小型多囊性、发育不良性肾脏疾病肾脏。例如，一项回顾性研究确定，40% 具有 JAG1 突变的 ALGS 患者患有 60% 的 ALGS 患者患有某种形式的肾脏疾病，其中肾脏发育不良，伴或不伴囊肿患有肾脏疾病。 Notch 信号传导确保的潜在细胞和分子机制人们对正常肾脏的发育和维护知之甚少。我们知道 JAG1 可以充当配体激活 NOTCH2，并假设经典的 Notch 信号通路可预防肾脏疾病与 ALGS 相关。然而，其他典型 Notch 信号通路成分的突变与 ALGS 没有关联。此外，肾脏疾病表现的高度可变性 ALGS 患者的情况令人费解。在我们的小鼠模型中，多囊肾疾病的严重程度增加随着发育中肾上皮中失活的 Notch1 和 Notch2 等位基因数量的增加。在这里我们建议确定肾上皮切迹信号网络 (RENSN) 并执行全外显子组对 ALGS 患者进行测序，以确定除了 JAG1 和/或之外，RENSN 基因是否存在其他变异 NOTCH2 决定 ALGS 中多囊性/发育不良性肾脏的发生。此外，我们将测试是否与 ALGS 相关的 NOTCH2 突变改变肾上皮细胞分裂、初级纤毛的方向不同肾上皮细胞培养物中肾上皮Notch靶基因的结构和表达。我们将还使用以下方法确定野生型与 ALGS 相关 NOTCH2 变体的近端相互作用蛋白生物识别号。此外，我们将应用三种策略来分子干预和防止Notch信号传导- 肾虚缺陷。这些研究将建立肾脏的潜在细胞和分子机制与 ALGS 相关的疾病，并确定增强肾细胞类型中 Notch 信号传导的方法通过确定Notch信号传导和肾上皮功能之间的分子界面。