Notch3 signaling in small-artery-diseases

小动脉疾病中的 Notch3 信号传导

基本信息

批准号：
7350196
负责人：
ANNE JOUTEL
金额：
$ 16.52万
依托单位：
NAT'L INST OF HLTH/MED RES INSERM-PARIS
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7350196
关键词：
Address Adult Affect Alleles Arterial Disorder Arteries Binding Sites Biological Blood Vessels Brain CADASIL Cell membrane Cell surface Characteristics Computer Simulation Cysteine Cytoskeleton Defect Dementia Deposition Depth Development Disease Electron Microscopy Exhibits Extracellular Domain Gene Expression Profile Gene Targeting Genes Goals Hand Histology Impaired cognition In Situ Hybridization In Vitro Infarction Knockout Mice LacZ Genes Laser-Doppler Flowmetry Maintenance Microarray Analysis Microscopy Missense Mutation Modeling Mus Mutant Strains Mice Mutation Nature Notch Signaling Pathway Numbers Optics Organ Pathway interactions Patients Phenotype Play Process RNA Reporter Research Resistance Resolution Role Shapes Signal Pathway Signal Transduction Smooth Muscle Myocytes Stereotyping Stroke System Tail Therapeutic Therapeutic Intervention Transgenic Mice Transgenic Organisms Vascular Dementia base cerebrovascular gain of function in vivo mouse Smc1l1 protein mouse Smc1l2 protein mutant notch protein null mutation programs promoter receptor research study therapeutic target tissue culture transcription factor white matter change

项目摘要

Small-vessel-diseases are a leading cause of Vascular Dementia. Our long-term goal is to elucidate the signaling pathways that control the structural and functional integrity of small arteries and understand the connections between these pathways and the development of small-vessel-diseases, as a prerequisite to the development of therapeutics. The research we propose is focused on the NotchS signaling pathway. Notch signaling is an evolutionary conserved pathway that plays a central role in the development and maturation of most vertebrate organs. We identified NotchS as the causative gene of CADASIL, an increasingly recognized autosomal dominant form of systemic small-vessel-disease causing stroke and dementia. We previously showed that 1¿) CADASIL patients carry highly stereotyped missense mutations leading to an odd number of cysteine residues within the extracellular domain of NotchS; 2¿) NotchS expression is largely confined to small arteries and vascular Smooth Muscle Cells (vSMC) and 3¿) Mice expressing an archetypal CADASIL mutation (R90C) targeted in vSMC develop features of the CADASIL arteriopathy. Our specific hypothesis is that appropriate level of NotchS activity is critical for structural and functional integrity of small arteries by modulating an RBP-JK dependent, Hes/HEY independent pathway. That hypothesis is supported by our recent findings: 1 ¿) In adult NotchS null mice, small arteries exhibit structural defects and cerebrovascular reactivity is strongly defective. Notably, NotchS null and CADASIL phenotypes are very different; 2¿) RBP-JK dependent activity is abolished in arteries of NotchS null mice but expression level of Hes/HEY genes is unaffected. Here we propose three specific aims to further our understanding of NotchS- dependent small-vessel-diseases and NotchS signaling and address a major unresolved issue: the extent to which CADASIL mutations impair NotchS activity. We will construct and analyze NotchS gain-of-function mutant mice to determine effect of increasing NotchS activity in small arteries (Aim #1). We will determine the gene expression signature of loss and gain-of-function alleles of NotchS and identify direct target genes of NotchS by microarray analysis on isolated small arteries (Aim # 2). We will investigate effect of the archetypal R90C CADASIL mutation on NotchS wildtype activity in vivo using our Notch3R90C mice, NotchS null mice as a "rescue" system and transgenic RBP-JK reporter mice (Aim # 3).

小血管疾病是血管性痴呆的主要原因。我们的长期目标是阐明其病因。控制小动脉结构和功能完整性的信号通路并了解这些途径与小血管疾病的发展之间的联系，作为我们建议的研究重点是NotchS信号通路。信号传导是一种进化保守的途径，在发育和成熟中发挥核心作用我们确定 NotchS 是 CADASIL 的致病基因，而 CADASIL 是一种越来越多的疾病。我们认识到导致中风和痴呆的系统性小血管疾病的常染色体显性遗传形式。之前表明 1¿ ) CADASIL 患者携带高度刻板的错义突变，导致奇怪的 NotchS 2¿胞外域内半胱氨酸残基的数量； )NotchS表达很大程度上是仅限于小动脉和血管平滑肌细胞 (vSMC) 和 3¿ ）表达原型的小鼠针对 vSMC 的 CADASIL 突变 (R90C) 形成了我们特有的 CADASIL 动脉病特征。假设认为，适当水平的 NotchS 活性对于小分子的结构和功能完整性至关重要。该假设得到了支持。根据我们最近的发现：1 ¿ ) 在成年 NotchS 缺失小鼠中，小动脉表现出结构缺陷，并且脑血管反应性存在严重缺陷，值得注意的是，NotchS null 和 CADASIL 表型非常严重。不同; 2¿ ) RBP-JK 依赖性活性在 NotchS 缺失小鼠的动脉中被消除，但表达水平 Hes/HEY 基因不受影响。在这里，我们提出了三个具体目标来进一步了解 NotchS- 相关的小血管疾病和 NotchS 信号传导并解决了一个未解决的主要问题：哪些 CADASIL 突变会损害 NotchS 活性，我们将构建并分析 NotchS 功能获得。突变小鼠以确定增加小动脉中 NotchS 活性的效果（目标#1）。 NotchS 功能丧失和获得功能等位基因的基因表达特征并识别直接目标基因通过对孤立的小动脉进行微阵列分析来检测 NotchS（目标#2）。使用我们的 Notch3R90C 小鼠 NotchS 对 NotchS 野生型活性进行体内原型 R90C CADASIL 突变无效小鼠作为“救援”系统和转基因 RBP-JK 报告小鼠（目标#3）。