CNS angiogenesis and blood-brain barrier regulation by the Wnt inhibitor Apcdd1

Wnt 抑制剂 Apcdd1 对 CNS 血管生成和血脑屏障的调节

基本信息

批准号：
9074367
负责人：
Dritan Agalliu
金额：
$ 36.42万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9074367
关键词：
Adult Autoimmune Diseases Biological Assay Blood Blood - brain barrier anatomy Blood Vessels Blood capillaries Brain CNS processing Carrier Proteins Cell Differentiation process Cell Line Cell Maturation Cell membrane Cells Central Nervous System Diseases Development Diffusion Disease Endothelial Cells Endothelium Etiology Gene Targeting Genes Health Homeostasis Immune In Vitro Knockout Mice Ligands Light Liver Maintenance Measures Mediating Methods Molecular Mouse Strains Multiple Sclerosis Mus Mutant Strains Mice Names Nervous System Physiology Neuraxis Organ Outcome Pathology Pathway interactions Pericytes Peripheral Process Property Proteins Regulation Resistance Role Signal Pathway Signal Transduction Stroke Structure Testing Tight Junctions Toxin Tracer Vascular Diseases Vascularization angiogenesis capillary cell motility gain of function in vivo inhibitor/antagonist novel novel therapeutic intervention overexpression pathogen postnatal prevent retinal angiogenesis selective expression solute therapeutic target transcytosis

项目摘要

DESCRIPTION (provided by applicant): The coordination of angiogenesis with the acquisition of specific properties of endothelial cells that line the blood vessels in distinct organs is essenial for their proper function. An important example of this principle is found in the brain endothelium, namely the blood-brain barrier (BBB), that restricts paracellular diffusion of molecules into the brain by forming high resistance tight junctions between endothelial cells. Despite its importance for the central nervous system (CNS) function, the mechanisms that regulate angiogenesis and development of this barrier remain poorly characterized. We have previously identified that Wnt/�-catenin signaling is active in brain but not liver endothelium during CNS angiogenesis and barrier formation. Moreover, this pathway is essential for CNS angiogenesis and acquisition of some barrier properties by endothelial cells. In addition, Apcdd1, a downstream effector of Wnt/�-catenin signaling, is highly expressed in CNS endothelial cells after angiogenesis when endothelial cells acquire BBB properties. Apcdd1 is selectively expressed in CNS, but not, peripheral endothelial cells. Apccd1 is present in CNS endothelium until postnatal day 20 (P20), but it is extinguished in the adult CNS blood vessels when angiogenesis is complete and BBB is fully formed. The protein is localized within the plasma membrane and the secretory pathway and it inhibits the activation of Wnt/�-catenin signaling in a cell-autonomous manner when overexpressed in cells that receive Wnt signaling. We propose that Apcdd1 inhibits Wnt/�-catenin signaling in CNS endothelium to allow cells to mature and acquire BBB properties. In this proposal, we will investigate the role of Apcdd1 in CNS angiogenesis and BBB formation. We will first examine if Apcdd1 interacts with Wnt ligands (e.g. Wnt7a/7b) that induce barrier properties in endothelial cells. Then we will test if Apcdd1 is necessary and sufficient to induce various aspects of angiogenesis and barrier properties in endothelial cells in vitro. We have generated Apcdd1 knockout mice using gene targeting methods and mice that overexpress Apcdd1 in CNS endothelial cells in an inducible manner. These mice will allow us to test the requirement and sufficiency of Apcdd1 for CNS angiogenesis and BBB formation in vivo. Understanding the development of CNS angiogenesis and BBB will shed light on the mechanisms of tight junction formation within CNS endothelial cells, the etiology of pathological CNS conditions associated with abnormal CNS angiogenesis and BBB breakdown, and help to develop novel therapeutic approaches to regulate these processes.

描述（由应用提供）：血管生成与采集在不同器官中排列血管的内皮细胞的特定特性对其适当功能至关重要。该原理的一个重要例子是在脑内皮中发现的，即血脑屏障（BBB），通过形成内皮细胞之间的高电阻紧密连接来限制分子向大脑的细胞细胞扩散。尽管它对中枢神经系统（CNS）功能的重要性，但调节该屏障的血管生成和发育的机制仍然很差。我们先前已经确定，在中枢神经系统血管生成和屏障形成期间，Wnt/Catenin信号在大脑中活跃，但在大脑中没有活性。此外，该途径对于中枢神经系统的血管生成和从希腊细胞对某些屏障特性的获取至关重要。另外，Wnt/Catenin信号的下游效应子APCDD1在血管生成后在CNS内皮细胞中高度表达，当内皮细胞获得BBB特性时。选择APCDD1在中枢神经系统中表达，但不能在周围内皮细胞中表达。 APCD1存在于中枢神经系统内皮中，直到产后第20天（P20），但是当血管生成完整并且BBB完全形成时，它在成年CNS血管中熄灭。该蛋白质位于质膜和秘密途径中，当在接收Wnt信号传导的细胞中过表达时，它以细胞自主的方式抑制Wnt/Catenin信号的激活。我们建议APCD1抑制CNS内皮中的Wnt/Catenin信号传导，以使细胞成熟并获得BBB性质。在此提案中，我们将研究APCD1在CNS血管生成和BBB形成中的作用。我们将首先检查APCDD1是否与诱导内皮细胞中屏障特性的Wnt配体（例如Wnt7a/7b）相互作用。然后，我们将测试APCDD1是否需要且足以诱导体外内皮细胞中血管生成和屏障特性的各个方面。我们已经使用基因靶向方法和小鼠以诱导的方式在中枢神经系统内皮细胞中过表达APCDD1的小鼠生成了APCDD1基因敲除小鼠。这些小鼠将使我们能够测试APCD1对CNS血管生成和体内BBB形成的需求和安全性。了解中枢神经系统血管生成和BBB的发展将阐明中枢神经系统内皮细胞内紧密连接形成的机制，病理中枢神经系统条件的病因学与中枢神经系统血管生成和BBB异常有关，并有助于开发新的治疗方法来调节这些过程。