Oligo-Vascular Signaling in Stroke

中风中的寡血管信号传导

基本信息

批准号：
8495431
负责人：
Ken Arai
金额：
$ 35.98万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8495431
关键词：
Antibodies Apoptosis Area Astrocytes Biological Biological Assay Blood Vessels Blood flow Brain Brain-Derived Neurotrophic Factor CSPG4 gene Cell Communication Cell Culture Techniques Cell Death Cell Proliferation Cell Survival Cell physiology Cells Central Nervous System Diseases Cerebral Ischemia Cerebrum Cessation of life Conditioned Culture Media Corpus Callosum Corpus striatum structure Coupling Data Disodium Salt Nitroprusside Dissection Endothelial Cells Endothelial Growth Factors Endothelin-1 Endothelium Event Fibroblast Growth Factor Functional disorder Gene Delivery Glucose Growth Factor Growth Factor Overexpression Growth Factor Receptors Homeostasis IGF1 gene In Situ Nick-End Labeling In Vitro Injection of therapeutic agent Insulin-Like Growth Factor I Intercellular Adhesion Molecule 2 Ischemia Lasers Liposomes Methods Modeling Molecular Monomeric GTP-Binding Proteins Mus Myelin Myelin Basic Proteins Neurons Oligodendroglia Oligonucleotides Oxidative Stress Oxygen Platelet-Derived Growth Factor Play Plumbing Production Protein Tyrosine Kinase Rattus Research Role Signal Pathway Signal Transduction Signaling Molecule Small Interfering RNA Staining method Stains Stress Stroke Surgical sutures Testing Time Vascular Dementia Western Blotting brain cell caspase-3 cell injury cerebrovascular deprivation in vivo inhibitor/antagonist intercellular communication killings mouse model novel oligodendrocyte precursor precursor cell prevent promoter protective effect public health relevance receptor expression research study small hairpin RNA vector white matter white matter damage white matter injury

项目摘要

DESCRIPTION (provided by applicant): Oligo-Vascular Signaling in Stroke White matter injury is a central event in the pathophysiology of diverse CNS disorders, including stroke and vascular dementia. But studies that investigate white matter are relatively uncommon in cerebrovascular research. Accumulating evidence suggest that cerebral endothelial cells have multiple functions in addition to conducting blood flow. Here, we hypothesize that cerebral endothelial cells secrete trophic factors to maintain oligodendrocyte (OL) and oligodendrocyte precursor cell (OPC) survival and function. Our 3 aims are: Aim 1: Dissect the cellular mechanisms of oligo-protection by endothelial-derived growth factors. Cultured rat OL/OPC will be subjected to oxygen-glucose deprivation. Endothelial conditioned media (Endo-CM) will be used to test whether growth factors from endothelial cells can protect OL/OPC cultures. We will investigate how Endo-CM promotes survival signaling (e.g. Akt) and decreases apoptosis in OL/OPC. Aim 2: Show that oxidative stress decreases endothelial growth factor production. We hypothesize that even without overt cell death, oxidatively-stressed endothelial cells will have reduced growth factor production. Cerebral endothelial cells will be exposed to oxidative stress, and we compare conditioned media from healthy endothelial cells vs oxidatively-stressed endothelial cells. Conditioned media from oxidatively-stressed endothelial cells should have less growth factors and thus, be less protective for OL/OPC against insults. Aim 3: Demonstrate that endothelial growth factors are important for OL/OPC in vivo. In various mouse models of cerebral ischemia, we will assess OL/OPC integrity in corpus callosum and striatum (i.e. white matter damage). Endothelial-specific gene delivery methods (liposome, scAAV9) will be used to modulate endothelial trophic factors in vivo. We predict that selectively increasing endothelial trophic factors protect white matter, whereas decreasing endothelial trophic factors makes white matter more vulnerable. Our pilot data (Arai and Lo, J Neurosci 2009) suggest that Endo-CM supports OPC proliferation via specific signaling pathways and oxidative stress interferes with oligovascular coupling. In this proposal, we will build on these initial findings to show that Endo-CM can truly prevent cell death in OL/OPC. And most importantly, we aim to show that oligovascular coupling protects against stroke in vivo. These experiments should provide evidence to show that trophic coupling between endothelium and OL/OPC maintains and protects white matter.

描述（由申请人提供）：中风中的寡血管信号转导白质损伤是多种中枢神经系统疾病（包括中风和血管性痴呆）的病理生理学的中心事件。但在脑血管研究中，研究白质的研究相对较少。越来越多的证据表明，脑内皮细胞除了传导血流外还具有多种功能。在这里，我们假设脑内皮细胞分泌营养因子来维持少突胶质细胞（OL）和少突胶质细胞前体细胞（OPC）的存活和功能。我们的 3 个目标是：目标 1：剖析内皮源性生长因子寡核苷酸保护的细胞机制。培养的大鼠 OL/OPC 将受到氧-葡萄糖剥夺。内皮条件培养基 (Endo-CM) 将用于测试内皮细胞生长因子是否可以保护 OL/OPC 培养物。我们将研究 Endo-CM 如何促进 OL/OPC 中的生存信号（例如 Akt）并减少细胞凋亡。目标 2：证明氧化应激会降低内皮生长因子的产生。我们假设即使没有明显的细胞死亡，氧化应激的内皮细胞也会减少生长因子的产生。脑内皮细胞将暴露于氧化应激，我们比较了来自健康内皮细胞和氧化应激内皮细胞的条件培养基。来自氧化应激内皮细胞的条件培养基应含有较少的生长因子，因此对 OL/OPC 免受损伤的保护作用较小。目标 3：证明内皮生长因子对于体内 OL/OPC 很重要。在各种脑缺血小鼠模型中，我们将评估胼胝体和纹状体中 OL/OPC 的完整性（即白质损伤）。内皮特异性基因递送方法（脂质体，scAAV9）将用于调节体内内皮营养因子。我们预测选择性增加内皮营养因子可以保护白质，而减少内皮营养因子则使白质更加脆弱。我们的试验数据（Arai 和 Lo，J Neurosci 2009）表明 Endo-CM 通过特定信号通路支持 OPC 增殖，并且氧化应激干扰少血管耦合。在本提案中，我们将在这些初步发现的基础上证明 Endo-CM 可以真正预防 OL/OPC 中的细胞死亡。最重要的是，我们的目标是证明寡血管耦合可以在体内预防中风。这些实验应该提供证据表明内皮细胞和 OL/OPC 之间的营养耦合维持和保护白质。