RhoA Signaling and Stroke

RhoA 信号传导和中风

基本信息

批准号：
10058968
负责人：
Zygmunt S Derewenda
金额：
$ 43.9万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10058968
关键词：
ARHGEF10 gene Affect Agonist Aneurysm Architecture Arteries Asians Binding Blood Platelets Blood Vessels Blood flow Brain Brain hemorrhage C-terminal Cell-Cell Adhesion Cellular biology Cerebral Aneurysm Cerebral hemisphere hemorrhage Cerebrovascular system Cerebrum Chemicals Code Crystallization DH Domain Data Development Disease Endothelial Cells Endothelium Etiology European Family Family history of Family member Future Genes Genetic Genetic Polymorphism Goals Guanine Nucleotide Exchange Factors Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Health Heavy Drinking Homologous Gene Human Human Genome Hypertension Impairment Individual Intracranial Aneurysm Investigation Ischemic Stroke Knockout Mice Knowledge Lead Light Link Measurement Membrane Messenger RNA Mission Molecular Molecular Structure Monitor Mus Mutate Mutation Nucleotides Outcome PH Domain Patients Permeability Phosphatidylinositols Physiological Plant Roots Play Population Proteins Proteome Public Health Reaction Regulation Regulatory Pathway Research Resistance Risk Risk Factors Role Rupture Ruptured Aneurysm Sequence Analysis Signal Pathway Signal Transduction Smoking Smooth Muscle Smooth Muscle Myocytes Specificity Stroke Structure Structure-Activity Relationship Subarachnoid Hemorrhage Testing Thrombus United States National Institutes of Health Vasodilation WD Repeat Work cerebral artery cerebrovascular cohort genetic regulatory protein high risk improved member model building monolayer novel pressure protein function response rho rho GTP-Binding Proteins small hairpin RNA vasoconstriction

项目摘要

Hemorrhagic stroke accounts for ~13% of all stroke cases. One of the main underlying cases of this type of stroke is the rupture of intracranial aneurysm (IA), with consequent subarachnoid hemorrhage and critically reduced blood flow downstream of the affected vessel. A recent study of both familial and sporadic IA cases identified mutations in the ArhGEF17 gene as associated with increased risk for IA in both European and Asian populations. ArhGEF17 codes for a Dbl-family GEF (guanine nucleotide exchange factor) specific for RhoA. The molecular architecture of ArhGEF17 is unusual for a Dbl-family GEF, in that it is predicted to harbor an unusual split PH-domain (sPH) downstream of the DH domain, and a predicted β-propeller (WD40) domain. Interestingly, the human genome has two homologues of ArhGEF17 with this architecture: ArhGEF10 and GRINCH-GEF (ArhGEF10L) (see Fig 1). A polymorphism in the ArhGEF10L gene has also been associated with increased risk to ischemic stroke. The structures of these three proteins are not known, and the specific functions of the putative sPH and WD40 domains are not understood. We propose an exploratory project aimed at structural and functional characterization of ArhGEF17/10. We will characterize the structural features of the ArhGEF17/10 family, with the overarching goal to gain understanding of the structure-function relationships in these proteins. Our ultimate objective is to gain detailed understanding of the structure and functions of the three globular domains, and to determine if and how are the GEFs regulated by the supramodular architecture. Focusing on the physiological function of ArhGEF17, we want to determine if it regulates cerebral vascular tone, by down regulating its expression in smooth muscle in mouse mid cerebral arteries, and in endothelial cells, using adenoviral shRNAs. RhoA activity and vasodilation will be monitored under basal conditions and in response to increases in intraluminal pressure and agonists in the arteries. Membrane resistance measurements in endothelial cells will monitor changes in permeability. This will lay the ground work for the future development of a conditional ArhGEF17 knockout mouse to determine whether deletion of ArhGEF17 in either smooth muscle or endothelial cells (or both) leads to increased cerebral vessel dilation and/or increased endothelial permeability, and increases the propensity for cerebral hemorrhage.

出血性中风占所有中风病例的约13％。这种类型的主要基础案例之一中风是颅内动脉瘤（IA）的破裂，随之而来的蛛网膜下腔出血受影响血管下游的血流减少。最近对家庭和零星病例的研究鉴定出ARHGEF17基因中的突变与欧洲和亚洲的IA风险增加有关人群。 ARHGEF17代码为RhoA的DBL家族GEF（鸟嘌呤核交换因子）代码。这 ARHGEF17的分子体系结构对于DBL家庭GEF是不寻常的，因为它可以预测具有不寻常的 DH结构域下游的pH域（SPH）和一个预测的β-螺旋桨（WD40）域。有趣的是，人类基因组具有与此结构的两个Arhgef17的同源物：Arhgef10和Grinch-GEF （ARHGEF10L）（见图1）。 ARHGEF10L基因的多态性也与风险增加有关缺血性中风。这三种蛋白质的结构尚不清楚，并且推定的SPH和WD40域尚不清楚。我们提出了一个探索性项目，旨在旨在ARHGEF17/10的结构和功能表征。我们将表征Arhgef17/10家族的结构特征，并具有获得的总体目标了解这些蛋白质中的结构功能关系。我们的最终目标是获取细节了解三个全球域的结构和功能，并确定是否以及如何以及如何 GEFS由超模型结构调节。专注于Arhgef17的身体功能，我们想要通过下调调节其在平滑肌中的表达来确定它是否调节脑血管张力小鼠使用腺病毒shRNA的脑动脉和内皮细胞中的小鼠。 RhoA活性和血管舒张将在基本条件下监测，并响应于腔内压力的增加和激动剂动脉。内皮细胞中的膜电阻测量将监测渗透率的变化。这将为有条件的ARHGEF17敲除鼠标的未来开发奠定基础工作以确定在平滑肌还是内皮细胞中删除ARHGEF17（或两者）是否导致大脑增加血管扩张和/或增加内皮渗透性，并增加了脑出血的希望。