KRIT1 in Vascular Development and Dysfunction

KRIT1 在血管发育和功能障碍中的作用

基本信息

批准号：
8533040
负责人：
KEVIN J WHITEHEAD
金额：
$ 31.45万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8533040
关键词：
Abnormal Cell Abnormal Endothelial Cell Accounting Address Affect Alleles Animal Model Apical Apoptosis Biological Assay Biological Markers Biomechanics Birth Blood Vessels Blood flow Brain Brain hemorrhage CCM1 gene CDC42 gene Caliber Cavernous Malformation Cell Communication Cell Culture Techniques Cell Line Cell Polarity Cell Size Cell model Cells Cellular Stress Cellular biology Cerebrum Cessation of life Clinical Clinical Treatment Controlled Environment Cytoskeleton Defect Development Disease Disease Marker Disease Outcome Disease Progression Effectiveness Embryo Endothelial Cells Endothelium Excision Family Functional disorder Gene Targeting Genes Genetic Guanosine Triphosphate Phosphohydrolases Hemorrhage Hispanics Histologic Human Individual Inherited Intercellular Junctions Knock-out Lesion Life Longitudinal Studies Loss of Heterozygosity MAP Kinase Gene Magnetic Resonance Imaging Measures Medical Modeling Morphogenesis Morphology Mus Mutation Natural History Neuraxis New Mexico Observational Study Operative Surgical Procedures Outcome Pathogenesis Pathologic Pathway interactions Patient observation Patients Penetrance Permeability Pharmaceutical Preparations Physiological Physiology Population Prevention Proteins Relative (related person)Reporting Retinal Rho-associated kinase Risk Role Rupture Seizures Shapes Signal Pathway Signal Transduction Small Interfering RNA Southwestern United States Specimen Staging Stress Stress Fibers Stroke Syndrome System Tamoxifen Telangiectasis Testing Tissues Vascular Diseases Vascular Permeabilities Work angiogenesis cell motility disorder control efficacy testing fluid flow in vivo inhibitor/antagonist insight malformation monolayer mouse model mutant neurosurgery outcome forecast preclinical study research study response rho shear stress tool

Abnormal Cell Abnormal Endothelial Cell Accounting Address Affect Alleles Animal Model Apical Apoptosis Biological Assay Biological Markers Biomechanics Birth Blood Vessels Blood flow Brain Brain hemorrhage CCM1 gene CDC42 gene Caliber Cavernous Malformation Cell Communication Cell Culture Techniques Cell Line Cell Polarity Cell Size Cell model Cells Cellular Stress Cellular biology Cerebrum Cessation of life Clinical Clinical Treatment Controlled Environment Cytoskeleton Defect Development Disease Disease Marker Disease Outcome Disease Progression Effectiveness Embryo Endothelial Cells Endothelium Excision Family Functional disorder Gene Targeting Genes Genetic Guanosine Triphosphate Phosphohydrolases Hemorrhage Hispanics Histologic Human Individual Inherited Intercellular Junctions Knock-out Lesion Life Longitudinal Studies Loss of Heterozygosity MAP Kinase Gene Magnetic Resonance Imaging Measures Medical Modeling Morphogenesis Morphology Mus Mutation Natural History Neuraxis New Mexico Observational Study Operative Surgical Procedures Outcome Pathogenesis Pathologic Pathway interactions Patient observation Patients Penetrance Permeability Pharmaceutical Preparations Physiological Physiology Population Prevention Proteins Relative (related person)Reporting Retinal Rho-associated kinase Risk Role Rupture Seizures Shapes Signal Pathway Signal Transduction Small Interfering RNA Southwestern United States Specimen Staging Stress Stress Fibers Stroke Syndrome System Tamoxifen Telangiectasis Testing Tissues Vascular Diseases Vascular Permeabilities Work angiogenesis cell motility disorder control efficacy testing fluid flow in vivo inhibitor/antagonist insight malformation monolayer mouse model mutant neurosurgery outcome forecast preclinical study research study response rho shear stress tool

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项目摘要

DESCRIPTION (provided by applicant): Cerebral Cavernous Malformations (CCM) are common vascular lesions of the central nervous system found in 1 in 200 people. Lesions consist of thin-walled enlarged vascular spaces prone to rupture leading to hemorrhagic strokes, seizures, and death. The disease can be hereditary, in which case affected individuals tend to have multiple lesions, placing them at even greater risk. A large population with hereditary CCM disease is found in New Mexico and the Southwestern United States in which the disease is caused by mutations in the gene, KRIT1. At present, treatment of the disease consists of choosing between neurosurgery to remove lesions, or conservative "watchful waiting". Meaningful medical treatment of the disease will require an understanding of the underlying disease mechanisms, and will be greatly aided by animal models for use in pre-clinical trials of treatment. Recent studies have suggested a potential role for increased RhoA GTPase activity in the pathogenesis of the disease. The Ras family GTPases including RhoA, Rac1, and CDC42 regulate the cellular cytoskeleton, cell-cell interactions, and are involved in the cellular response to biomechanical stress. Other reports have suggested the importance of KRIT1 - Rap1 interactions in regulating cell junction proteins and endothelial cell apical - basal polarity. We hypothesize that KRIT1 is an important component of a mechanosensory apparatus that controls RhoA signaling with downstream effects on endothelial polarity. In this project we propose to use cell biology with physiologic levels of shear stress, and a newly developed animal model of CCM disease in mice to explore the role of KRIT1 in RhoA activation and cell polarity in vascular development and disease. We will describe the response of KRIT1 deficient endothelial cells when exposed to flow, as in living blood vessels, and study the response of signaling pathways in these cells. Using conditional gene targeting we will confirm that the underlying defect is found in the endothelial cells that line the CCM lesions. We will use an endothelial specific, drug inducible Cre system to knockout Krit1 from mice at birth. We have found that this approach leads to CCM lesions that can be seen and followed by MRI. We will use this system to test the efficacy of proposed treatments of CCM. In all of these aims we will evaluate the relative importance of RhoA activation and abnormal cell polarity. Further, we will develop and characterize non-invasive biomarkers of disease activity in CCM lesions, using the exaggerated progression of disease in this model and the access to tissue for mechanistic analysis to full advantage.

描述（由申请人提供）：脑海绵状畸形（CCM）是中枢神经系统的常见血管病变，其中1人中有1人。病变由薄壁的稀有血管空间组成，容易发生破裂，导致出血性中风，癫痫发作和死亡。该疾病可能是遗传性的，在这种情况下，受影响的个体往往会有多种病变，使其面临更大的风险。在新墨西哥州和美国西南部发现了遗传性CCM疾病的大量人群，该疾病是由基因中的突变KRIT1引起的。目前，该疾病的治疗包括在神经外科手术之间进行选择以清除病变或保守的“注意等待”。对该疾病的有意义的医学治疗将需要了解潜在的疾病机制，并将受到动物模型在临床前治疗试验中的极大帮助。最近的研究表明，增加了RhoA GTPase活性在疾病发病机理中的潜在作用。 RAS家族GTPases在内的RHOA，RAC1和CDC42调节细胞细胞骨架，细胞 - 细胞相互作用，并参与细胞对生物力学应激的反应。其他报告表明，KRIT1 -RAP1相互作用在调节细胞连接蛋白和内皮细胞顶端 - 基础极性中的重要性。我们假设KRIT1是机械感觉设备的重要组成部分，该设备控制RhoA信号传导，对内皮极性有下游影响。在这个项目中，我们建议将细胞生物学具有剪切应力的生理水平，以及在小鼠中新开发的CCM疾病动物模型来探索KRIT1在RhoA激活和细胞极性在血管发育和疾病中的作用。我们将描述在暴露于流动的血管中的KRIT1缺乏的内皮细胞的反应，并研究这些细胞中信号通路的响应。使用条件基因靶向，我们将确认在CCM病变的内皮细胞中发现了潜在缺陷。我们将使用内皮特异性的，药物诱导的CRE系统来敲除小鼠的krit1。我们发现，这种方法会导致CCM病变，而MRI可以看到并随后看到。我们将使用该系统测试CCM拟议治疗的功效。在所有这些目标中，我们将评估RhoA激活和异常细胞极性的相对重要性。此外，我们将利用该模型中疾病的夸大进展，并访问组织以进行机械分析以充分优势，从而发展并表征CCM病变中疾病活性的非侵入性生物标志物。