Molecular and cellular etiology of cerebral cavernous malformations

脑海绵状血管瘤的分子和细胞病因学

基本信息

批准号：
7990816
负责人：
Jun Zhang
金额：
$ 22.28万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
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：
2010-05-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7990816
关键词：
Affect Affinity Amino Acids Angiogenic Proteins Animal Experimentation Animal Model Apoptosis Area Arginine Binding Proteins Biological Assay Blood Vessels Brain CCM1 gene CD29 Antigen Cardiovascular system Cavernous Malformation Cell Culture Techniques Cell Proliferation Cell membrane Cell physiology Cells Cerebrum Chromatography Clinical Complex Data Defect Detergents Development Discontinuous Capillary Disease Drug Formulations Electrospray Ionization Embryo Endothelial Cells Ethnic group Etiology Focal Adhesion Kinase 1 Fractionation Gene Expression Genes Genetic Genetic Transcription Genomics Goals Hispanics Homeostasis Homozygote Human In Vitro Integrin beta Chains Integrins Intracranial Hemorrhages Isotopes Knowledge Label Light Liquid Chromatography Lysine Mass Spectrum Analysis Mediating Mediator of activation protein Membrane Methods Modeling Molecular Mutate Nuclear Pathogenesis Performance Population Process Protein Biosynthesis Proteins Proteome Proteomics Public Health RNA Regulation Research Research Project Grants Role Seizures Signal Pathway Signal Transduction Small Interfering RNA Spinal Cord Stable Isotope Labeling Stroke Syndrome System Techniques Testing Therapeutic Transgenic Animals Transgenic Organisms Vascular Endothelial Cell Venous Zebrafish angiogenesis base cell motility genome-wide in vivo integrin-linked kinase malformation migration mutant offspring public health relevance stable isotope tandem mass spectrometry two-dimensional vasculogenesis

项目摘要

DESCRIPTION (provided by applicant): This project aims to elucidate pathogenetic mechanisms in cerebral cavernous malformations (CCMs), a clinical syndrome in the brain and spinal cord marked by venous sinusoids that predispose to intracranial hemorrhage. Three genes have been identified as causes of CCMs; KRIT1 (CCM1), MGC4607 (CCM2), and PDCD10 (CCM3), which when mutated leads to the formation of neurovascular malformations. We propose that CCM genes regulate beta1 integrin-mediated signaling pathways during angiogenesis, remodeling, and/or homeostasis of the microvasculature and that perturbation of this process contributes to the development of CCMs. We have specifically silenced krit1 expression in cultured human vascular endothelial cells and observed dramatic perturbations in cellular proliferation, motility, and survival. We have also demonstrated: 1) interaction of krit1 with icap1alpha, a known mediator of beta1 integrin signaling; 2) interaction of krit1 with MGC4607; 3) perturbation of cultured endothelial cell proliferation, migration, and survival accompanied by reduced signaling through focal adhesion kinase (FAK), Rac1, and integrin-linked kinase (ILK) cascades, respectively, after siRNA-mediated silencing of either krit1 or icap1alpha; 4) enhancement of multiple beta1 integrin signaling cascades through targeting icap1 to the cell membrane; and 5) failed nuclear localization and degradation of icap1alpha and MGC4607 after silencing of krit1; 6) well documented cardiovascular defects of Ccm1 (santa, san) and Ccm2 (valentine, vtn) zebrafish, and 7) perturbation of vasculatures due to increased vascular endothelial cell apoptosis in san zebrafish. In this research project, we will focus on three critical areas: 1). Isolation of the GFP-labeled vascular endothelial cells during pathogenesis of zebrafish Ccms; 2). The elucidation of the role of Ccm genes in the angiogenesis/vasculogenesis with multiple genomic and proteomic analysis of the vascular endothelial cells during pathogenesis of zebrafish CCMs; and 3) identification of other cellular factors interacting of krit1, MGC4607, and PDCD10 during microvascular angiogenesis. This animal research will provide detailed knowledge regarding the cellular role of CCM genes and their complex in vascular endothelial cells during angiogenesis, a critical first step in the development of nonsurgical therapeutic strategies for microvascular malformations. PUBLIC HEALTH RELEVANCE: Cerebral cavernous malformations (CCMs) is a vascular syndrome in the brain that causes stroke. CCMs occur in approximately 0.5% of the population and represent 10% of all vascular malformations. Since CCM is the major cause for seizures among Hispanic ethnic group which contributes to over 85% population in El Paso, our proposed genomic and proteomic study on animal model is not only essential for understanding the molecular pathogenesis of this disease, but also extremely important for regional public health.

描述（由申请人提供）：该项目旨在阐明脑海绵状畸形（CCMS）中的致病机制，这是大脑中的临床综合征和脊髓和脊髓，标志着静脉正弦曲线，可易感颅内出血。三个基因被确定为CCM的原因。 KRIT1（CCM1），MGC4607（CCM2）和PDCD10（CCM3），当突变时会导致神经血管畸形的形成。我们建议CCM基因在微脉管系统的血管生成，重塑和/或稳态期间调节BetA1整合素介导的信号通路，并且该过程的扰动有助于CCM的发展。我们已经在培养的人血管内皮细胞中特异性沉默了KRIT1的表达，并观察到细胞增殖，运动性和生存性的戏剧性扰动。我们还证明了：1）KRIT1与ICAP1Alpha的相互作用，ICAP1Alpha是Beta1整合素信号传导的已知介体； 2）KRIT1与MGC4607的相互作用； 3）分别通过局灶性粘附激酶（FAK），RAC1和整合素链接激酶（ILK）cascades降低信号传导的培养内皮细胞增殖，迁移和存活，伴随着信号的扰动。 4）通过将ICAP1靶向细胞膜来增强多个Beta1整合素信号级联反应； 5）krit1沉默后，ICAP1Alpha和MGC4607的核定位和降解失败； 6）CCM1（圣诞老人，SAN）和CCM2（Valentine，VTN）斑马鱼的心血管缺陷和7）由于血管内皮细胞细胞凋亡增加而导致血管扰动的心血管缺陷。在该研究项目中，我们将重点关注三个关键领域：1）。斑马鱼CCM的发病机理中GFP标记的血管内皮细胞的分离； 2）。在斑马鱼CCM的发病机理期间，对CCM基因在血管生成/血管生成中的作用阐明了血管内皮细胞的多种基因组和蛋白质组学分析； 3）在微血管血管生成过程中鉴定KRIT1，MGC4607和PDCD10的其他细胞因子相互作用。这项动物研究将提供有关CCM基因及其在血管生成过程中血管内皮细胞中复合物的细胞作用的详细知识，这是对微血管畸形的非外科治疗策略发展的关键第一步。公共卫生相关性：脑海绵状畸形（CCMS）是大脑中引起中风的一种血管综合征。 CCM大约发生在大约0.5％的人群中，占所有血管畸形的10％。由于CCM是西班牙族族中癫痫发作的主要原因，在El Paso中贡献了超过85％的人口，因此我们提出的关于动物模型的基因组和蛋白质组学研究不仅对了解该疾病的分子发病机理，而且对区域公共卫生非常重要。