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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：
8067037
负责人：
Jun Zhang
金额：
$ 18.19万
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 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8067037
关键词：
Affect Affinity Amino Acids Angiogenic Proteins Animal Experimentation Animal Model Apoptosis Area Arginine 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血管瘤（CCM）的发病机制，脑海绵状血管瘤是一种大脑和脊髓的临床综合征，以容易导致颅内出血的静脉窦为特征。三种基因已被确定为导致 CCM 的原因； KRIT1 (CCM1)、MGC4607 (CCM2) 和 PDCD10 (CCM3) 突变时会导致神经血管畸形的形成。我们认为，CCM 基因在血管生成、重塑和/或微脉管系统稳态过程中调节 β1 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。我们专门沉默了培养的人血管内皮细胞中的 krit1 表达，并观察到细胞增殖、运动和存活的显着扰动。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质； 2) krit1与MGC4607的相互作用； 3) 在 siRNA 介导的 krit1 或 icap1alpha 沉默后，培养的内皮细胞增殖、迁移和存活受到干扰，并分别通过粘着斑激酶 (FAK)、Rac1 和整合素连接激酶 (ILK) 级联信号减弱； 4) 通过将icap1靶向细胞膜来增强多个β1整合素信号级联； 5) krit1沉默后icap1alpha和MGC4607的核定位和降解失败； 6) 有据可查的 Ccm1 (santa, san) 和 Ccm2 (valentine, vtn) 斑马鱼的心血管缺陷，以及 7) 由于 san 斑马鱼血管内皮细胞凋亡增加而引起的脉管系统扰动。在这个研究项目中，我们将重点关注三个关键领域：1）。斑马鱼 Ccms 发病过程中 GFP 标记的血管内皮细胞的分离； 2）。通过对斑马鱼 CCM 发病机制中血管内皮细胞的多重基因组和蛋白质组分析，阐明 Ccm 基因在血管生成中的作用； 3) 鉴定微血管生成过程中 krit1、MGC4607 和 PDCD10 相互作用的其他细胞因子。这项动物研究将提供有关 CCM 基因及其复合物在血管生成过程中在血管内皮细胞中的细胞作用的详细知识，这是开发微血管畸形非手术治疗策略的关键的第一步。公共卫生相关性：脑海绵状血管瘤 (CCM) 是一种导致中风的大脑血管综合征。 CCM 发生于大约 0.5% 的人群，占所有血管畸形的 10%。由于 CCM 是西班牙裔群体癫痫发作的主要原因，该群体占埃尔帕索人口的 85% 以上，因此我们提出的动物模型基因组和蛋白质组学研究不仅对于了解这种疾病的分子发病机制至关重要，而且对于区域公共卫生也极其重要。