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Regulation and Function of the Matricellular Protein CCN1 in Ischemic Retinopathy

基质细胞蛋白CCN1在缺血性视网膜病变中的调控和功能

基本信息

批准号：
8776306
负责人：
BRAHIM CHAQOUR
金额：
$ 38.12万
依托单位：
SUNY DOWNSTATE MEDICAL CENTER
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：
2011-12-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8776306
关键词：
Affect Apoptosis Astrocytes Automobile Driving Behavior Binding Biological Biological Process Blindness Blood Vessels Cell Adhesion Cell Differentiation process Cell Surface Receptors Cell Therapy Cell physiology Cells Childhood Cues Data Development Disease Elements Endothelial Cells Environment Event Extracellular Matrix Proteins Fostering Future Gelatinase A Gene Expression Gene Targeting Genes Goals Growth Growth Factor Growth Factor Receptors Healthcare Systems Human Hyperoxia Immediate-Early Genes Impairment In Vitro Integrins Ions Ischemia Knowledge Laboratories Ligands Modeling Molecular Morphogenesis Mus Mutant Strains Mice Oxygen Parents Pathologic Neovascularization Pathologic Processes Pathology Pattern Formation Peptide Hydrolases Pericytes Pharmacotherapy Phase Phenotype Physiological Physiological Processes Prevention Proliferating Protein Isoforms Proteins Regulation Retina Retinal Retinal Diseases Retinopathy of Prematurity Rodent Model Role Signal Transduction Specificity Stress TNF gene Testing Therapeutic Vascular Diseases Vascular Endothelial Growth Factors Vascularization Vision Visual impairment Wnt proteins age group angiogenesis base cell behavior cell type cyr61 protein cytokine density design effective therapy extracellular gain of function gene repression improved in vivo insight loss of function migration neovascularization notch protein novel prevent promoter protein expression protein function receptor response retina blood vessel structure screening small molecule

项目摘要

DESCRIPTION (provided by applicant): Disorders of retinal vessel growth and function are responsible for vision loss in retinopathy of prematurity (ROP), a leading cause of vision impairment and blindness in childhood. ROP develops as a result of excessive growth of abnormal pre-retinal blood vessels, a compensatory mechanism that overcomes an earlier phase of hyperoxia-induced vaso-obliteration. Elucidation of the molecular bases of angiogenic cell function and behavior in physiological and pathological processes will have important therapeutic implications for the treatment of human retinal vascular diseases. The long term goal of our laboratory is to uncover the in vivo functions of the matricellular protein CCN1, also known as cysteine-rich protein 61, and the functional consequences of its expression, or lack thereof, in development and ischemic retinopathy. The CCN1 protein is an inducible immediate-early gene-encoded extracellular matrix (ECM) protein required for proper vascular development. In our preliminary studies, we have found that over-expression of CCN1 in the retina via either gene- or cell-based therapy, enhanced normal retinal vascularization and reduced pathological angiogenesis in the model of oxygen-induced retinopathy (OIR). In vitro data showed that CCN1 functions primarily through direct binding to specific integrins and ECM proteins, and/or indirectly through modulation of growth factor and Wnt protein expression and/or activity, thereby triggering signaling events that culminate in the regulation of cell adhesion, migration, proliferation, gene expression, differentiation, and survival. Our hypothesis is that CCN1 normalizes the biological mechanisms of retinal vessel formation during development and following OIR and overrides those leading to abnormal vessel formation. In Specific Aim 1, we will use mutant mice with inducible conditional inactivation of the CCN1 gene to determine how loss of CCN1 in endothelial cells (ECs) causes defective retinal vessel growth. We will identify interactions with Wnt- and Notch-derived signals known to influence functional specialization of ECs (tip and stalk cell phenotypes) and sprouting angiogenesis. We will further determine whether and how forced expression of CCN1 in ECs only, allows normal retinal vessel formation in OIR. In Specific Aim 2, we will define the molecular interactions of CCN1 with astrocytes, the primary proangiogenic cells responsible for retinal vessel formation and patterning during development, and we will determine the functional significance of CCN1 loss in astrocytes (and/or in ECs) in mutant mice on astrocyte activation state and behavior (e.g., migration, density and ensheathment) during development and in OIR. In Specific Aim 3, we will define the dynamics of the CCN1 promoter activity and identify the functional elements responsible for CCN1 gene modulation both in cultured retinal ECs subjected to hyperoxic stress and in retinas of OIR mice. This proposal will provide new insights into the molecular mechanisms of CCN1 activities in vivo and may foster future safer, less destructive, and more effective therapies to harness ischemia-induced neovascularization in ROP.

描述(申请人提供)：视网膜血管生长和功能障碍是早产儿视网膜病变(ROP)视力丧失的原因，ROP是儿童视力受损和失明的主要原因。ROP的发生是异常视网膜前血管过度生长的结果，这种代偿机制克服了高氧诱导血管闭塞的早期阶段。阐明血管生成细胞在生理和病理过程中的功能和行为的分子基础，将对人类视网膜血管疾病的治疗具有重要的治疗意义。我们实验室的长期目标是揭示基质细胞蛋白CCN1，也被称为富含半胱氨酸的蛋白61在体内的功能，以及它的表达或缺失在发育和缺血性视网膜病变中的功能后果。CCN1蛋白是一种可诱导的即刻早期基因编码的细胞外基质(ECM)蛋白，是血管正常发育所必需的。在我们的初步研究中，我们发现，在氧诱导视网膜病变(OIR)模型中，通过基因或细胞治疗在视网膜中过表达CCN1，促进了正常视网膜血管的形成，减少了病理性血管的生成。体外研究表明，CCN1主要通过直接与特定的整合素和ECM蛋白结合，和/或间接通过调节生长因子和Wnt蛋白的表达和/或活性发挥作用，从而触发最终调节细胞黏附、迁移、增殖、基因表达、分化和生存的信号事件。我们的假设是，在发育过程中和OIR后，CCN1使视网膜血管形成的生物学机制正常化，并覆盖那些导致异常血管形成的机制。在具体目标1中，我们将使用CCN1基因可诱导的条件性失活的突变小鼠来确定内皮细胞(ECs)中CCN1的丢失如何导致视网膜血管生长缺陷。我们将识别与Wnt和Notch衍生信号的相互作用，这些信号已知会影响内皮细胞的功能特化(尖端和茎细胞表型)和萌芽血管生成。我们将进一步确定仅在内皮细胞中强制表达CCN1是否以及如何允许OIR中正常的视网膜血管形成。在特定的目标2中，我们将定义CCN1与星形胶质细胞的分子相互作用，星形胶质细胞是在发育过程中负责视网膜血管形成和图案形成的主要血管生成细胞，我们将确定CCN1在突变小鼠的星形胶质细胞(和/或内皮细胞)中丢失对星形胶质细胞的激活状态和行为(例如，迁移、密度和包膜)在发育和OIR中的功能意义。在具体目标3中，我们将定义CCN1启动子活性的动态变化，并确定在高氧应激下培养的视网膜内皮细胞和OIR小鼠视网膜中负责CCN1基因调控的功能元件。这一建议将为体内CCN1活性的分子机制提供新的见解，并可能促进未来更安全、更具破坏性和更有效的治疗方法，以利用缺血诱导的ROP新生血管。