Cyr61/CCN1-Induced Angiogenesis and Vasculogenesis in the Retina

Cyr61/CCN1 诱导的视网膜血管生成

基本信息

批准号：
7740294
负责人：
BRAHIM CHAQOUR
金额：
$ 24.98万
依托单位：
SUNY DOWNSTATE MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7740294
关键词：
Ablation Acute Address Adhesions Adhesives Age related macular degeneration Anoikis Apoptosis Binding Proteins Biological Biological Process Blood Vessels Blood capillaries Bone Marrow C-terminal CD34 gene Cell Culture Techniques Cell Proliferation Cells Cessation of life Characteristics Complex Cultured Cells Cystine Cystine Knot Motifs Defect Development Diabetic Retinopathy Disease Embryo Endothelial Cells Ensure Environment Event Extracellular Matrix Extracellular Matrix Proteins Eye diseases Family Gene Expression Gene Family Genes Growth Hemorrhage Heparin In Vitro Individual Inflammation Injection of therapeutic agent Injury Insulin-Like Growth-Factor-Binding Proteins Integrin Binding Integrins Laboratories Lentivirus Vector Link Mediating Modeling Molecular Mononuclear Mus N-terminal Neonatal Oxygen Pathogenesis Pathologic Neovascularization Pericytes Pharmacotherapy Phenotype Population Process Property Protein Binding Recovery Retina Retinal Retinal Diseases Retinopathy of Prematurity Role SCID Mice Signal Transduction Site Source Stem cells Structure-Activity Relationship Tertiary Protein Structure Thrombospondin 1 Undifferentiated Vascular Diseases Vascularization Venous Work alternative treatment angiogenesis base capillary capillary bed cell growth cell type central retinal vein occlusion cyr61 protein design expectation extracellular improved in vivo member migration mouse model neonate neovascular neovascularization novel polypeptide postnatal precursor cell progenitor proliferative diabetic retinopathy public health relevance receptor repaired response retina blood vessel structure retinal apoptosis retinal progenitor cell therapeutic angiogenesis vasculogenesis von Willebrand Factor

项目摘要

DESCRIPTION (provided by applicant): Development and remodeling of retinal blood vessels occurs through the complex and still poorly understood processes of vasculogenesis and angiogenesis. These processes are recapitulated, though imperfectly, in the context of ischemic injury associated with numerous ocular diseases, (e.g., retinopathy of prematurity, age-related macular degeneration, diabetic retinopathy, etc). The pathogenesis of these retinal neovascular disorders is driven by a pathological angiogenesis leading to the formation of abnormal vessels, which break into the vitreous, and an insufficient/limited vasculogenesis in which the cellular components of the new vessel complex originate in part, from bone-marrow (BM) derived circulating vascular progenitor cells. Components of the vascular extracellular matrix (ECM) coordinate the execution of multiple angiogenic and vasculogenic activities. Recent work in the PI's laboratory have revealed that the cysteine-rich protein 61 (Cyr61), a novel ECM-associated, heparin- and integrin-binding protein, stimulates the angiogenic phenotype of the BM-derived hematopoeitic progenitors, CD34+ cells. However, when exposed to cultured retinal pericytes, which sheath and regulate the growth of retinal capillaries, Cyr61 induces their death by anoikis suggesting a role of Cyr61 in pathological angiogenesis. The current proposal consists of a strategy to tease out these disparate activities of the Cyr61 protein both in vitro and in vivo. We will aim to determine that the multiple activities of Cyr61 are linked to its multi-modular organization consisting of 4 distinct domains: an insulin-like growth factor binding protein (IGFBPs) domain, the von Willebrand factor type C (vWFC) repeat, the thrombospondin type I (TSP1) repeat and a C-terminal (CT) cystine-knot motif. We predict that the N-terminal region containing IGFBP and/or vWFC domains with integrin-binding capabilities retains the angiogenic potential of Cyr61 vis-`-vis the CD34+ cells and promote their adhesion, migration, differentiation, and angiogenic activity. Conversely, the C-terminal region containing the TSP1 and/or CT domains induces apoptosis of retinal pericytes by virtue of its anti-adhesive properties. We will reveal these activities, in Specific Aim 1 using undifferentiated hematopoeitic CD34+ cells and cultured retinal pericytes exposed to recombinantly produced domains of Cyr61 or lentiviral vectors expressing the N-terminal or C- terminal regions. In Specific Aim 2, we will utilize the neonate mouse model of oxygen-induced retinopathy characterized by vaso-obliteration of the central retinal capillary bed to determine whether administration of either Cyr61 or Cyr61 modules promotes vascular repair by enhancing recruitment of EPCs to retinal sites of ischemic injury. Similarly, we will establish whether, BM-derived CD34+ cells, when primed with the N- terminal domains of Cyr61, integrate into damaged retinal vessels and improve vascular recovery. Our studies will help establish treatment alternatives for the diminished/limited vasculogenic capability in ischemic retinopathy and the specific contribution of truncated ECM proteins to EPC mobilization and recruitment. PUBLIC HEALTH RELEVANCE: Neovascularization of the retina is a highly prevalent and potentially blinding disorder characteristic of a variety of ocular diseases including diabetic retinopathy, age-related macular degeneration, central retinal vein occlusion, and retinopathy of permaturity. Our studies address the role of a specific component of the extracellular environment in retinal vessels and its ability to either facilitate or antagonize retinal vascular repair/rescue. Results of our studies will determine how specific portions of such extracellular compound can be used/modified to allow the formation of normally functioning retinal blood vessels and improve the pharmacotherapy of several retinal diseases.

描述（由申请人提供）：视网膜血管的发育和重塑是通过复杂且仍知之甚少的血管发生和血管生成过程发生的。这些过程在与许多眼部疾病（例如，早产儿视网膜病变、年龄相关性黄斑变性、糖尿病性视网膜病变等）相关的缺血性损伤的背景下得到了概括，尽管并不完美。这些视网膜新生血管疾病的发病机制是由病理性血管生成驱动的，导致异常血管的形成，异常血管闯入玻璃体，以及血管生成不足/有限，其中新血管复合体的细胞成分部分源自骨-骨髓（BM）衍生的循环血管祖细胞。血管细胞外基质（ECM）的成分协调多种血管生成和血管生成活动的执行。 PI 实验室最近的工作表明，富含半胱氨酸的蛋白 61 (Cyr61) 是一种新型 ECM 相关肝素和整合素结合蛋白，可刺激 BM 来源的造血祖细胞 CD34+ 细胞的血管生成表型。然而，当暴露于培养的视网膜周细胞（其包裹并调节视网膜毛细血管的生长）时，Cyr61 会通过失巢凋亡诱导其死亡，这表明 Cyr61 在病理性血管生成中发挥作用。目前的提案包括一项策略，旨在梳理 Cyr61 蛋白在体外和体内的这些不同活性。我们的目标是确定 Cyr61 的多种活性与其由 4 个不同结构域组成的多模块组织相关：胰岛素样生长因子结合蛋白 (IGFBP) 结构域、C 型冯维勒布兰德因子 (vWFC) 重复序列、 I 型血小板反应蛋白 (TSP1) 重复序列和 C 端 (CT) 胱氨酸结基序。我们预测，含有具有整联蛋白结合能力的 IGFBP 和/或 vWFC 结构域的 N 末端区域保留了 Cyr61 相对于 CD34+ 细胞的血管生成潜力，并促进其粘附、迁移、分化和血管生成活性。相反，含有 TSP1 和/或 CT 结构域的 C 端区域凭借其抗粘附特性诱导视网膜周细胞凋亡。我们将在具体目标 1 中使用未分化的造血 CD34+ 细胞和培养的视网膜周细胞来揭示这些活性，这些细胞暴露于重组产生的 Cyr61 结构域或表达 N 末端或 C 末端区域的慢病毒载体。在具体目标 2 中，我们将利用以视网膜中央毛细血管床血管闭塞为特征的氧诱导视网膜病变新生小鼠模型，以确定 Cyr61 或 Cyr61 模块的给药是否通过增强 EPC 向视网膜部位的募集来促进血管修复。缺血性损伤。同样，我们将确定 BM 衍生的 CD34+ 细胞在用 Cyr61 的 N 末端结构域引发时是否会整合到受损的视网膜血管中并改善血管恢复。我们的研究将有助于针对缺血性视网膜病变中血管生成能力减弱/受限以及截短的 ECM 蛋白对 EPC 动员和募集的具体贡献建立治疗替代方案。公众健康相关性：视网膜新生血管形成是一种非常普遍且可能致盲的疾病，是多种眼部疾病的特征，包括糖尿病性视网膜病变、年龄相关性黄斑变性、视网膜中央静脉阻塞和早产儿视网膜病变。我们的研究探讨了视网膜血管中细胞外环境的特定成分的作用及其促进或拮抗视网膜血管修复/挽救的能力。我们的研究结果将确定如何使用/修饰这种细胞外化合物的特定部分，以形成功能正常的视网膜血管并改善几种视网膜疾病的药物治疗。