Investigation of endomucin as a novel regulator of angiogenesis

内粘蛋白作为血管生成的新型调节剂的研究

• 批准号: 9414681
• 负责人: Patricia Ann D'Amore
• 金额: $ 49.25万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9414681
• 关键词: Address; Adult; Age related macular degeneration; Binding; Biological Assay; Birth; Blindness; Blood Vessels; Blood capillaries; Cell Communication; Cell physiology; Cell surface; Cells; Choroidal Neovascularization; Cytoplasmic Tail; Development; Diabetic Retinopathy; Disease; Endothelial Cells; Equilibrium; Event; Extracellular Domain; Eye; Glycoproteins; Growth; Growth Factor; Histology; Human; ICAM2 gene; Impairment; In Vitro; Investigation; KDR gene; Lasers; Lead; Legal Blindness; Life; Light; Macular degeneration; Measures; Mediating; Messenger RNA; Modeling; Molecular; Mucins; Mus; Mutation; Ocular Pathology; Oxygen; Pathologic; Pathologic Neovascularization; Pathology; Permeability; Persons; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Process; Proteins; Regulation; Reporting; Reproduction; Retinal; Retinal Diseases; Retinal Neovascularization; Retinopathy of Prematurity; Role; Signal Transduction; Small Interfering RNA; Subfamily lentivirinae; Surface; System; Testing; Therapeutic; Tube; Tyrosine Phosphorylation Site; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors; Vascular Permeabilities; Vascularization; Veins; Wound Healing; angiogenesis; antiangiogenesis therapy; crosslink; density; design; endothelial-specific sialomucin; in vivo; inhibitor/antagonist; intravitreal injection; knock-down; loss of function; migration; mutant; new growth; novel; novel therapeutics; ocular neovascularization; overexpression; postnatal; proliferative diabetic retinopathy; promoter; retina blood vessel structure; retinal angiogenesis; small hairpin RNA

ABSTRACT Angiogenesis, the growth of new blood vessels, is an important component of development, wound healing, and reproduction as well as a number of pathologies. Abnormal retinal angiogenesis leads to vision loss in a range of ocular pathologies including diabetic retinopathy, wet macular degeneration, and the retinopathy of prematurity. Endomucin (EMCN) is a mucin-like glycoprotein specifically expressed by endothelial cells of veins and capillaries. Our preliminary studies reveal a role for EMCN in the regulation of VEGF-induced endothelial functions in vitro and in normal angiogenesis in vivo. Reduction of EMCN expression using siRNA led to decreased VEGF-stimulated endothelial proliferation, migration and tube formation in vitro, and impaired retinal vascularization in vivo. This application proposes to test the hypothesis that EMCN plays a role in VEGF-induced pathologic neovascularization and that EMCN influences VEGF-stimulated functions by modulating its signaling. Studies are outlined to examine the role of EMCN in pathological angiogenesis using models of abnormal retinal and choroidal blood vessel growth as well as to understand the molecular mechanism that underlies EMCN's effects on VEGF-induced endothelial cell functions using in vitro systems. Studies in Aim 1 will examine the role of EMCN in the development of pathologic ocular neovascularization in vivo. The oxygen-induced model of retinal neovascularization and the laser-induced model of choroidal neovascularization will be utilized in conjunction with both gain and loss of function studies, using both siRNA and lentivirally-delivered shEMCN under the control of ICAM2, an endothelial-specific promoter, as well as adEMCN under the control of ICAM2. The effect of EMCN knockdown and overexpression on pathologic vessel growth will be quantified. Aim 2 proposes to elucidate the molecular mechanism of EMCN's role in the regulation of VEGF-induced endothelial functions. Studies using EMCN deletion mutants will be employed to determine if the effects of EMCN are mediated at the cell surface. The role of EMCN at the cell surface will be further investigated by examining its effect on VEGF binding and VEGFR2 stabilization/internalization. Intracellular effects will focus on a possible role of EMCN phosphorylation, the effect of EMCN on VEGFR2 phosphatase activity as well as the identification of potential EMCN intracellular binding partners. Results of these proposed studies will provide a better understanding of the role of EMCN in normal and pathologic angiogenesis and may lead to the development of novel endothelial cell-specific therapies for the suppression of vessel growth. !

抽象的 血管生成是新血管的生长，是发育，伤口愈合的重要组成部分 和繁殖以及许多病理。视网膜血管生成异常导致A中的视力丧失 眼病理范围包括糖尿病性视网膜病，湿黄斑变性和视网膜病变 早熟。内核素（EMCN）是一种粘蛋白样糖蛋白，该糖蛋白特异性由内皮细胞表达 静脉和毛细血管。我们的初步研究揭示了EMCN在VEGF诱导的调节中的作用 体外和正常血管生成的内皮功能在体内。使用siRNA降低EMCN表达 导致VEGF刺激的内皮增殖，迁移和管的体外形成，并受损 体内视网膜血管化。该申请提出了测试EMCN在 VEGF诱导的病理新血管形成，EMCN影响VEGF刺激的功能 调节其信号。概述研究以检查EMCN在使用病理血管生成中的作用 视网膜和脉络膜血管生长的异常模型以及了解分子的模型 使用体外系统对EMCN对VEGF诱导的内皮细胞功能的影响的机制。 AIM 1的研究将检查EMCN在病理眼的发展中的作用 体内新血管化。氧诱导的视网膜新血管形成和激光诱导的模型 脉络膜新生血管形成模型将与功能研究的增益和丧失一起使用， 在ICAM2的控制下，同时使用siRNA和慢病毒交付的Shemcn（一种内皮特异性 在ICAM2控制下的启动子以及ADEMCN。 EMCN敲低和 将量化对病理血管生长的过表达。 AIM 2提议阐明分子 EMCN在VEGF诱导的内皮功能调节中的作用机制。使用的研究 将使用EMCN缺失突变体来确定EMCN的作用是否在细胞表面介导。 EMCN在细胞表面的作用将通过检查其对VEGF结合的影响和 VEGFR2稳定/内在化。细胞内影响将集中于EMCN的可能作用 磷酸化，EMCN对VEGFR2磷酸酶活性的影响以及电势的鉴定 EMCN细胞内结合伙伴。这些拟议的研究的结果将更好地理解 EMCN在正常和病理血管生成中的作用，可能导致新内皮的发展 细胞特异性抑制血管生长的疗法。 呢