Effect of Maspin on Corneal Heme-and lymph- angiogenesis

Maspin 对角膜血红素和淋巴管生成的影响

• 批准号: 8187367
• 负责人: Sally S. Twining
• 金额: $ 34.43万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8187367
• 关键词: Adhesions; Alleles; Angiogenesis Inhibitors; Angiogenic Proteins; Animal Experiments; Animals; Antibodies; Blood; Blood Vessels; C-terminal; Cell Culture Techniques; Cells; Chemical Burns; Chimeric Proteins; Chronic; Conjunctival Pterygium; Contact Lenses; Cornea; Corneal Neovascularization; Data; Development; Disease; Endothelial Cells; Eye; Fibroblasts; Generations; Goals; Graft Rejection; Growth; Heme; Human; Hypoxia; Immune; In Vitro; Injury; Keratitis; Lead; Light; Lymph; Maintenance; Malignant Epithelial Cell; Methods; Modality; Modeling; Mus; N-terminal; Organ Culture Techniques; Ovalbumin; Peptides; Phosphorylation; Play; Post-Translational Protein Processing; Prevention; Process; Property; Proteins; Public Health; Research Personnel; Retina; Role; Screening procedure; Serpins; Stevens-Johnson Syndrome; Testing; Therapeutic; Transgenic Mice; Tube; Vascularization; Vision; angiogenesis; cell type; corneal epithelium; in vivo; light transmission; maspin; migration; mutant; neovascular; reconstitution; research study; skills

DESCRIPTION (provided by applicant): The cornea serves as a barrier for the eye and its transparency is required for its function to refract and transmit light to the retina. The normal cornea is avascular. Examples of conditions associated with corneal angiogenesis include chemical burns, chronic contact lens hypoxia, Stevens-Johnson syndrome, pterygia, graft rejection and infectious keratitis (2-4). The normal cornea synthesizes a number of anti-angiogenic molecules, one of which is maspin, a non-inhibitory Serpin that regulates adhesion, migration and invasion of corneal fibroblasts as well as other cell types, including carcinoma cells. Maspin also inhibits in-growth of vessels in the corneal micropocket model and inhibits tube formation, proliferation and migration of microvascular endothelial cells. The overall goal of this project is to understand the mechanism of maspin inhibition of corneal angiogenesis and to develop forms of maspin for the prevention and treatment of angiogenesis. The Specific Objectives are: 1) To test the hypothesis that maspin alters heme and lymph-angiogenesis by multiple mechanisms involving both direct effects on vascular and lymph endothelial cells and indirect effects on corneal cells. The ability of maspin to alter lymph-angiogenesis will be determined and compared with its activity against heme-angiogenesis. The effect of maspin on the levels of pro-and anti-angiogenic proteins synthesized by corneal epithelial and stromal fibroblast cells and heme- and lymph-endothelial cells will be determined. 2) To test the hypothesis that the phosphorylated form of the N-terminal and/or C-terminal region of maspin inhibits angiogenesis, with peptides of these regions more potent than the intact molecule, and to develop maspin peptides that have potential as anti-angiogenic drugs. Identification of the region(s) and post-translational modifications of maspin required for its anti-angiogenic properties will utilize chimeric proteins constructed from maspin and ovalbumin, phosphorylation mimics, phosphorylation null forms and maspin peptides. An organ culture model of angiogenesis will be developed to test the maspin forms. 3) To test the hypothesis that maspin plays a major in vivo regulatory role of heme- and lymph- angiogenesis in the cornea. The effect of loss of one maspin allele on heme- and lymph-angiogenesis will be tested using maspin mice. Maspin reconstitution and depletion experiments with antibodies will be used. The proposed studies focus on understanding heme- and lymph-angiogenesis, a major public health issue that threatens vision, and aims to generate new treatment methods. The studies will also extend our understanding of factors that control corneal angiogenesis through identification of the mechanisms by which the anti-angiogenic molecule maspin alters angiogenesis and will characterize the properties of maspin required for this activity. The results will guide the generation and initial testing of maspin mutants and peptides as potential modalities for the treatment of neovascular diseases. PUBLIC HEALTH RELEVANCE: A critical feature of the cornea is its transparency, which is required for the passage and focusing of light onto the retina for normal vision. Injury and disease can cause the inappropriate formation of blood and lymph vessels in the cornea that can block light transmission. Maspin is a protein that inhibits vessel growth in the cornea. Studies will be conducted towards developing this protein as a therapeutic for the prevention of corneal vascularization.

描述（由申请人提供）：角膜充当眼睛的障碍，其功能需要折射光和将光传递到视网膜所需的透明度。正常的角膜是无血管的。与角膜血管生成相关的疾病的例子包括化学灼伤，慢性隐形眼镜缺氧，史蒂文斯 - 约翰逊综合征，pterygia，移植排斥和感染性角膜炎（2-4）。正常的角膜合成了许多抗血管生成分子，其中一种是maspin，一种非抑制性塞抗物，可调节角膜成纤维细胞的粘附，迁移和侵袭以及其他细胞类型，包括癌细胞。 Maspin还抑制了角膜微量源模型中血管的增长，并抑制微血管内皮细胞的管形，增殖和迁移。该项目的总体目的是了解马斯普蛋白抑制角膜血管生成的机制，并开发出MASPIN的形式，以预防和治疗血管生成。 具体目标是：1）测试Maspin通过多种机制来改变血红素和淋巴血管生成的假设，这些机制既涉及对血管和淋巴内皮细胞的直接影响，又对角膜细胞的间接影响。将确定MASPIN改变淋巴血管生成的能力，并与其对血红素血管生成的活性进行比较。将确定MASPIN对由角膜上皮和基质成纤维细胞以及血红素和淋巴性内皮细胞合成的抗血管生成蛋白水平的影响。 2）测试以下假设：Maspin的N末端和/或C末端区域的磷酸化形式抑制了血管生成，这些区域的肽比完整的分子更有效，并发展具有抗血管疾病药物的潜在的Maspin肽。鉴定其抗血管生成特性所需的Maspin区域和翻译后修饰将利用由Maspin和ovalbumin构建的嵌合蛋白，磷酸化模拟，磷酸化，磷酸化null形式和Maspin肽。将开发一种血管生成的器官培养模型来测试maspin形式。 3）测试Maspin在角膜中血红素和淋巴血管生成的体内调节作用的主要调节作用。将使用MASPIN小鼠测试一个Maspin等位基因对血红素和淋巴血管生成的影响。将使用Maspin重建和用抗体进行耗尽实验。 拟议的研究集中于理解血红素和淋巴血管生成，这是威胁视力的主要公共卫生问题，旨在产生新的治疗方法。研究还将扩展我们对控制角膜血管生成的因素，通过鉴定抗血管生成分子Maspin改变血管生成的机制，并将表征这种活性所需的Maspin的性质。结果将指导Maspin突变体和肽的产生和初始测试，作为治疗新生血管疾病的潜在方式。 公共卫生相关性：角膜的关键特征是它的透明度，这是将光线通过并将其集中在视网膜上以实现正常视力所必需的。损伤和疾病会导致角膜中血液和淋巴管的不适当形成，从而阻塞光传播。 maspin是一种蛋白质，可抑制角膜的血管生长。将进行研究，以开发该蛋白作为预防角膜血管形成的治疗性。