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

基本信息

批准号：
8500301
负责人：
Sally S. Twining
金额：
$ 36.34万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
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：
2011-08-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8500301
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 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 screening skills

项目摘要

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.

描述（由申请人提供）：角膜作为眼睛的屏障，其透明度是其将光线折射和传输到视网膜的功能所必需的。正常角膜是无血管的。与角膜血管生成相关的病症的例子包括化学烧伤、慢性隐形眼镜缺氧、Stevens-Johnson 综合征、翼状胬肉、移植物排斥和感染性角膜炎 (2-4)。正常角膜合成许多抗血管生成分子，其中之一是maspin，一种非抑制性丝氨酸蛋白酶抑制剂，可调节角膜成纤维细胞以及其他细胞类型（包括癌细胞）的粘附、迁移和侵袭。 Maspin 还抑制角膜微囊模型中血管的向内生长，并抑制微血管内皮细胞的管形成、增殖和迁移。该项目的总体目标是了解 maspin 抑制角膜血管生成的机制，并开发用于预防和治疗血管生成的 maspin 形式。具体目标是： 1) 检验 maspin 通过多种机制改变血红素和淋巴管生成的假设，包括对血管和淋巴内皮细胞的直接影响以及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抗血红素血管生成的活性进行比较。将确定maspin对角膜上皮和基质成纤维细胞以及血红素和淋巴内皮细胞合成的促血管生成蛋白和抗血管生成蛋白水平的影响。 2) 测试 maspin N 端和/或 C 端区域的磷酸化形式抑制血管生成的假设，这些区域的肽比完整分子更有效，并开发具有抗血管生成潜力的 maspin 肽血管生成药物。抗血管生成特性所需的 maspin 区域和翻译后修饰的鉴定将利用由 maspin 和卵清蛋白、磷酸化模拟物、磷酸化无效形式和 maspin 肽构建的嵌合蛋白。将开发血管生成的器官培养模型来测试 maspin 形式。 3) 检验 maspin 对角膜血红素和淋巴管生成发挥主要体内调节作用的假设。将使用 maspin 小鼠测试丢失一个 maspin 等位基因对血红素和淋巴管生成的影响。将使用抗体的 Maspin 重建和消耗实验。拟议的研究重点是了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。这些研究还将通过识别抗血管生成分子 maspin 改变血管生成的机制来扩展我们对控制角膜血管生成的因素的理解，并将表征该活动所需的 maspin 的特性。结果将指导 maspin 突变体和肽的生成和初步测试，作为治疗新生血管疾病的潜在方式。