MT1-MMP-based Animal Model of Age-related Macular Degeneration (AMD)

基于 MT1-MMP 的年龄相关性黄斑变性 (AMD) 动物模型

• 批准号: 7481783
• 负责人: GEORGE INANA
• 金额: $ 14.73万
• 依托单位: ITHERAPEUTICS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7481783
• 关键词: Advanced Development; Affect; Age related macular degeneration; Age-Years; Animal Model; Animals; Basement membrane; Biological; Biological Assay; Blindness; Blood Vessels; Candidate Disease Gene; Cell Line; Cells; Cessation of life; Characteristics; Choroidal Neovascularization; Collaborations; Complex; Custom; Daily; Development; Disease; Doxycycline; Drusen; Elderly; Employee Strikes; Endothelial Cells; Environmental Risk Factor; Extracellular Matrix; Eye; Functional disorder; Gelatinase A; Generations; Genes; Homeostasis; Human; Injection of therapeutic agent; Intellectual Property; International; Lipofuscin; MMP14 gene; MMP2 gene; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Membrane; Metalloproteases; Modeling; Molecular; Molecular Profiling; Names; Neural Retina; Numbers; Pathogenesis; Pathology; Patients; Phagocytosis; Phase; Phenotype; Photoreceptors; Play; Population; Process; Production; Proteins; Public Health; RNA; Rattus; Resources; Retinal Degeneration; Rights; Role; Safety; Severities; Small Business Technology Transfer Research; Sorsby' s fundus dystrophy; Stress; Structure of retinal pigment epithelium; Testing; Tetanus Helper Peptide; Therapeutic; Therapeutic Agents; Thinking; Transgenic Model; Transgenic Organisms; Ursidae Family; Vascular Endothelial Growth Factors; aminotris(methylenephosphonato)diamminoplatinum (II); base; bevacizumab; cell motility; concept; day; drinking water; gene function; human MMP14 protein; in vivo; membrane-type matrix metalloproteinase; migration; mouse model; neovascularization; normal aging; novel; novel therapeutics; photoreceptor degeneration; prevent; research and development; therapeutic target

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is the number one cause of blindness for the elderly population over 60. The primary damage in AMD occurs in the retinal pigment epithelium (RPE). There are two forms of AMD, the dry and the wet form, the latter being associated with choroidal neovascularization (CNV) and responsible for 90% of the blindness. AMD is a complex disease involving multiple genes. Expression profiling is a powerful approach to complex multi-gene diseases, when biological concepts are combined to identify genes fulfilling multiple criteria. Since daily phagocytosis of photoreceptor outer segments (OS) is a key function of RPE, and genes that play a role in the pathogenesis of AMD would be expected to show expression changes, we used a custom expression profiling strategy called CHANGE to identify candidate AMD genes fulfilling both criteria. Membrane-type matrix metalloproteinase I (MT1-MMP) was a gene identified by this strategy that showed 1) involvement in OS phagocytosis, 2) increase that correlated with the degree of pathology in AMD, and 3) increase in another retinal degeneration, the RCS rat model. A Tet-On conditional over-expression transgenic mouse model of MT1-MMP was constructed to confirm the pathogenic potential of this candidate gene. Induction of MT1-MMP over-expression by doxycycline administration in the model demonstrated dramatic vacuolar degeneration, proliferation, and migration of RPE, leading to CNV, resembling characteristics seen in AMD, within days of induction. MT1-MMP is a plausible candidate for AMD since 1) it is the primary activator of MMP2 which has been shown to be increased in the interphotoreceptor matrix and chorioneovascular membranes of AMD patients, 2) it is an important mediator of cellular migration for endothelial cells in blood vessel formation, making it relevant to CNV, 3) it up-regulates VEGF which has been shown to be important in wet AMD, and 4) it has activities against various other matrix components, and the importance of matrix has been demonstrated in AMD-like Sorsby's fundus dystrophy. Thus, we believe MT1-MMP to be an important new therapeutic target for AMD. This STTR application represents the Phase I of a project to develop in collaboration with iTherapeutics the potential dual commercial benefit of our discovery, namely 1) a model of AMD choroidal neovascularization in which the CNV can be easily and quickly induced by a simple administration of doxycycline for testing of various therapeutics against the wet form of AMD, and 2) anti-MT1-MMP agents as potential therapeutics for AMD. The aims for Phase I will focus on 1) above with Specific Aim 1, to select the best line of the transgenic model among the 3 selected lines for induction of MT1-MMP over-expression and consistency of phenotypic expression, and Specific Aim 2, to confirm the phenotype of progressive degeneration of RPE and CNV in the model through histological, immunological, and molecular analyses. For Phase II, we will pursue 2) above, using our model as a template. PUBLIC HEALTH RELEVANCE: Age-related macular degeneration, especially the wet form, is the number one cause of blindness for the elderly population over 60. This project whose aim is to develop a powerful animal model of AMD that can be used for testing of therapeutics for wet AMD and to use it to identify a new effective therapeutic for wet AMD is highly relevant to public health of the elderly.

描述（由申请人提供）：老年性黄斑变性（AMD）是60岁以上老年人失明的头号原因。AMD中的原发性损害发生在视网膜色素上皮（RPE）中。有两种形式的AMD，干性和湿性形式，后者与脉络膜新生血管形成（CNV）有关，并导致90%的失明。AMD是一种涉及多个基因的复杂疾病。表达谱分析是一个强大的方法来复杂的多基因疾病，当生物学概念相结合，以确定满足多个标准的基因。由于光感受器外节（OS）的日常吞噬作用是RPE的关键功能，并且预期在AMD发病机制中起作用的基因将显示表达变化，因此我们使用称为CHANGE的定制表达谱分析策略来鉴定满足这两个标准的候选AMD基因。膜型基质金属蛋白酶I（MT 1-MMP）是通过该策略鉴定的基因，其显示1）参与OS吞噬作用，2）与AMD中的病理学程度相关的增加，和3）另一种视网膜变性（RCS大鼠模型）中的增加。建立了MT 1-MMP的Tet-On条件性过表达转基因小鼠模型，以证实该候选基因的致病潜力。在模型中通过多西环素给药诱导MT 1-MMP过表达，在诱导的几天内表现出RPE的显著空泡变性、增殖和迁移，导致CNV，类似于在AMD中观察到的特征。MT 1-MMP是AMD的合理候选物，因为1）它是MMP 2的主要激活剂，已经显示MMP 2在AMD患者的感光器间基质和绒毛膜血管膜中增加，2）它是血管形成中内皮细胞的细胞迁移的重要介质，使其与CNV相关，3）它上调VEGF，已经显示VEGF在湿性AMD中是重要的，和4）它对各种其他基质成分具有活性，并且基质的重要性已在AMD样Sorsby眼底营养不良中得到证实。因此，我们认为MT 1-MMP是一个重要的新的治疗AMD的目标。该STTR申请代表了与iTherapeutics合作开发我们发现的潜在双重商业益处的项目的I期，即1）AMD脉络膜新血管形成的模型，其中CNV可以通过简单施用多西环素容易且快速地诱导，用于测试针对湿性AMD的各种治疗剂，和2）抗MT 1-MMP剂作为AMD的潜在治疗剂。I期的目标将集中于1）上述具体目标1，在3个选择的品系中选择诱导MT 1-MMP过表达和表型表达一致性的转基因模型的最佳品系，以及具体目标2，通过组织学、免疫学和分子分析确认模型中RPE和CNV进行性变性的表型。对于第二阶段，我们将使用我们的模型作为模板，继续上面的2）。公共卫生相关性：视网膜黄斑变性，特别是湿性黄斑变性，是60岁以上老年人失明的头号原因。该项目的目的是开发一种强大的AMD动物模型，可用于测试湿性AMD的治疗方法，并使用它来确定一种新的有效的湿性AMD治疗方法，该项目与老年人的公共健康高度相关。