Functional Analysis of Novel Genes in Eye Development an

眼睛发育中新基因的功能分析

• 批准号: 7322440
• 负责人: graeme j wistow
• 金额: --
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7322440
• 关键词: Functional; Analysis; Novel; Genes; Eye

Through use of genomics and bioinformatics approaches we have identified many novel genes and alternative gene products that are expressed in human and animal model eye tissues. Several of these have been selected for functional analysis using a broad range of functional and structural approaches to determine their roles in the eye and their implications for normal vision and for disease. These include: 1: Platelet-derived growth factor D (PDGFD) belongs to the PDGF/VEGF family. In the anterior segment PDGFD, it is localized to iris and ciliary body, whereas in the retina, it is restricted to the outer plexiform layer. We have shown that PDGFD has a key role in control of lens growth and that anti-PDGFD strongly inhibits lens epithelial cell proliferation This finding suggests a major in vivo role for PDGFD in the mechanisms of coordinated growth of eye tissues. Intervention in the PDGFD pathway in the eye, perhaps by antibody or blocking peptide, could be useful in the treatment of certain cataracts, including post-operative secondary cataract. PDGFD is also expressed in photoreceptors and in RPE, suggesting possible roles in controls of photoreceptors survival and control of retinal vascularization. A functional promoter of the PDGFD gene has been identified and correctly directs expression of a GFP reporter to eye tissues. A conditional knock out of PDGFD in mouse is nearing completion. 2: Lengsin is a novel member of the glutamine synthetase (GS) superfamily that is specific for the vertebrate eye lens. It has ATP-binding activity but, so far, no demonstrable enzyme activity. Lengsin is expressed specifically in the layer of terminally differentiating secondary fiber cells in which nuclei and other organelles are lost and cytoskeleton and intracellular junctions are reorganized. A knock out model of lengsin is completed and is currently in breeding for homozygosity. 3: Recent publications have shown an important role for complement factor H (CFH) in the development of age-related macular degeneration (AMD). We have constructed yeast 2-hybrod libraries for aged human RPE/choroid and retina. Using constructs for the AMD-related domain 7 of CFH as bait we have identified potentially important targets for CFH binding in human RPE/choroid. These are being further evaluated by techniques including surface plasmon resonance spectroscopy. Interaction of CFH with RPE/choroid proteins could have local effects on its role in the complement system and might provide opportunities for therapeutic intervention.

通过使用基因组学和生物信息学的方法，我们已经确定了许多新的基因和替代的基因产物，在人类和动物模型眼组织中表达。其中几个已经被选择用于功能分析，使用广泛的功能和结构方法来确定它们在眼睛中的作用及其对正常视力和疾病的影响。其中包括： 1：血小板衍生生长因子D（PDGFD）属于PDGF/VEGF家族。在眼前段PDGFD中，它局限于虹膜和睫状体，而在视网膜中，它局限于外丛状层。我们已经表明PDGFD在控制透镜生长中具有关键作用，并且抗PDGFD强烈抑制透镜上皮细胞增殖。这一发现表明PDGFD在眼组织协调生长机制中的主要体内作用。通过抗体或阻断肽干预眼睛中的PDGFD通路，可能有助于治疗某些白内障，包括术后继发性白内障。PDGFD也在光感受器和RPE中表达，表明在光感受器存活的控制和视网膜血管化的控制中可能起作用。PDGFD基因的功能性启动子已被鉴定，并正确指导GFP报告基因在眼组织中的表达。小鼠中PDGFD的条件性敲除接近完成。 2：Lengsin是谷氨酰胺合成酶（GS）超家族的新成员，对脊椎动物眼透镜具有特异性。它具有ATP结合活性，但到目前为止，没有可证实的酶活性。Lengsin在终末分化的次生纤维细胞层中特异性表达，其中细胞核和其他细胞器丢失，细胞骨架和细胞内连接重组。一个敲除的模型已经完成，目前正在培育纯合性。 3：最近的出版物表明，补体因子H（CFH）在年龄相关性黄斑变性（AMD）的发展中起着重要作用。我们构建了老年人RPE/脉络膜和视网膜的酵母2-hybrod文库。使用CFH的AMD相关结构域7的构建体作为诱饵，我们已经确定了人RPE/脉络膜中CFH结合的潜在重要靶点。这些正在进一步评估的技术，包括表面等离子体共振光谱。CFH与RPE/脉络膜蛋白的相互作用可能对其在补体系统中的作用产生局部影响，并可能为治疗干预提供机会。