The role of ADAM9 in retinal health and disease

项目摘要/摘要 这项建议的总体目标是确定光感受器的病理生理机制。 人和犬ADAM9(去整合素和金属蛋白酶9)基因突变引起的变性 病人。先前在小鼠和狗模型中的研究表明，ADAM9的丢失会导致大脑中 光感受器外节和视网膜色素上皮(RPE)之间的界面。初步数据 这一建议揭示了光感受器间基质中细胞外物质在 此界面。鉴于ADAM蛋白既是蛋白酶又是细胞黏附分子的公认角色， 这个项目将探索两个不相互排斥的假设：(1)ADAM9作为一种调节 光感受器间基质的组合物和(2)ADAM9作为细胞黏附分子形成 光感受器外节与RPE之间的接触。这些假设将使用各种不同的 转基因ADAM9小鼠模型。在指导阶段的目标1中，全球ADAM9基因敲除小鼠 将使用各种技术进行全面表征，这些技术将提供新的培训机会 候选人。在指导阶段的目标2中，候选人将接受高端蛋白质组学技术培训 从定性和定量两个水平分析光感受器间基质的组成。 在独立阶段，这种训练将被用来检验两个关于 视网膜中的ADAM9。目的3将探索ADAM9蛋白分解光感受器间基质的假说 而Aim 4将探索ADAM9作为细胞黏附分子发挥作用的假设。vt.给出 ADAM9既由光感受器表达，也由RPE表达，Aim 5中的实验将识别细胞类型 主要负责ADAM9相关的病理。综上所述，拟议的研究将指导未来 对携带ADAM9突变的视锥-视杆细胞营养不良患者的治疗努力。此外，考虑到关键的 光感受器间基质在支持外视网膜的完整性和视网膜与视网膜的粘连中的作用 RPE，这项建议将为视网膜疾病提供更广泛的见解，如视网膜脱离和一些 各种形式的视网膜色素变性。拟议研究提供的实验方法培训将 辅以杜克大学的职业发展培训，包括正规课程，在这两个方面 沟通能力，以及指导和领导能力。候选人将由瓦迪姆博士指导 阿尔沙夫斯基是视网膜细胞生物学领域的领导者，也是一位非常有成就的导师，他成功地 启动了十几名教职员工的职业生涯，他们中的大多数都是由NIH资助的。归根结底，这 培训将使候选人实现成为一名独立调查员的长期目标 研究视觉系统生物学和视网膜疾病的病理生理学。

PROJECT SUMMARY/ABSTRACT The overall goal of this proposal is to identify the pathophysiological mechanisms underlying photoreceptor degeneration caused by mutations in ADAM9 (A Disintegrin And Metalloproteinase 9) in human and canine patients. Previous work in mouse and canine models has revealed that loss of ADAM9 leads to disruptions in the interface between the photoreceptor outer segments and the retinal pigment epithelium (RPE). Preliminary data for this proposal reveal a substantial accumulation of extracellular material from the interphotoreceptor matrix at this interface. Given the well-established roles of ADAM proteins as both proteases and cell adhesion molecules, this project will explore two not mutually exclusive hypotheses: (1) ADAM9 functions as a protease regulating the composition of the interphotoreceptor matrix and (2) ADAM9 functions as a cell adhesion molecule forming contacts between the photoreceptor outer segments and RPE. These hypotheses will be tested using a variety of genetically modified ADAM9 mouse models. In Aim 1 of the mentored phase, the global ADAM9 knockout mouse will be comprehensively characterized using a variety of techniques that will provide new training opportunities to the candidate. In Aim 2 of the mentored phase, the candidate will train in high-end proteomic techniques to analyze the composition of the interphotoreceptor matrix on both qualitative and quantitative levels. In the independent phase, this training will be applied to test two complementary hypotheses on the function of ADAM9 in the retina. Aim 3 will explore the hypothesis that ADAM9 proteolyzes interphotoreceptor matrix components, whereas Aim 4 will explore the hypothesis that ADAM9 functions as a cell adhesion molecule. Given that ADAM9 is expressed by both photoreceptors and the RPE, the experiments in Aim 5 will identify the cell type primarily responsible for ADAM9-associated pathology. Taken together, the proposed studies will guide future therapeutic efforts for cone-rod dystrophy patients bearing ADAM9 mutations. Additionally, given the critical roles of the interphotoreceptor matrix in supporting the integrity of the outer retina and retinal adhesion to the RPE, this proposal will provide broader insights into retinal diseases such as retinal detachment and some forms of retinitis pigmentosa. The training in experimental approaches provided by the proposed research will be supplemented with career development training at Duke University, including formal coursework, in both communication skills as well as mentoring and leadership skills. The candidate will be mentored by Dr. Vadim Arshavsky, a leader in the field of retinal cell biology and a highly accomplished mentor who has successfully launched the careers of over a dozen faculty members, most of whom are funded by the NIH. Ultimately, this training will allow the candidate to achieve his long-term goal of becoming an independent investigator studying the biology of the visual system and pathophysiology of retinal disease.