The role of ADAM9 in retinal health and disease

ADAM9 在视网膜健康和疾病中的作用

• 批准号: 10424849
• 负责人: Tylor Robert Lewis
• 金额: $ 12.6万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10424849
• 关键词: 3-Dimensional; Address; Adhesions; Affect; Biology; Blindness; Canis familiaris; Cell Adhesion; Cell Adhesion Molecules; Cellular biology; Communication; Data; Disease; Disintegrins; Electroretinography; Exhibits; Extracellular Matrix; Faculty; Functional disorder; Funding; Future; Gene Delivery; Gene Expression; Genes; Goals; Health; Human; Immunoprecipitation; Knock-out; Knockout Mice; Lead; Leadership; Light; Location; Mass Spectrum Analysis; Mentors; Metalloproteases; Methods; Molecular; Mus; Mutation; Pathologic; Pathology; Patients; Peptide Hydrolases; Phase; Phenotype; Photoreceptors; Preparation; Protein Family; Proteins; Proteolysis; Proteome; Proteomics; Research; Research Personnel; Retina; Retinal Degeneration; Retinal Detachment; Retinal Diseases; Retinitis Pigmentosa; Role; Site; Structural defect; Structure of retinal pigment epithelium; Techniques; Testing; Therapeutic; Therapeutic Intervention; Training; Transgenic Organisms; United States National Institutes of Health; Universities; Visual system structure; Wild Type Mouse; Work; aged; canine model; career; career development; cell type; cone-rod dystrophy; electron tomography; experimental study; extracellular; insight; member; mouse model; photoreceptor degeneration; prevent; protein function; skills; training opportunity

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.

项目概要/摘要 该提案的总体目标是确定光感受器潜在的病理生理机制 人类和犬类 ADAM9（解整合素和金属蛋白酶 9）突变引起的退化 患者。先前在小鼠和犬类模型中的研究表明，ADAM9 的缺失会导致 光感受器外节和视网膜色素上皮（RPE）之间的界面。初步数据 对于这一提议，揭示了来自光感受器间基质的细胞外物质的大量积累 这个界面。鉴于 ADAM 蛋白作为蛋白酶和细胞粘附分子的既定作用， 该项目将探索两个并不相互排斥的假设：(1) ADAM9 作为一种蛋白酶调节 光感受器间基质的组成和 (2) ADAM9 作为细胞粘附分子形成 感光器外节和 RPE 之间的接触。这些假设将使用各种方法进行检验 转基因 ADAM9 小鼠模型。在指导阶段的目标 1 中，全局 ADAM9 敲除小鼠 将使用各种技术进行全面表征，这将为 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，以 在定性和定量水平上分析光感受器间基质的组成。 在独立阶段，该训练将用于测试关于函数的两个互补假设 视网膜中的 ADAM9。目标 3 将探讨 ADAM9 蛋白水解感光间基质的假设 成分，而目标 4 将探索 ADAM9 作为细胞粘附分子的假设。给定 ADAM9 由光感受器和 RPE 表达，目标 5 中的实验将识别细胞类型 主要负责 ADAM9 相关病理学。总而言之，拟议的研究将指导未来 针对携带 ADAM9 突变的视锥杆营养不良患者的治疗工作。此外，考虑到关键 光感受器间基质在支持外视网膜完整性和视网膜与视网膜粘附中的作用 RPE，该提案将为视网膜疾病（例如视网膜脱离和一些视网膜疾病）提供更广泛的见解 色素性视网膜炎的形式。拟议研究提供的实验方法培训将 在杜克大学获得职业发展培训的补充，包括正式课程， 沟通技巧以及指导和领导技巧。候选人将由 Vadim 博士指导 Arshavsky 是视网膜细胞生物学领域的领军人物，也是一位卓有成就的导师，他成功地 启动了十多名教员的职业生涯，其中大多数由美国国立卫生研究院资助。最终，这 培训将使候选人能够实现成为独立调查员的长期目标 研究视觉系统的生物学和视网膜疾病的病理生理学。