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: 10627840
• 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/10627840
• 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; United States National Institutes of Health; Universities; Visual System; 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.

项目总结/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 在人类和犬中由ADAM 9（解整合素和金属蛋白酶9）突变引起的变性 患者先前在小鼠和犬模型中的研究表明，ADAM 9的缺失会导致 视网膜色素上皮细胞（RPE）是感光细胞外节和视网膜色素上皮细胞（RPE）之间的界面。初步数据 这一建议揭示了大量的细胞外物质从感光细胞间基质的积累， 这个接口。鉴于ADAM蛋白作为蛋白酶和细胞粘附分子的公认作用， 本项目将探讨两个不相互排斥的假设：（1）ADAM 9作为蛋白酶调节细胞的功能， （2）ADAM 9作为细胞粘附分子形成的功能， 光感受器外节和RPE之间的接触。这些假设将使用各种 基因修饰的ADAM 9小鼠模型。在指导阶段的目标1中，整体ADAM 9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的培训， 在定性和定量水平上分析感光细胞间基质的组成。 在独立阶段，这种培训将用于测试两个互补的假设， 视网膜中的ADAM 9目的3探讨ADAM 9蛋白水解感光细胞间基质的假说 Aim 4将探索ADAM 9作为细胞粘附分子发挥功能的假设。给定 ADAM 9由光感受器和RPE表达，目标5中的实验将鉴定细胞类型 主要负责ADAM 9相关的病理。总的来说，拟议的研究将指导未来的 对携带ADAM 9突变的锥-杆营养不良患者的治疗努力。此外，鉴于关键的 光感受器间基质在支持外层视网膜的完整性和视网膜粘附到视网膜中的作用 RPE，这项建议将提供更广泛的见解视网膜疾病，如视网膜脱离和一些 视网膜色素变性的形式。拟议研究提供的实验方法培训将 在杜克大学接受职业发展培训，包括正式课程， 沟通技巧以及指导和领导技能。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，也是一位成功地 启动了十几名教职员工的职业生涯，其中大部分由NIH资助。最终这 培训将使候选人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。

项目成果

Absolute Quantification of Photoreceptor Outer Segment Proteins.

• DOI: 10.1021/acs.jproteome.3c00267
• 发表时间: 2023-08-04
• 期刊: JOURNAL OF PROTEOME RESEARCH
• 影响因子: 4.4
• 作者: Skiba, Nikolai P. P.;Lewis, Tylor R. R.;Spencer, William J. J.;Castillo, Carson M. M.;Shevchenko, Andrej;Arshavsky, Vadim Y. Y.
• 通讯作者: Arshavsky, Vadim Y. Y.