The differential behavior of Prph2/Rom1 in rods and cone contributes to Prph2-associated disease heterogeneity

Prph2/Rom1 在视杆细胞和视锥细胞中的差异行为导致与 Prph2 相关的疾病异质性

• 批准号: 10701039
• 负责人: Larissa Ikelle
• 金额: $ 4.15万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-28 至 2026-09-27
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701039
• 关键词: Address; Affect; Affinity Chromatography; Animal Model; Attention; Backcrossings; Behavior; Binding; Biochemical; Bypass; Clinical; Co-Immunoprecipitations; Complex; Cone; Data; Development; Disease; Disease model; Electron Microscopy; Etiology; Exhibits; Future; Goals; Golgi Apparatus; Health; Homo; Homologous Gene; Imaging Techniques; Impairment; In Vitro; Inherited; Investigation; Knock-out; Knowledge; Leucine Zippers; Macular degeneration; Maintenance; Membrane Microdomains; Membrane Proteins; Methods; Modeling; Modification; Molecular Conformation; Morphogenesis; Mutation; Nature; Neural Retina; Outcome Assessment; Outcome Study; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Pattern; Phenotype; Photoreceptors; Process; Proteins; Psammomys; Rattus; Research; Resolution; Retina; Retinal Diseases; Retinitis Pigmentosa; Rod; Rod Outer Segments; Rodent Model; Role; Sand Rats; Severity of illness; Silicon Dioxide; Structure; Testing; Therapeutic; Three-dimensional analysis; Vertebrate Photoreceptors; cell type; conditional knockout; disease heterogeneity; disease phenotype; effective therapy; in vivo; in vivo Model; innovation; interest; macular dystrophy; microscopic imaging; mutant; overexpression; pattern dystrophies; peripherin; photoreceptor degeneration; syntaxin; therapeutically effective; trafficking

PROJECT SUMMARY Mutations in Peripherin 2 (PRPH2) account for a significant portion of inherited retinal diseases (IRD). Delineating the role of PRPH2 in photoreceptor morphogenesis and rim formation is vital to understand how mutations in a single protein could contribute to such diverse group of pathologies, ranging from retinitis pigmentosa (RP) to pattern dystrophy (PD) and macular degeneration (MD). Even patients with intrafamilial mutations can exhibit a notable degree of phenotypic variability, attesting to the complex nature in which PRPH2 functions. My objective in this application is to advance our current knowledge of the role of PRPH2 in outer segment (OS) formation in order to have a better understanding of disease pathogeneses and for future development of effective therapeutic strategies. Over the years, our lab has developed an extensive toolbox of in vitro and in vivo mutant models, and are currently exploring innovative imaging techniques to further our understanding of the role of Prph2 in OS landscape. Thus far, it has been determined that the association between Prph2 and its homologue, Rod OS Protein 1 (Rom1), contributes significantly to the phenotypic variability observed in patients and that lowering Rom1 levels could shift a sever Prph2-assocaited PD phenotype to milder RP. Furthermore, it is know that the proper function of Prph2 at the disc rim is contingent upon proper complex assembly of Prph2-oligomers and Prph2/Rom1-complexes. The central focus of this investigation is to understand how Prph2 and Rom1 function differently in rods and cones with particular interest in Prph2 homo- and Prph2/Rom1 hetero-complex formation/trafficking, and the effects of Prph2 mutations on these processes. We propose two aims to address these goals: Aim 1 will assess the role of Rom1 in cones and the mechanism by which it modulates Prph2-disease phenotypes. By eliminating or overexpressing Rom1 in Prph2-disease models, we will conduct careful biochemical, structural, and functional analyses to assess phenotypic differences, as they pertain to complex assembly and OS trafficking. Aim 2 will investigate the role of interacting partners in the differential behavior of Prph2 in rods vs cones. In order to properly traffic and function in the OS, Prph2 forms membrane microdomains to organize and house a network of proteins that are necessary/specific for the formation of the closed rims in rods and open lamellae/rims in cones. In this aim, we will identify critical Prph2 binding partners and determine how these interactions differ between rods and cones. The outcomes of these studies will elucidate the specific function of Prph2 and Rom1 in rods and cones, determine how modulating Rom1 affects disease phenotypes, and will further our understanding of the role of Prph2 in the closed rim structure in rods and the open rim structures in cones. Given the lack of effective treatments for Prph2-associated diseases, these studies are essential and bring attention to the role of Rom1 as a common disease modulator for future development of effective therapeutic strategies.

项目摘要 外围蛋白2（PRPH2）中的突变解释了遗传性视网膜疾病（IRD）的很大一部分。 描述PRPH2在光感受器形态发生和边缘形成中的作用对于了解如何了解如何 单个蛋白质中的突变可能导致这种多样化的病理学，范围从视网膜炎 色素（RP）到模式营养不良（PD）和黄斑变性（MD）。即使患有家庭内部的患者 突变可以表现出显着的表型变异性，证明了PRPH2的复杂性质 功能。我在此应用程序中的目标是提高我们当前对PRPH2在外部作用的了解 片段（OS）形成以更好地了解疾病病原体和将来 制定有效的治疗策略。多年来，我们的实验室开发了一个广泛的工具箱 体外和体内突变模型，目前正在探索创新的成像技术以进一步 了解PRPH2在OS景观中的作用。到目前为止，已经确定该关联 在PRPH2及其同源物之间，ROD OS蛋白1（ROM1）对表型显着贡献 在患者中观察到的可变性，并且降低ROM1水平可能会改变pRPH2促成的PD表型 到RP温和的RP。此外，众所周知，光盘边缘处的PRPH2的适当功能取决于正确 PRPH2-OLIGOMERS和PRPH2/ROM1复合物的复杂组装。这项调查的主要重点 是了解PRPH2和ROM1在对棒和锥中的功能有所不同，对此特别感兴趣 PRPH2 HOMO-和PRPH2/ROM1杂音形成/运输以及PRPH2突变的影响 在这些过程中。我们提出了两个目标来解决这些目标的两个目标：AIM 1将评估ROM1的作用 锥体和调节PRPH2-疾病表型的机制。通过消除或过表达 ROM1在PRPH2-DISESE模型中，我们将进行仔细的生化，结构和功能分析 评估表型差异，因为它们与复杂的组装和OS运输有关。 AIM 2将调查 相互作用伙伴在杆锥与锥体中PRPH2差异行为中的作用。为了正确流量 PRPH2在OS中的功能形成膜微域，以组织和容纳一个蛋白质网络 对于杆中的闭合边缘和锥体中的薄片/轮辋形成是必要/特定的。在这个目标中 我们将确定关键的PRPH2结合伙伴，并确定这些相互作用在ROD和 锥。这些研究的结果将阐明杆和锥体中PRPH2和ROM1的特定功能， 确定调节ROM1如何影响疾病表型，并将进一步了解我们对 杆中的封闭边缘结构中的prph2和锥体中的开放边缘结构。鉴于缺乏有效 与PRPH2相关疾病的治疗方法，这些研究至关重要，并将ROM1的作用引起关注 一种常见的疾病调节剂，用于未来开发有效的治疗策略。