Molecular mechanisms of translational control in mice with inherited retinal degeneration

遗传性视网膜变性小鼠翻译控制的分子机制

• 批准号: 10360454
• 负责人: Marina Gorbatyuk
• 金额: $ 36.01万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10360454
• 关键词: AKT inhibition; Acute; Address; Affect; Animal Model; Attenuated; Basic Science; Binding; Binding Proteins; Blindness; Cell Death; Cells; Cellular biology; Chronic; Chronic stress; Clinical; Complex; Data; Development; Disease; Down-Regulation; EIF4EBP1 gene; Endoplasmic Reticulum; Eukaryotic Initiation Factor-4E; Eukaryotic Initiation Factors; FRAP1 gene; Genes; Genetic Translation; Genetically Modified Animals; Glucose; Heat shock proteins; Homologous Gene; Human; Individual; Inherited; Investigational Therapies; Leber' s amaurosis; Literature; Mediating; Mediator of activation protein; Methods; Molecular; Molecular Target; Mus; Mutation; Pathogenesis; Pathologic; Pathway interactions; Pharmacology; Phosphorylation; Phosphotransferases; Photoreceptors; Protein Biosynthesis; Protein Kinase; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Recovery; Repression; Retina; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Retinitis Pigmentosa; Rhodopsin; Role; Signal Transduction; Sphingolipids; Stress; System; Testing; Therapeutic; Therapeutic Intervention; Time; Translational Regulation; Translations; Up-Regulation; Viral Genes; Work; arm; attenuation; base; biological adaptation to stress; cytotoxic; effective therapy; endoplasmic reticulum stress; experience; gene replacement; gene therapy; human model; improved; inherited retinal degeneration; lipid metabolism; mRNA capping; mouse model; protein kinase R; response; response biomarker; restoration; retinal rods; targeted treatment; therapeutic development; therapeutically effective; translational impact; viral gene delivery

Inherited retinal degeneration (IRD) diseases are a clinically and genetically diverse group of retinal dystrophies including Leber congenital amaurosis (LCA) and autosomal dominant retinitis pigmentosa (ADRP). Despite significant progress in the basic science and clinical aspects of many retinopathies, the cellular mechanisms responsible for inherited vision loss require further investigation, and effective therapies are still under development. Therefore, we focused on the elucidation of the IRD mechanism and the development of therapeutic strategies. In particular, we address the role of translational attenuation in retinal pathogenesis of mice-mimicking human ADRP and LCA and propose therapeutic approaches based on the restoration of protein synthesis in affected photoreceptors. The proposal is supported by strong scientific premises created by our published and preliminary data, as well as the works of others. They suggest that mice with IRD experience persistent activation of the unfolded protein response (UPR) and translational attenuation in the retinas. In addition, our preliminary data demonstrate that PERK (protein kinase R [PKR]-like endoplasmic reticulum kinase) UPR arm is responsible for the sustained translational suppression in stressed photoreceptors. To support this, we will investigate three PERK-induced targets that independently regulate the consensual work of translational machinery. First, we plan to evaluate the impact of translational attenuation on retinal degeneration provided by phosphorylated eIF2a, a major hallmark of translational block. Second, we plan to assess the role of TRB3 in retinal degeneration by modifying its expression, thereby testing the hypothesis that TRB3 increase during IRD progression is cytotoxic, while its down-regulation enhances protein synthesis and retards IRD progression. Finally, we plan to test the hypothesis that stabilization of the 5'cap mRNA recognition complex by deactivation of eukaryotic initiation factor 4E (4E-BP), a negative translational regulator slows the onset of IRD through augmentation of protein synthesis. Genetically modified mice expressing altered levels of UPR and mRNA translation-associated markers; mice-modeling human IRD; pharmacological approaches altering levels of UPR markers; and adeno-associated viral gene delivery will be used in the study to regulate protein synthesis and explore therapeutic approaches. To our knowledge, this is the first comprehensive study of translational regulation in healthy and diseased photoreceptors. If successful, this study will not only enhance our molecular understanding of how degenerating PRs die but also create a platform for therapeutic intervention to halt IRD progression in humans. PERK pathway-targeted therapies could be used as a mutation- independent treatment for retinas experiencing chronic ER stress or as a supplemental strategy to enhance pre-existing gene replacement approaches. They would be particularly attractive for mutations in large genes that cannot be treated using conventional AAV-mediated gene therapy.

遗传性视网膜变性(IRD)疾病是一组临床上和遗传上多样化的视网膜疾病 营养不良包括Leber先天性黑色素症(LCA)和常染色体显性遗传性视网膜色素变性(ADRP)。 尽管许多视网膜病变在基础科学和临床方面取得了重大进展，但细胞 遗传性视力丧失的机制需要进一步研究，有效的治疗方法仍然存在。 正在开发中。因此，我们集中阐述税务局的机制和税务局的发展 治疗策略。特别是，我们讨论了翻译衰减在视网膜发病机制中的作用。 小鼠模仿人类ADRP和LCA，提出基于蛋白质恢复的治疗方法 受影响的光感受器的合成。这项建议得到了强大的科学前提的支持，这是由我们的 出版的和初步的数据，以及其他人的作品。他们建议有IRD经历的小鼠 视网膜中未折叠蛋白反应(UPR)的持续激活和翻译衰减。在……里面 此外，我们的初步数据表明，PERK(蛋白激酶R[PKR]样内质网状激酶) UPR ARM负责应激光感受器的持续翻译抑制。为了支持这一点， 我们将研究三个由PERK诱导的靶点，它们独立地调节翻译的共识工作 机械设备。首先，我们计划评估平移衰减对视网膜变性的影响 磷酸化的eIF2a，翻译块的一个主要标志。第二，我们计划评估TRB3在 通过改变其表达，从而检验TRB3在IRD期间增加的假设 进展是细胞毒性的，而它的下调促进了蛋白质的合成，延缓了IRD的进展。 最后，我们计划检验通过失活来稳定5‘帽mna识别复合体的假设。 在真核细胞起始因子4E(4E-BP)中，负翻译调控通过以下途径延缓IRD的发生 蛋白质合成的增强。表达UPR和mRNA水平改变的转基因小鼠 翻译相关标记物；小鼠模拟人类IRD；药理学方法改变UPR水平 标记物；以及腺相关病毒基因传递将在研究中用于调节蛋白质合成和 探索治疗方法。据我们所知，这是第一次对翻译进行全面研究 对健康和患病的光感受器的调节。如果成功，这项研究不仅将增强我们的 对退行性前列腺癌如何死亡的分子理解，也为治疗干预创造了一个平台 以阻止人类IRD的进展。PERK途径靶向治疗可能被用作突变- 对经历慢性内质网应激的视网膜进行独立治疗或作为补充策略 加强现有的基因替换方法。它们对基因突变特别有吸引力 使用传统的AAV介导的基因治疗无法治疗的大基因。

项目成果

Posttranslational modifications of proteins in diseased retina.

• DOI: 10.3389/fncel.2023.1150220
• 发表时间: 2023
• 期刊: FRONTIERS IN CELLULAR NEUROSCIENCE
• 影响因子: 5.3
• 作者: Starr, Christopher R.;Gorbatyuk, Marina S.
• 通讯作者: Gorbatyuk, Marina S.

Proteomic analysis of diabetic retinas.

糖尿病视网膜的蛋白质组学分析。

• DOI: 10.3389/fendo.2023.1229089
• 发表时间: 2023
• 期刊: Frontiers in endocrinology
• 影响因子: 5.2

Retinal injury mouse model and pathophysiological assessment of the effect of arsenical vesicants.

• DOI: 10.1016/j.exer.2022.109354
• 发表时间: 2023-01
• 期刊: EXPERIMENTAL EYE RESEARCH
• 影响因子: 3.4
• 作者: Zhylkibayev, Assylbek;Srivastava, Ritesh;Anantharam, Poojya;Crotch, Claire;Athar, Mohammad;Gorbatyuk, Marina
• 通讯作者: Gorbatyuk, Marina

GADD34 Ablation Exacerbates Retinal Degeneration in P23H RHO Mice.

• DOI: 10.3390/ijms232213748
• 发表时间: 2022-11-09
• 期刊: International journal of molecular sciences
• 影响因子: 5.6