Mechanistic Basis for Non-Canonical Translation in Neurological Disease

神经系统疾病非规范翻译的机制基础

• 批准号: 10591832
• 负责人: Hannah Shorrock
• 金额: $ 9.92万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10591832
• 关键词: Biology; CAG repeat; Cell Line; Cell Shape; Clustered Regularly Interspaced Short Palindromic Repeats; Coupled; Data; Detection; Disease; Disease model; Genetic; Goals; Human; Human Genome; Immunologics; Induced pluripotent stem cell derived neurons; Initiator Codon; Knowledge; Mass Spectrum Analysis; Mediating; Methods; Microsatellite Repeats; Modeling; Molecular; Mutagenesis; Pathogenesis; Pathogenicity; Peptide Initiation Factors; Play; Process; Production; Proteins; Proteome; RNA; Reading Frames; Reporter; Research; Ribosomes; Role; Sedimentation process; Small Interfering RNA; Structure; System; Testing; Therapeutic; Therapeutic Intervention; Tissues; Transcript; Translations; Treatment Efficacy; Untranslated RNA; Western Blotting; comparative; design; genome wide screen; genome-wide; innovation; interest; molecular modeling; nervous system disorder; novel; permissiveness; polyglutamine; polysome profiling; ran GTP-Binding Protein; therapy development; translation factor

Project Summary CAG repeat expansion diseases are the most common class of microsatellite expansion disorders. While canonical translation of the CAG repeat leads to the production of polyglutamine proteins, these expansions can also undergo non-canonical repeat associated non-AUG (RAN) translation. In the later process, translation is initiated across multiple reading frames in the absence of AUG- or AUG-like start codons. While a few factors that influence either canonical translation across microsatellite expansions or RAN translation have been identified, our understanding of the underlying molecular mechanisms responsible for translation in multiple reading frames is extremely limited. Despite evidence that RNA structure plays a key role in RAN translation our knowledge of the structural features that facilitate RAN translation is extremely limited. To examine the role of RNA structure in non-canonical translation, I used a structure-forward approach to screen for multi-frame translation, through which I identified several long non-coding RNAs (lncRNAs) that support translation in multiple reading frames in the absence of both microsatellite repeat expansions and AUG-start codons. This data suggests multiple avenues for RNA transcripts to support multi-frame translation, which has important implications in biology, disease, and therapeutics. The overarching goal of this proposal is to determine the RNA structures, protein networks and therapeutically relevant modifiers that govern various forms of multi-frame translation (MFT). To determine the proportion of the human genome capable of undergoing MFT, I will utilize a novel MFT reporter system to perform a genome-wide screen. Utilising candidate MFT sequences and CAG repeat expansions along with my already identified MFT lncRNAs, I will determine the cellular RNA structures and protein factors that facilitate non-canonical multi-frame translation. I will also employ a first-of-its-kind strategy to identify novel, therapeutically relevant modifiers of co-occurring canonical and non-canonical translation in multiple reading frames. These studies will enable a better understanding of the mechanism of multi-frame translation and identify therapeutic strategies that have the potential to provide therapeutic efficacy across multiple CAG repeat expansion diseases and be applicable to a wider range of neurological diseases. Together this research provides an innovative approach to understand a novel paradigm for translation diversity and will enable a better understanding of disease pathogenesis in repeat expansion diseases as well as elucidating the aspects of MFT most suitable to therapeutic intervention.

项目摘要 CAG重复膨胀疾病是最常见的微卫星扩张障碍。尽管 CAG重复的规范翻译导致多谷氨酰胺蛋白的产生，这些膨胀可以 还经历非典型重复相关的非aug（ran）翻译。在以后的过程中，翻译为 在没有八月或八月的启动密码子的情况下，跨多个读取帧启动。虽然有几个因素 这种影响在微卫星扩张之间的典型翻译或翻译的影响已经 确定的是，我们对负责翻译的基本分子机制的理解 读取帧非常有限。尽管有证据表明RNA结构在翻译中起着关键作用 了解促进翻译的结构特征非常有限。检查 RNA结构在非典型翻译中，我使用了一种结构前线方法来筛选多框架 翻译，通过这些翻译，我确定了支持翻译的几个长期非编码RNA（LNCRNA） 在没有微卫星重复扩展和AUG启动密码子的情况下，多读帧。这 数据提出了RNA转录本的多种途径，以支持多帧翻译，这很重要 生物学，疾病和治疗学的影响。该提案的总体目标是确定RNA 结构，蛋白质网络和具有治疗相关的修饰符，这些修饰符控制各种形式的多框架 翻译（MFT）。为了确定能够接受MFT的人类基因组的比例，我将利用一个 新型MFT报告基因系统以执行全基因组屏幕。利用候选MFT序列和CAG 重复扩展以及我已经识别的MFT lncrNA，我将确定细胞RNA结构 和促进非典型多框架翻译的蛋白质因子。我还将采用首个 识别同时存在的规范和非典型的新颖，治疗相关的修饰符的策略 在多个读取帧中翻译。这些研究将使人们能够更好地了解 多帧翻译并确定具有提供治疗功效的潜力的治疗策略 在多种CAG重复膨胀疾病中，适用于更广泛的神经系统疾病。 这项研究共同提供了一种创新的方法来了解翻译多样性的新颖范例 并将更好地了解重复膨胀疾病中的疾病发病机理以及 阐明最适合治疗干预的MFT方面。