Molecular mechanism of dipeptide repeat protein production from hexanucleotide repeats in C9ORF72-related ALS and FTD

C9ORF72相关ALS和FTD中六核苷酸重复产生二肽重复蛋白的分子机制

基本信息

批准号：
10378768
负责人：
Shuying Sun
金额：
$ 35.82万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10378768
关键词：
Affect Amyotrophic Lateral Sclerosis Antisense RNA Biochemical C9ORF72 CRISPR screen CRISPR/Cas technology Candidate Disease Gene Cell Nucleus Cells Cytoplasm Cytoplasmic Inclusion Cytosol Dipeptides Disease Event Frontotemporal Dementia Gene-Modified Genes Genetic Genetic Screening Genomics Goals Human Induced pluripotent stem cell derived neurons Inherited Introns Knock-out Link Mediating Molecular Nerve Degeneration Neurodegenerative Disorders Nuclear Export Nuclear RNA Nucleotides Pathogenesis Pathogenicity Pathologic Pathway interactions Patients Phenotype Process Production Proteins RNA RNA Processing RNA Splicing Reading Frames Regulation Regulatory Element Reporter Series System Techniques Technology Testing Therapeutic Tissues Toxic effect Translating Translations Untranslated RNA base frontotemporal lobar dementia-amyotrophic lateral sclerosis genome-wide induced pluripotent stem cell insight mRNA Precursor novel nucleocytoplasmic transport prevent protein expression research and development screening stem cell model therapeutic target tool

项目摘要

PROJECT SUMMARY Hexanucleotide repeat expansion in a non-coding region of C9orf72 is the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontal temporal dementia (FTD). One potential pathogenic mechanism is the aberrant accumulation of dipeptide repeat (DPR) proteins produced by repeat-associated non-AUG (RAN) translation in all six reading frames (poly-GA, poly-GR, poly-PA, poly-PR and poly-PG) of both sense and antisense RNAs. Abnormal cytoplasmic inclusions of these DPR proteins have been found in C9ORF72 patient tissues and cells. Several lines of evidence indicate that some forms of the DPR proteins can be pathogenic, yet little is known how the DPRs are generated from the expanded repeats. The goal of this proposal is to understand the molecular mechanisms and identify genetic modifiers of DPR production from both sense and antisense repeats, combining the genome-scale CRISPR/Cas9 knockout screening technology with biochemical and molecular approaches. We will primarily focus on two major steps of repeat RNA processing that could influence the final protein expression level: RNA nuclear export and RAN translation. We will dissect the molecular pathways of nuclear export for both sense and antisense repeats, what factors mediate this process, and how this is affected by C9-mediated toxicity on nucleocytoplasmic transport. We will decipher the molecular mechanisms of C9 RAN translation, and examine whether there are common modifiers between sense and antisense repeats. Altogether, these studies can provide novel insights on how DPRs are produced from C9 repeats and what approaches can be taken to prevent it. This will guide the development of research tools to understand how DRPs contribute to the disease pathogenesis. It will also possibly provide potential therapeutic targets to reduce DPR-mediated toxicity by inhibiting their production.

项目摘要在C9orf72的非编码区域中的六核苷酸重复膨胀是两者的最常见遗传原因肌萎缩性侧索硬化症（ALS）和额叶颞痴呆（FTD）。一种潜在的致病机制是由重复相关的非aug产生的二肽重复（DPR）蛋白的异常积累（ran）在所有六个含义的六个读取帧（poly-ga，poly-gr，poly-pa，poly-pg和poly-pg）中的翻译和反义RNA。这些DPR蛋白的胞质夹杂物异常已在C9orf72中发现患者组织和细胞。几条证据表明，某些形式的DPR蛋白可以是致病性，但鲜为人知的是如何从扩展的重复序列中产生DPR。目标的目标建议是了解分子机制，并从中鉴定出DPR产生的遗传修饰符感觉和反义重复，结合了基因组尺度CRISPR/CAS9敲除筛查技术采用生化和分子方法。我们将主要关注重复RNA的两个主要步骤可能影响最终蛋白质表达水平的处理：RNA核输出和RAN翻译。我们将剖析感官和反义重复的核出口的分子途径，什么因素介导这一过程，以及如何受到C9介导的毒性转运的毒性的影响。我们将破译C9的分子机制进行了翻译，并检查是否有常见的修饰符在感官和反义重复之间。总之，这些研究可以提供有关DPR的新见解通过C9重复产生，可以采取哪种方法来防止它。这将指导发展了解DRP如何促进疾病发病机理的研究工具。它也可能会提供潜在的治疗靶标可通过抑制其产量来降低DPR介导的毒性。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Measuring Repeat-Associated Non-AUG (RAN) Translation.

测量重复相关的非 AUG (RAN) 翻译。

DOI：
10.1007/978-1-0716-1975-9_8
发表时间：
2022
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Wang,Shaopeng;Sun,Shuying
通讯作者：
Sun,Shuying

DDX3X overexpression decreases dipeptide repeat proteins in a mouse model of C9ORF72-ALS/FTD.

DDX3X 过表达会减少 C9ORF72-ALS/FTD 小鼠模型中的二肽重复蛋白。

DOI：
10.1016/j.expneurol.2024.114768
发表时间：
2024
期刊：
Experimental neurology
影响因子：
5.3
作者：
Fu,Xiujuan;Zhang,Zhe;Hayes,LindseyR;Wright,Noelle;Asbury,Julie;Li,Shelley;Ye,Yingzhi;Sun,Shuying
通讯作者：
Sun,Shuying

Translation dysregulation in neurodegenerative diseases: a focus on ALS.