Investigating proteostasis in facioscapulohumeral muscular dystrophy

研究面肩肱型肌营养不良症的蛋白质稳态

基本信息

批准号：
10430945
负责人：
YI-WEN CHEN
金额：
$ 18.03万
依托单位：
CHILDREN'S RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10430945
关键词：
4q35 Affect Aftercare Algorithms Antisense Oligonucleotide Therapy Antisense Oligonucleotides Cells Chromosomes Complex D4Z4 Data Defect Deuterium Oxide Disease Disease Progression Double Effect Enzyme-Linked Immunosorbent Assay Epigenetic Process Facioscapulohumeral Muscular Dystrophy Gene Expression Genes Genetic Genetic Transcription Goals Homeobox Homeodomain Proteins Hour Human In Vitro Individual Inherited Isotope Labeling Label Liquid Chromatography Metabolic Molecular Mus Muscle Muscle Cells Muscular Dystrophies Mutation Myoblasts Myopathy Pathology Patients Peptides Performance Post-Transcriptional Regulation Process Protein Biosynthesis Proteins Regulation Reporting Repression Research Role Siblings Testing Time Transcription Process Treatment Efficacy Western Blotting Work Xenograft procedure disease phenotype efficacy evaluation improved in vivo insight knock-down mouse model muscular dystrophy mouse model nanoscale protein degradation protein profiling proteostasis response tandem mass spectrometry therapeutic development therapeutic target treatment effect

项目摘要

ABSTRACT Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant muscle disorder caused by complex genetic and epigenetic mechanisms. Previous studies showed that transcription de- repression of double homeobox protein 4 (DUX4) due to epigenetic changes in the D4Z4 region causes FSHD. The epigenetic changes are caused by either contraction of the D4Z4 array from 11-150 repeat units in unaffected individuals to 1-10 repeat units in roughly 95% of patients (FSHD1), or mutations in epigenetic regulators of the D4Z4 region (FSHD2). The expression of DUX4 leads to downstream molecular and cellular changes, which contribute to disease progression. Previous studies reported defects in protein degradation process in FSHD, however what proteins are affected and whether protein synthesis is affected is not clear. To study the dynamic changes in protein abundance in affected muscle cells, the research team conducted a preliminary study using liquid chromatography - tandem mass spectrometry in combination with metabolic labelling. The study identified distinct protein profiles between the myoblasts from individuals affected by FSHD and their unaffected siblings. We hypothesize that DUX4 expression is responsible for the disturbance of protein homeostasis, which contributes to the downstream molecular and cellular defects observed in FSHD. In the proposed study, we will first study the protein synthesis and degradation in immortalized FSHD myoblasts before and after a DUX4-reducing treatment. In aim 2, we will validate the in vitro findings using a xenograft mouse model. This project will discover proteins that are mis-regulated by degradative processes and synthetic processes in FSHD, which may open new opportunities for therapeutic development. The findings will also provide new insights of the disease mechanisms and evaluate the efficacy of a DUX4-reducing treatment using antisense oligonucleotides targeting the DUX4.

抽象的面肩肱型肌营养不良症 (FSHD) 是一种常染色体显性肌肉疾病由复杂的遗传和表观遗传机制引起。先前的研究表明转录去由于 D4Z4 区域的表观遗传变化而抑制双同源盒蛋白 4 (DUX4) FSHD。表观遗传变化是由 D4Z4 阵列从 11-150 次重复收缩引起的未受影响个体中的 1-10 个重复单位到大约 95% 的患者 (FSHD1) 中的 1-10 个重复单位，或者 D4Z4 区域 (FSHD2) 的表观遗传调节因子。 DUX4的表达导致下游分子和细胞变化，导致疾病进展。先前的研究报道 FSHD 中蛋白质降解过程存在缺陷，但是哪些蛋白质受到影响以及是否受到影响蛋白质合成是否受到影响尚不清楚。研究蛋白质丰度的动态变化研究小组利用液相色谱法进行了初步研究—— 串联质谱与代谢标记相结合。该研究确定了不同的蛋白质受 FSHD 影响的个体与其未受影响的兄弟姐妹的成肌细胞之间的概况。我们假设 DUX4 表达导致蛋白质稳态紊乱，这导致 FSHD 中观察到的下游分子和细胞缺陷。在拟议的研究中，我们首先研究永生化FSHD成肌细胞中蛋白质的合成和降解。经过 DUX4 还原处理后。在目标 2 中，我们将使用异种移植小鼠验证体外研究结果模型。该项目将发现被降解过程和合成过程错误调节的蛋白质 FSHD 的过程，这可能为治疗发展开辟新的机会。研究结果将还提供了疾病机制的新见解并评估了 DUX4 减少的功效使用针对 DUX4 的反义寡核苷酸进行治疗。