Biomarker Discovery in Muscles from FSHD Patients

FSHD 患者肌肉中生物标志物的发现

• 批准号: 7917471
• 负责人: LOUIS M KUNKEL
• 金额: $ 33.04万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7917471
• 关键词: 4q35; Affect; Biochemical; Biological Markers; Biopsy; Brazil; Candidate Disease Gene; Cell Culture Techniques; Cell Line; Chromosomes; Chromosomes, Human, Pair 4; Clinical; Clinical Trials; Complement; D4Z4; Data; Development; Disease; Family; Gene Expression; Genes; Goals; Human; Individual; Lead; Membrane; Messenger RNA; MicroRNAs; Molecular; Molecular Profiling; Muscle; Muscle Cells; Muscle Proteins; Muscle Weakness; Muscular Dystrophies; Mutation; Myoblasts; Patients; Pattern; Preparation; Proteins; Proteome; Proteomics; RNA; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Severities; Severity of illness; Skeletal Muscle; Tissues; Work; chemokine; design; follow-up; mRNA Expression; protein expression; research study; success; therapy design

FSHD is one of the more common forms of muscular dystrophy in humans with a very unusual and poorly understood biochemical, developmental and molecular underpinning. What is clear is that the D4Z4 repeat deletion in some way influences the expression of genes in muscle in a dominant fashion to cause a variable degree of myofiber degeneration and muscle weakness in different patients. We have established by both proteomic and RNA profiling of muscle biopsies from FSHD patient that both proteins and RNA change expression patterns in diseased tissue. In addition, we have observed in 5 FSHD families from Brazil that some asymptomatic carriers of D4Z4 deletions substantially increase expression of 12 genes including 2 chemokines encoded on chromosome 4 which are only modestly changed in symptomatic patients. We propose to follow up on these findings and use protein, microRNA and mRNA expression profiling as an approach to understanding the differences in disease severity in different individuals with the D4Z4 deletion with the hope that this understanding might lead to the discovery of biomarkers which will be useful in evaluating potential treatments of FSHD. We propose to accomplish this goal according to the following specific aims: 1) Continue to profile mRNA from FSHD patients, control muscle and cell lines generated from differentially affected muscles and confirm existing and new array data by RT-PCR. 2) Confirm our observation on the differential expression of certain miRNAs in FSHD muscle and look at the change in expression of the predicted targets of our observed miRNAs in the mRNA expression arrays from aim 1. We will also ablate these candidate miRNAs in myogenic cell lines to see if we can recapitulate in culture the gene expression changes we see in skeletal muscle of FSHD patients. 3) Lastly, we will continue parallel experiments to define the changes in the proteome in FSHD muscles and myogenic cell lines.

FSHD是人类肌肉萎缩症的一种更常见的形式，具有非常不寻常和差的 了解生物化学，发育和分子基础。很明显，D4 Z4重复 缺失以某种方式影响肌肉中基因的表达，以显性方式导致肌肉中基因的缺失。 不同患者存在不同程度的肌纤维变性和肌无力。我们建立 通过来自FSHD患者的肌肉活检的蛋白质组学和RNA分析， 改变患病组织中的表达模式。此外，我们还观察到5个FSHD家庭， 一些D4 Z4缺失的无症状携带者大幅增加了12个基因的表达 包括在4号染色体上编码的2种趋化因子，其在有症状的 患者我们建议跟进这些发现，并使用蛋白质，microRNA和mRNA表达 作为了解不同个体疾病严重程度差异的方法， D4 Z4缺失，希望这种理解可能会导致生物标志物的发现， 可用于评估FSHD的潜在治疗方法。我们建议按照 1）继续分析来自FSHD患者、对照肌肉和细胞系的mRNA 从差异受影响的肌肉中产生，并通过RT-PCR确认现有和新的阵列数据。（二） 确认我们对FSHD肌肉中某些miRNA差异表达的观察，并观察FSHD肌肉中的miRNA表达。 在mRNA表达阵列中，我们观察到的miRNA的预测靶点表达的变化， 目标1.我们还将在肌源性细胞系中消除这些候选miRNAs，看看我们是否能在 培养我们在FSHD患者骨骼肌中看到的基因表达变化。3)最后，我们将继续 平行实验来确定FSHD肌肉和肌源性细胞系中蛋白质组的变化。