镶边空泡肌病的基因表型与蛋白组学的相关性研究

Rimmed vacuoles myopathy (MRV) is defined as a group of myopathies with characteristic histopathological features of rimmed vacuoles in muscle fibers.The disorder including genetic myopathy and acquired myopathy with significant differences in clinical manifestations, pathogenic gene and treatment exist.Most rimmed vacuoles myopathy are accompanied with abnormal protein degradation pathway. This study based on the genetic resources of Chinese myopathy patients and families. We will collect the muscle and DNA samples of MRV patients, and perform mutation screenings and causative gene lacating by the second generation of gene chip technology and whole-genome sequencing. At the same time, we will perform two-dimensional gel electrophoresis - mass spectrometry and Western blot to investigate the differential protein expression in rimmed vacuoles myopathy. The results of this study will further expand the clinical and genetic spectrum of rimmed vacuoles myopathies, find specific protein markers closely associated with development and morbidity of rimmed vacuoles myopathies, which in turn may provide new clues for undersanding the relationship between the gene phenotype and proteome of the disease.

镶边空泡性肌病（myopathy with rimmed vacuoles，MRV）是指在病理学上观察到肌纤维内出现一种特殊的空泡为表现的肌肉病，包括有遗传性肌病和后天获得性肌病，临床表现、致病基因及治疗存在显著差异，多数镶边空泡肌病都伴有蛋白降解途径的异常。本课题通过肌肉病理检查筛选出伴镶边空泡病理改变的患者，采集这些患者及家系的肌肉及DNA样本，利用二代基因芯片技术、全外显子测序技术，进行致病基因筛查及基因定位研究；同时，采用二维凝胶电泳-质谱和Western Blot方法，检测筛选镶边空泡性肌病的差异表达蛋白。以期进一步扩展现有镶边空泡性肌病基因表型谱和寻找镶边空泡性肌病可能的生物学标志物提供依据，探讨基因表型与蛋白组学的相关性，也为进一步了解镶边空泡产生的可能蛋白降解途径机制提供基础。

背景：镶边空泡肌病(myopathy with rimmed vacuoles，MRV)是指在病理学上观察到肌纤维内出现一种特殊的空泡为表现的肌肉病，包括有遗传性肌病和后天获得性肌病，临床表现、致病基因及治疗存在显著差异，多数镶边空泡肌病都伴有蛋白降解途径的异常。.主要研究内容：通过肌肉病理检查筛选出伴镶边空泡病理改变的患者，采集这些患者及家系的肌肉及 DNA样本，利用二代基因芯片技术、全外显子测序技术，进行致病基因筛查及基因定位研究;同时，采用二维凝胶电泳-质谱和Western Blot方法，检测筛选镶边空泡性肌病的差异表达蛋白。.重要结果及关键数据：20例GNE肌病患者共检测到24种基因突变，16种为已报道的致病突变，8种在HGMDpro数据库未见报道，分别为c.1619-2A>C、c.1559A>G、c.1711_1712del、c.556T>C、c.1634-1G>C、c.1897G>T、c.1205dupT以及c.733A>G。其中，c.1619-2A>C、c.1634-1G>C为剪切位点突变，c.1711_1712del为缺失突变，c.1205dupT为重复突变。与正常对照相比，s-IBM有124种差异蛋白，其中87种上调蛋白，37种下调蛋白。这些蛋白主要富集于细胞质、细胞骨架、线粒体及肌小节，具有结合、酶活性及细胞骨架支撑等分子功能，参与肌肉收缩及分解代谢等生物学过程，作用于糖酵解、细胞骨架调节以及炎症反应等多种通路。与正常对照相比，GNE肌病有70种差异蛋白，其中38种上调蛋白，32种下调蛋白。这些蛋白主要富集于收缩纤维、肌原纤维及肌小节等结构，具有蛋白结合、钙素结合及肌肉结构成分等分子功能，参与肌肉收缩及分解代谢等生物学过程，作用于糖酵解、细胞骨架调节等多种通路。.科学意义：发现8种GNE肌病新发突变，丰富了镶边空泡肌病基因突变谱。针对镶边空泡肌病蛋白质组学特点进行系统定量分析、筛选和生物信息学分析，完成了细胞定位、生物学功能、信号通路等的分析，提出差异蛋白分子特征，为进一步揭示镶边空泡肌病的发病机制提供了研究基础。