FHL1-related reducing-body myopathy and other genetic forms of myofibrillar myopathies: composition of pathological protein aggregates revealed by laser capture microdissection and subsequent proteomic analysis

四个半LIM结构域基因-（FHL 1）相关肌病（FHL 1 pathies）的谱包括至少两种不同的肌病，还原体肌病（RBM）和X染色体肌病伴姿势性肌萎缩（XMPMA）。两者都是进行性肌病，骨骼肌组织中存在特异性胞浆内包涵体（胞质和还原体）。严重的RBM往往导致幼儿死亡。申请人独立地参与发现FHL 1突变是两种疾病的病因1，18。此外，在RBM肌肉组织标本中使用胞质内包涵体和聚集体的激光显微切割，然后通过纳米流LC-MS/MS结合蛋白质组学分析，为发现罕见肌病中的潜在基因突变与骨骼肌结构异常提供了一种新的方法。除了特定的夹杂物，FHL 1病变的组织病理学与肌原纤维肌病的谱重叠，有颗粒丝状物质和Z线改变的证据。因此，这些肌病必须包括在组的肌原纤维肌病。明确FHL 1病的临床、组织病理学和突变谱及其对疾病机制的影响; 2.定义来自遗传分类的肌原纤维肌病和参与该聚生体的转基因动物的病理蛋白质聚集体的蛋白质组学谱; 3.通过激光捕获显微切割（LCM）病理性蛋白质聚集体，随后进行蛋白质组学和随后的遗传分析，在未分类形式的MFM中确定新的致病基因。

The spectrum of four and a half LIM domain gene- (FHL1) associated myopathies (FHL1opathies) encompasses at least two distinct myopathies, the reducing body myopathy (RBM) and the X-chromosomal myopathy with postural muscle atrophy (XMPMA). Both are progressive myopathies with pathognomonic intracytoplasmic inclusions (cytoplasmic and reducing bodies) in the skeletal muscle tissue. The severe form of RBM often leads to death in early childhood. The applicants were independently involved in the discovery of FHL1 mutations to be causative for both diseases1,18. Furthermore, using laser microdissection of the intracytoplasmic inclusions and aggregates followed by nanoflow LC-MS/MS coupled with proteomic analysis in RBM muscle tissue specimens, a new approach was set for the discovery of underlying gene mutations in rare myopathies with structural abnormalities of skeletal muscle. In addition to specific inclusions, the histopathology of FHL1opathies overlaps with the spectrum of myofibrillar myopathies, with evidence of granulofilamentous material and Z-line alterations. Therefore these myopathies have to be included in the group of myofibrillar myopathies.The aims of our project are: 1. to define the clinical, histopathological, and mutational spectrum of FHL1opathies and their consequences for the disease mechanisms; 2. to define proteomic spectra of pathological protein aggregates from genetically classified myofibrillar myopathies and transgenic animals involved in this consortium; 3. to define new causative genes in unclassified forms of MFM by laser capture microdissection (LCM) pathological protein aggregates followed by proteomic and subsequent genetic analysis.