Proteomics profiling of urine reveals specific titin fragments as biomarkers of Duchenne muscular dystrophy
Proteomics profiling of urine reveals specific titin fragments as biomarkers of Duchenne muscular dystrophy
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DOI：
10.1016/j.nmd.2014.03.012
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发表时间：
2014-07-01
影响因子：
2.8
通讯作者：
Svinartchouk, Fedor
中科院分区：
文献类型：
--
作者：
Rouillon, Jeremy;Zocevic, Aleksandar;Svinartchouk, Fedor
肌营养不良症的诊断目前是基于侵入性方法，需要肌肉活检或血液检查。本研究的目的是确定尿生物标志物作为肌营养不良症的诊断工具。在这里，杜氏肌营养不良症（DMD）患者和健康供体的尿蛋白质组进行了比较与自下而上的蛋白质组学方法。对1100多个已鉴定的蛋白质的无标记分析显示，其中32个在健康对照和DMD患者之间差异表达。在这32种蛋白质中，肌联蛋白在健康受试者和DMD患者之间显示出最高的倍数变化。有趣的是，大多数测序的肽属于肌联蛋白的N-末端和C-末端部分，并且通过Western印迹分析证实DMD患者尿液中存在相应片段。对DMD患者和年龄匹配的对照（总共104个个体，年龄从3岁到20岁）的大队列的分析证实了除了两个患者之外的所有患者中存在N-ter片段。在两名年龄为16岁和20岁的DMD患者中，该片段检测不到，而两名16岁和19岁的血清CK >800 IU/L的健康对照显示出低水平的片段。N-和C-末端肌联蛋白片段也检测到尿液中的其他肌营养不良症，如贝克肌营养不良症和肢带型肌营养不良症（1D，2D和2 J型），但没有在神经源性脊髓性肌萎缩症。它们也存在于肌营养不良蛋白缺陷动物模型（GRMD犬和mdx小鼠）的尿液中。肌联蛋白是第一个尿生物标志物，它提供了开发一种简单，非侵入性和易于使用的测试用于肌营养不良症的预筛选的可能性，并且也可能被证明对治疗中的DMD患者的非侵入性随访有用。(C)2014爱思唯尔有限公司版权所有。
Diagnosis of muscular dystrophies is currently based on invasive methods requiring muscle biopsies or blood tests. The aim of the present study was to identify urinary biomarkers as a diagnostic tool for muscular dystrophies. Here, the urinary proteomes of Duchenne muscular dystrophy (DMD) patients and healthy donors were compared with a bottom-up proteomic approach. Label-free analysis of more than 1100 identified proteins revealed that 32 of them were differentially expressed between healthy controls and DMD patients. Among these 32 proteins, titin showed the highest fold change between healthy subjects and DMD patients. Interestingly, most of the sequenced peptides belong to the N-terminal and C-terminal parts of titin, and the presence of the corresponding fragments in the urine of DMD patients was confirmed by Western blot analysis. Analysis Of a large cohort of DMD patients and age-matched controls (a total of 104 individuals aged from 3 to 20 years) confirmed presence of the N-ter fragment in all but two patients. In two DMD patients aged 16 and 20 years this fragment was undetectable and two healthy controls of 16 and 19 years with serum CK >800 IU/L demonstrated a low level of the fragment. N- and C-terminal titin fragments were also detected in urine from patients with other muscular dystrophies such as Becker muscular dystrophy and Limb-girdle muscular dystrophy (type 1D, 2D and 2J) but not in neurogenic spinal muscular atrophy. They were also present in urine of dystrophin-deficient animal models (GRMD dogs and mdx mice). Titin is the first urinary biomarker that offers the possibility to develop a simple, non-invasive and easy-to-use test for pre-screening of muscular dystrophies, and may also prove to be useful for the non-invasive follow up of DMD patients under treatment. (C) 2014 Elsevier B.V. All rights reserved.