Institute of Neurology, University College London, London, UK.
Cell Biology and Genetics Research Centre, St. George's University of London, London, UK.
Mol Cell Biochem. 2019 Jun;456(1-2):157-165. doi: 10.1007/s11010-019-03501-2. Epub 2019 Jan 29.
Stroke is a common disorder with significant morbidity and mortality, and complex aetiology involving both environmental and genetic risk factors. Although some of the major risk factors for stoke, such as smoking and hypertension, are well-documented, the underlying genetic and detailed molecular mechanisms remain elusive. Exploring the relevant biochemical pathways may contribute to the clinical diagnosis of stroke and shed light on its aetiology. A comparative proteomic analysis of blood serum of a pair of monozygotic (MZ) twins discordant for ischaemic stroke (IS) was performed using a label-free quantitative proteomics approach. To overcome the limit of reproducibility in the serum preparation, two separate runs were performed, each consisting of three technical replicates per sample. Biological processes associated with proteins differentially expressed between the twins were explored with gene ontology (GO) classification using the functional analysis tool g:Profiler. ANOVA test performed in Progenesis LC-MS identified 179 (run 1) and 209 (run 2) proteins as differentially expressed between the affected and unaffected twin (p < 0.05). Furthermore, the level of serum fibulin 1, an extracellular matrix protein associated with arterial stiffness, was on average 13.37-fold higher in the affected twin. Each dataset was then analysed independently, and the proteins were classified according to GO terms. The categories overrepresented in the affected twin predominantly corresponded to stroke-relevant processes, including wound healing, blood coagulation and haemostasis, with a high proportion of the proteins overexpressed in the affected twin associated with these terms. By contrast, in the unaffected twin diagnosed with atopic dermatitis, there were increased levels of keratin proteins and GO terms associated with skin development. The identification of cellular pathways enriched in IS as well as the upregulation of fibulin 1 sheds new light on the underlying disease-causing mechanisms at the molecular level. Our findings of distinct proteomic signatures associated with IS and atopic dermatitis suggest proteomic profiling could be used as a general approach for improved diagnostic, prognostic and therapeutic strategies.
中风是一种常见的疾病，具有显著的发病率和死亡率，其病因复杂，涉及环境和遗传风险因素。尽管一些中风的主要危险因素，如吸烟和高血压，已经有充分的记录，但潜在的遗传和详细的分子机制仍然难以捉摸。探索相关的生化途径可能有助于中风的临床诊断，并揭示其病因。采用无标记定量蛋白质组学方法，对一对缺血性中风（IS）不一致的同卵双胞胎（MZ）的血清进行了比较蛋白质组分析。为了克服血清制备重复性的限制，进行了两次独立的运行，每个运行包括每个样本的三个技术重复。使用功能分析工具 g:Profiler 对基因本体（GO）分类进行了与双胞胎之间差异表达蛋白相关的生物学过程的探索。Progenesis LC-MS 中的 ANOVA 测试鉴定出 179 个（运行 1）和 209 个（运行 2）蛋白在受影响和未受影响的双胞胎之间差异表达（p < 0.05）。此外，在受影响的双胞胎中，血清纤维连接蛋白 1 的水平平均高出 13.37 倍，纤维连接蛋白 1 是一种与动脉僵硬相关的细胞外基质蛋白。然后分别分析每个数据集，并根据 GO 术语对蛋白进行分类。受影响的双胞胎中过度表达的蛋白主要与中风相关的过程相对应，包括伤口愈合、血液凝固和止血，其中很大一部分与这些术语相关的蛋白在受影响的双胞胎中过度表达。相比之下，在被诊断患有特应性皮炎的未受影响的双胞胎中，角蛋白蛋白和与皮肤发育相关的 GO 术语的水平升高。IS 中富含细胞通路的鉴定以及纤维连接蛋白 1 的上调，为分子水平上的潜在致病机制提供了新的见解。我们发现与 IS 和特应性皮炎相关的独特蛋白质组学特征表明，蛋白质组学分析可以作为一种通用方法，用于改善诊断、预后和治疗策略。
