Heart Failure and Cardiac Regeneration (ICREC) Research Program, Health Science Research Institute Germans Trias i Pujol (IGTP), Badalona, Spain.
Center of Regenerative Medicine in Barcelona, Barcelona, Spain.
Lab Invest. 2018 Oct;98(10):1291-1299. doi: 10.1038/s41374-018-0044-5. Epub 2018 Mar 14.
Dilated cardiomyopathy (DCM) remains a major cause of heart failure and carries a poor prognosis despite important advances in recent years. Better disease characterization using novel molecular techniques is needed to refine its progression. This study explored the proteomic signature of plasma-derived extracellular vesicles (EVs) obtained from DCM patients and healthy controls using size-exclusion chromatography (SEC). EV-enriched fractions were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Raw data obtained from LC-MS/MS were analyzed against the Uniprot human database using MaxQuant software. Additional analyses using Perseus software were based on the Intensity-Based Absolute Quantification (iBAQ) values from MaxQuant analyses. A total of 90.07 ± 21 proteins (227 different proteins) in the DCM group and 96.52 ± 17.91 proteins (183 different proteins) in the control group were identified. A total of 176 proteins (74.6%) were shared by controls and DCM patients, whereas 51 proteins were exclusive for the DCM group and 7 proteins were exclusive for the control group. Fibrinogen (α, β and γ chain), serotransferrin, α-1-antitrypsin, and a variety of apolipoprotein family members (C-I, C-III, D, H or β-2-glycoprotein, and J or clusterin) were clustered in SEC-EVs derived from DCM patients relative to controls (p < 0.05). Regarding Gene Ontology analysis, response to stress and protein activation-related proteins were enriched in DCM-EVs compared with controls. Thus, the present study reports the distinct proteomic signature of circulating DCM-EVs compared with control-EVs. Furthermore, we confirm that SEC obtains highly purified EV fractions from peripheral blood samples for subsequent use in determining disease-specific proteomic signatures.
扩张型心肌病（DCM）仍然是心力衰竭的主要原因，尽管近年来取得了重要进展，但预后仍然不佳。需要使用新型分子技术更好地描述疾病特征，以改善其进展。本研究使用尺寸排阻色谱法（SEC）探索了来自 DCM 患者和健康对照者的血浆衍生细胞外囊泡（EV）的蛋白质组学特征。使用液相色谱-质谱联用仪（LC-MS/MS）分析 EV 富集级分。从 LC-MS/MS 获得的原始数据使用 MaxQuant 软件针对 Uniprot 人类数据库进行分析。使用 Perseus 软件进行的其他分析基于 MaxQuant 分析的基于强度的绝对定量（iBAQ）值。DCM 组中共有 90.07 ± 21 种蛋白质（227 种不同蛋白质）和对照组中 96.52 ± 17.91 种蛋白质（183 种不同蛋白质）被鉴定。对照组和 DCM 患者共有 176 种蛋白质（74.6%），DCM 组特有的蛋白质有 51 种，对照组特有的蛋白质有 7 种。纤维蛋白原（α、β和γ链）、转铁蛋白、α-1-抗胰蛋白酶和多种载脂蛋白家族成员（C-I、C-III、D、H 或β-2-糖蛋白和 J 或聚集素）在 SEC-EVs 中聚集DCM 患者相对于对照组（p < 0.05）。关于基因本体论分析，与对照组相比，DCM-EVs 中富含与应激反应和蛋白质激活相关的蛋白质。因此，本研究报告了与对照-EV 相比，循环 DCM-EV 的独特蛋白质组学特征。此外，我们证实 SEC 从外周血样本中获得高度纯化的 EV 级分，用于随后确定疾病特异性蛋白质组学特征。
