Department of Cardiovascular Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
Sakakibara Heart Institute, Japan Research Promotion Society for Cardiovascular Diseases, Tokyo, Japan.
Sci Rep. 2021 Sep 23;11(1):18958. doi: 10.1038/s41598-021-98615-3.
Takayasu arteritis (TAK) is an autoimmune systemic arteritis of unknown etiology. Although a number of investigators have attempted to determine biomarkers for diagnosing TAK, there exist no specific serological markers of this intractable disease. We undertook the exploration of novel serological markers which could be useful for an accurate diagnosis of TAK using an unbiased proteomics approach. The purified plasma samples from untreated patients with TAK and healthy individuals were separated by two-dimensional electrophoresis. The differentially expressed protein spots were detected by gel comparison and identified using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MS). Next, we validated plasma concentrations of identified proteins by enzyme-linked immunosorbent assay (ELISA). Two-dimensional electrophoresis and numerical analysis revealed 19 spots and 3 spot clusters whose sum of the sample averages was ≥ 0.01, and the average concentrations were ≥ 1.5 times in the patient group compared with the control group. Among them, 10 spots and spot clusters that met the condition of the average spot concentration being 2.5 times more than that in the control group were selected. After processing these spots using MS and conducting MS/MS ion search, we identified 10 proteins: apolipoprotein C-2 (ApoC-2), actin, apolipoprotein A-1, complement C3, kininogen-1, vitronectin, α2-macroglobulin, 14-3-3 protein ζ/δ, complement C4, and inter-α-trypsin inhibitor heavy chain H4 isoform 1 precursor. Finally, ELISA demonstrated that plasma ApoC-2 level was significantly elevated in patients with TAK compared with that in healthy individuals. Thus, ApoC-2 would be a promising candidate biomarker for TAK diagnosis.
Takayasu 动脉炎(TAK)是一种病因不明的自身免疫性系统性动脉炎。虽然许多研究人员试图确定用于诊断 TAK 的生物标志物,但这种难治性疾病目前没有特异性的血清标志物。我们采用无偏蛋白质组学方法,探索有助于 TAK 准确诊断的新型血清标志物。未经治疗的 TAK 患者和健康个体的纯化血浆样本通过二维电泳分离。通过凝胶比较检测差异表达的蛋白质斑点,并使用基质辅助激光解吸/电离飞行时间/飞行时间质谱(MS)进行鉴定。接下来,我们通过酶联免疫吸附试验(ELISA)验证鉴定出的蛋白质的血浆浓度。二维电泳和数值分析显示,19 个斑点和 3 个斑点簇的样本平均值之和≥0.01,且患者组的平均浓度≥对照组的 1.5 倍。其中,有 10 个斑点和斑点簇符合平均斑点浓度是对照组的 2.5 倍以上的条件。用 MS 处理这些斑点并进行 MS/MS 离子搜索后,我们鉴定出 10 种蛋白质:载脂蛋白 C-2(ApoC-2)、肌动蛋白、载脂蛋白 A-1、补体 C3、激肽原-1、纤连蛋白、α2-巨球蛋白、14-3-3 蛋白 ζ/δ、补体 C4 和 α1-抗胰蛋白酶重链 H4 同工型 1 前体。最后,ELISA 表明 TAK 患者的血浆 ApoC-2 水平明显高于健康个体。因此,ApoC-2 可能是 TAK 诊断的有前途的候选生物标志物。