Research on plasma proteome of patients with severe coronary artery calcifications based on mass spectrometry and bioinformatics
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摘要: 目的 基于非标记蛋白组学方法筛选冠状动脉(冠脉)严重钙化病变患者外周血浆中的特异性蛋白质标志物。方法 采用数据非依赖采集质谱技术检测30例冠脉严重钙化病变患者与30例非钙化对照人群血浆，生物信息学软件进一步分析差异表达蛋白质数据。结果 冠脉严重钙化病变患者与非钙化人群血浆相比较，共筛选出表达量差异2倍以上的差异蛋白共28种(其中表达上调的蛋白20种，表达下调的蛋白8种)，基因本体论(GO)分析差异表达蛋白主要分布于细胞外区域、细胞外泌体和胞外细胞器部分；生物学过程主要涉及细胞过程、肌动蛋白细胞骨架和应激反应、白细胞介导的免疫和细胞活化；而其分子功能与蛋白质结合、信号受体结合和肌动蛋白结合相关。京都基因与基因组百科全书(KEGG)分析显示差异表达蛋白与补体和凝血级联、糖酵解/糖异生作用、细胞凋亡、HIF-1和Rap1信号通路相关。结论 冠脉严重钙化病变患者和非钙化人群的血浆蛋白质组学存在显著差异，这些差异表达蛋白质有望成为冠脉严重钙化病变鉴别诊断的新型生物标志物。Abstract: Objective Screening of specific plasma protein markers from patients with severe coronary artery calcifications using label free quantitative proteomics.Methods Plasmas were obtained from 30 patients with severe coronary artery calcifications and 30 non-calcification patients and DIA mass spectrometry-based proteomics were used to detect the proteins in plasma. Bioinformatics analysis tools were used to analysis the dysregulated proteomics data.Results A total of 28 plasma proteins were differentially expressed between the coronary calcification patients and controls, with 20 upregulated and 8 downregulated. Gene ontology(GO) analysis indicated that differentially expressed proteins were mainly distributed in extracellular region, extracellular region part, and extracellular organelle. Biological process was mainly involved in cellular process, response to stimulus, actin cytoskeleton organization, leukocyte mediated immunity and cell activation. Molecular function was primarily associated with protein binding, signaling receptor binding, and cytokine receptor binding. Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis revealed that these differentially expressed proteins were mainly involved in complement and coagulation cascade pathway, glycolysis/ gluconeogenesis, Rap1 signaling pathway, HIF-1 signaling pathway, and apoptosis.Conclusion This study demonstrated that significant differences in plasma proteomics were detected between patients with severe coronary artery calcifications and non-calcification controls. These dysregulated proteins would be novel biomarkers for differential diagnosis of severe coronary artery calcification.
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Key words:
- coronary calcification /
- plasma /
- proteome
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图 2 冠脉严重钙化病变患者与非钙化组人群血浆差异蛋白聚类热图
Figure 2. Clustering heat map of plasma differential proteins
图 3 冠脉严重钙化病变患者与非钙化组人群血浆差异蛋白GO富集分析结果
Figure 3. Results of GO enrichment analysis of plasma differential proteins
图 4 冠脉严重钙化病变患者与非钙化组人群血浆差异蛋白KEGG分析结果
Figure 4. Analysis of plasma differential protein KEGG
表 1 两组人群基线资料比较
Table 1. General data
X±S 参数 非钙化组(30例) 钙化组(30例) P值 年龄/岁 68.90±1.15 68.37±1.27 0.76 男性/例(%) 25(83.33) 25(83.33) 1.00 BMI/(kg·m-2) 23.88±0.49 23.87±0.67 0.99 高血压病史/例(%) 20(66.67) 21(70.00) 1.00 糖尿病史/例(%) 1(3.33) 3(10.00) 0.61 吸烟史/例(%) 13(43.33) 9(30.00) 0.42 饮酒史/例(%) 11(36.67) 8(26.67) 0.58 收缩压/mmHg 132.60±3.67 137.70±3.68 0.33 舒张压/mmHg 77.60±2.33 79.43±2.06 0.56 空腹血糖/(mmol·L-1) 5.60±0.26 5.51±0.27 0.81 胆固醇/(mmol·L-1) 3.85±0.18 3.92±0.23 0.83 低密度脂蛋白/(mmol·L-1) 2.08±0.15 2.09±0.17 0.96 高密度脂蛋白/(mmol·L-1) 1.08±0.04 1.06±0.05 0.73 甘油三酯/(mmol·L-1) 1.24±0.11 1.35±0.10 0.51 肌酐/(μmol·L-1) 80.29±3.83 74.46±3.87 0.29 
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表 2 冠脉严重钙化病变患者与非钙化组人群血浆中28种蛋白质的表达差异
Table 2. Differences in the expression of 28 proteins in plasma
编号 蛋白质序列号 蛋白信息描述 基因名称 表达倍数 P值 1 A0A2U8J8Y3 Ig heavy chain variable region IgH 5.201 0.017 2 A0A5C2GMF2 IG c998_light_IGKV4-1_IGKJ2 N/A 4.567 0.003 3 A0A5C2FZG2 IGL c1546_light_IGLV3-21_IGLJ3 N/A 4.147 0.007 4 P14618 Pyruvate kinase PKM PKM 3.273 0.002 5 Q6ZVM5 cDNA FLJ42083 fis，clone TCERX2000613 N/A 3.175 0.001 6 A0A5C2GUI4 IG c1185_light_IGKV1-6_IGKJ1 N/A 2.567 0.029 7 P67936 Tropomyosin alpha-4 chain TPM4 2.560 0.001 8 A0A024RDT4 Lymphocyte cytosolic protein 1(L-plastin)，isoform CRA_a LCP1 2.457 0.014 9 A0A5C2GK17 IGH + IGL c357_light_IGKV1D-39_IGKJ1 N/A 2.350 0.016 10 A0A5C2FZ57 IGL c2179_light_IGKV4-1_IGKJ1 N/A 2.321 0.027 11 A0A5C2G5C2 IGH c189_heavy__IGHV3-33_IGHD6-19_IGHJ5 N/A 2.137 0.025 12 A0A024RAG6 Complement C1q subcomponent subunit A ADIC 2.114 0.007 13 B4DMC3 cDNA FLJ60302，highly similar to Neurocan core protein N/A 2.098 0.001 14 A0A5C2FYH6 IGL c2011_light_IGKV1D-33_IGKJ4 N/A 2.081 0.018 15 A0A5C2GUT1 IG c1271_heavy_IGHV3-33_IGHD1-1_IGHJ4 N/A 2.076 0.047 16 Q7Z351 Uncharacterized protein DKFZp686N02209 DKFZp686N02209 2.079 0.016 17 A0A5C2GNX4 IG c870_heavy_IGHV3-66_IGHD3-16_IGHJ1 N/A 2.073 0.005 18 A0A5C2GVS9 IG c657_heavy_IGHV3-7_IGHD5-12_IGHJ4 N/A 2.059 0.024 19 A0A5C2G1H9 IGL c2982_light_IGKV4-1_IGKJ1 N/A 2.047 0.041 20 A0A024R6Q0 HCG2029544，isoform CRA_a hCG_2029544 2.000 0.025 21 P07737 Profilin-1 PFN1 0.483 0.002 22 A0A5C2GIJ0 IG c818_light_IGKV1D-33_IGKJ4 N/A 0.460 0.001 23 A0A5C2FY46 IGL c1564_light_IGKV4-1_IGKJ4 N/A 0.436 0.012 24 A0A5C2GIW4 IGH + IGL c645_heavy_IGHV3-23_IGHD2-21_IGHJ4 N/A 0.384 0.029 25 A0A3B3IRV3 Malignant T-cell-amplified sequence MCTS2P 0.316 0.004 26 A0A5C2GDY8 IGH + IGL c247_heavy_IGHV3-48_IGHD3-9_IGHJ6 N/A 0.287 0.041 27 A0A5C2GRG0 IG c1788_heavy_IGHV1-46_IGHD5-18_IGHJ3 N/A 0.254 0.012 28 A0A5C2G173 IGL c2300_light_IGKV1D-13_IGKJ4 N/A 0.226 0.002 N/A=not avaible(未提供)。
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