Department of Pediatrics Laboratory, The Affiliated Wuxi Children's Hospital of Jiangnan University, Wuxi 214023, Jiangsu Province, China.
Department of Clinical Laboratory, Huishan District Hospital, Wuxi 214187, Jiangsu Province, China.
Int Immunopharmacol. 2023 Jun;119:110249. doi: 10.1016/j.intimp.2023.110249. Epub 2023 May 3.
A lack of validated blood diagnostic markers presents an obstacle to asthma control. The present study sought to profile the plasma proteins of children with asthma and to determine potential biomarkers. Plasma samples from children in acute exacerbation (n = 4), in clinical remission (n = 4), and from healthy children (n = 4, control) were analyzed using a tandem mass tag (TMT)-labeling quantitative proteomics and the candidate biomarkers were validated using liquid chromatography-parallel reaction monitoring (PRM)/mass spectrometry (MS) with enzyme-linked immunosorbent assay (ELISA). We identified 347 proteins with differential expression between groups: 125 (50 upregulated, 75 downregulated) between acute exacerbation and control, 142 (72 upregulated, 70 downregulated) between clinical remission and control, and 55 (22 upregulated, 33 downregulated) between acute and remission groups (all between-group fold changes > 1.2; P < 0.05 by Student's t-test). Gene ontology analysis implicated differentially expressed proteins among children with asthma in immune response, the extracellular region, and protein binding. Further, KEGG pathway analysis of differentially expressed proteins identified complement and coagulation cascades and Staphylococcus aureus infection pathways as having the highest protein aggregation. Our analyses of protein interactions identified important node proteins, particularly KRT10. Among 11 differentially expressed proteins, seven proteins (IgHD, IgHG4, AACT, IgHA1, SAA, HBB, and HBA1) were verified through PRM/MS. Protein levels of AACT, IgA, SAA, and HBB were verified through ELISA and may be useful as biomarkers to identify individuals with asthma. In conclusion, our study presents a novel comprehensive analysis of changes in plasma proteins in children with asthma and identifies a panel for accessory diagnosis of pediatric asthma.
缺乏经过验证的血液诊断标志物是哮喘控制的障碍。本研究旨在分析儿童哮喘患者的血浆蛋白谱，并确定潜在的生物标志物。使用串联质量标签 (TMT) 标记定量蛋白质组学分析急性加重期 (n = 4)、临床缓解期 (n = 4) 和健康儿童 (n = 4，对照组) 的血浆样本，并使用液相色谱-平行反应监测 (PRM)/质谱 (MS) 和酶联免疫吸附测定 (ELISA) 验证候选生物标志物。我们在组间鉴定出 347 种具有差异表达的蛋白质：急性加重期与对照组之间有 125 种 (50 种上调，75 种下调)，临床缓解期与对照组之间有 142 种 (72 种上调，70 种下调)，急性期与缓解期之间有 55 种 (22 种上调，33 种下调) (所有组间折叠变化 > 1.2；学生 t 检验 P < 0.05)。GO 分析表明哮喘儿童差异表达蛋白参与免疫反应、细胞外区和蛋白结合。此外，差异表达蛋白的 KEGG 途径分析确定补体和凝血级联以及金黄色葡萄球菌感染途径具有最高的蛋白聚集。我们对蛋白质相互作用的分析确定了重要的节点蛋白，特别是 KRT10。在 11 种差异表达蛋白中，有 7 种蛋白 (IgHD、IgHG4、AACT、IgHA1、SAA、HBB 和 HBA1) 通过 PRM/MS 得到验证。通过 ELISA 验证了 AACT、IgA、SAA 和 HBB 的蛋白水平，它们可能是识别哮喘患者的有用生物标志物。总之，本研究对儿童哮喘患者血浆蛋白变化进行了全面分析，并确定了一组用于儿童哮喘辅助诊断的标志物。
