SARS-CoV-2感染与非感染人群血清1-10 ku蛋白质组差异性分析

Differential proteomic analysis of 1-10-ku proteins in serum from people infected with SARS-CoV-2 and people not infected with SARS-CoV-2

目的通过比较分析新型冠状病毒(SARS-CoV-2)感染人群与新冠肺炎疫情爆发前后非SARS-CoV-2感染人群血清的差异蛋白质组,从蛋白水平发现SARS-CoV-2感染人群1-10 ku的特异性血清生物标志物,并确定新冠肺炎疫情爆发前后非SARS-CoV-2感染人群1-10 ku血清蛋白表达的差异性。方法采集166例新冠肺炎爆发前非SARS-CoV-2感染人群血清、245例新冠肺炎爆发后非SARS-CoV-2感染人群血清及228例SARS-CoV-2感染人群血清,每组血清标本分为3份进行预混,形成3组生物学重复,弱阳离子磁珠富集后使用非标记蛋白质组定量技术对富集得到的1-10 ku小分子蛋白或多肽进行非标定量分析。结果蛋白质组学定量分析得到332个蛋白。差异分析显示,新冠肺炎疫情爆发前后非SARS-CoV-2感染人群血清蛋白组分构成差异无统计学意义(P>0.05),SARS-CoV-2感染人群血清与新冠肺炎疫情爆发前后非SARS-CoV-2感染人群血清相比,筛选到差异蛋白49个(上调表达蛋白20个,下调表达蛋白29个),其中19个差异蛋白已作为癌症、心脑血管等其他疾病的诊断或者预后生物标志物。IPA经典信号通路分析显示,差异蛋白显著富集到LXR/RXR途径、FXR/RXR途径、急性期反应信号传导、补体系统、巨噬细胞IL-12信号传导与产生、凝血系统、网格蛋白介导的内吞信号传导等通路。IPA疾病功能分析显示,差异蛋白富集到细胞妥协、炎症反应、传染性疾病、蛋白合成等。在血液中检测到PEBP4、SH3BGRL3基因产物,以及GNPTG、THBS1、HSPG2、MAN1A1、LGALS3BP、CLSTN1、IGFALS、PCOLCE、PGLYRP2、PRG4、QSOX1、SPARC、ATRN 13个未曾报道的与SARS-CoV-2感染相关的差异表达基因。结论新冠肺炎疫情爆发前后非SARS-CoV-2感染人群血清1-10 ku蛋白组分无显著不同,而SARS-CoV-2感染人群血清与非SARS-CoV-2感染人群血清相比存在多个差异蛋白,为新型冠状病毒病(COVID-19)的致病机制和治疗靶点研究提供了依据。

Objective To identify specific serum biomarkers of 1-10-ku proteins in people infected with SARS-CoV-2 at the protein level through comparative proteomic analysis of serum from people infected with SARS-CoV-2 and people not infected with SARS-CoV-2 and to determine the significance of protein expression profiles of 1-10-ku proteins in people not infected with SARS-CoV-2 before and after an outbreak of COVID-19. Methods Serum samples were collected from 166 people not infected with SARS-CoV-2 before a COVID-19 outbreak, 245 people not infected with SARS-CoV-2 after a COVID-19 outbreak, and 228 people infected with SARS-CoV-2. Serum samples from each group were premixed into 3 copies to form 3 groups of biological replicates. After enrichment using weak cationic magnetic beads, 1-10-ku small molecule proteins or peptides were analyzed using label-free quantitative proteomic analysis. Results Quantitative proteomic analysis yielded a total of 332 proteins. Differential analysis revealed no significant differences in serum profiles before and after the outbreak of COVID-19(P>0.05). A total of 49 differential proteins(20 up-regulated proteins and 29 down-regulated proteins) were screened in serum from people infected with SARS-CoV-2 compared to serum from people not infected with SARS-CoV-2. Of those, 19 differential proteins have been used as diagnostic or prognostic biomarkers for cancer, cardiovascular and cerebrovascular diseases, and other diseases. IPA classical signaling pathway analysis revealed significant enrichment in the LXR/RXR pathway, FXR/RXR pathway, acute phase response signaling, the coagulation system, the complement system, macrophage IL-12 signaling and production, the coagulation system, and clathrin-mediated endocytosis signaling. The diseases and functional modules enriched by differential proteins are related to cellular compromise, inflammatory response, infectious diseases, and protein synthesis. PEBP4 and SH3 BGRL3 gene products were detected in blood, along with 13 previously unreported differentially expressed gene products including GNPTG, THBS1, HSPG2, MAN1 A1, LGALS3 BP, CLSTN1, IGFALS, PCOLCE, PGLYRP2, PRG4, QSOX1, SPARC, and ATRN. Conclusion There were no significant differences in 1-10-ku proteins in serum from people not infected with SARS-CoV-2 before and after an outbreak of the SARS-CoV-2. The serum protein expression profile of people infected with SARS-CoV-2 differed significantly from that of people not infected with SARS-CoV-2. This finding has provided a basis for study of the mechanism and therapeutic targets of COVID-19.