广西医学

目的　通过双样本孟德尔随机化（MR）分析法研究血浆蛋白与HIV感染之间的因果效应，以探寻HIV感染的潜在治疗靶点。方法　基于冰岛癌症项目和deCODE 项目获得的全基因组关联研究（GWAS）数据集获取蛋白数量性状位点（pQTLs），从IEU OpenGWAS Project的GWAS数据库获取HIV感染样本。以血浆蛋白作为暴露因素，HIV感染作为结局，以与血浆蛋白的表达量相关的顺式pQTLs作为工具变量。筛选合格的变量工具后，采用逆方差加权进行双样本双向MR分析。采用Cochran's Q检验、MR⁃Egger截距测试、留一法分析进行敏感性分析。针对MR分析筛选出来的血浆蛋白，基于R语言软件进行基因本体论富集分析及京都基因和基因组百科全书通路富集分析，并利用DGIdb数据库分析其与药物之间的相互作用。将THP⁃1细胞分为实验组（加入HIV⁃1 89.6）和对照组，采用实时荧光定量PCR检测相关基因的mRNA相对表达量，以验证MR分析的结果。结果　共纳入与1 394个血浆蛋白相关的4 144个顺式pQTLs进行MR分析。MR分析结果提示18个血浆蛋白与HIV感染相关，其中9个为保护因素，9个为危险因素。敏感性分析结果提示上述血浆蛋白与HIV感染的因果效应不存在异质性和水平多效性。富集分析结果提示上述血浆蛋白在吞噬和识别、凋亡细胞清除等生物过程中显著富集，并在免疫性疾病相关性通路中显著富集（P<0.05）。成药性评估显示其中5种蛋白已被靶向用于药物开发。与对照组相比，实验组THP⁃1来源巨噬细胞中SIGLEC14的mRNA表达量上调，LEAP2、SERPINA4的mRNA表达量下调（P<0.05）。结论　本研究鉴定的HIV感染相关蛋白标志物，可作为潜在的诊断生物标志物和药物靶点，为HIV感染的治疗提供新视角。

Objective To study the causal effect between plasma proteins and HIV infection by two⁃sample Mendelian randomization (MR) analysis, so as to explore potential therapeutic targets for HIV infection. Methods Protein quantitative trait loci (pQTLs) were obtained from genome⁃wide association studies (GWAS) datasets acquired based on the Icelandic Cancer Project and the deCODE project, and HIV infection samples were obtained from GWAS database of the IEU OpenGWAS Project. Plasma proteins were used as the exposure factor, HIV infection as the outcome, and cis⁃pQTLs related to plasma protein expressions were used as instrumental variables. After screening for qualified variable instruments, the two⁃sample bidirectional MR analyses were performed by using inverse variance weighting. The Cochran's Q test, MR⁃Egger intercept test, and leave⁃one⁃out analysis were adopted to perform sensitivity analysis. For plasma proteins screened by MR analysis, Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed based on the R language software, and their interactions with drugs were analyzed by employing the DGIdb database. THP⁃1 cells were assigned to experiment group (adding HIV⁃189.6) or control group. The real⁃time fluorescent quantitative PCR was adopted to detect mRNA expressions of relevant genes for validating the results of MR analyses. Results A total of 4144 cis⁃pQTLs related to 1394 plasma proteins were enrolled for MR analyses. The results of MR analyses suggested that 18 plasma proteins were related to HIV infection, of which 9 were protective factors and 9 were risk factors. The results of sensitivity analysis revealed that there was no heterogeneity and horizontal pleiotropy of causal effect between plasma proteins as above and HIV infection. The results of enrichment analyses indicated that the aforementioned plasma proteins are significantly enriched in biological processes such as phagocytosis and recognition, apoptotic cell clearance, and in pathways related to immune diseases (P<0.05). Druggability assessment depicted that 5 of these proteins had been targeted for drug development. Compared to the control group, the experiment group exhibited up⁃regulated mRNA expression of SIGLEC14 in THP⁃1⁃derived macrophages, whereas down⁃regulated mRNA expressions of LEAP2 and SERPINA4 (P<0.05). Conclusion The identified HIV infection⁃related protein markers may use as potential diagnostic biomarkers and drug targets, and provide a new perspective for the treatment of HIV infection.