DataSheet_1_Novel Insights Into Gene Signatures and Their Correlation With Immune Infiltration of Peripheral Blood Mononuclear Cells in Behcet’s Disease.pdf

BackgroundBehcet’s disease (BD) is a chronic inflammatory disease that involves systemic vasculitis and mainly manifests as oral and genital ulcers, uveitis, and skin damage as the first clinical symptoms, leading to gastrointestinal, aortic, or even neural deterioration. There is an urgent need for effective gene signatures for BD’s early diagnosis and elucidation of its underlying etiology.
MethodsWe identified 82 differentially expressed genes (DEGs) in BD cases compared with healthy controls (HC) after combining two Gene Expression Omnibus datasets. We performed pathway analyses on these DEGs and constructed a gene co-expression network and its correlation with clinical traits. Hub genes were identified using a protein–protein interaction network. We manually selected CCL4 as a central hub gene, and gene-set enrichment and immune cell subset analyses were applied on patients in high- and low-CCL4 expression groups. Meanwhile, we validated the diagnostic value of hub genes in differentiating BD patients from HC in peripheral blood mononuclear cells using real-time PCR.
ResultsTwelve hub genes were identified, and we validated the upregulation of CCL4 and the downregulation of NPY2R mRNA expression. Higher expression of CCL4 was accompanied by larger fractions of CD8 + T cells, natural killer cells, M1 macrophages, and activated mast cells. Receiver operator characteristic curves showed good discrimination between cases and controls based on the expression of these genes.
ConclusionCCL4 and NPY2R could be diagnostic biomarkers for BD that reveal inflammatory status and predict vascular involvement in BD, respectively.

【背景】贝赫切特病（Behcet’s disease, BD）是一种以全身血管炎为主要病理特征的慢性炎症性疾病，首发临床表现以口腔与生殖器溃疡、葡萄膜炎及皮肤损害为主，病情可进展累及胃肠道、主动脉甚至神经系统，引发功能恶化。目前临床亟需有效的基因标志物以实现BD的早期诊断，并阐明其潜在发病机制。
【方法】本研究整合两项基因表达综合（Gene Expression Omnibus, GEO）数据集，对比贝赫切特病患者与健康对照（healthy controls, HC）的转录组数据，筛选得到82个差异表达基因（differentially expressed genes, DEGs）。随后对上述DEGs进行通路富集分析，构建基因共表达网络并分析其与临床表型的相关性；通过蛋白质-蛋白质相互作用（protein–protein interaction, PPI）网络筛选核心枢纽基因。本研究手动选取CCL4作为核心枢纽基因，分别对CCL4高、低表达组患者开展基因集富集分析与免疫细胞亚群分析。同时，本研究通过实时聚合酶链式反应（real-time PCR）在外周血单个核细胞中验证核心基因区分BD患者与健康对照的诊断价值。
【结果】本研究共筛选得到12个核心枢纽基因，并验证了CCL4的mRNA表达上调与NPY2R的mRNA表达下调。CCL4高表达组伴随CD8+T细胞、自然杀伤细胞、M1型巨噬细胞及活化肥大细胞的浸润比例显著升高。受试者工作特征（Receiver Operating Characteristic, ROC）曲线分析显示，基于上述核心基因的表达水平可较好地区分BD患者与健康对照。
【结论】CCL4与NPY2R可分别作为贝赫切特病的诊断生物标志物，其中CCL4可反映疾病的炎症状态，NPY2R可预测BD的血管受累情况。