Table12_Immune cell infiltration-related clinical diagnostic model for Ankylosing Spondylitis.XLSX

Background: The pathogenesis and diagnosis of Ankylosing Spondylitis (AS) has remained uncertain due to several reasons, including the lack of studies on the local and systemic immune response in AS. To construct a clinical diagnostic model, this study identified the micro RNA-messenger RNA (miRNA-mRNA) interaction network and immune cell infiltration-related hub genes associated with AS.Materials and Methods: Total RNA was extracted and purified from the interspinous ligament tissue samples of three patients with AS and three patients without AS; miRNA and mRNA microarrays were constructed using the extracted RNA. Bioinformatic tools were used to construct an miRNA-mRNA network, identify hub genes, and analyze immune infiltration associated with AS. Next, we collected the blood samples and clinical characteristics of 359 patients (197 with AS and 162 without AS). On the basis of the clinical characteristics and results of the routine blood tests, we selected immune-related cells whose numbers were significantly different in patients with AS and patients without AS. Univariate and multivariate logistic regression analysis was performed to construct a nomogram. Immunohistochemistry staining analysis was utilized to verify the differentially expression of LYN in AS and controls.Results: A total of 225 differentially expressed miRNAs (DE miRNAs) and 406 differentially expressed mRNAs (DE mRNAs) were identified from the microarray. We selected 15 DE miRNAs and 38 DE mRNAs to construct a miRNA-mRNA network. The expression of LYN, an immune-related gene, correlated with the counts of monocytes, neutrophils, and dendritic cells. Based on the independent predictive factors of sex, age, and counts of monocytes, neutrophils, and white blood cells, a nomogram was established. Receiver operating characteristic (ROC) analysis was performed to evaluate the nomogram, with a C-index of 0.835 and AUC of 0.855.Conclusion:LYN, an immune-related hub gene, correlated with immune cell infiltration in patients with AS. In addition, the counts of monocytes and neutrophils were the independent diagnostic factors for AS. If verified in future studies, a diagnostic model based on these findings may be used to predict AS effectively.

背景：由于多种原因，包括对强直性脊柱炎（AS）的局部和全身免疫反应研究不足，强直性脊柱炎的发病机制和诊断一直处于不确定状态。本研究旨在构建临床诊断模型，因此识别了与AS相关的miRNA-mRNA相互作用网络和免疫细胞浸润相关核心基因。材料与方法：从三位AS患者和三位非AS患者的椎间韧带组织样本中提取和纯化总RNA；利用提取的RNA构建了miRNA和mRNA微阵列。采用生物信息学工具构建miRNA-mRNA网络，识别核心基因，并分析与AS相关的免疫浸润。接下来，我们收集了359位患者的血液样本和临床特征（197位AS患者和162位非AS患者）。基于临床特征和常规血液检查结果，我们选择了在AS患者和非AS患者中数量显著不同的免疫相关细胞。通过单因素和多因素逻辑回归分析构建了评分系统。利用免疫组化染色分析验证了LYN在AS患者和对照组中的差异表达。结果：从微阵列中鉴定出225个差异表达miRNA（DE miRNA）和406个差异表达mRNA（DE mRNA）。我们选择了15个DE miRNA和38个DE mRNA来构建miRNA-mRNA网络。免疫相关基因LYN的表达与单核细胞、中性粒细胞和树突状细胞的数量相关。基于性别、年龄以及单核细胞、中性粒细胞和白细胞计数等独立预测因素，建立了评分系统。通过受试者工作特征（ROC）分析评估评分系统，C指数为0.835，AUC为0.855。结论：免疫相关核心基因LYN与AS患者的免疫细胞浸润相关。此外，单核细胞和中性粒细胞的数量是AS的独立诊断因素。如果在未来研究中得到验证，基于这些发现的诊断模型可能被有效地用于预测AS。