mRNA and miRNA Screening to Identify Treatment Responses in SLE Patients Receiving Belimumab

Systemic lupus erythematosus (SLE) is a complex autoimmune disease driven by autoreactive B cells and the cytokine BAFF (BLyS), which is vital for B-cell function. Despite advances in genomics, no clear profile exists to predict which patients benefit most from BLyS-targeted therapies. miRNAs, small noncoding RNAs, regulate numerous cellular activities and have emerged as potential biomarkers for autoimmune diseases like SLE. Dysregulated miRNAs in SLE T and B cells contribute to immune hyperactivation, promoting autoantibody production and heightened type 1 interferon levels. While miRNAs are promising diagnostic tools, their role in predicting responses to anti-BLyS therapy remains unexplored. This study examines miRNA-mRNA interaction networks in T cells, B cells, and myeloid cells from SLE patients before and after BLyS therapy, identifying differentially expressed miRNAs and mRNAs. Integrative analyses reveal novel regulatory networks and pathways associated with clinical improvement, offering new insights into the pharmacological mechanisms of anti-BLyS therapy. PBMCs were sorted using spectral cytometry to isolate B cells, T cells, and myeloid cell populations. RNA and miRNA were extracted from each subset, followed by a microarray analysis. Gene expression profiling was conducted using the Clariom S assay, and miRNA expression profiling was performed with GeneChip miRNA 4.0 arrays (Thermo Fisher Scientific, Inc.).

系统性红斑狼疮（Systemic lupus erythematosus, SLE）是一种由自身反应性B细胞及细胞因子BAFF（BLyS）驱动的复杂自身免疫性疾病，BAFF对B细胞功能至关重要。尽管基因组学领域已取得诸多进展，但目前尚无明确的特征谱可预测哪些患者能从BLyS靶向治疗中获益最多。
微小RNA（miRNAs, microRNAs）是一类小型非编码RNA，可调控多种细胞活动，现已成为系统性红斑狼疮等自身免疫性疾病的潜在生物标志物。SLE患者T细胞与B细胞中失调的miRNAs会加剧免疫过度活化，促进自身抗体产生并升高I型干扰素水平。尽管miRNAs具备成为诊断工具的潜力，但其在预测抗BLyS治疗应答方面的作用仍未得到探索。
本研究针对接受BLyS治疗前后的SLE患者的T细胞、B细胞及髓系细胞中的miRNA-mRNA互作网络展开分析，鉴定出差异表达的miRNAs与mRNAs。整合分析揭示了与临床改善相关的新型调控网络及通路，为抗BLyS治疗的药理学机制提供了全新见解。研究人员通过光谱流式细胞术对外周血单个核细胞（PBMCs, peripheral blood mononuclear cells）进行分选，以分离B细胞、T细胞及髓系细胞群。从每个细胞亚群中提取RNA与miRNA，随后进行微阵列分析。基因表达谱分析采用Clariom S检测试剂盒，miRNA表达谱分析则使用GeneChip miRNA 4.0芯片（赛默飞世尔科技公司，Thermo Fisher Scientific, Inc.）。