Evidence of Dynamically Dysregulated Gene Expression Pathways in Hyper-responsive B Cells from African American Lupus Patients. Homo sapiens

In this report we applied standard and original methods of analysis of gene expression microarray data to delineate differences in the molecular pathways impacted by stimulation of Epstein-Barr virus (EBV) transformed B cells derived from patients with systemic lupus erythematosus (SLE) and normal unrelated controls. In order to understand the dynamics and interconnections the B cell molecular pathways, the system was perturbed with a biologically relevant signal, co-crosslinking of the B cell antigen receptor (BCR) and FcγR2b and global gene expression changes were assessed at various timepoints post-stimulation. Both traditional analysis of differential gene expression and analysis of the dynamics of gene expression variations were performed. Combining these two methods in an analysis process we call Pathway Dysregulation Analysis allows us to establish model networks of functional gene expression important for B cell signaling responses and elucidate gene expression regulatory interconnections disrupted in B cells from individuals with lupus. Through this technique, we found two main groups of gene associations changed significantly with the disease phenotype, which included genes with established controlling function of the B cell activation, and genes involved in apoptosis initiation or prevention. Overall design: Epstein-Barr virus (EBV) transformed B cells derived from two patients with systemic lupus erythematosus (SLE) and two normal unrelated controls were stimulated with a biologically relevant signal, co-crosslinking of the B cell antigen receptor (BCR) and FcγR2b. Total RNA was isolated at various timepoints post-stimulation. Gene expression data were used for analysis of differential gene expression and analysis of the dynamics of gene expression variations.

本研究采用标准分析方法与原创分析手段对基因表达微阵列数据开展分析，旨在阐明系统性红斑狼疮（systemic lupus erythematosus, SLE）患者来源的Epstein-Barr病毒（Epstein-Barr virus, EBV）转化B细胞，与健康无关对照个体来源的同类细胞在受刺激后其分子通路的差异。为解析B细胞分子通路的动态变化与相互关联，我们通过生物学相关刺激信号——B细胞抗原受体（B cell antigen receptor, BCR）与FcγR2b共交联——对细胞系统施加扰动，并在刺激后的多个时间点评估全基因组基因表达变化。本研究同时开展了差异基因表达分析与基因表达变异动态分析。将这两种方法结合于我们命名为通路失调分析（Pathway Dysregulation Analysis）的分析流程中，可构建与B细胞信号转导应答密切相关的功能性基因表达模型网络，并阐明狼疮患者B细胞中被破坏的基因表达调控相互关联。通过该分析技术，我们发现与疾病表型显著相关的两大基因关联模块：其一为已证实具有B细胞活化调控功能的基因，其二为参与细胞凋亡启动或抑制过程的基因。实验整体设计：从2名系统性红斑狼疮（SLE）患者与2名健康无关对照个体体内分离B细胞并经EBV转化，随后通过BCR与FcγR2b共交联的生物学相关刺激信号进行处理。分别在刺激后的多个时间点提取总RNA，利用所得基因表达数据开展差异基因表达分析与基因表达变异动态分析。