DNA microarray-based approach for identifying new therapeutic targets in neuroimmunological diseases

使用定制的cDNA微阵列，我们分析了来自多发性硬化症（MS）的外周血T细胞的基因表达谱，这些患者未经治疗或用干扰素（IFN）-β治疗。本研究旨在深入了解MS的发病机制和IFN-β的作用机制。我们已经发现在MS T细胞中上调或下调的许多基因是与凋亡、炎症或DNA损伤调节相关的基因。在MS中上调的凋亡相关基因中，我们选择了Nurr 4A 2（Nurrl）并深入分析了其意义。已知NR 4A 2靶向骨桥蛋白（OPN），一种Th 1细胞因子，被认为参与MS的发病机制。我们假设NR 4A 2和OPN可能在MS中协同改变，但事实并非如此。相反，我们发现NR 4A 2的表达将与NF κ B靶向的IkB α和IkB β的表达或TNF α IP 3的表达强烈相关，TNF α IP 3可被TNF信号转导诱导。提示NF-κ B相关信号可能引起基因表达的改变，抗炎治疗可能纠正这种改变。我们还通过分析IFN治疗患者的血液研究了人类MS中的IFN应答基因。立即诱导的基因包含炎性细胞因子如IL-6和炎性趋化因子如CXCR 3配体，这将解释IFN-b在MS患者中的早期副作用，包括头痛和发热。这些结果可能提示抗炎治疗在MS中的重要性。

Using a custom cDNA microarray, we have analyzed gene expression profiling of peripheral blood T cells derived from multiple sclerosis (MS) who are untreated or treated with interferon (IFN)-β. This study was aimed at obtaining insights into the pathogenesis of MS and the mechanism of action of IFN-β. We have found that a number of genes up- or down-regulated in MS T cells, are those related to apoptosis, inflammation, or DNA damage-regulation. Among apoptosis-related genes up-regulated in MS, we have chosen Nurr4A2 (Nurrl) and analyzed the implication in depth. NR4A2 is known to target osteopontin (OPN), a Th1 cytokine, thought to be involved in the pathogenesis of MS. We hypothesized that NR4A2 and OPN might be coordinately altered in MS, but this was not the case. Rather, we found that expression of NR4A2 would strongly correlate with that of IkB alpha and IkB ipsilon, targeted by NFkB or that of TNFAIP3, that is inducible with TNF signaling. This suggests that NFkB linked signals would cause the change of gene expression, which is probably corrected by anti-inflamunatory therapy. We also investigated IFN-responsive genes in human MS by analyzing the blood from IFN-treated patients. Immediately induced genes contained inflammatory cytokines such as IL-6 and inflammatory chemokines such as CXCR3 ligand, which would account for the early side effects of IFN-b in MS patients, including headache and fever up. These results may suggest the importance of anti-inflammatory therapy in MS.