Replication data for: Potential Transcriptional Biomarkers to Guide Glucocorticoid Replacement in Autoimmune Addison’s Disease

Background: No reliable biomarkers exist to guide glucocorticoid (GC) replacement treatment in autoimmune Addison’s disease (AAD), leading to overtreatment with alarming and persistent side-effects or undertreatment, which could be fatal. Objective: To explore changes in gene expression following different GC replacement doses as a means of identifying candidate transcriptional biomarkers to guide GC replacement in AAD. Methods: Step 1: Global microarray expression analysis on RNA from whole blood before and after intravenous infusion of 100 mg hydrocortisone (HC) in 10 patients with AAD. In three of the most highly upregulated genes, we performed real-time PCR (rt-PCR) to compare gene expression levels before and two, four, and six hours after the HC infusion. Step 2: Rt-PCR to compare expression levels of 93 GC-regulated genes in normal versus very low morning cortisol levels in 27 patients with AAD. Results: Step 1: Two hours after infusion of 100 mg HC, there was a marked increase in FKBP5, MMP9, and DSIPI expression levels. MMP9 and DSIPI expression levels correlated with serum cortisol. Step 2: Expression levels of CEBPB, DDIT4, FKBP5, DSIPI, and VDR were increased and ADARB1, ARIDB5, and POU2F1 decreased in normal versus very low morning cortisol. Normal serum cortisol levels positively correlated with DSIPI, DDIT4, and FKBP5 expression. Conclusions: We introduce gene expression as a novel approach to guide GC replacement in AAD. We suggest that gene expression of DSIPI, DDIT4, and FKBP5 are particularly promising candidate biomarkers of GC replacement, followed by MMP9, CEBPB, VDR, ADARB1, ARID5B, and POU2F1.背景：目前尚无可靠的生物标志物可用于指导自身免疫性艾迪生病（autoimmune Addison’s disease, AAD）患者的糖皮质激素（glucocorticoid, GC）替代治疗，这可能导致治疗过度（伴随令人担忧且持续存在的不良反应）或治疗不足，甚至可能致命。研究目的：探索不同糖皮质激素替代剂量下的基因表达变化，以期筛选出可用于指导自身免疫性艾迪生病患者糖皮质激素替代治疗的潜在转录生物标志物。方法：步骤1：纳入10名自身免疫性艾迪生病患者，分别采集其静脉输注100mg氢化可的松（hydrocortisone, HC）前后的全血RNA，进行全基因组微阵列表达分析；选取上调幅度最高的3个基因，采用实时荧光定量PCR（real-time PCR, rt-PCR）对比输注前、输注后2、4、6小时的基因表达水平。步骤2：纳入27名自身免疫性艾迪生病患者，按照清晨皮质醇水平分为正常组与极低水平组，采用实时荧光定量PCR对比两组中93个糖皮质激素调控基因的表达水平。结果：步骤1结果：输注100mg氢化可的松后2小时，FKBP5、MMP9及DSIPI的表达水平显著升高；MMP9与DSIPI的表达水平与血清皮质醇水平呈显著相关。步骤2结果：相较于清晨皮质醇极低水平组，正常皮质醇组的CEBPB、DDIT4、FKBP5、DSIPI及VDR表达水平升高，而ADARB1、ARIDB5及POU2F1表达水平降低；血清皮质醇正常水平与DSIPI、DDIT4及FKBP5的表达呈正相关。结论：本研究提出将基因表达作为指导自身免疫性艾迪生病患者糖皮质激素替代治疗的全新策略；我们认为DSIPI、DDIT4及FKBP5的基因表达是极具潜力的糖皮质激素替代治疗生物标志物候选者，其次为MMP9、CEBPB、VDR、ADARB1、ARID5B及POU2F1。