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指导自身免疫性肾上腺皮质功能减退症（AAD）中的糖皮质激素（GC）替代治疗，导致过度治疗伴随令人担忧且持久的副作用，或治疗不足，可能危及生命。研究目标：探讨不同GC替代剂量下基因表达的变化，以识别潜在的转录生物标志物，从而指导AAD中的GC替代治疗。研究方法：第一步：对10例AAD患者的全血RNA进行全局微阵列表达分析，以比较在静脉注射100毫克氢化可的松（HC）前后基因表达水平。在三个上调最显著的基因中，我们通过实时PCR（rt-PCR）比较了注射HC前及注射后两、四、六小时的基因表达水平。第二步：对27例AAD患者的正常与极低早晨皮质醇水平进行比较，通过rt-PCR比较93个GC调控基因的表达水平。结果：第一步：注射100毫克HC后两小时，FKBP5、MMP9和DSIPI的表达水平显著升高。MMP9和DSIPI的表达水平与血清皮质醇水平相关。第二步：与极低早晨皮质醇水平相比，CEBPB、DDIT4、FKBP5、DSIPI和VDR的表达水平升高，而ADARB1、ARIDB5和POU2F1的表达水平降低。正常血清皮质醇水平与DSIPI、DDIT4和FKBP5的表达呈正相关。结论：本研究引入基因表达作为指导AAD中GC替代治疗的新方法。我们提出，DSIPI、DDIT4和FKBP5的基因表达是GC替代治疗的潜在生物标志物，其次为MMP9、CEBPB、VDR、ADARB1、ARID5B和POU2F1。