Table_6_Gene expression patterns associated with multidrug therapy in multibacillary leprosy.xls

Multidrug therapy (MDT) has been successfully used in the treatment of leprosy. However, although patients are cured after the completion of MDT, leprosy reactions, permanent disability, and occasional relapse/reinfection are frequently observed in patients. The immune system of multibacillary patients (MB) is not able to mount an effective cellular immune response against M. leprae. Consequently, clearance of bacilli from the body is a slow process and after 12 doses of MDT not all MB patients reduce bacillary index (BI). In this context, we recruited MB patients at the uptake and after 12-month of MDT. Patients were stratified according to the level of reduction of the BI after 12 doses MDT. A reduction of at least one log in BI was necessary to be considered a responder patient. We evaluated the pattern of host gene expression in skin samples with RNA sequencing before and after MDT and between samples from patients with or without one log reduction in BI. Our results demonstrated that after 12 doses of MDT there was a reduction in genes associated with lipid metabolism, inflammatory response, and cellular immune response among responders (APOBEC3A, LGALS17A, CXCL13, CXCL9, CALHM6, and IFNG). Also, by comparing MB patients with lower BI reduction versus responder patients, we identified high expression of CDH19, TMPRSS4, PAX3, FA2H, HLA-V, FABP7, and SERPINA11 before MDT. From the most differentially expressed genes, we observed that MDT modulates pathways related to immune response and lipid metabolism in skin cells from MB patients after MDT, with higher expression of genes like CYP11A1, that are associated with cholesterol metabolism in the group with the worst response to treatment. Altogether, the data presented contribute to elucidate gene signatures and identify differentially expressed genes associated with MDT outcomes in MB patients.

多药联合治疗（Multidrug therapy, MDT）已成功应用于麻风病的临床治疗。然而，尽管患者在完成多药联合治疗后可获得临床治愈，但仍有不少患者会出现麻风反应、永久性残疾，偶见复发或再感染病例。多菌型麻风患者（multibacillary patients, MB）的免疫系统无法针对麻风分枝杆菌（Mycobacterium leprae, M. leprae）产生有效的细胞免疫应答。因此，体内分枝杆菌的清除过程较为缓慢，且在完成12个剂量的多药联合治疗后，并非所有多菌型麻风患者的菌量指数（bacillary index, BI）都会出现下降。在此背景下，本研究招募了多菌型麻风患者，分别在其入组时以及完成12个月多药联合治疗后进行样本采集。研究人员根据患者在完成12个剂量的多药联合治疗后菌量指数的下降幅度对其进行分层：若患者的菌量指数下降至少一个对数级，则被归类为治疗应答者。本研究通过RNA测序技术，对多药联合治疗前后的皮肤样本，以及菌量指数下降与否的患者皮肤样本中的宿主基因表达模式进行了分析。研究结果显示，在完成12个剂量的多药联合治疗后，治疗应答者体内与脂质代谢、炎症反应及细胞免疫应答相关的基因表达出现下调，其中包括APOBEC3A、LGALS17A、CXCL13、CXCL9、CALHM6以及IFNG。此外，通过对比菌量指数下降幅度较低的多菌型麻风患者与治疗应答者，本研究发现，在多药联合治疗前，CDH19、TMPRSS4、PAX3、FA2H、HLA-V、FABP7以及SERPINA11在前者体内呈现高表达。通过对差异表达最显著的基因进行分析，我们发现多药联合治疗可调控多菌型麻风患者皮肤细胞中与免疫应答及脂质代谢相关的通路；在治疗应答最差的患者组中，与胆固醇代谢相关的CYP11A1等基因呈现高表达。综上，本研究提供的数据有助于阐明多菌型麻风患者多药联合治疗预后相关的基因特征，并筛选出与治疗结局相关的差异表达基因。