DataSheet_2_Gene expression patterns associated with multidrug therapy in multibacillary leprosy.pdf

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.

多药联合疗法（Multidrug therapy, MDT）已成功应用于麻风病的临床治疗。尽管患者在完成MDT疗程后可获得治愈，但仍有部分患者会频繁出现麻风反应、永久性残疾，偶见复发或再感染情况。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）发起有效的细胞免疫应答，因此机体清除病菌的进程极为缓慢，在完成12剂MDT疗程后，并非所有多菌型患者的菌量指数（bacillary index, BI）都能出现下降。
在此研究背景下，我们招募了多菌型麻风患者，分别在入组时以及接受12个月MDT疗程后采集皮肤样本。研究人员根据患者完成12剂MDT疗程后的菌量指数下降幅度进行分层：菌量指数至少下降1个对数单位的患者被划定为应答者。我们通过RNA测序（RNA sequencing）技术，对患者皮肤样本中的宿主基因表达模式进行了分析，对比了MDT治疗前后的样本，以及菌量指数出现/未出现1个对数单位下降的患者样本。
研究结果显示，在应答者群体中，完成12剂MDT疗程后，与脂质代谢、炎症反应及细胞免疫应答相关的基因（如APOBEC3A、LGALS17A、CXCL13、CXCL9、CALHM6及IFNG）的表达水平显著下调。此外，通过对比菌量指数下降幅度较低的患者与应答者，我们发现MDT治疗前，CDH19、TMPRSS4、PAX3、FA2H、HLA-V、FABP7及SERPINA11等基因在前者中呈现高表达。
从差异表达最显著的基因分析来看，MDT可调节多菌型患者皮肤细胞中与免疫应答及脂质代谢相关的通路；在治疗应答较差的群体中，CYP11A1等与胆固醇代谢相关的基因表达水平更高。总体而言，本研究呈现的数据有助于阐明多菌型麻风患者的基因特征谱，并识别出与MDT治疗结局相关的差异表达基因。