Division of Pulmonary and Critical Care Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan; Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.
Division of Pulmonary and Critical Care Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan; Graduate Institute of Clinical Medical Sciences, Taiwan.
J Infect. 2022 Nov;85(5):519-533. doi: 10.1016/j.jinf.2022.08.035. Epub 2022 Aug 31.
RNA therapeutics is an emerging field that widens the range of treatable targets and would improve disease outcome through bypassing the antibiotic bactericidal targets to kill Mycobacterium tuberculosis (M.tb).
We screened for microRNA with immune-regulatory functions against M.tb by next generation sequencing of peripheral blood mononuclear cells, followed by validation in an independent cohort.
Twenty three differentially expressed microRNAs were identified between 12 active pulmonary TB patients and 4 healthy subjects, and 35 microRNAs before and after 6-month anti-TB therapy. Enriched predicted target pathways included proteoglycan, HIF-1 signaling, longevity-regulating, central carbon metabolism, and autophagy. We validated miR-431-3p down-regulation and miR-1303 up-regulation accompanied with corresponding changes in their predicted target genes in an independent validation cohort of 46 active TB patients, 30 latent TB infection subjects, and 24 non-infected healthy subjects. In vitro experiments of transfections with miR-431-3p mimic/miR-1303 short interfering RNA in THP-1 cells under ESAT-6 stimuli showed that miR-431-3p and miR-1303 were capable to augment and suppress autophagy/apoptosis/phagocytosis of macrophage via targeting MDR1/MMP16/RIPOR2 and ATG5, respectively.
This study provides a proof of concept for microRNA-based host-directed immunotherapy for active TB disease. The combined miR-431-3p over-expression and miR-1303 knock-down revealed new vulnerabilities of treatment-refractory TB disease.
RNA 疗法是一个新兴领域，它拓宽了可治疗靶标的范围，并通过绕过抗生素杀菌靶标来杀死结核分枝杆菌（M.tb），从而改善疾病结局。
我们通过对外周血单核细胞进行下一代测序筛选具有免疫调节功能的 microRNA，然后在独立队列中进行验证。
在 12 例活动性肺结核患者和 4 例健康对照者之间，我们鉴定出 23 个差异表达的 microRNA，以及在 6 个月抗结核治疗前后，鉴定出 35 个 microRNA。富集的预测靶标途径包括蛋白聚糖、HIF-1 信号、长寿调节、中心碳代谢和自噬。我们在 46 例活动性肺结核患者、30 例潜伏性结核感染患者和 24 例非感染健康对照者的独立验证队列中验证了 miR-431-3p 的下调和 miR-1303 的上调，以及它们预测的靶基因的相应变化。在 ESAT-6 刺激下，用 miR-431-3p 模拟物/miR-1303 短发夹 RNA 转染 THP-1 细胞的体外实验表明，miR-431-3p 和 miR-1303 能够通过靶向 MDR1/MMP16/RIPOR2 和 ATG5 分别增强和抑制巨噬细胞的自噬/凋亡/吞噬作用。
本研究为基于 microRNA 的针对活动性结核病的宿主导向免疫治疗提供了概念验证。联合 miR-431-3p 过表达和 miR-1303 敲低揭示了治疗耐药性结核病的新弱点。
