Next generation sequencing reveals miR-431-3p and miR-1303 as immune-regulatory microRNA for active pulmonary tuberculosis through targeting MDR1/MMP16/RIPOR2 and ATG5 signaling

Abstract

<b>Background:</b> Host-directed therapy, such as RNA therapeutics, in combination with current standard antimicrobial treatments would improve disease outcome through bypassing the antibiotic bactericidal targets to kill Mycobacterium tuberculosis (M.tb). <b>Methods:</b> We sought to screen for microRNA with immune-regulatory functions against M.tb by using next generation sequencing. <b>Results:</b> 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 based on biological process included cell apoptosis, pyruvate metabolic, macroautophagy. We verified miR-431-3p down-regulation and miR-1303 up-regulation accompanied with corresponding changes in their 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 and miR-1303&nbsp;Si RNA in THP-1 cells under ESAT-6 stimuli showed that miR-431-3p and miR-1303 were capable to augment and suppress apoptosis/phagocytosis of macrophage via up regulating MDR1/MMP16/RIPOR2 and down regulating ATG5, respectively. <b>Conclusions:</b> This provides a proof of concept for microRNA-based therapy for active TB disease.