Differential Targeting of c-Maf, Bach-1, and Elmo-1 by microRNA-143 and microRNA-365 Promotes the Intracellular Growth of Mycobacterium tuberculosis in Alternatively IL-4/IL-13 Activated Macrophages

• Published in: Frontiers in immunology Vol. 10; p. 421
• Main Authors: Tamgue, Ousman, Gcanga, Lorna, Ozturk, Mumin, Whitehead, Lauren, Pillay, Shandre, Jacobs, Raygaana, Roy, Sugata, Schmeier, Sebastian, Davids, Malika, Medvedeva, Yulia A, Dheda, Keertan, Suzuki, Harukazu, Brombacher, Frank, Guler, Reto
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 19-03-2019
• Subjects: Adaptor Proteins, Signal Transducing - immunology; alternative activated macrophages; Animals; Basic-Leucine Zipper Transcription Factors - immunology; c-Maf; CAGE transcriptomics; Immunology; Interleukin-13 - pharmacology; Interleukin-4 - pharmacology; Macrophages - drug effects; Macrophages - immunology; Macrophages - microbiology; Male; Mice, Inbred BALB C; microRNA-143; microRNA-365; MicroRNAs - immunology; Mycobacterium tuberculosis; Mycobacterium tuberculosis - growth & development; Proto-Oncogene Proteins c-maf - immunology; Tuberculosis - genetics; Tuberculosis - immunology; Tuberculosis - microbiology; c-Maf; microRNA-143; microRNA-365; CAGE transcriptomics; Mycobacterium tuberculosis; alternative activated macrophages; Bach-1; Elmo-1
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2019.00421

(Mtb) can subvert the host defense by skewing macrophage activation toward a less microbicidal alternative activated state to avoid classical effector killing functions. Investigating the molecular basis of this evasion mechanism could uncover potential candidates for host directed therapy against tuberculosis (TB). A limited number of miRNAs have recently been shown to regulate host-mycobacterial interactions. Here, we performed time course kinetics experiments on bone marrow-derived macrophages (BMDMs) and human monocyte-derived macrophages (MDMs) alternatively activated with IL-4, IL-13, or a combination of IL-4/IL-13, followed by infection with Mtb clinical Beijing strain HN878. MiR-143 and miR-365 were highly induced in Mtb-infected M(IL-4/IL-13) BMDMs and MDMs. Knockdown of miR-143 and miR-365 using antagomiRs decreased the intracellular growth of Mtb HN878, reduced the production of IL-6 and CCL5 and promoted the apoptotic death of Mtb HN878-infected M(IL-4/IL-13) BMDMs. Computational target prediction identified c-Maf, Bach-1 and Elmo-1 as potential targets for both miR-143 and miR-365. Functional validation using luciferase assay, RNA-pulldown assay and Western blotting revealed that c-Maf and Bach-1 are directly targeted by miR-143 while c-Maf, Bach-1, and Elmo-1 are direct targets of miR-365. Knockdown of c-Maf using GapmeRs promoted intracellular Mtb growth when compared to control treated M(IL-4/IL-13) macrophages. Meanwhile, the blocking of Bach-1 had no effect and blocking Elmo-1 resulted in decreased Mtb growth. Combination treatment of M(IL-4/IL-13) macrophages with miR-143 mimics or miR-365 mimics and c-Maf, Bach-1, or Elmo-1 gene-specific GapmeRs restored Mtb growth in miR-143 mimic-treated groups and enhanced Mtb growth in miR-365 mimics-treated groups, thus suggesting the Mtb growth-promoting activities of miR-143 and miR-365 are mediated at least partially through interaction with c-Maf, Bach-1, and Elmo-1. We further show that knockdown of miR-143 and miR-365 in M(IL-4/IL-13) BMDMs decreased the expression of HO-1 and IL-10 which are known targets of Bach-1 and c-Maf, respectively, with Mtb growth-promoting activities in macrophages. Altogether, our work reports a host detrimental role of miR-143 and miR-365 during Mtb infection and highlights for the first time the role and miRNA-mediated regulation of c-Maf, Bach-1, and Elmo-1 in Mtb-infected M(IL-4/IL-13) macrophages.