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

Front Immunol. 2019 Mar 19;10:421. doi: 10.3389/fimmu.2019.00421. eCollection 2019.

Abstract

Mycobacterium tuberculosis (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.

Keywords: Bach-1; CAGE transcriptomics; Elmo-1; Mycobacterium tuberculosis; alternative activated macrophages; c-Maf; microRNA-143; microRNA-365.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / immunology*
  • Animals
  • Basic-Leucine Zipper Transcription Factors / immunology*
  • Interleukin-13 / pharmacology
  • Interleukin-4 / pharmacology
  • Macrophages / drug effects
  • Macrophages / immunology
  • Macrophages / microbiology*
  • Male
  • Mice, Inbred BALB C
  • MicroRNAs / immunology*
  • Mycobacterium tuberculosis / growth & development*
  • Proto-Oncogene Proteins c-maf / immunology*
  • Tuberculosis / genetics
  • Tuberculosis / immunology
  • Tuberculosis / microbiology

Substances

  • Adaptor Proteins, Signal Transducing
  • Bach1 protein, mouse
  • Basic-Leucine Zipper Transcription Factors
  • ELMO1 protein, mouse
  • Interleukin-13
  • MIRN365 microRNA, mouse
  • Maf protein, mouse
  • MicroRNAs
  • Mirn143 microRNA, mouse
  • Proto-Oncogene Proteins c-maf
  • Interleukin-4