The N6-methyladenosine (m6A)-forming enzyme METTL3 facilitates M1 macrophage polarization through the methylation of STAT1 mRNA

Am J Physiol Cell Physiol. 2019 Oct 1;317(4):C762-C775. doi: 10.1152/ajpcell.00212.2019. Epub 2019 Jul 31.

Abstract

Compelling evidence indicates that epigenetic regulations orchestrate dynamic macrophage polarization. N6-methyladenosine (m6A) methylation is the most abundant epigenetic modification of mammalian mRNA, but its role in macrophage polarization is still completely unknown. Here, we show that the m6A-catalytic enzyme methyltransferase like 3 (METTL3) is specifically upregulated following the M1 polarization of mouse macrophages. Furthermore, METTL3 knockdown through siRNA transfection markedly inhibited M1, but enhanced M2, macrophage polarization. Conversely, its overexpression via plasmid transfection greatly facilitated M1, but attenuated M2, macrophage polarization. Further methylated RNA immunoprecipitation and in vitro m6A methylation assays suggested that METTL3 directly methylates mRNA encoding signal transducer and activator of transcription 1 (STAT1), a master transcription factor controlling M1 macrophage polarization, at its coding sequence and 3'-untranslated regions. In addition, METTL3-mediated STAT1 mRNA methylation significantly increased mRNA stability and subsequently upregulated STAT1 expression. In conclusion, METTL3 drives M1 macrophage polarization by directly methylating STAT1 mRNA, potentially serving as an anti-inflammatory target.

Keywords: METTL3; m6A methylation; macrophage polarization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / pharmacology
  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Gene Expression Regulation / drug effects
  • Macrophage Activation / drug effects
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Male
  • Methylation / drug effects
  • Methyltransferases / drug effects*
  • Mice, Inbred C57BL
  • RNA, Messenger / metabolism
  • STAT1 Transcription Factor / drug effects

Substances

  • Anti-Inflammatory Agents
  • RNA, Messenger
  • STAT1 Transcription Factor
  • N-methyladenosine
  • Methyltransferases
  • Mettl3 protein, mouse
  • Adenosine