Pooled CRISPR screening identifies m6A as a positive regulator of macrophage activation

Sci Adv. 2021 Apr 28;7(18):eabd4742. doi: 10.1126/sciadv.abd4742. Print 2021 Apr.

Abstract

m6A RNA modification is implicated in multiple cellular responses. However, its function in the innate immune cells is poorly understood. Here, we identified major m6A "writers" as the top candidate genes regulating macrophage activation by LPS in an RNA binding protein focused CRISPR screening. We have confirmed that Mettl3-deficient macrophages exhibited reduced TNF-α production upon LPS stimulation in vitro. Consistently, Mettl3 flox/flox;Lyzm-Cre mice displayed increased susceptibility to bacterial infection and showed faster tumor growth. Mechanistically, the transcripts of the Irakm gene encoding a negative regulator of TLR4 signaling were highly decorated by m6A modification. METTL3 deficiency led to the loss of m6A modification on Irakm mRNA and slowed down its degradation, resulting in a higher level of IRAKM, which ultimately suppressed TLR signaling-mediated macrophage activation. Our findings demonstrate a previously unknown role for METTL3-mediated m6A modification in innate immune responses and implicate the m6A machinery as a potential cancer immunotherapy target.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / metabolism
  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Lipopolysaccharides
  • Macrophage Activation* / genetics
  • Methyltransferases* / genetics
  • Methyltransferases* / metabolism
  • Mice

Substances

  • Lipopolysaccharides
  • Methyltransferases
  • Adenosine