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

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.

SUBMITTER: Tong J

PROVIDER: S-EPMC8081357 | biostudies-literature |

REPOSITORIES: biostudies-literature

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