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The m⁶A methyltransferase METTL3 drives neuroinflammation and neurotoxicity through stabilizing BATF mRNA in

Xun Wu¹, Haixiao Liu¹, Jin Wang²

¹Department of Neurosurgery, Tangdu Hospital, the Fourth Military Medical University, Xi'an, 710032, Shaanxi, China.

Cell Death and Differentiation

|June 20, 2024

Abstract:

Persistent neuroinflammation and progressive neuronal loss are defining features of acute brain injury including traumatic brain injury (TBI) and cerebral stroke. Microglia, the most abundant type of brain-resident immune cells, continuously surveil the environment and play a central role in shaping the inflammatory state of the central nervous system (CNS). In the study, we discovered that the protein expression of METTL3 (a m⁶A methyltransferase) was upregulated in inflammatory microglia independent of increased Mettl3 gene transcription following TBI in both human and mouse subjects. Subsequently, we identified TRIP12, a HECT-domain E3 ubiquitin ligase, as a negative regulator of METTL3 protein expression by facilitating METTL3 K48-linked polyubiquitination. Importantly, selective ablation of Mettl3 inhibited microglial pathogenic activities, diminished neutrophil infiltration, rescued neuronal loss and facilitated functional recovery post-TBI. Using MeRIP-seq and CUT&Tag sequencing, we identified that METTL3 promoted the expression of Basic Leucine Zipper Transcriptional Factor ATF-Like (BATF), which in turn directly bound to a cohort of characteristic inflammatory cytokines and chemokine genes. Enhanced activities of BATF in microglia elicited TNF-dependent neurotoxicity and can also promote neutrophil recruitment through releasing CXCL2. Pharmacological inhibition of METTL3 using a BBB-penetrating drug-loaded nano-system showed satisfactory therapeutic effects in both TBI and stroke mouse models. Collectively, our findings identified METTL3-m⁶A-BATF axis as a potential therapeutic target for terminating detrimental neuroinflammation and progressive neuronal loss following acute brain injury. METTL3 protein is significantly up-regulated in inflammatory microglia due to the decreased proteasomal degradation mediated by TRIP12 and ERK-USP5 pathways. METTL3 stabilized BATF mRNA stability and promoted BATF expression through the m⁶A-IGF2BP2-dependent mechanism. Elevated expression of BATF elicits a pro-inflammatory gene program in microglia, and aggravates neuroinflammatory response including local immune responses and peripheral immune cell infiltration. Genetic deletion or pharmaceutically targeting METTL3-BATF axis suppressed microglial pro-inflammatory activities and promoted neurological recovery following TBI and stroke.

Insights

Researchers found METTL3 protein, not gene, increases in microglia after brain injury. Targeting the METTL3-BATF pathway reduces neuroinflammation and aids recovery in traumatic brain injury and stroke models.

Area of Science:

Neuroscience and Immunology
Molecular Biology and Genetics
Biochemistry and Pharmacology

Background:

Acute brain injuries like TBI and stroke cause persistent neuroinflammation and neuronal loss.
Microglia, the brain's immune cells, are central to CNS inflammatory responses.
METTL3 (a methyltransferase) protein levels are elevated in microglia post-TBI.

Purpose of the Study:

To investigate the role of METTL3 in microglial activation and neuroinflammation following acute brain injury.
To identify regulators of METTL3 protein expression and downstream targets.
To evaluate METTL3 inhibition as a therapeutic strategy for TBI and stroke.

Main Methods:

Analysis of METTL3 protein and gene expression in human and mouse brain injury models.
Identification of TRIP12 as an E3 ubiquitin ligase regulating METTL3 degradation.
MeRIP-seq and CUT&Tag sequencing to identify METTL3 targets, including BATF.
Genetic ablation and pharmacological inhibition of METTL3 in TBI and stroke models.
Assessment of microglial activity, immune cell infiltration, neuronal loss, and functional recovery.

Main Results:

METTL3 protein, but not gene, is upregulated in inflammatory microglia post-TBI.
TRIP12 facilitates METTL3 polyubiquitination and degradation; METTL3 stabilizes BATF mRNA.
METTL3 promotes BATF expression, leading to pro-inflammatory cytokine release and neurotoxicity.
Selective METTL3 ablation or pharmacological inhibition improves neurological outcomes in TBI and stroke models.
Targeting the METTL3-BATF axis reduces neuroinflammation and promotes neuronal survival and functional recovery.

Conclusions:

The METTL3-m⁶A-BATF signaling pathway is a key driver of detrimental neuroinflammation in acute brain injury.
METTL3 protein upregulation, mediated by reduced degradation, drives microglial pro-inflammatory responses.
Pharmacological inhibition of METTL3 offers a promising therapeutic avenue for treating TBI and stroke.

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