USP1/UAF1-stabilized METTL3 promotes reactive astrogliosis and improves functional recovery after spinal cord injury through m6A modification of YAP1 mRNA

RNA N⁶-methyladenosine (m⁶A) modification is involved in diverse biological processes. However, its role in spinal cord injury (SCI) is poorly understood. The m⁶A level increases in injured spinal cord, and METTL3, which is the core subunit of methyltransferase complex, is upregulated in reactive astrocytes, and further stabilized by the USP1/UAF1 complex after SCI. The USP1/UAF1 complex specifically binds to and subsequently removes K48-linked ubiquitination of the METTL3 protein to maintain its stability after SCI. Moreover, conditional knockout of astrocytic METTL3 in both sexes of mice significantly suppressed reactive astrogliosis after SCI, thus resulting in widespread infiltration of inflammatory cells, aggravated neuronal loss, hampered axonal regeneration, and impaired functional recovery. Mechanistically, the YAP1 transcript was identified as a potential target of METTL3 in astrocytes. METTL3 could selectively methylate the 3'-UTR region of the YAP1 transcript, which subsequently maintains its stability in an IGF2BP2-dependent manner. In vivo, YAP1 overexpression by adeno-associated virus-YAP1 injection remarkably contributed to reactive astrogliosis and partly reversed the detrimental effects of METTL3 knockout on functional recovery after SCI. Furthermore, we found that the methyltransferase activity of METTL3 plays an essential role in reactive astrogliosis and motor repair, whereas METTL3 mutant without methyltransferase function failed to promote functional recovery after SCI. Our study reveals the previously unreported role of METTL3-mediated m⁶A modification in SCI and might provide a potential therapy for SCI.SIGNIFICANCE STATEMENT:Spinal cord injury is a devastating trauma of the central nervous system involving motor and sensory impairments. However, epigenetic modification in spinal cord injury is still unclear. Here, we propose an m⁶A regulation effect of astrocytic METTL3 following spinal cord injury and we further characterize its underlying mechanism, which might provide promising strategies for spinal cord injury treatment.