Modulating the m6A Modified Transcription Factor GATA6 Impacts Epithelial Cytokines in Acute Lung Injury.

Abstract

The methylation of the N6-position of adenosine (m6A) has been found to be associated with the inflammatory response. We hypothesize that m6A modification plays a role in the inflammation of airway epithelial cells during lung inflammation. However, the precise changes and functions of m6A modification in airway epithelial cells in acute lung injury (ALI) are not well understood. Here we report that RNA Methyltransferase 3 (METTL3)-mediated m6A of GATA6 mRNA inhibits ALI and the secretion of pro-inflammatory cytokine in airway epithelial cells. The expression of METTL3 and m6A levels decrease in lung tissues of ALI-mice. In co-cultures, peripheral blood monocytes secreted tumor necrosis factor α (TNFα) which reduces METTL3 and m6A levels in human bronchial epithelial cell line (BEAS-2B). Knockdown of METTL3 promotes IL-6 and TNFα release of BEAS-2B cells. Conversely, overexpression of METTL3 increases total RNA m6A level and reduces the levels of pro-inflammatory cytokines TNFα, transforming growth factor-β (TGFβ), and thymic stromal lymphopoietin (TSLP). Elevating METTL3 in mouse lungs prevented LPS-induced ALI and reduced the synthesis of pro-inflammatory cytokines. Mechanistically, sequencing and functional analysis show that METTL3 catalyzes m6A in the 3'UTR of GATA6 read by YTH N6-Methyladenosine RNA Binding Protein 2 (YTHDF2) and triggers mRNA degradation. GATA6 knockdown rescues TNFα induced inflammatory cytokine secretion of epithelial cells, indicating that GATA6 is a main substrate of METTL3 in airway epithelial cells. Overall, this study provides evidence of a novel role for METTL3 in the inflammatory cytokine release of epithelial cells and provide an innovative therapeutic target for ALI.