METTL14 depletion induces trophoblast cell dysfunction by inhibiting miR-21-5p processing in an m6A-dependent manner.

Spontaneous abortion (SA) is a devastating, but common outcome for expectant parents and their families. However, the mechanism of SA occurrence remains mostly unknown. Herein, we examined human SA villi samples and found decreased N6-methyladenosine (m6A) levels and methyltransferase-like protein 14 (METTL14) expression compared with those in healthy women. Knockdown of METTL14 in trophoblast HTR8 cells induced cellular dysfunction. We identified candidate differentially expressed microRNAs and found that METTL14 accelerated miR-21-5p processing by modulating its m6A modification level. Exogenous miR-21-5p expression attenuated METTL14 knockdown-induced cellular dysfunction. Subsequently, we found that SMAD family member 7 (SMAD7) expression is inhibited by miR-21-5p and that knockdown of SMAD7 rescued the trophoblast cell dysfunction induced by miR-21-5p inhibitors. Then, we revealed that METTL14 can regulate the SMAD7 pathway by modulating miR-21-5p. Finally, we found that exposing pregnant mice to an m6A inhibitor caused embryo loss and reduced expression levels of Mettl14 and miR-21-5p while increasing Smad7 levels. Taken together, this study establishes the involvement of m6A in SA and identified a novel SA signaling pathway. These results reveal the underlying molecular mechanisms of trophoblast cell dysfunction induced by m6A modification and provide new strategies to identify and mitigate SA.