N¹-methyladenosine (m¹A) is one of the important post-transcriptional modifications in RNA and plays an important role in promoting translation or decay of m¹A-methylated messenger RNA (mRNA), but the “reader” protein and the exact biological role of m¹A remain to be determined. Here, we identified that nine potential m¹A “reader” proteins including YTH domain family and heterogeneous nuclear ribonucleoprotein by mass spectrometry, and among them, YTH domain-containing protein 3 (YTHDF3), could bind directly to m¹A-carrying RNA. YTHDF3 was then identified to negatively regulate invasion and migration of trophoblast. Mechanistically, we found that the m¹A “reader” YTHDF3 bound to certain m¹A-methylated transcripts, such as insulin-like growth factor 1 receptor (IGF1R), with the combination of iCLIP-seq (individual-nucleotide resolution ultraviolet crosslinking and immunoprecipitation high-throughput sequencing) and m¹A-seq. Furthermore, YTHDF3 could promote IGF1R mRNA degradation and thus inhibit IGF1R protein expression along with its downstream matrix metallopeptidase 9 signaling pathway, consequently decreasing migration and invasion of trophoblast. Thus, we demonstrated that YTHDF3 as an m¹A reader decreased invasion and migration of trophoblast by inhibiting IGF1R expression. Our study outlines a new m¹A epigenetic way to regulate the trophoblast activity, which suggests a novel therapeutic target for trophoblast-associated pregnancy disorders.

N ¹ -甲基腺苷（m ¹ A）是RNA中重要的转录后修饰之一，在促进m ¹ A甲基化信使RNA（mRNA）的翻译或衰变中发挥重要作用，但“阅读器”蛋白和m ¹ A 的确切生物学作用仍有待确定。在这里，我们通过质谱鉴定了九种潜在的m ¹ A“阅读器”蛋白，包括YTH结构域家族和异质核核糖核蛋白，其中，含有YTH结构域的蛋白3（YTHDF3）可以直接与携带m 1 A的^RNA结合。随后发现 YTHDF3 可以负向调节滋养层的侵袭和迁移。从机制上讲，我们发现 m ^{1 A“阅读器”YTHDF3 与某些 m 1} A 甲基化转录本结合，例如胰岛素样生长因子 1 受体 (IGF1R)，并结合 iCLIP-seq（单个核苷酸分辨率紫外交联）和免疫沉淀高通量测序）和 m ¹ A-seq。此外，YTHDF3可以促进IGF1R mRNA降解，从而抑制IGF1R蛋白及其下游基质金属肽酶9信号通路的表达，从而减少滋养层的迁移和侵袭。因此，我们证明YTHDF3作为m ¹ A阅读器通过抑制IGF1R表达来减少滋养层的侵袭和迁移。我们的研究概述了一种新的 m ¹ A 表观遗传方式来调节滋养层活性，这为滋养层相关妊娠疾病提供了一个新的治疗靶点。