The m6A demethylase ALKBH5 controls trophoblast invasion at the maternal-fetal interface by regulating the stability of CYR61 mRNA.,Theranostics

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The m6A demethylase ALKBH5 controls trophoblast invasion at the maternal-fetal interface by regulating the stability of CYR61 mRNA.
Theranostics ( IF 12.4 ) Pub Date : 2019-01-01 , DOI: 10.7150/thno.31868
Xiao-Cui Li ₁ , Feng Jin ₂ , Bei-Ying Wang ₁ , Xiang-Jie Yin ₁ , Wei Hong ₁ , Fu-Ju Tian _{3,

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Affiliation

N6-Methyladenosine (m6A) is the most prevalent internal modification in mammalian mRNAs. Although m6A is important in many biological processes, its roles in the placenta are unclear. Methods: Levels of global mRNA m6A methylation and ALKBH5 expression in recurrent miscarriage (RM) patients were determined using quantitative reverse transcription-PCR (qRT-PCR), m6A RNA methylation quantification, and immunohistochemical methods. Using ALKBH5 overexpression and knockdown methods, we determined the role of ALKBH5 in trophoblast invasion at the maternal interface through trophoblasts and an extravillous explant culture experiments. Furthermore, the regulation of CYR61 by ALKBH5 was explored by RNA-sequencing coupled with methylated RNA immunoprecipitation. Results: We found that the level of global mRNA m6A methylation was significantly decreased in placental villous tissue from RM patients, while ALKBH5 expression was specifically unregulated. Furthermore, we demonstrated that ALKBH5 knockdown in human trophoblast promoted trophoblast invasion. Conversely, overexpression of ALKBH5 inhibited cell invasion. ALKBH5 knockdown promoted trophoblast invasion in villous explant culture experiments, while overexpression of ALKBH5 repressed these effects. Furthermore, we clarified that ALKBH5 inhibited trophoblast invasion by regulating CYR61 mRNA stability, and this RNA regulation is m6A dependent. Mechanistic analyses showed that decreased ALKBH5 in trophoblast increased the half-life of CYR61 mRNA and promoted steady-state CYR61 mRNA expression levels. Conclusions: We elucidated the functional roles of ALKBH5 and mRNA m6A methylation in trophoblast and identified a novel RNA regulatory mechanism, providing a basis for further exploration of broad RNA epigenetic regulatory patterns in RM diseases.

中文翻译：

m6A 去甲基化酶 ALKBH5 通过调节 CYR61 mRNA 的稳定性来控制母胎界面的滋养层侵袭。

N6-甲基腺苷 (m6A) 是哺乳动物 mRNA 中最普遍的内部修饰。尽管 m6A 在许多生物过程中很重要，但它在胎盘中的作用尚不清楚。方法：使用定量逆转录 PCR (qRT-PCR)、m6A RNA 甲基化定量和免疫组织化学方法确定复发性流产 (RM) 患者的整体 mRNA m6A 甲基化和 ALKBH5 表达水平。使用 ALKBH5 过表达和敲低方法，我们通过滋养层和绒毛外植体培养实验确定了 ALKBH5 在母体界面滋养层侵袭中的作用。此外，通过 RNA 测序结合甲基化 RNA 免疫沉淀探索了 ALKBH5 对 CYR61 的调节。结果：我们发现 RM 患者的胎盘绒毛组织中整体 mRNA m6A 甲基化水平显着降低，而 ALKBH5 表达特别不受调节。此外，我们证明了人类滋养层细胞中 ALKBH5 的敲低促进了滋养层细胞的侵袭。相反，ALKBH5 的过表达抑制细胞侵袭。在绒毛外植体培养实验中，ALKBH5 敲低促进了滋养层侵袭，而 ALKBH5 的过表达抑制了这些影响。此外，我们阐明 ALKBH5 通过调节 CYR61 mRNA 稳定性来抑制滋养层侵袭，并且这种 RNA 调节是 m6A 依赖性的。机制分析表明，滋养层细胞中 ALKBH5 的减少增加了 CYR61 mRNA 的半衰期并促进了稳态 CYR61 mRNA 表达水平。结论：

更新日期：2019-01-01

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