The pathophysiology of polycystic ovary syndrome (PCOS) is characterized by granulosa cell (GC) dysfunction. m⁶A modification affects GC function in patients with premature ovarian insufficiency (POI), but the role of m⁶A modification in PCOS is unknown. The purpose of the prospective comparative study was to analyse the m⁶A profile of the luteinized GCs from normovulatory women and non‐obese PCOS patients following controlled ovarian hyperstimulation. RNA m⁶A methylation levels were measured by m⁶A quantification assay in the luteinized GCs of the controls and PCOS patients. Then, m⁶A profiles were analysed by methylated RNA immunoprecipitation sequencing (MeRIP‐seq). We reported that the m⁶A level was increased in the luteinized GCs of PCOS patients. Comparative analysis revealed differences between the m⁶A profiles from the luteinized GC of the controls and PCOS patients. We identified FOXO3 mRNA with reduced m⁶A modification in the luteinized GCs of PCOS patients. Selectively knocking down m⁶A methyltransferases or demethylases altered expression of FOXO3 in the luteinized GCs from the controls, but did not in PCOS patients. These suggested an absence of m⁶A‐mediated transcription of FOXO3 in the luteinized GCs of PCOS patients. Furthermore, we demonstrated that the involvement of m⁶A in the stability of the FOXO3 mRNA that is regulated via a putative methylation site in the 3’‐UTR only in the luteinized GCs of the controls. In summary, our findings showed that altered m⁶A modification was involved in up‐regulated expression of FOXO3 mRNA in the luteinized GCs from non‐obese PCOS patients following controlled ovarian hyperstimulation.

中文翻译：

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改变的 m6 A 修饰参与非肥胖多囊卵巢综合征患者黄素化颗粒细胞中 FOXO3 的上调表达。

多囊卵巢综合征 (PCOS) 的病理生理特点是颗粒细胞 (GC) 功能障碍。m ⁶ A 修饰影响卵巢早衰 (POI) 患者的 GC 功能，但 m ⁶ A 修饰在 PCOS 中的作用尚不清楚。前瞻性比较研究的目的是分析正常女性和非肥胖 PCOS 患者在受控卵巢过度刺激后黄素化 GC的 m ⁶ A 谱。RNA m ⁶ A 甲基化水平通过 m ⁶ A 定量测定在对照和 PCOS 患者的黄素化 GC中测量。然后，m ⁶通过甲基化RNA免疫沉淀测序（MeRIP-seq）分析A谱。我们报告说，PCOS 患者的黄素化 GCs 中的 m ⁶ A 水平增加。比较分析揭示了来自对照和 PCOS 患者的黄素化 GC的 m ⁶ A 谱之间的差异。我们在 PCOS 患者的黄素化 GC 中鉴定了具有减少的 m ⁶ A 修饰的FOXO3 mRNA 。选择性敲低 m ⁶ A 甲基转移酶或去甲基化酶会改变对照组黄素化 GC 中 FOXO3 的表达，但在 PCOS 患者中没有。这些表明不存在 m ⁶ A 介导的FOXO3转录在 PCOS 患者的黄体化 GC 中。此外，我们证明 m ⁶ A参与FOXO3 mRNA的稳定性，该稳定性仅在对照的黄素化 GC 中通过 3'-UTR 中的假定甲基化位点进行调节。总之，我们的研究结果表明，改变的 m ⁶ A 修饰与受控卵巢过度刺激后非肥胖 PCOS 患者的黄素化 GC中FOXO3 mRNA 的表达上调有关。