miR-340-3p 修饰的骨髓间充质干细胞衍生的外泌体通过 METTL3 介导的 HMOX1 的 mA 修饰抑制铁死亡，从而促进受损大鼠子宫的恢复。
miR-340-3p-modified bone marrow mesenchymal stem cell-derived exosomes inhibit ferroptosis through METTL3-mediated mA modification of HMOX1 to promote recovery of injured rat uterus.
机构信息
Department of Medical Genetics, Naval Medical University, Shanghai, 200433, China.
Department of Anatomy, Institute of Biomedical Engineering, Naval Medical University, Shanghai, 200433, China.
出版信息
Stem Cell Res Ther. 2024 Jul 29;15(1):224. doi: 10.1186/s13287-024-03846-6.
BACKGROUND
Ferroptosis is associated with the pathological progression of hemorrhagic injury and ischemia-reperfusion injury. According to our previous study, exosomes formed through bone marrow mesenchymal stem cells modified with miR-340-3p (MB-exos) can restore damaged endometrium. However, the involvement of ferroptosis in endometrial injury and the effect of MB-exos on ferroptosis remain elusive.
METHODS
The endometrial injury rat model was developed. Exosomes were obtained from the supernatants of bone marrow mesenchymal stromal cells (BMSCs) and miR-340/BMSCs through differential centrifugation. We conducted RNA-seq analysis on endometrial tissues obtained from the PBS and MB-exos groups. Ferroptosis was induced in endometrial stromal cells (ESCs) by treating them with erastin or RSL3, followed by treatment with B-exos or MB-exos. We assessed the endometrial total mA modification level after injury and subsequent treatment with B-exos or MB-exos by methylation quantification assay. We performed meRIP-qPCR to analyze mA modification-regulated endogenous mRNAs.
RESULTS
We reveal that MB-exos facilitate the injured endometrium to recover by suppressing ferroptosis in endometrial stromal cells. The injured endometrium showed significantly upregulated N-methyladenosine (mA) modification levels; these levels were attenuated by MB-exos through downregulation of the methylase METTL3. Intriguingly, METTL3 downregulation appears to repress ferroptosis by stabilizing HMOX1 mRNA, thereby potentially elucidating the mechanism through which MB-exos inhibit ferroptosis in ESCs. We identified YTHDF2 as a critical mA reader protein that contributes to HMOX1 mRNA degradation. YTHDF2 facilitates HMOX1 mRNA degradation by identifying the mA binding site in the 3'-untranslated regions of HMOX1. In a rat model, treatment with MB-exos ameliorated endometrial injury-induced fibrosis by inhibiting ferroptosis in ESCs. Moreover, METTL3 short hairpin RNA-mediated inhibition of mA modification enhanced the inhibitory effect of MB-exos on ferroptosis in endometrial injury.
CONCLUSIONS
Thus, these observations provide new insights regarding the molecular mechanisms responsible for endometrial recovery promotion by MB-exos and highlight mA modification-dependent ferroptosis inhibition as a prospective therapeutic target to attenuate endometrial injury.
背景
铁死亡与出血性损伤和缺血再灌注损伤的病理进展有关。根据我们之前的研究，通过 miR-340-3p（MB-exos）修饰的骨髓间充质干细胞形成的外泌体可以修复受损的子宫内膜。然而，铁死亡在子宫内膜损伤中的作用以及 MB-exos 对铁死亡的影响仍不清楚。
方法
建立子宫内膜损伤大鼠模型。通过差速离心从骨髓间充质基质细胞（BMSCs）和 miR-340/BMSCs 的上清液中获得外泌体。我们对 PBS 和 MB-exos 组获得的子宫内膜组织进行 RNA-seq 分析。用 erastin 或 RSL3 诱导子宫内膜基质细胞（ESCs）发生铁死亡，然后用 B-exos 或 MB-exos 处理。通过甲基化定量分析评估损伤后和随后用 B-exos 或 MB-exos 处理后的子宫内膜总 mA 修饰水平。我们进行 meRIP-qPCR 分析 mA 修饰调节的内源性 mRNAs。
结果
我们揭示 MB-exos 通过抑制子宫内膜基质细胞中的铁死亡促进受损子宫内膜的恢复。损伤的子宫内膜表现出明显上调的 N-甲基腺苷（mA）修饰水平；这些水平通过下调甲基转移酶 METTL3 被 MB-exos 减弱。有趣的是，METTL3 的下调似乎通过稳定 HMOX1 mRNA 来抑制铁死亡，从而可能阐明 MB-exos 抑制 ESCs 中铁死亡的机制。我们确定 YTHDF2 作为一种关键的 mA 阅读蛋白，有助于 HMOX1 mRNA 的降解。YTHDF2 通过识别 HMOX1 3'-UTR 中的 mA 结合位点促进 HMOX1 mRNA 的降解。在大鼠模型中，MB-exos 治疗通过抑制 ESCs 中的铁死亡改善了子宫内膜损伤引起的纤维化。此外，METTL3 短发夹 RNA 介导的 mA 修饰抑制增强了 MB-exos 对子宫内膜损伤中铁死亡的抑制作用。
结论
因此，这些观察结果为 MB-exos 促进子宫内膜恢复的分子机制提供了新的见解，并强调 mA 修饰依赖性铁死亡抑制作为减轻子宫内膜损伤的有前途的治疗靶点。
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