Myometrial oxidative stress drives MED12 mutations in leiomyoma.
Myometrial oxidative stress drives MED12 mutations in leiomyoma.
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DOI：
10.1186/s13578-022-00852-0
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发表时间：
2022-07-22
影响因子：
7.5
通讯作者：
Wei, Jian-Jun
中科院分区：
文献类型：
--
作者：
Li, Yinuo;Xu, Xiuhua;Asif, Huma;Feng, Yue;Kohrn, Brendan F.;Kennedy, Scott R.;Kim, J. Julie;Wei, Jian-Jun
超过70%的肌瘤(LM)存在MED12突变，主要发生在第二外显子中的C.130-131(GG)。肌层细胞中MED12突变的原因在很大程度上仍不清楚。我们假设，增加的ROS通过鸟嘌呤核苷酸的氧化促进子宫肌层细胞的MED12突变，随后是修复错误。用免疫组织化学方法检测8-OHdG在体外和体内的变化。Sanger测序和深度测序检测MED12基因突变。用RNA-seq方法检测有无光镜下子宫肌层的转录组，以及用ROS处理的原代子宫肌层细胞。用CRISPR/Cas9对8-OHdG介导的错误修复进行分析。高LM负荷的子宫MED12基因突变率显著高于低LM负荷的子宫。令人信服的数据表明，子宫通常会产生反应性氧化物种(ROS)以响应压力，而LM中的ROS水平由于代谢缺陷而升高。我们发现基因组氧化鸟嘌呤(8-OHdG)在有多个LM的子宫肌层中的水平显著高于没有LM的子宫肌层。转录组和通径分析检测到子宫肌层中的ROS应激。CRISPR/Cas9通过CRISPR/Cas9在MED12 c.130用8-OHdG靶向替换鸟嘌呤显著增加G>；T的错误修复。体外原代子宫肌层细胞暴露在氧化应激下通过双链测序检测到错误修复/突变的增加。总而言之，我们的数据确定了子宫肌层氧化应激增加和MED12高突变率之间的明显联系，这可能是育龄妇女发生LM的风险和严重程度的基础。网上版载有补充材料，可在10.1186/s13578-022-00852-0查阅。MED12突变率高和肌瘤负担明显与子宫肌层氧化应激有关。通过体外和体内的深度测序分析发现，MED12突变是由位于C.130-C.131的8-OHdG的错误修复所驱动的。网上版载有补充材料，可在10.1186/s13578-022-00852-0查阅。
More than 70% of leiomyomas (LM) harbor MED12 mutations, primarily in exon 2 at c.130-131(GG). The cause of MED12 mutations in myometrial cells remains largely unknown. We hypothesized that increased ROS promotes MED12 mutations in myometrial cells through the oxidation of guanine nucleotides followed by misrepair. Genomic oxidative burden (8-OHdG) was evaluated in vitro and in vivo by immunohistochemistry. MED12 mutations were examined by Sanger sequencing and deep sequencing. Transcriptome examined by RNA-seq was performed in myometrium with and without LM, in primary myometrial cells treated with ROS. 8-OHdG mediated misrepair was analyzed by CRISPR/Cas9. Uteri with high LM burden had a significantly higher rate of MED12 mutations than uteri with low LM burden. Compelling data suggest that the uterus normally produces reactive oxidative species (ROS) in response to stress, and ROS levels in LM are elevated due to metabolic defects. We demonstrated that genomic oxidized guanine (8-OHdG) was found at a significantly higher level in the myometrium of uteri that had multiple LM compared to myometrium without LM. Transcriptome and pathway analyses detected ROS stress in myometrium with LM. Targeted replacement of guanine with 8-OHdG at MED12 c.130 by CRISPR/Cas9 significantly increased the misrepair of G>T. Exposure of primary myometrial cells to oxidative stress in vitro increased misrepair/mutations as detected by duplex sequencing. Together, our data identified a clear connection between increased myometrial oxidative stress and a high rate of MED12 mutations that may underlie the risk of LM development and severity in women of reproductive age. The online version contains supplementary material available at 10.1186/s13578-022-00852-0. High rate of MED12 mutations and leiomyoma burden are clearly associated with myometrial oxidative stress. MED12 mutations are driven by misrepair of 8-OHdG at c.130-c.131 detected by deep sequencing analysis both in vitro and in vivo. The online version contains supplementary material available at 10.1186/s13578-022-00852-0.
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DOI：
10.1073/pnas.1208715109
发表时间：
2012-09-04
影响因子：
11.1
作者：
Schmitt, Michael W.;Kennedy, Scott R.;Loeb, Lawrence A.
通讯作者：
Loeb, Lawrence A.
影响因子：
2.9
作者：
Fletcher, Nicole M.;Saed, Mohammed G.;Saed, Ghassan M.
通讯作者：
Saed, Ghassan M.
影响因子：
7.4
作者：
Foksinski, M;Kotzbach, R;Olinski, R
通讯作者：
Olinski, R
影响因子：
14.9
作者：
Amente, Stefano;Di Palo, Giacomo;Majello, Barbara
通讯作者：
Majello, Barbara
影响因子：
8
作者：
Sekiguchi, M;Tsuzuki, T
通讯作者：
Tsuzuki, T