Myometrial oxidative stress drives MED12 mutations in leiomyoma

Yinuo Li, Xiuhua Xu, Huma Asif, Yue Feng, Brendan F. Kohrn, Scott R. Kennedy, J. Julie Kim*, Jian Jun Wei*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Background: 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. Methods: 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. Results: 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. Conclusions: 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. Graphical Abstract: [Figure not available: see fulltext.]

Original languageEnglish (US)
Article number111
JournalCell and Bioscience
Volume12
Issue number1
DOIs
StatePublished - Dec 2022

Keywords

  • 8-OHdG
  • Duplex sequencing
  • Leiomyoma
  • MED12 mutation
  • Myometrium
  • Reactive oxidative species (ROS)

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

Fingerprint

Dive into the research topics of 'Myometrial oxidative stress drives MED12 mutations in leiomyoma'. Together they form a unique fingerprint.

Cite this