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EET Reduction Contributes to Granulosa Cell Senescence And Endometriosis-Associated Infertility Via the PI3K/AKT/mTOR Signaling Pathway

GSE304488 Mus musculus (mouse) RNA-seq (gene expression) 16 samples Submitted 2025/08/05 Platform GPL34328
At a glance

RNA-seq (gene expression) · mouse · 16 samples

What it is
RNA-seq (gene expression)Gene activity measured by sequencing. The summary does not mention single-cell work, so this is most likely bulk: one average per sample, the usual starting point for comparing groups.
Organism
Mus musculus (mouse)
Samples
16 samples: 5 × mGCs from sham surgery group, vs 5 × mGCs from EM group, vs 3 × mGCs from EM group, cultured in DMEM/F12 with 5% FBS for 24 hours, vs 3 × mGCs from EM group, treated additionally with 100 nM 14, 15-EET for 24 hours,Groups are read from the sample names, not from the paper. Check the list below.
Data you can get
A processed table (series matrix) and supplementary files from NCBI; the raw sequencing reads from ENA. Which of these hold counts depends on the study, so check the file list.
Summary (written by the submitters)
Oxidative stress is involved in endometriosis (EM)-related infertility via various mechanisms, such as accelerating ovarian granulosa cell (GC) senescence, inhibiting ovulation, and interfering with lipid metabolism. In this study, we aimed to examine abnormal oxidative lipid levels and their related mechanisms in EM-associated infertility.
Read the full summary ↓
Through liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis, we observed that epoxyeicosatrienoic acids (EETs), which have antioxidant and anti-senescence effects, had decreased levels in follicular fluid samples from EM patients. The EET levels were positively correlated with in vitro fertilization outcomes. Using qPCR and western blot experiments, we found elevated soluble epoxide hydrolase (EPHX2) expression levels in EM-GCs. This enzyme metabolizes EETs into dihydroxyeicosatrienoic acids (DHETs), explaining the decreased 14, 15-EET levels in EM patients. Lower 14, 15-EET concentrations led to decreased GC antioxidant capacity, reduced ATP production, reactive oxygen species (ROS) accumulation in oocytes, and abnormal cumulus-oocyte complex (COC) expansion, which ultimately resulted in decreased fertility. Inhibiting EPHX2 in vivo or in vitro could reverse these observed abnormalities by upregulating 14, 15-EET. Mechanistic studies showed that 14, 15-EET alleviated GC senescence and improved fertility by inhibiting excessive PI3K/AKT/mTOR signaling pathway activation in EM-GCs. Moreover, the PI3K/AKT/mTOR pathway inhibitor BEZ-235 significantly alleviated ROS-induced cell senescence and abnormal COC expansion. Further work showed that an oxidative stress-induced decrease in EZH2/H3K27Me3 histone methylation led to the elevated EPHX2 expression patterns in EM-GCs. In addition, the decreased 14, 15-EET levels led to ROS accumulation, decreased EZH2 enzymatic activity, less EPHX2/H3K27Me3 histone methylation, and increased EPHX2 protein expression levels, which further reduced 14, 15-EET levels as part of a vicious feedback loop. Using LC-MS/MS, viral infection, transcriptomic sequencing, knockout mice, and EM mouse model experiments, we elucidated the molecular mechanisms of 14, 15-EET in EM-associated infertility and the mechanism controlling its decreased levels.
This dataset
Open GSE304488 in finder → Find more mouse RNA-seq datasets →
Download · which file do I need?

Direct links to NCBI and ENA, no account and no request form. Pick by what you want to do: take the small processed table to look, the supplementary files to re-analyse, or the original raw data to start over.

1 · Just have a look

Processed table

The series matrix: one table of values per sample as the submitters processed it. The smallest download, and it opens in Excel or R. Start here to see what the data look like.

Series matrix ↓
2 · Re-analyse it

Supplementary files

The count tables and other files the authors uploaded. Take only the samples you need, or the whole study as GSE304488_RAW.tar (it can run to tens of gigabytes).

All supplementary files ↗ Whole study · RAW.tar ↓
3 · Start from the raw reads

Raw sequencing reads

The original FASTQ files, straight from the sequencer, held at ENA. Large, and you align them yourself, so only choose this if you need to re-process the study from scratch.

Raw reads on ENA ↗

What the files usually look like: For RNA-seq the counts are usually a table among the supplementary files (look for 'counts', 'expression', 'FPKM' or 'TPM' in the file name). The series matrix of a sequencing study is sometimes nearly empty.

Also filed as BioProject PRJNA1301370 and SRA study SRP606336. Searching any of these in the dataset finder brings you back here.

NCBI GEO page ↗
Study design
5 × mGCs from sham surgery group, vs 5 × mGCs from EM group, vs 3 × mGCs from EM group, cultured in DMEM/F12 with 5% FBS for 24 hours, vs 3 × mGCs from EM group, treated additionally with 100 nM 14, 15-EET for 24 hours,

Supports a case/control comparison: 3 samples read as cases, 5 as controls.

How we read this: 4 replicated groups read from 16 sample titles; they account for 16 of them. These groups are guessed from the sample names, not taken from the paper, so check them against the sample list below before relying on them.

Samples in this study
Datasets on a similar topic

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