P–129 Follicular extracellular vesicles of women with polycystic ovarian syndrome inhibit oocyte maturation
Authors: Liu C , Wang M , Yao H , Cui M , Gong X , Zhang H , Sui C 
Published: August 2021

Abstract

Study question
Does follicular extracellular vesicles of women with polycystic ovarian syndrome (PCOS-EVs) interfere with the quality of oocytes?

Summary answer
PCOS-EVs induced oxidative stress in the oocytes and inhibited oocyte maturation by increasing the abnormal mitochondria distribution and abnormal spindle rates.

What is known already
Polycystic ovarian syndrome (PCOS) is a common endocrine disorder in women of reproductive age, with a prevalence up to 10%. Women with PCOS are characterized by a clustering of features, including hyperandrogenism, polycystic ovarian morphology, and notably, anovulation. Although international guidelines recommend assisted reproduction techniques to be an effective resort for PCOS patients to conceive. However, even after overcoming ovulatory dysfunction via ovulation induction, the pregnancy outcomes of patients with PCOS were far from satisfying with lower fertilization, cleavage, and implantation rates, implicating that theoocyte quality of these patients are affected. Whereas the mechanisms have not been elucidated yet.

Study design, size, duration
Follicular fluid of PCOS patients (n = 10) and healthy controls (n = 10) were collected and used for extracellular vesicles (EVs) isolation via ultracentrifugation. Germinal vesicle (GV) oocytes collected from female ICR mice were cocultured with RIF-EVs or FER-EVs, respectively, and PBS served as a blank control. GV breakdown (GVBD) rate and maturation rate were calculated at two-hour and fourteen-hour of co-culture, respectively. Besides, oocyte mitochondria distribution, meiosis spindle morphology, and oxidative status were assessed in different groups.

Participants/materials, setting, methods
EVs were determined by western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Fluorescence labeled EVs were used to visualize internalization by oocytes. Oocytes mitochondria and mitosis spindles were stained with fluorescence, and abnormal mitochondria rate or abnormal spindle rate was calculated. Reactive oxygen species (ROS) level was detected in the differently treated oocytes. Moreover, the expression of CAT, GSS, and SOD was determined in the oocytes using quantitative reverse transcription polymerase chain reaction.

Main results and the role of chance
Both PCOS-EVs and CTRL-EVs are bilayered vesicles, ranging from 100 to 150 nm, and enriched in Alix, TSG101, and CD9. EVs could be internalized by oocytes within one hour. After coculture, the GVBD rate was similar among the three groups; whereas the maturation rate was significantly decreased in the PCOS-EV group compared with CTRL-EV group or PBS group. In addition, the abnormal mitochondria distribution rate or abnormal spindle rate were significantly increased in the PCOS-EV group compared with PBS or CTRL-EV group. The ROS level was increased in the PCOS-EV group compared with CTRL-EV group, and the expression of CAT, GSS, and SOD was increased in the PCOS-EV-treated oocytes.

Limitations, reasons for caution
Our study did not identify the contents of PCOS-EVs and CTRL-EVs, and the molecular mechanisms of dysregulations induced by PCOS-EVs need further researches to investigate.
Wider implications of the findings: This work confirmed that EV-conducted cellular communication played an important role in oocyte development in women with PCOS. The dysregulation of oocytes induced by PCOS-EVs might be related to the poor oocyte quality of women with PCOS, which may provide a novel target to improve pregnancy outcomes of these patients.

Trial registration number
Not applicable