High-Throughput Sequencing Profiles About lncRNAs and mRNAs of Ovarian Granulosa Cells in Polycystic Ovary Syndrome.

Yanjun Zheng,Wei Chen,Yuehong Bian,Ping Li,Xiao Yang,Shigang Zhao,Linlin Fu,Yuhua Shi,Richao Wu,Ying Wang

doi:10.3389/fmed.2021.741803

Abstract

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in women of reproductive age, which is characterized by ovulatory dysfunction, clinical and/or biochemical androgen excess, polycystic ovaries on ultrasound and genetic heterogeneity. It was well-accepted that many lncRNAs and mRNAs were associated with PCOS, however, remain unclear. Therefore, the purpose of our study was to examine different expression profiles of lncRNAs and mRNAs in ovarian granulosa cells (GCs) in PCOS and Controls, and identify the correlation between lncRNAs, mRNAs and clinical parameters. Sixty five PCOS patients and 65 Controls were enrolled in this study and adopted standard long agonist protocols or GnRH antagonist protocols. Then 6 GCs samples in each group were subjected to high-thoughput sequencing and the remaining samples were used for the further verification by quantitative real-time PCR (qRT-PCR). Gene Oncology (GO), Kyoto Encyclopedia Genes and Genomes (KEGG) enrichment analysis were performed. We predicted the relationship between lncRNAs and mRNAs by Cytoscape software. According to the expression level of lncRNAs, mRNAs and the clinical parameters, we also explored their relationship and evaluate their predictive values for embryos quality and PCOS. We identified 1,049 differential expressed lncRNAs and 3,246 mRNAs (fold-change ≥2, p-value < 0.05). Seven lncRNAs (NONHSAT101926.2, NONHSAT136825.2, NONHSAT227177.1, NONHSAT010538.2, NONHSAT191377.1, NONHSAT230904.1, ENST00000607307) and 3 mRNAs (EREG, ENTPD6, YAP1) were validated consistent with sequence profile. Seven lncRNAs were related to hormone level and follicle counts, 3 mRNAs had connections with lipid metabolism. The area under curve (AUC) of 7 lncRNAs were valuable in distinguishing patients with PCOS from Controls. The AUC of NONHSAT230904.1 and NONHSAT227177.1 were 0.6807 and 0.6410, respectively, for distinguishing whether the rate of high-quality embryos exceeds 50%. Our study showed that the GCs lncRNAs and mRNAs were involved in the occurrence and development of PCOS, which contribute to clarify the pathogenesis mechanism of PCOS.

Highlights

Polycystic ovary syndrome (PCOS) is a common endocrine disorder affecting 8–13% of women of reproductive age [1]
Ovarian granulosa cells (GCs) were collected from 65 polycystic ovary syndrome (PCOS) patients (PCOS group) and 65 patients with an indication of male factor infertility serving as Controls (Control group), all of whom underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) at the Center for Reproductive Medicine, Shandong University, between December 2018 and December 2019
54,615 Long non-coding RNAs (lncRNAs) and 123,141 mRNAs were detected in six PCOS patients and six Control patients by the highthroughput sequencing of lncRNAs

Summary

Introduction

Polycystic ovary syndrome (PCOS) is a common endocrine disorder affecting 8–13% of women of reproductive age [1]. Many studies have revealed that lncRNAs exhibit certain characteristics of mRNAs. For instance, lncRNAs are transcribed by RNA polymerase II, equipped with a 3′ poly (A) tail and a 5′ cap, and contain a promoter and multiple exons [8, 9]. LncRNAs are transcribed by RNA polymerase II, equipped with a 3′ poly (A) tail and a 5′ cap, and contain a promoter and multiple exons [8, 9] These features enable lncRNAs to interact with DNA, RNA, and proteins to directly regulate several biological processes, including chromatin modification, RNA transcription, pre-mRNA splicing, mRNA translation, and gene expression [10, 11]. Several studies have shown a wide range of interactions among mRNAs, microRNAs, and lncRNAs [15]

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