Expression of long noncoding RNAs in the ovarian granulosa cells of women with diminished ovarian reserve using high-throughput sequencing

Abstract

Background Infertility is a global reproductive-health problem, and diminished ovarian reserve (DOR) is one of the common causes of female infertility. Long noncoding RNAs (lncRNAs) are crucial regulators of numerous physiological and pathological processes in humans. However, whether lncRNAs are involved in the development of DOR remains to be elucidated. Methods Ovarian granulosa cells (OGCs) extracted from infertile women with DOR and from women with normal ovarian reserve (NOR) were subjected to high-throughput sequencing. Comprehensive bioinformatics analysis was conducted to identify the differential expression of messenger RNAs (mRNAs) and lncRNAs. Sequencing results were validated by the selection of lncRNAs and mRNAs using real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results Compared with the NOR group, a total of 244 lncRNAs were upregulated (53 known and 191 novel), and 222 lncRNAs were downregulated (36 known and 186 novel) in the DOR group. Similarly, 457 mRNAs had differential expression between the two groups. Of these, 169 were upregulated and 288 were downregulated. Bioinformatics analysis revealed that the differentially expressed genes of mRNA and lncRNAs were considerably enriched in “cell adhesion and apoptosis”, “steroid biosynthesis”, and “immune system”. A co-expression network comprising lncRNAs and their predicted target genes revealed the possible involvement of the “thyroid hormone signaling pathway” and “protein binding, digestion and absorption” in DOR pathogenesis. The expression of SLC16A10 was positively regulated by multiple lncRNAs. After RT-qPCR validation of seven differentially expressed lncRNAs and mRNAs, respectively, the expression of lncRNA NEAT1, GNG12, ZEB2-AS1, and mRNA FN1, HAS3, RGS4, SUOX were in accordance with RNA-sequencing. Conclusions We presented the first data showing that the expression profiles of lncRNA and mRNA in OGCs between NOR and DOR patients using RNA sequencing. The lncRNAs and mRNAs that we identified may serve as novel diagnostic biomarkers for patients with DOR.

Influence of Long Non-Coding RNAs on Human Oocyte Development

“…According to bioinformatics analysis, “cell adhesion and apoptosis”, “steroid biosynthesis”, “immune system”, and other biological processes were considerably enriched in the differentially expressed lncRNAs genes. 68 Patients with DOR had lower expression of lnc-GULP1-2:1, but patients with PCOS had significantly higher reserve. 69 Duan et al discovered 78 lncRNAs differentially expressed in GCs of patients with premature ovarian failure (POF), among these, the significantly down-regulated lncRNA ZNF674-AS1 was positively correlated with serum basal estradiol and AMH levels, and negatively correlated with basal FSH levels.…”

Follicular development and ovary aging: single-cell studies

“…Several investigations frequently utilize overexpression, knockdown, RNA immunoprecipitation, luciferase reporter [28], real-time quantitative polymerase chain reaction (RT-qPCR) [29], and flow cytometry [30]. The data generated using these conventional techniques are predominantly unidirectional, focusing on changes in cell and protein quantities or a subset of genes and their activities.…”

“…Next-generation sequencing (NGS), also known as highthroughput sequencing, was invented in 2005 and has resulted in a more comprehensive knowledge of follicular development in both healthy and pathological circumstances [29,[36][37][38]. In addition, several noncoding RNAs have been subsequently and persistently investigated.…”

Metabolomic Analysis Reveals Association between Decreased Ovarian Reserve and In Vitro Fertilization Outcomes

“…Owing to the deleterious impact of environmental pollution, psychological stress, and unhealthy lifestyles, infertility has witnessed a marked uptick in recent years and emerged as a pressing global health issue [ 1 , 2 , 3 , 4 ]. In a recent report published by the World Health Organization in 2022, it was estimated that the global prevalence of infertility stands at 17.5%, with the Western Pacific region exhibiting the highest rates, peaking at 23.2% [ 5 , 6 ].…”