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Changes in the transcriptomic profile of cumulus cells under the influence of pre-incubation of human oocytes in patients treated in ART cycles

ABSTRACT: In this study, we tried to find out if the pre-incubation of the cumulus-oocyte complex can lead to better cumulus cell (CCs) function and higher oocyte quality by changing the transcriptomic profile of CCs. For this purpose, 140 cumulus cell samples were isolated from 12 participants and divided into two groups based on pre-incubation time. In the T0 group, the cumulus-oocyte complex was immediately dissected to separate the CCs from around the oocytes. In the T2 group, dissection and preparation of CCs were done after 2 hours of incubation. Then, the transcriptomic profile of the CCs of the non-pre-incubation group (T0) was compared to the 2-hour pre-incubation group (T2). 70 samples were placed in each group. 40 samples from each group were used for RNA-Sequencing and the remaining 30 samples of each group were used to confirm RNA-Sequencing results via qRT‑PCR. The CCs transcriptome analysis showed 17 genes were downregulated and 22 genes upregulated in the T2 group compared to the T0 group. Also, the pathways related to ATP production (oxidative phosphorylation, electron transport chain, and Mitochondrial complex I assembly model OXPHOS system), TNF-alpha signaling pathway, and glucocorticoid receptor pathway enriched in the T2 group compared to the T0 group. Also, the TGF-β pathway was decreased in the T2 group compared to the T0 group. This study showed that 2 hours’ pre-incubation leads to changes in important pathways in CCs, which has a positive effect on oocyte quality. 2 hours’ pre-incubation may lead to a better support of the CCs to the oocyte and thus to a more complete maturation of the oocyte.

ORGANISM(S): Homo sapiens

PROVIDER: GSE252038 | GEO | 2024/06/22

REPOSITORIES: GEO

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Project description:Purpose: The goal of this study was to identify differentially expressed genes and pathways between the cumulus of compact/unstimulated cumulus-oocyte-complex (COC) and the cumulus of expanded/stimulated COC. Methods: mRNA profiles of Compact/unstimulated cumulus cells (CCs) from germinal vesicle (GV) COC obtained from two patients undergoing unstimulated IVM procedure and expanded/stimulated CCs from metaphase 2 (MII) COC obtained from three patients undergoing IVF/ICSI were generated by deep sequencing using Illumina HiSeq 2000. The sequence reads that passed quality filters were mapped to the human genome (hg19) using Tophat software. Differential expression analysis was done using DESeq bioconductor package. qRT–PCR validation was performed using SYBR Green assays. Results: A total of 40-80 million sequence reads per sample were mapped to the human genome (hg19). A total of 1746 differentially expressed genes between compact and expanded CCs with fold change > 2, and adjusted p value < 0.05 were identified. Gene ontology analysis of differentially expressed genes revealed a number of cellular processes regulated during the periovulatory interval including cellular movement, inflammatory response, immune cell trafficking, tissue development, lipid metabolism, tissue morphology, DNA replication and cell cycle. A total of 116 of the differentially expressed genes were annotated as long non-coding RNAs, 10 of them coded from introns of genes known to be involved in granulosa cell processes suggesting that unique non coding RNA transcripts may contribute to the regulation of cumulus expansion and oocyte maturation. Results were validated using qRT-PCR. Conclusions: Using global transcriptome sequencing we identified new important genes and non coding RNAs involved in COC maturation and cumulus expansion, which may contribute to improve the process of in vitro maturation of immature oocytes utilized in IVM cycles mRNA profiles of Compact/unstimulated cumulus cells (CCs) from germinal vesicle (GV) COC obtained from two patients undergoing unstimulated IVM procedure and expanded/stimulated CCs from metaphase 2 (MII) COC obtained from three patients undergoing IVF/ICSI were generated by deep sequencing using Illumina HiSeq 2000.

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Changes in the transcriptomic profile of cumulus cells under the influence of pre-incubation of human oocytes in patients treated in ART cycles

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