Transcriptome analysis of human cumulus cells reveals hypoxia as the main determinant of follicular senescence.
Can RNA sequencing of human cumulus cells (CC) reveal molecular pathways involved in the physiology of reproductive aging?



Senescent but not young CC activate gene pathways associated with hypoxia and oxidative stress.



Shifts in socioeconomic norms are resulting in larger numbers of women postponing childbearing. The reproductive potential is sharply decreased with aging, and the reasons are poorly understood. Since CCs play an integral role in oocyte maturation and direct access to human oocytes is limited, we used whole transcriptome analysis of these somatic cells to gain insights into the molecular mechanisms playing a role in follicular senescence.



Twenty CC samples (from a total of 15 patients) were obtained from oocytes of either male factor or egg donor patients. RNA sequencing and bioinformatic tools were used to identify differentially expressed genes between CCs from seven aged and eight young patients (<35 (years old) y.o. vs >40 y.o.). Quantitative-PCR and immunoflourescent staining were used for validation.



RNA sequencing identified 11 572 genes expressed in CC of both age cohorts, 45 of which were differentially expressed. In CC collected from patients >40 y.o., genes involved in the hypoxia stress response (NOS2, RORA and NR4A3), vasculature development (NR2F2, PTHLH), glycolysis (RALGAPA2 and TBC1D4) and cAMP turnover (PDE4D) were significantly overexpressed when compared with CC of patients younger than 35 y.o.



This study focused almost exclusively on assessing the genetic differences in CC transcriptome between young and older women. These genetic findings were not fully correlated with embryonic development and clinical outcome.



Our data provide a new hypothesis-follicular hypoxia-as the main mechanism leading to ovarian follicular senescence and suggest a link between cumulus cell aging and oocyte quality decay. If specific molecular findings of hypoxia would be confirmed also in oocytes, genetic platforms could screen CC for hypoxic damage and identify healthier oocytes. Protocols of ovarian stimulation in older patients could also be adjusted to diminish oocyte exposure time to hypoxic follicles.



GEO accession number: GSE81579 STUDY FUNDING AND COMPETING INTERESTS: Funded in part by EMD Serono Grant for Fertility Innovation (GFI).
Department of Obstetrics, Gynecology and Reproductive Sciences, Yale Fertility Center, Yale University, New Haven, CT 065112, USA.
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Molinari
Department of Statistics, University of Connecticut, Storrs, CT 06269, USA.
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Bar
Department of Molecular, Cellular and Developmental Biology and Howard Huges Medical Institute, Yale University, New Haven, CT 06511, USA.
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Pyle
Department of Obstetrics, Gynecology and Reproductive Sciences, Yale Fertility Center, Yale University, New Haven, CT 065112, USA pasquale.patrizio@yale.edu.
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Patrizio
D000328:Adult; D015687:Cell Hypoxia / Q000235:genetics / Q000502:physiology*; D054885:Cumulus Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D006080:Ovarian Follicle / Q000166:cytology* / Q000378:metabolism*; D010062:Ovulation Induction; D011247:Pregnancy; D017423:Sequence Analysis, RNA; D059467:Transcriptome / Q000235:genetics
RNA sequencing; WGCNA; cumulus cells transcriptome; oocyte senescence