{
    "abstract": "Is the microRNA (miRNA) expression pattern of cumulus oophorus cells (COCs) in women undergoing medically assisted reproduction (MAR) procedures differentially modulated according to patient age and gonadotropin treatment strategy?\nMaternal age is an independent factor impacting miRNA expression in COCs while gonadotropin treatment may affect follicular miRNA expression and IVF efficacy.\nEpigenetic mechanisms in female infertility are complex and poorly studied. DNA methylation, histone modifications, miRNAs and nucleosome positioning influence cellular machinery through positive and negative feedback mechanisms either alone or interactively. miRNAs are important regulators during oogenesis, spermatogenesis and early embryogenesis, and are reported to play a role in regulating crosstalk between the oocyte and COCs. Although miRNome analysis has been performed in female human reproductive tissues (endometrium, myometrium, cervix and ovaries), epigenetic modifications in women with infertility have not been explored in detail. In addition, the impact of gonadotropin treatments during MAR on miRNA expression in COCs has not been fully investigated.\nThis study was carried out in 53 COC samples obtained from mature metaphase II (MII) oocytes in 53 women undergoing MAR treatment. A total of 38 samples for assay development were pooled by maternal age and gonadotropin treatment into four predetermined subgroups: \u226536\u2009years and recombinant human FSH (r-hFSH), n\u2009=\u200910; \u226536\u2009years and r-hFSH+ recombinant human-luteinizing hormone (r-hLH), n\u2009=\u200910; \u226435\u2009years and r-hFSH, n\u2009=\u20099; \u226435\u2009years and r-hFSH+r-hLH, n\u2009=\u20099. miRNome profiles were determined and compared between subgroups. Expression of defined miRNAs was validated in the remaining fifteen samples, representative of each subgroup, by quantitative polymerase chain reaction (PCR).\nCOCs were processed for miRNA-enriched total RNA extraction and pooled in homogeneous subgroups to obtain a sufficient amount and quality of starting material to perform the analysis. Each pooled sample underwent miRNA profiling using PCR assay system to examine expression of 752 human miRNAs without pre-amplification. Data were analyzed using the delta-delta Ct method for relative quantitation and prediction of target genes (with at least four algorithms predicting the same miRNA-gene interaction pair (HIT)>4). The miRSystem database provided functional annotation enrichment (raw P-value <0.05) of co-expressed miRNAs.\nWe found distinctive miRNA expression profiles in each subgroup correlating with age and MAR stimulation. In addition, a number of selective and co-expressed miRNAs were revealed by comparative analysis. A cluster of 37 miRNAs were commonly but differentially expressed in all four pools. Significant differences were observed in expression regulation of 37 miRNAs between age groups (\u226435 or \u226536) in women receiving r-hFSH+r-hLH compared to those receiving r-hFSH alone. Higher concentrations and increased numbers of miRNAs were recorded in younger than in older patients, regardless of treatment. Functional and expression studies performed to retrieve common miRNome profiles revealed an enrichment of biological functions in oocyte growth and maturation, embryo development, steroidogenesis, ovarian hyperstimulation, apoptosis and cell survival, glucagon and lipid metabolism, and cell trafficking. The highest scored pathways of target genes of the 37 common miRNAs were associated with mitogen-activated protein kinase (MAPK) signaling pathways, G alpha signaling, transcription regulation, tight junctions, RNA polymerase I and III, and mitochondrial transcription. We identified a potential age- and MAR stimulation-dependent signature in the miRNA landscape of COCs.\nWe cannot rule out the possibility that other unknown individual genetic or clinical factors may have interfered with the reported results. Since miRNA profiling was conducted with a predefined array of target probes, other miRNA molecules, potentially modulated by age and hormonal stimulation, may have been missed in this study.\nmiRNA expression in COCs is modulated by gonadotropin treatment and correlates strongly with age. A better understanding of the expression patterns and functions of miRNAs may lead to the development of novel therapeutics to treat ovarian dysfunction and improve fertility in older women.\nThis study was funded by Merck KGaA, Darmstadt, Germany. All authors declared no competing interest, except SL and TD who are fully employed by Merck KGaA.\nN/A.",
    "authors": [
        {
            "affiliation": "Department of Precision Medicine, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.\nInstitute of Experimental Endocrinology and Oncology 'Gaetano Salvatore' (IEOS)-National Research Council (CNR), Naples 80131, Italy.",
            "firstname": "C",
            "initials": "C",
            "lastname": "Dell'Aversana"
        },
        {
            "affiliation": "EPI-C S.r.l., Naples 80138, Italy.",
            "firstname": "F",
            "initials": "F",
            "lastname": "Cuomo"
        },
        {
            "affiliation": "Merck KGaA, Darmstadt 64293, Germany.",
            "firstname": "S",
            "initials": "S",
            "lastname": "Longobardi"
        },
        {
            "affiliation": "Merck KGaA, Darmstadt 64293, Germany.",
            "firstname": "T",
            "initials": "T",
            "lastname": "D'Hooghe"
        },
        {
            "affiliation": "Outpatient Fertility Unit, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.",
            "firstname": "F",
            "initials": "F",
            "lastname": "Caprio"
        },
        {
            "affiliation": "Department of Precision Medicine, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.\nDepartment of Medicine, Surgery and Dentistry \"ScuolaMedicaSalernitana\", University of Salerno, Baronissi, SA 84081, Italy.",
            "firstname": "G",
            "initials": "G",
            "lastname": "Franci"
        },
        {
            "affiliation": "Department of Woman, Child and General and Special Surgery, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.",
            "firstname": "M",
            "initials": "M",
            "lastname": "Santonastaso"
        },
        {
            "affiliation": "Outpatient Fertility Unit, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.\nDepartment of Woman, Child and General and Special Surgery, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.",
            "firstname": "N",
            "initials": "N",
            "lastname": "Colacurci"
        },
        {
            "affiliation": "Department of Assisted Reproduction, Versilia Hospital, Lido di Camaiore, Lucca 55049, Italy.",
            "firstname": "S",
            "initials": "S",
            "lastname": "Barone"
        },
        {
            "affiliation": "Department of Reproductive Medicine, International Evangelical Hospital, Genoa 16122, Italy.",
            "firstname": "V",
            "initials": "V",
            "lastname": "Pisaturo"
        },
        {
            "affiliation": "Merck Serono S.p.A, Rome 00176, Italy.\nInstitute of Genetic Research (IRG), Naples 80143, Italy.",
            "firstname": "D",
            "initials": "D",
            "lastname": "Valerio"
        },
        {
            "affiliation": "Department of Precision Medicine, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy.",
            "firstname": "L",
            "initials": "L",
            "lastname": "Altucci"
        }
    ],
    "conclusions": null,
    "copyrights": "\u00a9 The Author(s) 2021. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology.",
    "doi": "10.1093/humrep/deaa364",
    "journal": "Human reproduction (Oxford, England)",
    "keywords": [
        "aging",
        "cumulus oophorus cells",
        "epigenetics",
        "gonadotropin stimulation",
        "medically assisted reproduction",
        "miRNA"
    ],
    "methods": null,
    "publication_date": "2021-01-18",
    "pubmed_id": "33454781\n20215419\n30207252\n28007692\n15381691\n29747587\n28071684\n24419497\n30008696\n22451503\n27158401\n25191020\n30588778\n16648150\n27578177\n28723025\n31115532\n29259413\n24882616\n15967989\n19357131\n15035550\n26198660\n26707514\n26692146\n26558383\n28188594\n11925399\n28152035\n26209829\n21245905\n19584175\n30315515\n15013804\n26458360\n30561694\n31356852\n27102646\n30710230\n16580310\n11835587\n29170163\n31207900\n25188034\n23660593\n29228146\n23632103\n22796494\n30353018\n24828505\n28729464",
    "results": null,
    "title": "Age-related miRNome landscape of cumulus oophorus cells during controlled ovarian stimulation protocols in IVF cycles.",
    "xml": "<Element 'PubmedArticle' at 0x7885032da1b0>"
}