{"paper_id":"6e760cce-b858-4679-8ded-0d32368fb747","body_text":"Abstract\nPurpose\nThe objective of this experimental study was to compare the global gene expression profile of CC of mature oocytes in 18 patients with severe endometriosis and CC in 18 control patients affected by a severe male factor.\nMethods\nFor each group, the CC were pooled, RNA was extracted and a microarray performed. For validating the microarray, a quantitative real-time PCR was performed in the CC of an independent set of patients with endometriosis (n = 5) and controls (n = 7).\nResults\n595 differentially expressed genes (320 down-regulated, 275 up-regulated, p < 0.05, fold change ≥1.5) were identified. The most significant changes were observed in genes involved in the chemokine signaling and cell-cell or cell-extracellular matrix adhesion pathways. Several genes of these pathways were down-regulated in endometriosis. Individual RT-PCR assays confirmed the microarray for ten genes.\nConclusions\nSeveral genes involved in the chemokine mediated-signaling pathway and in the functional cross-talk between CC and the oocyte are down-regulated in endometriosis CC. The impairment of these processes could explain the reduction of oocyte competence in endometriosis. This preliminary knowledge could be the starting point for a more detailed elucidation of the relationship between endometriosis and oocyte competence.\nSimilar content being viewed by others\nReferences\nBarnhart K, Dunsmoor-Su R, Coutifaris C. Effect of endometriosis on in vitro fertilization. Fertil Steril. 2002;77:1148–55.\nKennedy S, Bergqvist A, Chapron C, D’Hooghe T, Dunselman G, Greb R, et al. On behalf of the ESHRE special interest group for endometriosis and endometrium guideline development group. ESHRE guideline for the diagnosis and treatment of endometriosis. Hum Reprod. 2005;20:2698–704.\nThe Practice Committee of the American Society for Reproductive Medicine. Endometriosis and infertility: a committee opinion. Fertil Steril. 2012;98:591–8.\nLin X, Wei M, Tong X, Xu W, Zhou F, Huang Q, et al. Outcome of in vitro fertilization in endometriosis-associated infertility: a 5-year database cohort study. Chin Med J. 2012;125:2688–93.\nSociety for Assisted Reproductive Technology, the American Society for Reproductive Medicine Assisted reproductive technology in the United States: 2010 results generated from the American Society for Reproductive Medicine/Society for Assisted Reproduction registry. Available at: www.sart.org\nDunselman G, Vermeulen N, Becker C, Calhaz-Jorge C, D'Hooghe T, De Bie B, et al. ESHRE guideline: management of women with endometriosis. Hum Reprod. 2014;29:400–12.\nSimón C, Gutierrez A, Vidal A, Tarin JJ, Remohí J, Pellicer A. Outcome of patients with endometriosis in assisted reproduction: results from in-vitro fertilization and oocyte donation. Hum Reprod. 1994;9:725–9.\nGarrido N, Navarro J, García-Velasco J, Remohí J, Pellicer A, Simón C. The endometrium versus embryonic quality in endometriosis-related infertility. Hum Reprod Update. 2002;8:95–103.\nAdriaenssens T, Wathlet S, Segers I, Verheyen G, De Vos A, Van der Elst J, et al. Cumulus cell gene expression is associated with oocyte developmental quality and influenced by patient and treatment characteristics. Hum Reprod. 2010;25:1259–70.\nAssou S, Haouzi D, Mahmoud K, Aouacheria A, Guillemin Y, Pantesco V, et al. A non-invasive test for assessing embryo potential by gene expression profiles of human cumulus cells: a proof of concept study. Mol Hum Reprod. 2008;14:711–9.\nFeuerstein P, Cadoret V, Dalbies-Tran R, Guerif F, Bidault R, Royere D. Gene expression in human cumulus cells: one approach to oocyte competence. Hum Reprod. 2007;22:3069–77.\nHamel M, Dufort I, Robert C, Gravel C, Leveille MC, Leader A, et al. Identification of differentially expressed markers in human follicular cells associated with competent oocytes. Hum Reprod. 2008;23:1118–27.\nMcKenzie LJ, Pangas SA, Carson SA, Kovanci E, Cisneros P, Buster JE, et al. Human cumulus granulosa cell gene expression: a predictor of fertilization and embryo selection in women undergoing IVF. Hum Reprod. 2004;19:2869–74.\nvan Montfoort AP, Geraedts JP, Dumoulin JC, Stassen AP, Evers JL, Ayoubi TA. Differential gene expression in cumulus cells as a prognostic indicator of embryo viability: a microarray analysis. Mol Hum Reprod. 2008;14:157–68.\nWathlet S, Adriaenssens T, Segers I, Verheyen G, Van de Velde H, Coucke W, et al. Cumulus cell gene expression predicts better cleavage-stage embryo or blastocyst development and pregnancy for ICSI patients. Hum Reprod. 2011;26:1035–51.\nBrannian J, Eyster K, Mueller BA, Bietz MG, Hansen K. Differential gene expression in human granulosa cells from recombinant FSH versus human menopausal gonadotropin ovarian stimulation protocols. Reprod Biol Endocrinol. 2010;8:25.\nGrondahl ML, Borup R, Lee YB, Myrhoj V, Meinertz H, Sorensen S. Differences in gene expression of granulosa cells from women undergoing controlled ovarian hyperstimulation with either recombinant follicle-stimulating hormone or highly purified human menopausal gonadotropin. Fertil Steril. 2009;91:1820–30.\nSaito H, Seino T, Kaneko T, Nakahara K, Toya M, Kurachi H. Endometriosis and oocyte quality. Gynecol Obstet Invest. 2002;53 Suppl 1:46–51.\nSifer C, Benifla JL, Bringuier AF, Porcher R, Blanc-Layrac G, Madelenat P, et al. Could induced apoptosis of human granulosa cells predict in vitro fertilization-embryo transfer outcome? A preliminary study of 25 women. Eur J Obstet Gynecol Reprod Biol. 2002;103:150–3.\nAmerican Society for Reproductive Medicine. Revised american society for reproductive medicine classification of endometriosis: 1996. Fertil Steril. 1997;67:817–21.\nVolpes A, Sammartano F, Coffaro F, Mistretta V, Scaglione P, Allegra A. Number of good quality embryos on day 3 is predictive for both pregnancy and implantation rates in in vitro fertilization/intracytoplasmic sperm injection cycles. Fertil Steril. 2004;82:1330–6.\nPatrat C, Kaffel A, Delaroche L, Guibert J, Jouannet P, Epelboin S, et al. Optimal timing for oocyte denudation and intracytoplasmic sperm injection. Obstet Gynecol Int. 2012;2012:403531.\nGautier L, Cope L, Bolstad BM, Irizarry RA. affy--analysis of Affymetrix GeneChip data at the probe level. Bioinformatics. 2004;20:307–15.\nKanehisa M, Goto S. KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 2000;28:27–30.\nIrizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 2003;4:249–64.\nSmyth GK. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol. 2004; 3:Article3\nBenjamini Y, Drai D, Elmer G, Kafkafi N, Golani I. Controlling the false discovery rate in behavior genetics research. Behav Brain Res. 2001;125:279–84.\nWettenhall JM, Simpson KM, Satterley K, Smyth GK. affylmGUI: a graphical user interface for linear modeling of single channel microarray data. Bioinformatics. 2006;22:897–9.\nPellicer A, Albert C, Mercader A, Bonilla-Musoles F, Remohi J, Simón C. The follicular and endocrine environment in women with endometriosis: local and systemic cytokine production. Fertil Steril. 1998;70:425–31.\nYanushpolsky EH, Best CL, Jackson KV, Clarke RN, Barbieri RL, Hornstein MD. Effects of endometriomas on oocyte quality, embryo quality, and pregnancy rates in in vitro fertilization cycles: a prospective, case-controlled study. J Assist Reprod Genet. 1998;15:193–7.\nBuccione R, Schroeder AC, Eppig JJ. Interactions between somatic cells and germ cells throughout mammalian oogenesis. Biol Reprod. 1990;43:543–7.\nMotta PM, Makabe S, Naguro T, Correr S. Oocyte follicle cells association during development of human ovarian follicle. A study by high resolution scanning and transmission electron microscopy Arch Histol Cytol. 1994;57:369–94.\nAlbertini DF, Combelles CM, Benecchi E, Carabatsos MJ. Cellular basis for paracrine regulation of ovarian follicle development. Reproduction. 2001;121:647–53.\nAssidi M, Dufort I, Ali A, Hamel M, Algriany O, Dielemann S, et al. Identification of potential markers of oocyte competence expressed in bovine cumulus cells matured with follicle-stimulating hormone and/or phorbol myristate acetate in vitro. Biol Reprod. 2008;79:209–22.\nAssou S, Anahory T, Pantesco V, Le Carrour T, Pellestor F, Klein B, et al. The human cumulus–oocyte complex gene-expression profile. Hum Reprod. 2006;21:1705–19.\nMay-Panloup P, Ferre-L’Hotellier V, Moriniere C, Marcaillou C, Lemerle S, Malinge MC, et al. Molecular characterization of corona radiata cells from patients with diminished ovarian reserve using microarray and microfluidic-based gene expression profiling. Hum Reprod. 2012;27:829–43.\nBettegowda A, Patel OV, Lee KB, Park KE, Salem M, Yao J, et al. Identification of novel bovine cumulus cell molecular markers predictive of oocyte competence: functional and diagnostic implications. Biol Reprod. 2008;79:301–9.\nAssidi M, Montag M, Van Der Ven K, Sirard MA. Biomarkers of human oocyte developmental competence expressed in cumulus cells before ICSI: a preliminary study. J Assist Reprod Genet. 2011;28:173–88.\nGatta V, Tatone C, Ciriminna R, Vento M, Franchi S, Aurora M, et al. Gene expression profiles of cumulus cells obtained from women treated with recombinant human luteinizing hormone + recombinant human follicle-stimulating hormone or highly purified human menopausal gonadotropin versus recombinant human follicle-stimulating hormone alone. Fertil Steril. 2013;99:2000–8.\nFritz JH, Girardin SE. How Toll-like receptors and Nod-like receptors contribute to innate immunity in mammals. J Endotoxin Res. 2005;11:390–4.\nLiu Z, Shimada M, Richards JS. The involvement of the Toll-like receptor family in ovulation. J Assist Reprod Genet. 2008;25:223–8.\nLiu Z, de Matos DG, Fan HY, Shimada M, Palmer S, Richards JS. Interleukin-6: an autocrine regulator of the mouse cumulus cell-oocyte complex expansion process. Endocrinology. 2009;150:3360–8.\nRichards JS, Russell DL, Robker RL, Dajee M, Alliston TN. Molecular mechanisms of ovulation and luteinization. Mol Cell Endocrinol. 1998;145:47–54.\nGerard N, Caillaud M, Martoriati A, Goudet G, Lalmanach AC. The interleukin-1 system and female reproduction. J Endocrinol. 2004;180:203–12.\nBuscher U, Chen FC, Kentenich H, Schmiady H. Cytokines in the follicular fluid of stimulated and non-stimulated human ovaries; is ovulation a suppressed inflammatory reaction? Hum Reprod. 1999;14:162–6.\nSarapik A, Velthut A, Haller-Kikkatalo K, Faure GC, Bene MC, de Carvalho Bittencourt M, et al. Follicular proinflammatory cytokines and chemokines as markers of IVF success. Clin Dev Immunol. 2012;2012:606459.\nMehrad B, Keane MP, Strieter RM. Chemokines as mediators of angiogenesis. Thromb Haemost. 2007;97:755–62.\nSchindler R, Nilsson E, Skinner MK. Induction of ovarian primordial follicle assembly by connective tissue growth factor CTGF. PLoS One. 2010;5:e12979.\nFulop C, Szanto S, Mukhopadhyay D, Bardos T, Kamath RV, Rugg MS, et al. Impaired cumulus mucification and female sterility in tumor necrosis factor-induced protein-6 deficient mice. Development. 2003;130:2253–61.\nMilner CM, Day AJ. TSG-6: a multifunctional protein associated with inflammation. JCell Sci. 2003;116:1863–73.\nCampbell S, Swann HR, Aplin JD, Seif MW, Kimber SJ, Elstein M. CD44 is expressed throughout pre-implantation human embryo development. Hum Reprod. 1995;10:425–30.\nOhta N, Saito H, Kuzumaki T, Takahashi T, Ito MM, Saito T, et al. Expression of CD44 in human cumulus and mural granulosa cells of individual patients in in-vitro fertilization programmes. Mol Hum Reprod. 1999;5:22–8.\nFamiliari G, Verlengia C, Nottola SA, Renda T, Micara G, Aragona C, et al. Heterogeneous distribution of fibronectin, tenascin-C, and laminin immunoreactive material in the cumulus-corona cells surrounding mature human oocytes from IVF-ET protocols–evidence that they are composed of different subpopulations: an immunohistochemical study using scanning confocal laser and fluorescence microscopy. Mol Reprod Dev. 1996;43:392–402.\nKonno R, Fujiwara H, Netsu S, Odagiri K, Shimane M, Nomura H, et al. Gene expression profiling of the rat endometriosis model. Am J Reprod Immunol. 2007;58:330–43.\nPeng H, Chang B, Lu C, Su J, Wu Y, Lv P, et al. Nlrp2, a maternal effect gene required for early embryonic development in the mouse. PLoS One. 2012;7:e30344.\nDeclaration of interest\nThe Authors of this Paper declare there to be no conflict of interest which could be perceived as prejudicing the impartiality of the research reported herein.\nFunding\nThis research did not receive any specific grant from any funding agency in the public, commercial or not-for-profit sector; the funds were provided directly by the ANDROS Day Surgery Clinic, Palermo, Italy.\nAuthor information\nAuthors and Affiliations\nCorresponding author\nAdditional information\nCapsule Several genes involved in the chemokine mediated-signaling pathway and in the cross-talk between cumulus cells and the oocyte are down-regulated in the cumulus cells of patients with endometriosis.\nThe first two authors contributed equally to this work\nElectronic supplementary material\nBelow is the link to the electronic supplementary material.\nESM 1 (download DOC )\n(DOC 30 kb)\nRights and permissions\nAbout this article\nCite this article\nAllegra, A., Raimondo, S., Volpes, A. et al. The gene expression profile of cumulus cells reveals altered pathways in patients with endometriosis. J Assist Reprod Genet 31, 1277–1285 (2014). https://doi.org/10.1007/s10815-014-0305-1\nReceived:\nAccepted:\nPublished:\nIssue date:\nDOI: https://doi.org/10.1007/s10815-014-0305-1","source_license":"CC0","license_restricted":false}