Advertisement

Association between early embryo morphokinetics plus cumulus cell gene expression and assisted reproduction outcomes in polycystic ovary syndrome women

Published:December 14, 2018DOI:https://doi.org/10.1016/j.rbmo.2018.10.010

      Abstract

      Research question

      Can a combination of time-lapse morphokinetic parameters and cumulus cell gene expression in polycystic ovary syndrome (PCOS) women be used to predict assisted reproductive treatment outcome?

      Design

      A total of 547 embryos from 100 intracytoplasmic sperm injection (ICSI) cycles were evaluated. Fifty women with PCOS and 50 women who were categorized as tubal factor infertility were recruited. Time-lapse records were annotated for time to pronuclear fading (tPNf), time to 2 to 8 cells (t2–t8), reverse cleavage, direct cleavage and also for the presence of multinucleation. Expression levels of three genes involved in mitotic divisions, diaphanous-related formin 2 (DIAPH2), nibrin (NBN) and NIMA-related protein kinase (NEK4), were measured in 100 associated cumulus cell samples using quantitative real-time polymerase chain reaction.

      Results

      Expression of DIAPH2 and NBN was significantly higher in the embryos of PCOS patients that resulted in implantation, biochemical and clinical pregnancies as well as live birth compared with embryos that were negative for these outcomes (P <0.01). However, in the tubal factor group, NBN gene expression was significantly higher in embryos resulting in biochemical pregnancy, clinical pregnancy and live birth (P <0.01) only. Multivariate logistic regression analysis showed that tPNf together with DIAPH2 gene expression were independent prognostic factors of clinical pregnancy rate and live birth in both groups.

      Conclusions

      Some time-lapse embryo parameters may be related to cumulus gene expression and clinical outcome. Furthermore, the expressions of cumulus cell genes involved in mitotic divisions are significantly associated with ICSI outcome using Day 3 embryo transfer.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Reproductive BioMedicine Online
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Adamson G.D.
        • Abusief M.E.
        • Palao L.
        • Witmer J.
        • Palao L.M.
        • Gvakharia M.
        Improved implantation rates of day 3 embryo transfers with the use of an automated time-lapse-enabled test to aid in embryo selection.
        Fertil Steril. 2016; 105 (e6): 369-375
        • Anderson R.A.
        • Sciorio R.
        • Kinnell H.
        • Bayne R.A.
        • Thong K.J.
        • De Sousa P.A.
        • Pickering S.
        Cumulus gene expression as a predictor of human oocyte fertilization, embryo development and competence to establish a pregnancy.
        Reproduction. 2009; 138: 629-637
        • Assou S.
        • Haouzi D.
        • De Vos J.
        • Hamamah S.
        Human cumulus cells as biomarkers for embryo and pregnancy outcomes.
        Mol Hum Reprod. 2010; 16: 531-538
        • Assou S.
        • Haouzi D.
        • Mahmoud K.
        • Aouacheria A.
        • Guillemin Y.
        • Pantesco V.
        • Reme T.
        • Dechaud H.
        • De Vos J.
        • Hamamah S.
        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: 711-719
        • Azzarello A.
        • Hoest T.
        • Mikkelsen A.L.
        The impact of pronuclei morphology and dynamicity on live birth outcome after time-lapse culture.
        Hum. Reprod. 2012; 27: 2649-2657
        • Balakier H.
        • Sojecki A.
        • Motamedi G.
        • Librach C.
        Impact of multinucleated blastomeres on embryo developmental competence, morphokinetics, and aneuploidy.
        Fertil. Steril. 2016; 106 (e2): 608-614
        • Bellver J.
        • Mifsud A.
        • Grau N.
        • Privitera L.
        • Meseguer M.
        Similar morphokinetic patterns in embryos derived from obese and normoweight infertile women: a time-lapse study.
        Hum. Reprod. 2013; 28: 794-800
        • Boomsma C.M.
        • Macklon N.S.
        What can the clinician do to improve implantation?.
        Reprod. Biomed. Online. 2007; 14 Spec No 1: 27-37
        • Borup R.
        • Thuesen L.L.
        • Andersen C.Y.
        • Nyboe-Andersen A.
        • Ziebe S.
        • Winther O.
        • Grondahl M.L.
        Competence Classification of Cumulus and Granulosa Cell Transcriptome in Embryos Matched by Morphology and Female Age.
        PLoS. One. 2016; 11e0153562
        • Braga D.P.
        • Setti A.S.
        • Lo Turco E.G.
        • Cordeiro F.B.
        • Cabral E.C.
        • Cortezzi S.S.
        • Ono E.
        • Figueira R.C.
        • Eberlin M.N.
        • Borges JR, E.
        Protein expression in human cumulus cells as an indicator of blastocyst formation and pregnancy success.
        J. Assist. Reprod. Genet. 2016; 33: 1571-1583
        • Broer S.L.
        • Van Disseldorp J.
        • Broeze K.A.
        • Dolleman M.
        • Opmeer B.C.
        • Bossuyt P.
        • Eijkemans M.J.
        • Mol B.W.
        • Broekmans F.J.
        • Group I.S.
        Added value of ovarian reserve testing on patient characteristics in the prediction of ovarian response and ongoing pregnancy: an individual patient data approach.
        Hum. Reprod. Update. 2013; 19: 26-36
        • Burnik Papler T.
        • Vrtacnik Bokal E.
        • Lovrecic L.
        • Kopitar A.n.
        • Maver A.
        No specific gene expression signature in human granulosa and cumulus cells for prediction of oocyte fertilization and embryo implantation.
        PLoS. One. 2015; 10e0115865
        • Conaghan J.
        • Chen A.A.
        • Willman S.P.
        • Ivani K.
        • Chenette P.E.
        • Boostanfar R.
        • Baker V.L.
        • Adamson G.D.
        • Abusief M.E.
        • Gvakharia M.
        • Loewke K.E.
        • Shen S.
        Improving embryo selection using a computer-automated time-lapse image analysis test plus day 3 morphology: results from a prospective multicentre trial.
        Fertil. Steril. 2013; 100 (e5): 412-419
        • Coticchio G.
        • Mignini Renzini M.
        • Novara P.V.
        • Lain M.
        • De Ponti E.
        • Turchi D.
        • Fadini R.
        • Dal Canto M.
        Focused time-lapse analysis reveals novel aspects of human fertilization and suggests new parameters of embryo viability.
        Hum. Reprod. 2018; 33: 23-31
        • Dal Canto M.
        • Coticchio G.
        • Mignini Renzini M.
        • De Ponti E.
        • Novara P.V.
        • Brambillasca F.
        • Comi R.
        • Fadini R.
        Cleavage kinetics analysis of human embryos predicts development to blastocyst and implantation.
        Reprod. Biomed. Online. 2012; 25: 474-480
        • Davar R.
        • Oskouian H.
        • Ahmadi S.
        • Firouzabadi R.D.
        GnRH antagonist/letrozole versus microdose GnRH agonist flare protocol in poor responders undergoingin vitro fertilization.
        Taiwan J Obstet Gynecol. 2010; 49: 297-301
        • Desai N.
        • Ploskonka S.
        • Goodman L.
        • Attaran M.
        • Goldberg J.M.
        • Austin C.
        • Falcone T.
        Delayed blastulation, multinucleation, and expansion grade are independently associated with live-birth rates in frozen blastocyst transfer cycles.
        Fertil. Steril. 2016; 106: 1370-1378
        • Desai N.
        • Ploskonka S.
        • Goodman L.R.
        • Austin C.
        • Goldberg J.
        • Falcone T.
        Analysis of embryo morphokinetics, multinucleation and cleavage anomalies using continuous time-lapse monitoring in blastocyst transfer cycles.
        Reprod. Biol. Endocrinol. 2014; 12: 54
        • Desch L.
        • Bruno C.
        • Luu M.
        • Barberet J.
        • Choux C.
        • Lamotte M.
        • Schmutz E.
        • Sagot P.
        • Fauque P.
        Embryo multinucleation at the two-cell stage is an independent predictor of intracytoplasmic sperm injection outcomes.
        Fertil. Steril. 2017; 107 (e4): 97-103
        • Devjak R.
        • Burnik Papler T.
        • Verdenik I.
        • Fon Tacer K.
        • Vrtacnik Bokal E.
        Embryo quality predictive models based on cumulus cells gene expression.
        Balkan J. Med. Genet. 2016; 19: 5-12
        • Eftekhar M.
        • Aflatoonian A.
        • Mohammadian F.
        • Eftekhar T.
        Adjuvant growth hormone therapy in antagonist protocol in poor responders undergoing assisted reproductive technology.
        Arch. Gynecol. Obstet. 2013; 287: 1017-1021
        • Feuerstein 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: 3069-3077
        • Fishel S.
        • Campbell A.
        • Montgomery S.
        • Smith R.
        • Nice L.
        • Duffy S.
        • Jenner L.
        • Berrisford K.
        • Kellam L.
        • Smith R.
        • D’cruz I.
        • Beccles A.
        Live births after embryo selection using morphokinetics versus conventional morphology: a retrospective analysis.
        Reprod. Biomed. Online. 2017; 35: 407-416
        • Fragouli E.
        • Lalioti M.D.
        • Wells D.
        The transcriptome of follicular cells: biological insights and clinical implications for the treatment of infertility.
        Hum. Reprod. Update. 2014; 20: 1-11
        • Gebhardt K.M.
        • Feil D.K.
        • Dunning K.R.
        • Lane M.
        • Russell D.L.
        Human cumulus cell gene expression as a biomarker of pregnancy outcome after single embryo transfer.
        Fertil. Steril. 2011; 96 (e2): 47-52
        • Gleicher N.
        • Weghofer A.
        • Barad D.H.
        Anti-Mullerian hormone (AMH) defines, independent of age, low versus good live-birth chances in women with severely diminished ovarian reserve.
        Fertil. Steril. 2010; 94: 2824-2827
        • Goodman L.R.
        • Goldberg J.
        • Falcone T.
        • Austin C.
        • Desai N.
        Does the addition of time-lapse morphokinetics in the selection of embryos for transfer improve pregnancy rates? A randomized controlled trial.
        Fertil. Steril. 2016; 105 (e10): 275-285
        • Gurbuz A.S.
        • Gode F.
        • Uzman M.S.
        • INce B.
        • Kaya M.
        • Ozcimen N.
        • Ozcimen E.E.
        • Acar A.
        GnRH agonist triggering affects the kinetics of embryo development: a comparative study.
        J. Ovarian Res. 2016; 9: 22
        • Hammond E.R.
        • Stewart B.
        • Peek J.C.
        • Shelling A.N.
        • Cree L.M.
        Assessing embryo quality by combining non-invasive markers: early time-lapse parameters reflect gene expression in associated cumulus cells.
        Hum. Reprod. 2015; 30: 1850-1860
        • Hardarson T.
        • Bungum M.
        • Conaghan J.
        • Meintjes M.
        • Chantilis S.J.
        • Molnar L.
        • Gunnarsson K.
        • Wikland M.
        Noninferiority, randomized, controlled trial comparing embryo development using media developed for sequential or undisturbed culture in a time-lapse setup.
        Fertil. Steril. 2015; 104 (e1–4): 1452-1459
        • Heijnen E.M.
        • Eijkemans M.J.
        • Hughes E.G.
        • Laven J.S.
        • Macklon N.S.
        • Fauser B.C.
        A meta-analysis of outcomes of conventional IVF in women with polycystic ovary syndrome.
        Hum. Reprod. Update. 2006; 12: 13-21
        • Huang X.
        • Hao C.
        • Shen X.
        • Liu X.
        • Shan Y.
        • Zhang Y.
        • Chen L.
        Differences in the transcriptional profiles of human cumulus cells isolated from MI and MII oocytes of patients with polycystic ovary syndrome.
        Reproduction. 2013; 145: 597-608
        • Khader A.
        • Lloyd S.M.
        • McConnachie A.
        • Fleming R.
        • Grisendi V.
        • La Marca A.
        • Nelson S.M.
        External validation of anti-Mullerian hormone based prediction of live birth in assisted conception.
        J. Ovarian Res. 2013; 6: 3
        • Kirkegaard K.
        • Kesmodel U.S.
        • Hindkjaer J.J.
        • Ingerslev H.J.
        Time-lapse parameters as predictors of blastocyst development and pregnancy outcome in embryos from good prognosis patients: a prospective cohort study.
        Hum. Reprod. 2013; 28: 2643-2651
        • La Marca A.
        • Nelson S.M.
        • Sighinolfi G.
        • Manno M.
        • Baraldi E.
        • Roli L.
        • Xella S.
        • Marsella T.
        • Tagliasacchi D.
        • D'amico R.
        • Volpe A.
        Anti-Mullerian hormone-based prediction model for a live birth in assisted reproduction.
        Reprod. Biomed. Online. 2011; 22: 341-349
        • Lee T.H.
        • Liu C.H.
        • Huang C.C.
        • Hsieh K.C.
        • Lin P.M.
        • Lee M.S.
        Impact of female age and male infertility on ovarian reserve markers to predict outcome of assisted reproduction technology cycles.
        Reprod. Biol. Endocrinol. 2009; 7: 100
        • Meseguer M.
        • Herrero J.
        • Tejera A.
        • Hilligsoe K.M.
        • Ramsing N.B.
        • Remohi J.
        The use of morphokinetics as a predictor of embryo implantation.
        Hum. Reprod. 2011; 26: 2658-2671
        • Milewski R.
        • Czerniecki J.
        • Kuczynska A.
        • Stankiewicz B.
        • Kuczynski W.
        Morphokinetic parameters as a source of information concerning embryo developmental and implantation potential.
        Ginekol. Pol. 2016; 87: 677-684
        • Motato Y.
        • De Los Santos M.J.
        • Escriba M.J.
        • Ruiz B.A.
        • Remohi J.
        • Meseguer M.
        Morphokinetic analysis and embryonic prediction for blastocyst formation through an integrated time-lapse system.
        Fertil. Steril. 2016; 105 (e9): 376-384
        • Mutlu M.F.
        • Erdem M.
        • Erdem A.
        • Yildiz S.
        • Mutlu I.
        • Arisoy O.
        • Oktem M.
        Antral follicle count determines poor ovarian response better than anti-Mullerian hormone but age is the only predictor for live birth inin vitro fertilization cycles.
        J. Assist. Reprod. Genet. 2013; 30: 657-665
        • Nikmard F.
        • Aflatoonian B.
        • Hosseini E.
        • Aflatoonian A.
        • Bakhtiyari M.
        • Aflatoonian R.
        A comparative study on the results of agonist and antagonist protocols based on serum AMH levels in patients undergoing intracytoplasmic sperm injection.
        Int. J. Reprod. Biomed. (Yazd). 2016; 14: 769-776
        • Pfaffl M.W.
        A new mathematical model for relative quantification in real-time RT-PCR.
        Nucleic Acids Res. 2001; 29: e45
        • Pribenszky C.
        • Nilselid A.M.
        • Montag M.
        Time-lapse culture with morphokinetic embryo selection improves pregnancy and live birth chances and reduces early pregnancy loss: a meta-analysis.
        Reprod. Biomed. Online. 2017; 35: 511-520
        • Ramezanali F.
        • Ashrafi M.
        • Hemat M.
        • Arabipoor A.
        • Jalali S.
        • Moini A.
        Assisted reproductive outcomes in women with different polycystic ovary syndrome phenotypes: the predictive value of anti-Mullerian hormone.
        Reprod Biomed. Online. 2016; 32: 503-512
        • Rotterdam E.A.
        -S.P.C.W.G. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome (PCOS).
        Hum. Reprod. 2004; 19: 41-47
        • Rubio I.
        • Galan A.
        • Larreategui Z.
        • Ayerdi F.
        • Bellver J.
        • Herrero J.
        • Meseguer M.
        Clinical validation of embryo culture and selection by morphokinetic analysis: a randomized, controlled trial of the EmbryoScope.
        Fertil. Steril. 2014; 102 (e5): 1287-1294
        • Rubio I.
        • Kuhlmann R.
        • Agerholm I.
        • Kirk J.
        • Herrero J.
        • Escriba M.J.
        • Bellver J.
        • Meseguer M.
        Limited implantation success of direct-cleaved human zygotes: a time-lapse study.
        Fertil. Steril. 2012; 98: 1458-1463
        • Wathlet S.
        • Adriaenssens T.
        • Segers I.
        • Verheyen G.
        • Janssens R.
        • Coucke W.
        • Devroey P.
        • Smitz J.
        New candidate genes to predict pregnancy outcome in single embryo transfer cycles when using cumulus cell gene expression.
        Fertil. Steril. 2012; 98 (e1–4): 432-439
        • Wathlet S.
        • Adriaenssens T.
        • Segers I.
        • Verheyen G.
        • Van de Velde H.
        • Coucke W.
        • Ron El R.
        • Devroey P.
        • SMITZ J.
        Cumulus cell gene expression predicts better cleavage-stage embryo or blastocyst development and pregnancy for ICSI patients.
        Hum. Reprod. 2011; 26: 1035-1051
        • Wei L.N.
        • Liang X.Y.
        • Fang C.
        • Zhang M.F.
        Abnormal expression of growth differentiation factor 9 and bone morphogenetic protein 15 in stimulated oocytes during maturation from women with polycystic ovary syndrome.
        Fertil. Steril. 2011; 96: 464-468
        • Wissing M.L.
        • Bjerge M.R.
        • Olesen A.I.
        • Hoest T.
        • Mikkelsen A.L.
        Impact of PCOS on early embryo cleavage kinetics.
        Reprod. Biomed. Online. 2014; 28: 508-514
        • Wong C.C.
        • Loewke K.E.
        • Bossert N.L.
        • Behr B.
        • De Jonge C.J.
        • Baer T.M.
        • Reijo Pera R.A.
        Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage.
        Nat. Biotechnol. 2010; 28: 1115-1121
        • Wood J.R.
        • Dumesic D.A.
        • Abbott D.H.
        • Strauss 3rd, J.F.
        Molecular abnormalities in oocytes from women with polycystic ovary syndrome revealed by microarray analysis.
        J. Clin. Endocrinol. Metab. 2007; 92: 705-713

      Biography

      Nasim Tabibnejad was born on August 1975 and obtained her Medical Doctor degree in 2002. She has published more than 30 papers in the field of reproductive biology. She is MD, PhD, and a faculty member of Yazd Reproductive Sciences Institute.
      Key message
      Some embryo time-lapse kinetics parameters may be related to cumulus cell gene expression and clinical outcome. The expression of cumulus cell gene DIAPH2 can predict intracytoplasmic sperm injection outcome following Day 3 embryo transfer. Embryo assessment and selection by time-lapse imaging and cumulus cell gene expression should be prospectively studied.