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Cutting-edge techniques provide insights regarding repeated implantation failure patients

Published:December 20, 2022DOI:https://doi.org/10.1016/j.rbmo.2022.12.013

      Abstract

      Research question

      Can time-lapse parameters and the transcriptional profile of cumulus cells be used to achieve a more stringent and non-invasive method of embryo assessment and to identify possible factors affecting the embryo's ability to implant in repeated implantation failure (RIF) patients?

      Design

      A total of 190 embryos from 18 oocyte donors and 145 embryos from 15 RIF patients were evaluated based on time-lapse parameters. Three morphokinetic parameters including T5 (time to reach five cells), T3 (time to reach three cells) and CC2 (time to two to three cells) were recorded for all embryos. Embryos that had all three parameters in the normal range were graded as high quality and comparison between these parameters were compared in high-quality embryos between two groups. The transcriptional profile of cumulus cells related to high-quality embryos of both groups were analysed by RNA sequencing and compared. Finally, the possible relationship between differentially expressed genes and time-lapse parameters was examined.

      Results

      T5 was significantly lower in the RIF group than the donor group (P = 0.011). The cumulus cell transcriptome analysis showed 193 genes were down-regulated and 222 genes up-regulated. The mammalian target of rapamycin and the transforming growth factor beta pathways were significantly increased in the RIF group compared to the donor group (P = 0.007 and 0.01, respectively). Vitamin B12 and fatty acid beta-oxidation pathways were also significantly reduced in the RIF group compared to the donor group (P = 0.006 and 0.01, respectively).

      Conclusions

      Differences in the transcriptomic profiles of cumulus cells and some morphokinetic parameters may be one of the main factors contributing to unexplained RIF.

      Graphical Abstract

      Keywords

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      References

        • Adolfsson E.
        • Andershed A.N.
        Morphology vs morphokinetics: a retrospective comparison of inter-observer and intra-observer agreement between embryologists on blastocysts with known implantation outcome.
        JBRA Assisted Reproduction. 2018; 22: 228
        • Akino R.
        • Matsui D.
        • Kawahara-Miki R.
        • Amita M.
        • Tatsumi K.
        • Ishida E.
        • Kang W.
        • Takada S.
        • Miyado K.
        • Sekizawa A.
        • Saito T.
        • Kono T.
        • Saito H.
        Next-generation sequencing reveals downregulation of the Wnt signaling pathway in human dysmature cumulus cells as a hallmark for evaluating oocyte quality.
        Reproductive Medicine. 2020; 1: 205-215
        • Anderson R.
        • Sciorio R.
        • Kinnell H.
        • Bayne R.
        • Thong K.
        • De Sousa P.
        • Pickering S.
        Cumulus gene expression as a predictor of human oocyte fertilisation, embryo development and competence to establish a pregnancy.
        Reproduction. 2009; 138: 629-637
        • Auclair S.
        • Uzbekov R.
        • Elis S.
        • Sanchez L.
        • Kireev I.
        • Lardic L.
        • Dalbies-Tran R.
        • Uzbekova S.
        Absence of cumulus cells during in vitro maturation affects lipid metabolism in bovine oocytes.
        American Journal of Physiology-Endocrinology and Metabolism. 2013; 304: E599-E613
        • Aydos A.
        • Gurel A.
        • Oztemur Islakoglu Y.
        • Noyan S.
        • Gokce B.
        • Ecemis T.
        • Kaya C.
        • Tarik Aksu A.
        • Gur Dedeoglu B.
        Identification of polycystic ovary syndrome (PCOS) specific genes in cumulus and mural granulosa cells.
        PLoS One. 2016; 11e0168875
        • Basile N.
        • Vime P.
        • Florensa M.
        • Aparicio Ruiz B.
        • Garcia Velasco J.
        • Remohí J.
        • Meseguer M.
        The use of morphokinetics as a predictor of implantation: a multicentric study to define and validate an algorithm for embryo selection.
        Human Reproduction. 2015; 30: 276-283
        • Boachie J.
        • Adaikalakoteswari A.
        • Gázquez A.
        • Zammit V.
        • Larqué E.
        • Saravanan P.
        Vitamin B12 induces hepatic fatty infiltration through altered fatty acid metabolism.
        Cellular Physiology and Biochemistry. 2021; 55: 241-255
        • Bronet F.
        • Martínez E.
        • Gaytán M.
        • Liñán A.
        • Cernuda D.
        • Ariza M.
        • Nogales M.
        • Pacheco A.
        • San Celestino M.
        • Garcia-Velasco J.
        Sperm DNA fragmentation index does not correlate with the sperm or embryo aneuploidy rate in recurrent miscarriage or implantation failure patients.
        Human Reproduction. 2012; 27: 1922-1929
        • Caillaud M.
        • Dell'Aquila M.
        • De Santis T.
        • Nicassio M.
        • Lacalandra G.
        • Goudet G.
        • Gérard N.
        In vitro equine oocyte maturation in pure follicular fluid plus interleukin-1 and fertilization following ICSI.
        Animal Reproduction Science. 2008; 106: 431-439
        • Conaghan J.
        • Chen A.A.
        • Willman S.P.
        • Ivani K.
        • Chenette P.E.
        • Boostanfar R.
        • Baker V.
        • Adamson G.D.
        • Abusief M.E.
        • Gvakharia
        • Loewke K.E.
        Improving embryo selection using a computer-automated time-lapse image analysis test plus day 3 morphology: results from a prospective multicenter trial.
        Fertility and Sterility. 2013; 100 (L.M.) (412–419.e415)
        • Coughlan C.
        • Clarke H.
        • Cutting R.
        • Saxton J.
        • Waite S.
        • Ledger W.
        • Li T.
        • Pacey A.A.
        Sperm DNA fragmentation, recurrent implantation failure and recurrent miscarriage.
        Asian Journal of Andrology. 2015; 17: 681
        • Coughlan C.
        • Ledger W.
        • Wang Q.
        • Liu F.
        • Demirol A.
        • Gurgan T.
        • Cutting R.
        • Ong K.
        • Sallam H.
        • Li T.C.
        Recurrent implantation failure: definition and management.
        Reproductive Biomedicine Online. 2014; 28: 14-38
        • Dal Canto M.
        • Coticchio G.
        • Renzini M.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.
        Reproductive Biomedicine Online. 2012; 25: 474-480
        • Desquiret-Dumas V.
        • Clément A.
        • Seegers V.
        • Boucret L.
        • Ferré-L'Hotellier V.
        • Bouet P.E.
        • Descamps P.
        • Procaccio V.
        • Reynier P.
        • May-Panloup P
        The mitochondrial DNA content of cumulus granulosa cells is linked to embryo quality.
        Human Reproduction. 2017; 32: 607-614
        • Dunning K.R.
        • Robker R.L.
        Promoting lipid utilization with l-carnitine to improve oocyte quality.
        Animal Reproduction Science. 2012; 134: 69-75
        • Gardner D.K.
        • Lane M.
        • Stevens J.
        • Schlenker T.
        • Schoolcraft W.B.
        Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer.
        Fertility and Sterility. 2000; 73: 1155-1158
        • 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.
        Fertility and Sterility. 2011; 96 (47–52.e42)
        • Go K.J.
        Gardner DK, Weissman A, Howles CM, Shoham Z, editors. Textbook of Assisted Reproductive Techniques: Laboratory and Clinical Perspectives: Andover, Hampshire, United Kingdom: Thomas Publishing Services, 2001: 1–768. Price: $125.00.
        Fertility and sterility. 2002; 78: 1357-1358
        • Govahi A.
        • Amjadi F.
        • Nasr-Esfahani M.-H.
        • Raoufi E.
        • Mehdizadeh M.
        Accompaniment of time-lapse parameters and cumulus cell RNA-sequencing in embryo evaluation.
        Reproductive Sciences. 2022; 29: 395-409
        • Guo J.
        • Shi L.
        • Gong X.
        • Jiang M.
        • Yin Y.
        • Zhang X.
        • Li H.
        • Emori C.
        • Sugiura K.
        • Eppig J.J.
        • Su Y.Q.
        Oocyte-dependent activation of MTOR in cumulus cells controls the development and survival of cumulus–oocyte complexes.
        Journal of Cell Science. 2016; 129: 3091-3103
        • Hakimpour S.
        • Jelodar G.
        • Shabani R.
        • Pourheydar B.
        • Ajdary M.
        • Mehdizadeh M.
        Study of vitamin D3 formulation prepared by phytosolve technique and its effect on CTRP6 gene expression in PCOS model.
        Journal of Drug Delivery Science and Technology. 2022; 103489
        • 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.
        Human Reproduction. 2015; 30: 1850-1860
        • Huang Z.
        • Wells D
        The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome.
        Molecular Human Reproduction. 2010; 16: 715-725
        • Jääskeläinen M.
        • Prunskaite-Hyyryläinen R.
        • Naillat F.
        • Parviainen H.
        • Anttonen M.
        • Heikinheimo M.
        • Vaskivuo T.E.
        WNT4 is expressed in human fetal and adult ovaries and its signaling contributes to ovarian cell survival.
        Molecular and Cellular Endocrinology. 2010; 317: 106-111
        • Jiang Y.
        • Zhao Q.
        • Zhang Y.
        • Zhou L.
        • Lin J.
        • Chen Y.
        • Qian X.
        Treatment of G-CSF in unexplained, repeated implantation failure: a systematic review and meta-analysis.
        Journal of Gynecology Obstetrics and Human Reproduction. 2020; 49101866
        • Kirkegaard K.
        • Agerholm I.E.
        • Ingerslev H.J.
        Time-lapse monitoring as a tool for clinical embryo assessment.
        Human Reproduction. 2012; 27: 1277-1285
        • Krisher R.
        The effect of oocyte quality on development.
        Journal of Animal Science. 2004; 82: E14-E23
        • Li J.
        • Li C.
        • Liang D.
        • Lv F.
        • Yuan T.
        • The E.
        • Ma X.
        • Wu Y.
        • Zhen L.
        • Xie D.
        • Wang S.
        • Liu Y.
        • Huang J.
        • Shi D.
        • Xu L.
        • Lin L.
        • Peng L.
        • Cui J.
        • Zhu W.
        • Chen Y
        LRP6 acts as a scaffold protein in cardiac gap junction assembly.
        Nature Communications. 2016; 7: 1-11
        • Li Z.
        • Zhu W.
        • Xiong L.
        • Yu X.
        • Chen X.
        • Lin Q
        Role of high expression levels of STK39 in the growth, migration and invasion of non-small cell type lung cancer cells.
        Oncotarget. 2016; 7: 61366
        • Liu J.
        • Wu D.C.
        • Qu L.H.
        • Liao H.Q.
        • Li M.X.
        The role of mTOR in ovarian neoplasms, polycystic ovary syndrome, and ovarian aging.
        Clinical Anatomy. 2018; 31: 891-898
        • Liu Q.
        • Zhang J.
        • Wen H.
        • Feng Y.
        • Zhang X.
        • Xiang H.
        • Cao Y.
        • Tong X.
        • Ji Y.
        • Xue Z.
        Analyzing the transcriptome profile of human cumulus cells related to embryo quality via RNA sequencing.
        BioMed Research International. 2018; 20189846274
        • Lolicato F.
        • Brouwers J.F.
        • van de Lest C.H.A.
        • Wubbolts R.
        • Aardema H.
        • Priore P.
        • Roelen B.A.J.
        • Helms J.B.
        • Gadella B.M.
        The cumulus cell layer protects the bovine maturing oocyte against fatty acid-induced lipotoxicity.
        Biology of Reproduction. 2015; 92: 16
        • Luo Y.
        • Xia N.
        • Yang L.
        • Li Z.
        • Yang H.
        • Yu H.
        • Liu Y.
        • Lei H.
        • Zhou F.
        • Xie C.
        • Zhou Y.
        CTC1 increases the radioresistance of human melanoma cells by inhibiting telomere shortening and apoptosis.
        International Journal of Molecular Medicine. 2014; 33: 1484-1490
        • Meseguer M.
        • Herrero J.
        • Tejera A.
        • Hilligsøe K.M.
        • Ramsing N.B.
        • Remohí J.
        The use of morphokinetics as a predictor of embryo implantation.
        Human Reproduction. 2011; 26: 2658-2671
        • Mohammadi F.
        • Varanloo N.
        • Heydari Nasrabadi M.
        • Vatannejad A.
        • Amjadi F.
        • Javedani Masroor M.
        • Bajelan L.
        • Mehdizadeh M.
        • Aflatoonian R.
        • Zandieh Z.
        Supplementation of sperm freezing medium with myoinositol improve human sperm parameters and protects it against DNA fragmentation and apoptosis.
        Cell and Tissue Banking. 2019; 20: 77-86
        • Ogino M.
        • Tsubamoto H.
        • Sakata K.
        • Oohama N.
        • Hayakawa H.
        • Kojima T.
        • Shigeta M.
        • Shibahara H.
        Mitochondrial DNA copy number in cumulus cells is a strong predictor of obtaining good-quality embryos after IVF.
        Journal of Assisted Reproduction and Genetics. 2016; 33: 367-371
        • Oliveira J.B.A.
        • Cavagna M.
        • Petersen C.G.
        • Mauri A.L.
        • Massaro F.C.
        • Silva L.F.
        • Baruffi R.L.
        • Franco J.G.
        Pregnancy outcomes in women with repeated implantation failures after intracytoplasmic morphologically selected sperm injection (IMSI).
        Reproductive Biology and Endocrinology. 2011; 9: 1-7
        • Paczkowski M.
        • Silva E.
        • Schoolcraft W.B.
        • Krisher R.L.
        Comparative importance of fatty acid beta-oxidation to nuclear maturation, gene expression and glucose metabolism in mouse, bovine, and porcine cumulus oocyte complexes.
        Biology of Reproduction. 2013; 88: 111
        • Parikh F.
        • Agarwal S.
        • Naik D.
        • Katakdhond M.
        • Dhumal S.
        • Khandeparkar M.
        • Reddy S.
        • Mehta T.
        Intrauterine instillation of cumulus cells enhances implantation rates in women with previous failed implantation following embryo transfer.
        Fertility and Sterility. 2017; 108: e371-e372
        • Reznikoff-Etiévant M.F.
        • Zittoun J.
        • Vaylet C.
        • Pernet P.
        • Milliez J.
        Low vitamin B12 level as a risk factor for very early recurrent abortion.
        European Journal of Obstetrics and Gynecology and Reproductive Biology. 2002; 104: 156-159
        • Sanchez-Lazo L.
        • Brisard D.
        • Elis S.
        • Maillard V.
        • Uzbekov R.
        • Labas V.
        • Desmarchais A.
        • Papillier P.
        • Monget P.
        • Uzbekova S.
        Fatty acid synthesis and oxidation in cumulus cells support oocyte maturation in bovine.
        Molecular Endocrinology. 2014; 28: 1502-1521
        • Sheng J.G.
        • Jones R.A.
        • Zhou X.Q.
        • McGinness J.M.
        • Van Eldik L.J.
        • Mrak R.E.
        • Griffin W.S.T.
        Interleukin-1 promotion of MAPK-p38 overexpression in experimental animals and in Alzheimer's disease: potential significance for tau protein phosphorylation.
        Neurochemistry International. 2001; 39: 341-348
        • Shufaro Y.
        • Schenker J.G.
        Implantation failure, etiology, diagnosis and treatment.
        International Journal of Infertility and Fetal Medicine. 2011; 2: 1-7
        • Simon A.
        • Laufer N.
        Repeated implantation failure: clinical approach.
        Fertility and Sterility. 2012; 97: 1039-1043
        • Sutton-McDowall M.L.
        • Feil D.
        • Robker R.L.
        • Thompson J.G.
        • Dunning K.R.
        Utilization of endogenous fatty acid stores for energy production in bovine preimplantation embryos.
        Theriogenology. 2012; 77: 1632-1641
        • Swope D.
        • Cheng L.
        • Gao E.
        • Li J.
        • Radice G.L.
        Loss of cadherin-binding proteins β-catenin and plakoglobin in the heart leads to gap junction remodeling and arrhythmogenesis.
        Molecular and Cellular Biology. 2012; 32: 1056-1067
        • Taugourdeau A.
        • Desquiret-Dumas V.
        • Hamel J.-F.
        • Chupin S.
        • Boucret L.
        • Ferré-L'Hotellier V.
        • Bouet P.E.
        • Descamps P.
        • Reynier P.
        • May-Panloup P
        The mitochondrial DNA content of cumulus cells may help predict embryo implantation.
        Journal of Assisted Reproduction and Genetics. 2019; 36: 223-228
        • Wang H.-X.
        • Tekpetey F.R.
        • Kidder G.M.
        Identification of Wnt/β-catenin signaling pathway components in human cumulus cells.
        Molecular Human Reproduction. 2009; 15: 11-17
        • Wang Z.
        • Gerstein M.
        • Snyder M.
        RNA-Seq: a revolutionary tool for transcriptomics.
        Nature Reviews Genetics. 2009; 10: 57-63
        • Wong C.C.
        • Loewke K.E.
        • Bossert N.L.
        • Behr B.
        • De Jonge C.J.
        • Baer T.M.
        • Pera R.A.R.
        Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage.
        Nature Biotechnology. 2010; 28: 1115-1121
        • Yang S.C.
        • Yu E.J.
        • Park J.K.
        • Kim T.H.
        • Eum J.H.
        • Paek S.K.
        • Hwang Y.
        • Lyu S.W.
        • Kim J.Y.
        • Lee W.S.
        • Yoon T.K.
        • Song H.
        • Lee H.J.
        The ratio of mitochondrial DNA to genomic DNA copy number in cumulus cell may serve as a biomarker of embryo quality in IVF cycles.
        Reproductive Sciences. 2021; 28: 2495-2502

      Biography

      Dr Azam Govahi is a graduate of reproductive biology from the Iran University of Medical Sciences. She is currently working in the Endometriosis Research Center of the Iran University of Medical Sciences and conducting research in the field of female infertility.
      Key message
      Differences were found between the transcriptional profiles of cumulus cells and the morphokinetic parameter T5 of embryos from repeated implantation failure (RIF) and healthy fertile women. Differences in transcriptomic profiling of cumulus cells and some morphokinetic parameters may be factors contributing to implantation failure in unexplained RIF.