Cumulus-corona gene expression analysis combined with morphological embryo scoring in single embryo transfer cycles increases live birth after fresh transfer and decreases time to pregnancy
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Clinical pregnancy rate after IVF with eSET stagnates between 30 and 40%. In order to increase pregnancy and live birth rates, multiple embryo transfer is still common practice. Providing additional non-invasive tools to choose the competent embryo for transfer could avoid multiple pregnancy and improve time to pregnancy. Cumulus mRNA analysis with quantitative PCR (QPCR) is a non-invasive approach. However, so far, no gene sets have been validated in prospective interventional studies.
A prospective interventional single-center pilot study with two matched controls (day-3 and day-5 eSET) was performed in 96 patients consenting to the analysis of the cumulus-corona of their oocytes. All patients were super-ovulated for ICSI and eSET at day 3. All oocytes were denuded individually and cumulus was analyzed by quantitative PCR using three predictive genes (EFNB2, SASH1, CAMK1D) and two housekeeping genes (UBC and β2M). Patients (n = 62) with 2 or more day-3 embryos (good or excellent morphology) had their embryo chosen following the normalized expression of the genes.
Corona testing significantly increased the clinical pregnancy and live births rates (63% and 55%) compared to single embryo transfer (eSET) on day 3 (27% and 23%: p < 0.001) and day 5 (43% and 39%: p = 0.022 and p = 0.050) fresh transfer cycle controls with morphology-only selection. Time-to-pregnancy was significantly reduced, regardless of the number of good-quality embryos available on day 3.
Combining standard morphology scoring and cumulus/corona gene expression analysis increases day-3 eSET results and significantly reduces the time to pregnancy.
Trial registration number
This is not an RCT study and was only registered by the ethical committee of the University Hospital UZBRUSSEL of the Vrije Universiteit Brussel VUB (BUN: 143201318000).
KeywordsCumulus cells Gene expression Single embryo transfer Clinical pregnancy Non-invasive Oocyte quality
The authors would like to thank their colleagues of the Centre for Reproductive Medicine, UZ Brussel, for their cooperation in this clinical study, the clinical data manager Walter Meul, and Prof. Dr. André Rosenthal for critical reading and suggestions.
This study was funded by IWT/VLAIO Innovation Mandate 130327 and 140568 and by the Vrije Universiteit Brussel IOFPOC26.
- 4.Glujovsky D, Blake D, Farquhar C, Bardach A. Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology. Glujovsky D, editor. Cochrane database Syst Rev. Chichester, UK: John Wiley & Sons, Ltd; 2012;CD002118.Google Scholar
- 6.Heino A, Gissler M, Hindori-Mohangoo AD, Blondel B, Klungsøyr K, Verdenik I, et al. Variations in multiple birth rates and impact on perinatal outcomes in Europe. Baud O, editor. PLoS One. 2016;11:e0149252.Google Scholar
- 10.Capalbo A, Ubaldi FM, Cimadomo D, Noli L, Khalaf Y, Farcomeni A, et al. MicroRNAs in spent blastocyst culture medium are derived from trophectoderm cells and can be explored for human embryo reproductive competence assessment. Fertil Steril. 2016;105:225–35.e1–3.Google Scholar
- 13.Armstrong S, Arroll N, Cree LM, Jordan V, Farquhar C. Time-lapse systems for embryo incubation and assessment in assisted reproduction. Cochrane database Syst Rev. 2015;CD011320.Google Scholar
- 18.Munné S, Blazek J, Large M, Martinez-Ortiz PA, Nisson H, Liu E, et al. Detailed investigation into the cytogenetic constitution and pregnancy outcome of replacing mosaic blastocysts detected with the use of high-resolution next-generation sequencing. Fertil Steril. 2017;108:62–71.e8.CrossRefPubMedGoogle Scholar
- 20.Yang Z, Liu J, Collins GS, Salem SA, Liu X, Lyle SS, et al. Selection of single blastocysts for fresh transfer via standard morphology assessment alone and with array CGH for good prognosis IVF patients: results from a randomized pilot study. Mol Cytogenet. 2012;5:24.CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Simon AL, Kiehl M, Fischer E, Proctor JG, Bush MR, Givens C, et al. Pregnancy outcomes from more than 1,800 in vitro fertilization cycles with the use of 24-chromosome single-nucleotide polymorphism–based preimplantation genetic testing for aneuploidy. Fertil Steril. 2018;110:113–21.CrossRefPubMedGoogle Scholar
- 30.Wathlet S, Adriaenssens T, Segers I, Verheyen G, Van Landuyt L, Coucke W, et al. Pregnancy prediction in single embryo transfer cycles after ICSI using QPCR: validation in oocytes from the same cohort. Lambalk CB, editor. PLoS One. 2013;8:e54226.Google Scholar
- 33.Zegers-Hochschild F, Adamson GD, de Mouzon J, Ishihara O, Mansour R, Nygren K, et al. International Committee for Monitoring Assisted Reproductive Technology (ICMART) and the World Health Organization (WHO) revised glossary of ART terminology, 2009. Fertil Steril. 2009;92:1520–4.CrossRefPubMedGoogle Scholar
- 34.Devroey P, Pellicer A, Nyboe Andersen A, Arce J-C, Menopur in GnRH Antagonist Cycles with Single Embryo Transfer Trial Group. A randomized assessor-blind trial comparing highly purified hMG and recombinant FSH in a GnRH antagonist cycle with compulsory single-blastocyst transfer. Fertil Steril. 2012;97:561–71.CrossRefPubMedGoogle Scholar
- 37.Segers I, Mateizel I, Van Moer E, Smitz J, Tournaye H, Verheyen G, et al. In vitro maturation (IVM) of oocytes recovered from ovariectomy specimens in the laboratory: a promising "ex vivo" method of oocyte cryopreservation resulting in the first report of an ongoing pregnancy in Europe. J Assist Reprod Genet. 2015;32:1221–31.CrossRefPubMedPubMedCentralGoogle Scholar
- 38.Gardner D, Schoolcraft W. In-vitro culture of human blastocysts. In: Jansen R, Mortimer D, editors. Towar reprod certain fertil genet beyond 1999. Carnforth: Parthenon Press; 1999. p. 378–88.Google Scholar
- 40.Ziebe S, Lundin K, Janssens R, Helmgaard L, Arce J-C. MERIT (Menotrophin vs Recombinant FSH in vitro Fertilisation Trial) Group. Influence of ovarian stimulation with HP-hMG or recombinant FSH on embryo quality parameters in patients undergoing IVF. Hum Reprod. 2007;22:2404–13.CrossRefPubMedGoogle Scholar
- 42.Rienzi L, Gracia C, Maggiulli R, LaBarbera AR, Kaser DJ, Ubaldi FM, et al. Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance. Hum Reprod Update. 2017;23:139–55.PubMedGoogle Scholar
- 46.Grøndahl ML, Borup R, Lee YB, Myrhøj V, Meinertz H, Sørensen 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.CrossRefPubMedGoogle Scholar
- 48.Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, Treff NR, et al. The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening. Fertil Steril. 2014;101:656–663.e1.CrossRefPubMedGoogle Scholar