Transcriptome sequencing of endometrium revealed alterations in mRNAs and lncRNAs after ovarian stimulation
Using RNA-sequencing analysis, we investigated the relationship between ovarian stimulation and endometrial transcriptome profiles during the window of implantation (WOI) to identify candidate predictive factors for the WOI and to optimize timing for embryo transfer.
Twelve women with normal basal hormone levels and regular ovulation were randomly assigned into three groups based on sampling time: late-proliferate phase (P group), and receptive phase in natural cycles (LH+7, N group) and stimulated cycles (hCG+7, S group). Transcriptome profiles of mRNAs and long non-coding RNAs (lncRNAs) were then compared among the three groups. Validation was performed using real-time qPCR.
Comparison of transcriptome profiles between the natural and stimulated endometrium revealed 173 differentially expressed genes (DEGs), with a > 2-fold change (FC) and p < 0.05, under the influence of supraphysiological estradiol (E2) induced by ovarian stimulation. By clustering and KEGG pathway analysis, molecules and pathways associated with endometrial receptivity were identified. Of the 39 DEGs common to the three groups, eight genes were validated using real-time PCR. ESR1, MMP10, and HPSE were previously reported to be associated with endometrial receptivity. In addition, three novel genes (IL13RA2, ZCCHC12, SRARP) and two lncRNAs (LINC01060, LINC01104) were new potential endometrial receptivity-related markers.
Using mRNA and lncRNA sequencing, we found that supraphysiological E2 levels from ovarian stimulation had a marked impact upon endometrial transcriptome profiles and may result in a shift of the WOI. The precise mechanisms underlying the supraphysiological hormone-induced shift of the WOI require further research.
KeywordsOvarian stimulation Window of implantation Endometrial receptivity mRNAs lncRNAs
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 3.Rachel SMD. The receptivity of the endometrium to implementation. Journal of Obstetrics & Gynaecology of the British Empire. 2010;69(1):107–9.Google Scholar
- 6.Kumar P, Sharma A. Understanding implantation window, a crucial phenomenon. J Hum Reprod Sci. 2012;5:2. Available from: https://www.jhrsonline.org/text.asp?2012/5/1/2/97777 .PubMedPubMedCentralCrossRefGoogle Scholar
- 9.Ozkaya E, Kutlu T, Yayla CA, et al. Area under the curve of temporal estrogen and progesterone measurements during assisted reproductive technology: Which hormone is the main determinant of cycle outcome? Journal of Obstetrics and Gynaecology Research. 2018;44(2):263–9.PubMedCrossRefPubMedCentralGoogle Scholar
- 15.Boomsma CM, Kavelaars A, Eijkemans MJC, Fauser BCJM, Heijnen CJ, MacKlon NS. Ovarian stimulation for in vitro fertilization alters the intrauterine cytokine, chemokine, and growth factor milieu encountered by the embryo. Fertil Steril. Elsevier Ltd; 2010;94:1764–8. https://doi.org/10.1016/j.fertnstert.2009.10.044 PubMedCrossRefPubMedCentralGoogle Scholar
- 17.Thomas K, Thomson AJ, Sephton V, Cowan C, Wood S, Vince G, et al. The effect of gonadotrophic stimulation on integrin expression in the endometrium. Hum Reprod 2002;17:63–68. Available from: http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=11756363&retmode=ref&cmd=prlinks%5Cnpapers2://publication/uuid/D41C33EE-8D6D-4CFC-86A6-FAD09F5B5F19 (Accessed: 15 June 2017)
- 18.Bourgain C, Ubaldi F, Tavaniotou A, Smitz J, Van Steirteghem AC, Devroey P. Endometrial hormone receptors and proliferation index in the periovulatory phase of stimulated embryo transfer cycles in comparison with natural cycles and relation to clinical pregnancy outcome. Fertil Steril. 2002;78:237–44.PubMedCrossRefPubMedCentralGoogle Scholar
- 19.Altmae S, Martinez-Conejero JA, Salumets A, Simon C, Horcajadas JA, Stavreus-Evers A. Endometrial gene expression analysis at the time of embryo implantation in women with unexplained infertility. Mol Hum Reprod. 2010;16:178–87. Available from: https://academic.oup.com/molehr/article-lookup/doi/10.1093/molehr/gap102 .PubMedCrossRefPubMedCentralGoogle Scholar
- 20.Kuokkanen S, Chen B, Ojalvo L, Benard L, Santoro N, Pollard JW. Genomic profiling of microRNAs and messenger RNAs reveals hormonal regulation in microRNA expression in human endometrium. Biol Reprod. 2010;82:791–801. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19864316 .PubMedCrossRefPubMedCentralGoogle Scholar
- 24.Borthwick JM, Charnock-jones DS, Tom BD, Hull ML, Teirney R, Phillips SC, et al. Determination of the transcript pro ® le of human endometrium. Mol Hum Reprod. 2003;9:19–33. Available from: http://molehr.oxfordjournals.org/content/9/1/19.long .
- 25.Riesewijk A, Martin J, van Os R, Horcajadas JA, Polman J, Pellicer A, et al. Gene expression profiling of human endometrial receptivity on days LH+2 versus LH+7 by microarray technology. Mol Hum Reprod. 2003;9:253–64. Available from: http://molehr.oxfordjournals.org/cgi/content/abstract/9/5/253 .PubMedCrossRefPubMedCentralGoogle Scholar
- 26.Kao L, Tulac S, Lobo S, Imani B. Global gene profiling in human endometrium during the window of implantation. … [Internet]. 2002;143:2119–38. Available from: http://press.endocrine.org/doi/abs/10.1210/endo.143.6.8885 (Accessed: 23 October 2016)PubMedCrossRefPubMedCentralGoogle Scholar
- 27.Carson DD. Changes in gene expression during the early to mid-luteal (receptive phase) transition in human endometrium detected by high-density microarray screening. Mol Hum Reprod . 2002;8:871–879. Available from: http://molehr.oxfordjournals.org/content/8/9/871.short PubMedCrossRefPubMedCentralGoogle Scholar
- 29.Macklon NS, Van Der Gaast MH, Hamilton A, Fauser BCJM, Giudice LC. The Impact of Ovarian Stimulation With Recombinant. :357–65.Google Scholar
- 32.Horcajadas JA, Mínguez P, Dopazo J, Esteban FJ, Domínguez F, Giudice LC, et al. Controlled Ovarian Stimulation Induces a Functional Genomic Delay of the Endometrium with Potential Clinical Implications. J Clin Endocrinol Metab. 2008;93:4500–10. Available from: https://academic.oup.com/jcem/article-lookup/doi/10.1210/jc.2008-0588 .CrossRefGoogle Scholar
- 33.Simon C, Oberye J, Bellver J, Vidal C, Bosch E, Horcajadas JA, et al. Similar endometrial development in oocyte donors treated with either high- or standard-dose GnRH antagonist compared to treatment with a GnRH agonist or in natural cycles. Hum Reprod. 2005;20:3318–27.PubMedCrossRefPubMedCentralGoogle Scholar
- 42.Fluhr H, Spratte J, Bredow M, Heidrich S, Zygmunt M. Constitutive activity of Erk1/2 and NF-κB protects human endometrial stromal cells from death receptor-mediated apoptosis. Reprod Biol [Internet]. Society for Biology of Reproduction and; the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn; 2013;13:113–21. Available from: https://doi.org/10.1016/j.repbio.2013.03.001 PubMedCrossRefPubMedCentralGoogle Scholar
- 44.Jemt A, Sigurgeirsson B, Hanna A, Ujvari D, Westgren M, Lundeberg J. Comprehensive RNA sequencing of healthy human endometrium at two time points of the menstrual cycle. Biol Reprod. 2016;96:24–33. Available from: https://academic.oup.com/biolreprod/article/96/1/24/2725477/Comprehensive-RNA-sequencing-of-healthy-human .
- 45.Ganesh V, Venkatesan V, Koshy T, Nellapalli S, Muthumuthiah S, Franklin S, et al. Systems Biology in Reproductive Medicine Association of estrogen , progesterone and follicle stimulating hormone receptor polymorphisms with in vitro fertilization outcomes. Syst Biol Reprod Med. Taylor & Francis; 2018;64:260–5. https://doi.org/10.1080/19396368.2018.1482030 PubMedCrossRefPubMedCentralGoogle Scholar
- 46.Paskulin DD, Cunha-filho JS, Paskulin LD, Augusto C, Souza B, Ashton-prolla P. ESR1 rs9340799 Is Associated with Endometriosis-Related Infertility and In Vitro Fertilization Failure. 2013;35:907–13.Google Scholar
- 48.Biosciences M, Medicine V, Li S, Neill SRSO, Zhang Y, Holtzman MJ, et al. Estrogen receptor a is required for oviductal transport of embryos. :1595–607.Google Scholar
- 50.Boudjenah R, Molina-gomes D, Torre A, Bergere M, Bailly M, Wainer R, et al. Genetic Polymorphisms Influence the Ovarian Response to rFSH Stimulation in Patients Undergoing In Vitro Fertilization Programs with ICSI. 2012;7.Google Scholar
- 53.Quintero-Ronderos P, Mercier E, Fukuda M, Suarez C, Gonzalez R, Patarroyo M, et al. Novel genes and mutations in patients affected by recurrent spontaneous abortion. PLoS One. 2017;Submitted:1–14.Google Scholar
- 54.Kim MS, Yu JH, Lee MY, Kim AL, Jo MH, Kim MG, et al. Differential expression of extracellular matrix and adhesion molecules in fetalorigin amniotic epithelial cells of preeclamptic pregnancy. PLoS One. 2016;11:1–16.Google Scholar
- 55.Kaartokallio T, Cervera A, Kyllönen A, Laivuori K, Kere J, Laivuori H. Gene expression profiling of pre-eclamptic placentae by RNA sequencing. Sci Rep [Internet]. Nature Publishing Group; 2015;5:1–15. Available from: https://doi.org/10.1038/srep14107
- 59.Evans J, Salamonsen LA, Winship A, Menkhorst E, Nie G, Gargett CE, et al. Fertile ground : human endometrial programming and lessons in health and disease. Nat Publ Gr [Internet]. Nature Publishing Group; 2016;12:654–67. Available from: https://doi.org/10.1038/nrendo.2016.116 PubMedCrossRefPubMedCentralGoogle Scholar