Effects of Low Luteinizing Hormone During Ovarian Stimulation on Endometrial Gene Expression and Function — Transcriptome Analysis During the Implantation Window

This study explored the impact of low luteinizing hormone (LH) levels during ovarian stimulation on endometrial function. Based on previous studies by us and others, we divided the patients into low ( 10 IU/L) LH groups. The study utilized a comparison control group design with three groups of 10 patients. Gene set enrichment analysis (GSEA) was applied for functional annotation. By analyzing the exon differentially expressed genes in the endometrium of these three patient groups during the embryo implantation window, we found that when compared to the medium LH group, low LH downregulated endometrial cell metabolism, including mitochondrial-nicotinamide adenine dinucleotide (Normalized Enrichment Scores, NES = − 1.53) and glycolytic metabolism (NES = − 1.22), immune regulation, and autophagy (NES = − 1.58). Transcription factors were the main regulators of cell function. We found that MCM2 was probably involved in regulating the endometrial function induced by low LH. MCM2 target genes were enriched in low LH group, NES = − 1.54. Low LH, but not high LH, altered the endometrial receptivity assay gene expression in comparison to the medium LH. Our results indicated that low LH impacted the endometrial cell function, with a greater effect than high LH. This research provides timely and necessary data on the involvement of LH in important endometrial cellular processes and these data support further clinical development of endometrial receptivity. Similar content being viewed by others Availability of Data and Material The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Hillier SG. Current concepts of the roles of follicle stimulating hormone and luteinizing hormone in folliculogenesis. Hum Reprod. 1994;9:188–91. https://doi.org/10.1093/OXFORDJOURNALS.HUMREP.A138480. Weghofer A, Schnepf S, Barad D, Gleicher N. The impact of luteinizing hormone in assisted reproduction: a review. Curr Opin Obstet Gynecol. 2007;19:253–7. https://doi.org/10.1097/GCO.0B013E3280BAD843. Kumar P, Sait SF. Luteinizing hormone and its dilemma in ovulation induction. J Hum Reprod Sci. 2011;4:2. https://doi.org/10.4103/0974-1208.82351. Liu M, Liu S, Li L, Wang P, Li H, Li Y. LH levels may be used as an indicator for the time of antagonist administration in GnRH antagonist protocols—a proof-of-concept study. Front Endocrinol (Lausanne). 2019;10:67. https://doi.org/10.3389/FENDO.2019.00067. Stanger JD, Yovich JL. Reduced in-vitro fertilization of human oocytes from patients with raised basal luteinizing hormone levels during the follicular phase. BJOG An Int J Obstet Gynaecol. 1985;92:385–93. https://doi.org/10.1111/J.1471-0528.1985.TB01113.X. Seibel MM, Smith DM, Levesque L, Borten M, Taymor ML. The temporal relationship between the luteinizing hormone surge and human oocyte maturation. Am J Obstet Gynecol. 1982;142:568–72. https://doi.org/10.1016/0002-9378(82)90763-3. Humaidan P, Bungum L, Bungum M, Andersen CY. Ovarian response and pregnancy outcome related to mid-follicular LH levels in women undergoing assisted reproduction with GnRH agonist down-regulation and recombinant FSH stimulation. Hum Reprod. 2002;17:2016–21. https://doi.org/10.1093/HUMREP/17.8.2016. Tesarik J, Hazout A, Mendoza C. Luteinizing hormone affects uterine receptivity independently of ovarian function. Reprod Biomed Online. 2003;7:59–64. https://doi.org/10.1016/S1472-6483(10)61729-4. Ramezanali F, Khalili G, Arabipoor A, Lankarani NB, Moini A. Relationships between serum luteinizing hormone level, endometrial thickness and body mass index in polycystic ovary syndrome patients with and without endometrial hyperplasia. Int J Fertil Steril. 2016;10:36. https://doi.org/10.22074/IJFS.2016.4766. Luo Y, Liu S, Su H, Hua L, Ren H, Liu M, et al. Low serum LH levels during ovarian stimulation with GnRH antagonist protocol decrease the live birth rate after fresh embryo transfers but have no impact in freeze-all cycles. Front Endocrinol (Lausanne). 2021;12:405. https://doi.org/10.3389/FENDO.2021.640047. Lyga S, Volpe S, Werthmann RC, Götz K, Sungkaworn T, Lohse MJ, et al. Persistent cAMP signaling by internalized LH receptors in ovarian follicles. Endocrinology. 2016;157:1613–21. https://doi.org/10.1210/EN.2015-1945. Brar AK, Frank GR, Kessler CA, Cedars MI, Handwerger S. Progesterone-dependent decidualization of the human endometrium is mediated by cAMP. Endocrine. 1997;6:301–7. https://doi.org/10.1007/BF02820507. Puissant F, Van Rysselberge M, Barlow P, Deweze J, Leroy F. Embryo scoring as a prognostic tool in IVF treatment. Hum Reprod. 1987;2:705–8. https://doi.org/10.1093/OXFORDJOURNALS.HUMREP.A136618. Han S, Wei S, Wang X, Han X, Zhang M, Su M, et al. Enhanced intrinsic CYP3A4 activity in human hepatic C3A cells with optically controlled CRISPR/dCas9 activator complex. Integr Biol (United Kingdom). 2018;10:780–90. https://doi.org/10.1039/c8ib00109j. Lahoud R, Al-Jefout M, Tyler J, Ryan J, Driscoll G. A relative reduction in mid-follicular LH concentrations during GnRH agonist IVF/ICSI cycles leads to lower live birth rates. Hum Reprod. 2006;21:2645–9. https://doi.org/10.1093/HUMREP/DEL219. Shoham Z. The clinical therapeutic window for luteinizing hormone in controlled ovarian stimulation. Fertil Steril. 2002;77:1170–7. https://doi.org/10.1016/S0015-0282(02)03157-6. Lu X, Hong Q, Sun LH, Chen Q, Fu Y, Ai A, et al. Dual trigger for final oocyte maturation improves the oocyte retrieval rate of suboptimal responders to gonadotropin-releasing hormone agonist. Fertil Steril. 2016;106:1356–62. https://doi.org/10.1016/J.FERTNSTERT.2016.07.1068. Meyer L, Murphy LA, Gumer A, Reichman DE, Rosenwaks Z, Cholst IN. Risk factors for a suboptimal response to gonadotropin-releasing hormone agonist trigger during in vitro fertilization cycles. Fertil Steril. 2015;104:637–42. https://doi.org/10.1016/J.FERTNSTERT.2015.06.011. Benmachiche A, Benbouhedja S, Zoghmar A, Humaidan P. Low LH level on the day of GnRH agonist trigger is associated with reduced ongoing pregnancy and live birth rates and increased early miscarriage rates following IVF/ICSI treatment and fresh embryo transfer. Front Endocrinol. 2019;10:639. https://doi.org/10.3389/FENDO.2019.00639. Liu S, Liu M, Li L, Li H, Qu D, Ren H, et al. Patients with deep ovarian suppression following GnRH agonist long protocol may benefit from a modified GnRH antagonist protocol: a retrospective cohort study. Front Endocrinol. 2021;12:618580. https://doi.org/10.3389/FENDO.2021.618580. Ryu MJ, Seo BJ, Choi YJ, Han MJ, Choi Y, Chung MK, et al. Mitochondrial and metabolic dynamics of endometrial stromal cells during the endometrial cycle. Stem Cells Dev. 2020;29(1407):15. https://doi.org/10.1089/SCD.2020.0130. Czarnecka AM, Klemba A, Semczuk A, Plak K, Marzec B, Krawczyk T, et al. Common mitochondrial polymorphisms as risk factor for endometrial cancer. Int Arch Med. 2009;21(2):1–12. https://doi.org/10.1186/1755-7682-2-33. Cormio A, Guerra F, Cormio G, Pesce V, Fracasso F, Loizzi V, et al. The PGC-1α-dependent pathway of mitochondrial biogenesis is upregulated in type I endometrial cancer. Biochem Biophys Res Commun. 2009;390:1182–5. https://doi.org/10.1016/J.BBRC.2009.10.114. Liu Y, Luo C, Li T, Zhang W, Zong Z, Liu X, et al. Reduced nicotinamide mononucleotide (NMNH) potently enhances NAD+ and suppresses glycolysis, the TCA cycle, and cell growth. J Proteome Res. 2021;20:2596–606. https://doi.org/10.1021/ACS.JPROTEOME.0C01037. Rajman L, Chwalek K, Sinclair DA. Therapeutic potential of NAD-boosting molecules: the in vivo evidence. Cell Metab. 2018;27:529–47. https://doi.org/10.1016/J.CMET.2018.02.011. Katsyuba E, Romani M, Hofer D, Auwerx J. NAD+ homeostasis in health and disease. Nat Metab. 2020;21(2):9–31. https://doi.org/10.1038/s42255-019-0161-5. Schumacher A, Poloski E, Spörke D, Zenclussen AC. Luteinizing hormone contributes to fetal tolerance by regulating adaptive immune responses. Am J Reprod Immunol. 2014;71:434–40. https://doi.org/10.1111/AJI.12215. Taneja V. Sex hormones determine immune response. Front Immunol. 2018;9:1931. https://doi.org/10.3389/FIMMU.2018.01931. Kammerl IE, Meiners S. Proteasome function shapes innate and adaptive immune responses. Am J Physiol Lung Cell Mol Physiol. 2016;311:L328-36. https://doi.org/10.1152/AJPLUNG.00156.2016. Choi J, Jo M, Lee E, Oh YK, Choi D. The role of autophagy in human endometrium. Biol Reprod. 2012;86:70–1. https://doi.org/10.1095/BIOLREPROD.111.096206. Mei J, Zhu X-Y, Jin L-P, Duan Z-L, Li D-J, Li M-Q. Estrogen promotes the survival of human secretory phase endometrial stromal cells via CXCL12/CXCR4 up-regulation-mediated autophagy inhibition. Hum Reprod. 2015;30:1677–89. https://doi.org/10.1093/HUMREP/DEV100.

Firstly, I want to give a special thank to Wei-min Hou for his constant encouragement. Secondly, I wish to thank Hai-bo Li for his constant support. Funding This study was supported by 1351 Talent training Program of Beijing Chao-Yang Hospital (CYXX-2017–20); Capital Health Development Scientific Research Project (Independent Innovation, 2020–1-2039); Beijing Health Promotion Foundation (2019–09-05); 2020 Fertility Research Program of Young and Middle-aged Physicians-China Health Promotion Foundation; Beijing Hospitals Authority Youth Program (QML20200301); and Beijing Municipal Natural Science Foundation (No.7214234). Author information Authors and Affiliations Contributions S Han and S Liu carried out the experiments. S Han and MH Liu wrote the manuscript, with support from YS Lv, J Guo, and S Liu. S Liu helped supervise the project. Y Li and HY Ren supervised the project. All authors read and approved the final manuscript. Corresponding author Ethics declarations Ethics Approval This study was approved by the Ethics Review Board of Beijing Chao-Yang Hospital (2021-Science-45). Consent to Participate All the participants agreed to participate in this study and assigned the informed consent. Consent for Publication All the participants agreed to publish their data that came from individuals. Conflict of Interest The authors declare no competing interests. Supplementary Information Below is the link to the electronic supplementary material. Rights and permissions About this article Cite this article Han, S., Liu, Mh., Lv, Ys. et al. Effects of Low Luteinizing Hormone During Ovarian Stimulation on Endometrial Gene Expression and Function — Transcriptome Analysis During the Implantation Window. Reprod. Sci. 29, 1908–1920 (2022). https://doi.org/10.1007/s43032-022-00875-5 Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s43032-022-00875-5