Dose Of rFSH Research Articles

Embryo production by Holstein heifers superovulated with a recombinant long-acting follicle stimulating hormone analog.

This study aimed to evaluate embryo yield in Holstein heifers superovulated with a single injection of recombinant, long-acting human follicle-stimulating hormone (rFSH) versus multiple injections of pituitary-derived follicle-stimulating hormone (FSH). In Experiment 1, heifers were assigned randomly to one of 4 experimental groups: Control (280mg of pituitary derived FSH; six injections of 40mg and two injections of 20mg, each ~12h apart, n = 16); rFSH1 (50 μg of FSH analog protein, n = 16); rFSH2 (75 μg of FSH analog protein, n = 16) or; rFSH3 (100 μg of FSH analog protein, n = 16). The rFSH was administered as a single injection. Estrous cycles were presynchronized with gonadorelin acetate (GnRH) and an intra-vaginal progesterone insert (CIDR) on d -8, followed by cloprostenol sodium (PGF2α) on d -3 and d -2 with removal of the CIDR, and GnRH on d 0. On d 5, ovarian follicles ≥ 5mm in diameter were ablated by transvaginal ultrasound-guided aspiration. On d 6.5, heifers received a CIDR and the rFSH or the first injection of pituitary-derived FSH. On d 9, heifers received two injections of PGF2α, 12h apart. On d 10.5, the CIDR was removed, and on d 11, heifers received a GnRH injection. Heifers were AI-inseminated 12- and 24-hours post-GnRH injection, and uterine contents were flushed trans-cervically on day 18 (7 d after the GnRH injection). Ovarian follicles ≥ 5mm and corpora lutea were counted via ultrasound on days 5, 9, and 18. In Experiment 1, group did not affect (P = 0.52) the number of follicles ≥ 5mm (Control = 15.9 ± 1.2; rFSH1 = 17.5 ± 1.3; rFSH2 = 17.1 ± 1.3; rFHS 3 = 18.6 ± 1.4 follicles) or the number of corpora lutea (P = 0.96) on d 9 (Control = 1.1 ± 0.3; rFSH1 = 1.1 ± 0.3; rFSH2 = 1.1 ± 0.3; rFSH3 = 0.9 ± 0.2). Furthermore, there was no effect (P = 0.28) of rFSH dose on freezable embryos (grade 1 and 2 embryos) collected on d 18 (Control = 4.7 ± 1.1; rFSH1 = 4.7 ± 1.2; rFSH2 = 4.4 ± 1.1; rFSH3 = 2.6 ± 0.7 embryos). In Experiment 2, Control (n=8) and rFSH1 (n=16) groups were repeated in 3 replicates using the same protocols as Experiment 1. Consequently, Results showed that rFSH produced fewer total number of ova/embryos (Control = 9.9 ± 1.5 vs. rFHS1 = 5.9 ± 0.9, P = 0.04) and fewer freezable embryos (Control = 5.3 ± 1.0 vs. rFSH1 = 1.4 ± 0.3, P < 0.01). In conclusion, the single rFSH injection effectively induced superovulation; however, its repeated use reduced embryo production.

In Vitro Fertilization (IVF) Outcomes in Patients With Endometriosis Compared to Patients With Tubal Factor Infertility in Indonesia: A Retrospective Study.

Endometriosis is a disease characterized by dysmenorrhea, chronic pelvic pain, and infertility. The pathogenesis of endometriosis and its relationship to infertility are still not fully understood. In vitro fertilization (IVF) is considered an effective treatment for patients with endometriosis-associated infertility. This study compared the pregnancy rates of endometriosis patients who underwent IVF with those of patients with tubal obstruction. This was a retrospective cross-sectional study of 225 patients with endometriosis and tubal factor infertility who underwent IVF at Yasmin Clinic, Dr. Cipto Mangunkusumo National General Hospital, Jakarta, Indonesia, between January 2013 and August 2021. Demographic data, anti-Mullerian hormone (AMH) levels, the initial and total dose of recombinant follicle stimulating hormone (rFSH), the total dose of recombinant luteinizing hormone (rLH), stimulation duration, oocyte maturation rates, fertilization rates, embryo cleavage rates, and pregnancy rates (biochemical, clinical and ongoing) were obtained. AMH level, number of oocytes retrieved, and embryo cleavage rates were significantly lower in the endometriosis group. Initial and total doses of rFSH and total dose of rLH were higher, and the duration of stimulation was longer in the endometriosis group compared to the tubal factor group. In the endometriosis group, the biochemical (47.3% vs. 52.7%, p=0.375), clinical (43.1% vs. 56.9%, p=0.215), and ongoing (45.5% vs. 54.5%, p=0.511) pregnancy rates were lower than those in the tubal factor group. However, there were no statistically significant differences between the two groups. There was no significant difference in pregnancy rates between the short antagonist and ultra-long protocols (three months of downregulation). From multivariate analysis, only rLH supplementation was found to be significantly related to pregnancy outcomes in patients with endometriosis. AMH levels, number of oocytes retrieved, and embryo cleavage rates were lower in patients with endometriosis. However, pregnancy rates were not significantly different from patients with tubal factors. Supplementation with rLH may improve pregnancy rates in patients with endometriosis who have undergone IVF programs.

Significance of FSHR and LHCGR gene polymorphisms on clinical outcomes in gonadotropin-releasing hormone antagonist protocol with freeze-all strategy: A case-control study.

Follicle-stimulating hormone receptor (FSHR) and luteinizing hormone/choriogonadotropin receptor (LHCGR) are integral to ovarian function, facilitating follicle development and maturation through their respective hormonal interactions. The influence of receptor polymorphisms on the outcomes of freeze-all cycles remains unclear. This study investigates the impact of FSHR N680S and LHCGR N312S polymorphisms on clinical outcomes in freeze-all cycles. Women undergoing controlled ovarian stimulation for assisted reproductive technology participated in this study. They were administered a gonadotropin-releasing hormone antagonist protocol, with recombinant follicle-stimulating hormone (rFSH) dosages adjusted according to age, body mass index, antral follicle count, and individual hormonal responses. Additionally, human menopausal gonadotropin dosages were tailored based on the LHCGR N312S genetic variant. Analysis revealed no significant differences in age, body mass index, antral follicle count, or marital status across the genotypes of FSHR N680S and LHCGR N312S. However, notable differences were observed in the rFSH dosage required daily and in total among the FSHR polymorphism genotypes. Genotypes of the LHCGR polymorphism correlated with fewer stimulation days. A significant interaction was observed between the 2 polymorphisms concerning total rFSH dosage. The presence of serine in the FSHR polymorphism was associated with higher rFSH dosage requirements. Both FSHR N680S and LHCGR N312S polymorphisms significantly influenced clinical pregnancy and live birth outcomes in freeze-all cycles, underscoring the potential of a pharmacogenomic approach to optimize hormone supplementation in controlled ovarian stimulation protocols during assisted reproductive technology treatments.

Inositols and female reproduction disorders: a consensus statement from the working group of the Club of the Italian Society of Endocrinology (SIE)-Women's Endocrinology.

To provide the latest scientific knowledge on the efficacy of inositols for improving reproductive disorders in women with and without polycystic ovary syndrome (PCOS) and to reach a consensus on their potential use through a Delphi-like process. A panel of 17 endocrinologists and 1 gynecologist discussed 4 key domains: menses irregularity and anovulation, fertility, pregnancy outcomes, and neonatal outcomes. A total of eight consensus statements were drafted. Myo-inositol (Myo) supplementation can be used to improve menses irregularities and anovulation in PCOS. Myo supplementation can be used in subfertile women with or without PCOS to reduce the dose of r-FSH for ovarian stimulation during IVF, but it should not be used to increase the clinical pregnancy rate or live birth rate. Myo supplementation can be used in the primary prevention of gestational diabetes mellitus (GDM), but should not be used to improve pregnancy outcomes in women with GDM. Myo can be preconceptionally added to folic acid in women with a previous neural tube defects (NTD)-complicated pregnancy to reduce the risk of NTDs in newborns. Myo can be used during pregnancy to reduce the risk of macrosomia and neonatal hypoglycemia in mothers at risk of GDM. This consensus statement provides recommendations aimed at guiding healthcare practitioners in the use of inositols for the treatment or prevention of female reproductive disorders. More evidence-based data are needed to definitively establish the usefulness of Myo, the appropriate dosage, and to support the use of D-chiro-inositol (DCI) or a definitive Myo/DCI ratio.

P-592 Comparison of the cumulative live birth rate of different doses of rFSH in poor ovarian reserve women undergoing GnRH antagonist ART treatment: a Randomized Controlled Trail

Abstract Study question Does an increased FSH dosage result in higher cumulative live birth rates in advanced age women with poor ovarian reserve treated with IVF or ICSI? Summary answer In poor ovarian reserve women, FSH dose of 300 IU/day does not improve cumulative live birth rates as compared to 150 IU/day. What is known already Natural fecundity and pregnancy rate following ART decrease dramatically in women aged 35 years and above. An open-label multicenter RCT described no difference in the cumulative live birth rate between the increased FSH dose (225/450 IU/day) and standard dose (150 IU/day) in women with a predicted poor ovarian response. Nevertheless, it is still unclear whether advanced age women with poor ovarian reserve who are undergoing IVF/ICSI benefit from a higher gonadotrophin dose. Therefore, we aimed to assess whether rFSH 300 IU/day compared to 150 IU/day in advanced age women with poor ovarian reserve resulted in higher cumulative live birth rates. Study design, size, duration A single center prospective randomized controlled trail that included first in vitro fertilization cycle with a GnRH-ant protocol and subsequent fresh and frozen-thawed cycles from April 2019 to December 2022. The study group included 104 women that received rFSH 150 IU/day and 99 women received rFSH 300 IU/day in antagonist protocol. Participants/materials, setting, methods Women for their first stimulation cycle, aged 35 to 42 years with antral follicle count（AFC）≤ 5 or anti-mullerian hormone （AMH） ≤ 1.2 ng/ml were randomized. Study group and control group respectively received 300 IU and 150 IU rFSH from menstrual cycle day 2 or 3 till the day of trigger. The primary outcome was cumulative live birth rate per woman per stimulation over a 2-year period to account for the first live birth. Main results and the role of chance This study included 203 women with the GnRH antagonist protocol. Of them, 99 patients were in 300 IU/day rFSH group, and 104 patients were in 150 IU/day rFSH group, respectively. The two groups were comparable for all baseline characteristics. The cumulative live birth rate for increased rFSH dosing (300 IU) versus standard dosing (150 IU) was 13.13% (13/99) versus 25% (26/104) per stimulation cycle. (P = 0.049). There were no differences in cumulative pregnancy rates 27.27% (27/99) vs 35.58% (37/104) per stimulation cycle. (P = 0.262). The cumulative live birth rate and cumulative pregnancy rates per transfer cycle in 300 IU/day rFSH group vs 150 IU/day rFSH group presented no statistical difference. No significant difference was found in secondary outcomes including time to live birth, number of MII oocytes, number of transferable embryos, implantation rate, multiple pregnancy rate, ectopic pregnancy rate and miscarriage rate between the two groups. Limitations, reasons for caution As this study is open-label, potential selective cancelling of cycles in women could have influenced the cumulative results. Also, this study practiced in a single-center and specific patient population, the results may not be generalizable to a broader population. Wider implications of the findings In women 35 years or older with poor ovarian reserve, improving rFSH dosage to 300IU in GnRH-ant protocol did not improve cumulative live birth rate. rFSH 150IU/day may be recommended at the start of COS to achieve live birth outcomes while decrease costs. Trial registration number ’not applicable

O-303 What is the ideal dose of daily FSH after corifollitropin-alfa? A multicentre-randomized control trial

Abstract Study question Does the mean serum trigger-progesterone levels show significant variation based on the administered rFSH dose from day 8 onwards during ovarian stimulation using corifollitropin-alfa? Summary answer Lower rFSH doses from day 8 of ovarian stimulation onwards are linked to reduced serum progesterone (P4) levels without impact on cumulus-oocyte complexes. What is known already Progesterone elevation during ovulation trigger prompts a non-elective freeze-all strategy, given the lower live birth rates in fresh embryo transfers. Ongoing research explores methods to mitigate late follicular phase P4 rise to allow a fresh embryo transfer. Granulosa cells’ progesterone production is an FSH-driven and dose-dependent process, and FSH dose is furthermore pivotal for a good ovarian response. Balancing FSH dose is crucial for both optimal ovarian response and preventing undesirable P4 elevation while maintaining an adequate number of retrieved oocytes and allowing a fresh embryo transfer. Study design, size, duration This study is an interim-analysis of a randomized-controlled multi-center clinical-trial comparing three ovarian-stimulation-protocols: (A) corifollitropin alfa (CFA) followed by rFSH 50 IU/day; (B) CFA followed by rFSH 150 IU/day; (C) CFA followed by rFSH 250 IU/day.The primary outcome is the serum-progesterone-level on the day of hCG among the study groups, secondary outcomes included number of oocytes, clinical pregnancy rates and freeze-all cycles. 167 women (of 261) were randomized between July 2019-November 2023, with a 1:1allocation. Participants/materials, setting, methods Young (&amp;lt;40 years old), normo-ovulatory cycle (25-35 days), with AFC&amp;gt;7 and &amp;lt;20, and a BMI&amp;lt;29 Kg/m2 patients were recruited in the fertility clinic of three different tertiary university hospitals. On day 8 of ovarian stimulation (OS) following CFA administration, patients who needed additional rFSH were randomized into 3 groups (A-B-C). The OS was performed in a GnRH-antagonist protocol and the ovulation triggered with hCG. A fresh embryo-transfer was performed on day 5 following oocyte retrieval. Main results and the role of chance Overall, 167 patients were randomized, 60 in group A, 55 in group B and 52 in group C. The mean age and BMI were comparable between the groups. Mean AMH was respectively 2.2, 2.1 and 2.2 ng/mL, for group A, B and C (p = 0.95). Group A resulted in a significantly lower P4 level at trigger as compared with group B and C (respectively, 0.4ng/mL, 0.9 ng/mL and 1.4 ng/mL; p &amp;lt; 0.001). The mean number of retrieved oocytes was 10.7, 11 and 12.1 for group A, B and C, respectively (p = 0.47), while the mean number of mature oocytes was 7.3, 6.9 and 7.8 for group A, B and C, respectively (p = 0.7). A freeze-all approach was decided for 27%, 29% and 46% for group A, B and C respectively (p = 0.057). No significant differences were found for fresh-embryo-transfer outcomes (chemical pregnancy rates: 33%, 31% and 29%, respectively for A, B and C (p = 0.12).Of interest, comparisons between serum FSH at trigger showed a mean of 12.5, 17 and 20.5 IU/L for group A, B and C, respectively (p &amp;lt; 0.001) while the serum estradiol levels on day of trigger were comparable among the groups (1659, 2092 and 2098 ng/L for group A, B and C, respectively (p = 0.091). Limitations, reasons for caution This is an interim analysis planned to detect the mean difference in COC between groups. Therefore, the mean differences of the primary outcome should be interpreted as exploratory findings which warrant future investigation. Wider implications of the findings Administering a lower rFSH dose from day 8 onward during ovarian stimulation results in comparable numbers of COC but also leads to significantly lower serum P4 levels. Consequently, this finding has the potential to reduce the overall consumption of rFSH, thereby possibly increasing the likelihood of a fresh embryo transfer. Trial registration number NCT03686852

P-734 Low cost antagonist protocol in ivf-icsi cycles using oral ovulation induction agents in developing countries

Abstract Study question The study aimed at comparing the cost-effectiveness of a low cost antagonist protocol using oral-ovulation-induction agents with a standard long GnRH-Agonist protocol in IVF-ICSI cycles. Summary answer Low cost antagonist protocol using oral-ovulation-induction agents comparing with a standard long GnRH-Agonist protocol in IVF-ICSI cycles is cost-effectiveness in developing countries. What is known already IVF-ICSI had always been proved to be costly treatment. The additional expenditure is mainly due to use of high dose of gonadotrophins in order to get more number of eggs. To cut down the cost,low dose or mild stimulation protocol have been popularized all over the world in last few years. Use of aromatase inhibitors along with low doses of gonadotrophins have been an effective means of mild stimulation protocols for IVF cycles. Oral ovulogens, reduces the total dose of gonadotrophin required for inducing follicular maturation. We evaluated if incorporation of letrozole could be an effective alternative for low-cost IVF. Study design, size, duration A total of 304 patients under 40 years of age were studied between 2018 to 2022 Subsequently, they were randomly divided into two groups. The agonist group underwent standard long GnRH analogue protocol. The antagonist group received oral ovulation inducing drugs (CC/Letrozole) and 0.25mg antagonist injections upto the day of hCG injection, using a flexible start approach. Participants/materials, setting, methods A total of 304 patients under 40 years of age were studied between 2018 to 2022. All patients received low-dose OCP before starting treatment. Subsequently, they were randomly divided into two groups. The agonist-group underwent standard long GnRH analogue protocol. The antagonist group received oral ovulation inducing drugs (CC/Letrozole) and 0.25mg antagonist injections upto the day of hCG injection, using a flexible start approach. Follicular development was monitored by vaginal ultrasound and serum estradiol levels. Main results and the role of chance There was a significant difference in the gonadotropin usage between the two groups. As a result, the cost of the CC/Letrozole-gonadotropin-antagonist cycle was significantly lower than the long GnRH agonist-gonadotropin protocol. Though the mean number of oocytes retrieved was higher in the long protocol group, yet the clinical pregnancy rate per transfer was the same in both groups. While comparing for the primary outcomes pregnancy rate and miscarriage rate were comparable between two groups. It is of particular importance to note that despite 60%reduction of total dose of rFSH in the Let-rFSH group,which entailed significant reduction of the cost of treatment, the outcomes in all major aspects including pregnancy rate were comparable between the groups. However, numbers of matured follicles, retrieved oocytes and transferable embryos were not statistically different between study and control group in this study. This prospective study, designed to evaluate the efficacy of oral-ovulogens -FSH as a low-cost IVF protocol, involved an average number of patients; however, the results have been encouraging. We have been successful in reaching our objective to modify an expensive conventional down regulated IVF protocol into one in which the cost was reduced to a larger extent, without compromising the rate of success in developing countries. Limitations, reasons for caution Since last Cochrane review, GnRH-antagonist protocol was proved to be a short and simple protocol with good clinical outcome with significant reduction in incidence of severe OHSS and amount of gonadotrophins requirement. Although, we need more RCTs to come to a conclusion on regular use of such combined protocol. Wider implications of the findings Usage of GnRH-antagonist in combination with Oral-Ovulogens offers the advantage of a low-cost economical stimulation in IVF, as compared to an agonist-protocol, with similar pregnancy-rate in developing countries. It not only offers a cost-saving stimulation-protocol but also reduces side effects like OHSS and multiple pregnancies yet maintain a descent success-rate. Trial registration number not applicable

Ovarian response and embryo ploidy following oral micronized progesterone-primed ovarian stimulation versus GnRH antagonist protocol. A prospective study with repeated ovarian stimulation cycles.

Is there any difference in ovarian response and embryo ploidy following progesterone-primed ovarian stimulation (PPOS) using micronized progesterone or GnRH antagonist protocol? Pituitary downregulation with micronized progesterone as PPOS results in higher number of oocytes retrieved and a comparable number of euploid blastocysts to a GnRH antagonist protocol. Although the GnRH antagonist is considered by most the gold standard protocol for controlling the LH surge during ovarian stimulation (OS) for IVF/ICSI, PPOS protocols are being increasingly used in freeze-all protocols. Still, despite the promising results of PPOS protocols, an early randomized trial reported potentially lower live births in recipients of oocytes resulting following downregulation with medroxyprogesterone acetate as compared with a GnRH antagonist protocol. The scope of the current prospective study was to investigate whether PPOS with micronized progesterone results in an equivalent yield of euploid blastocysts to a GnRH antagonist protocol. In this prospective study, performed between September 2019 to January 2022, 44 women underwent two consecutive OS protocols within a period of 6 months in a GnRH antagonist protocol or in a PPOS protocol with oral micronized progesterone. Overall, 44 women underwent two OS cycles with an identical fixed dose of rFSH (225 or 300 IU) in both cycles. Downregulation in the first cycles was performed with the use of a flexible GnRH antagonist protocol (0.25 mg per day as soon as one follicle of 14 mm) and consecutively, after a washout period of 1 month, control of LH surge was performed with 200 mg of oral micronized progesterone from stimulation Day 1. After the completion of both cycles, all generated blastocysts underwent genetic analysis for aneuploidy screening (preimplantation genetic testing for aneuplody, PGT-A). Comparisons between protocols did not reveal differences between the duration of OS. The hormonal profile on the day of trigger revealed statistically significant differences between protocols in all the tested hormones except for FSH: with significantly higher serum E2 levels, more elevated LH levels and higher progesterone levels in PPOS cycles as compared with antagonist cycles, respectively. Compared with the GnRH antagonist protocol, the PPOS protocol resulted in a significantly higher number of oocytes (12.7 ± 8.09 versus 10.3 ± 5.84; difference between means [DBM] -2.4 [95% CI -4.1 to -0.73]), metaphase II (9.1 ± 6.12 versus 7.3 ± 4.15; DBM -1.8 [95% CI -3.1 to -0.43]), and 2 pronuclei (7.1 ± 4.99 versus 5.7 ± 3.35; DBM -1.5 [95% CI -2.6.1 to -0.32]), respectively. Nevertheless, no differences were observed regarding the mean number of blastocysts between the PPOS and GnRH antagonist protocols (2.9 ± 2.11 versus 2.8 ± 2.12; DBM -0.07 [95% CI -0.67 to 0.53]) and the mean number of biopsied blastocysts (2.9 ± 2.16 versus 2.9 ± 2.15; DBM -0.07 [95% CI -0.70 to 0.56]), respectively. Concerning the euploidy rates per biopsied embryo, a 29% [95% CI 21.8-38.1%] and a 35% [95% CI 26.6-43.9%] were noticed in the PPOS and antagonist groups, respectively. Finally, no difference was observed for the primary outcome, with a mean number of euploid embryos of 0.86 ± 0.90 versus 1.00 ± 1.12 for the comparison of PPOS versus GnRh antagonist. The study was powered to detect differences in the mean number of euploid embryos and not in terms of pregnancy outcomes. Additionally, per protocol, there was no randomization, the first cycle was always a GnRH antagonist cycle and the second a PPOS with 1 month of washout period in between. In case of a freeze-all protocol, clinicians may safely consider oral micronized progesterone to control the LH surge and patients could benefit from the advantages of a medication of oral administration, with a potentially higher number of oocytes retrieved at a lower cost, without any compromise in embryo ploidy rates. This research was supported by an unrestricted grant from Theramex. N.P.P. has received Research grants from Merck Serono, Organon, Ferring Pharmaceutical, Roche, Theramex, IBSA, Gedeon Richter, and Besins Healthcare; honoraria for lectures from: Merck Serono, Organon, Ferring Pharmaceuticals, Besins International, Roche Diagnostics, IBSA, Theramex, and Gedeon Richter; consulting fees from Merck Serono, Organon, Besins Healthcare, and IBSA. M.d.M.V., F.M., and I.R. declared no conflicts of interest. The study was registered at Clinical Trials Gov. (NCT04108039).

Can AMH levels predict the need to step up FSH dose for controlled ovarian stimulation following a long GnRH agonist protocol in PCOS women?

BackgroundTo explore the role of anti-Mullerian hormone (AMH) in predicting the need to step up recombinant FSH (rFSH) dose following long GnRH agonist protocol in IVF/ICSI cycles of polycystic ovarian syndrome (PCOS) women.MethodsThis is a retrospective cohort study of 825 PCOS women undergoing long GnRH agonist protocol enrolled from Jan 2019 to Dec 2021. The daily rFSH dose at which the first response to rFSH were recorded. The dose at which the first response to rFSH was based on folliculometry during follow up in which two or more follicles reached ≥ 11 mm. A receiver operating characteristic (ROC) curve analysis was done to investigate the ability of AMH to predict the need to step up initial rFSH dose.ResultsPCOS women who needed to step up initial rFSH dose had a significantly higher AMH compared with those didn’t step up initial rFSH dose (11.37 ± 3.25ng/ml vs. 8.69 ± 3.16ng/ml, p < 0.001). In multivariate logistic regression analysis, increased AMH level was an independent factor for the need to step up initial rFSH dose in PCOS patients after adjusted for confounding factors. ROC curve analysis showed AMH could predict the need to step up initial rFSH dose (AUC = 0.738, 95%CI: 0.704–0.773), having 75.4% specificity and 63% sensitivity when the threshold AMH concentration was 9.30ng/ml. 58.8% PCOS women with AMH > 9.30 ng/ml required increased rFSH dose compared to 18.8% of women with AMH ≤ 9.30ng/ml (p < 0.001). Although the clinical pregnancy rate and live birth rate were not significantly different, there was a higher incidence of OHSS among women with AMH > 9.30 ng/ml vs. AMH ≤ 9.30ng/ml (20.8% vs. 15.3%, p = 0.043).ConclusionPCOS women with AMH > 9.30 ng/ml were resistant to rFSH stimulation and require increased dose for the cycle recruitment of ovarian follicles.

The effect of letrozole as an adjunct in GnRH-antagonist protocol on IVF/ICSI outcome in women with endometriosis: a randomized clinical trial

BackgroundTo evaluate the effect of adding letrozole to the antagonist ovarian stimulation protocol (COS) on in-vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcome in endometriosis patients.MethodsThis randomized clinical trial was carried out in the department of infertility treatment at Arash Women’s Hospital from May 2019 to May 2021. The eligible women with normal ovarian reserve tests who had endometriosis diagnosis and underwent IVF/ICSI cycles were evaluated. A flexible regimen of GnRH-antagonist protocol was used for COS. In the experimental (n = 34), the patients received 5 mg letrozole daily for the first 5 days in combination with 150 IU of recombinant follicle-stimulating hormone (rFSH). In the control group (n = 30), the patients received only the same dose of rFSH. The treatment cycle was compared between groups.ResultsAnalysis of demographic characteristics, severity of endometriosis, and baseline hormonal tests of patients showed that the two groups were similar and comparable. The means of total used gonadotropins ampoules and serum E2 level on oocyte trigger day in the letrozole group were significantly lower than those of in the control group (P = 0.03 and P = 0.004, respectively). No statistically significant difference in terms of the total number of retrieved and MII oocytes as well as the total numbers of obtained and top-quality embryos, and cryopreserved embryos was found.ConclusionThe co-treatment of letrozole with gonadotropins during the antagonist protocol was associated with a reduction in the total dose of gonadotropins, although it had no effect on the oocyte or embryo yield in patients with endometriosis.Trial registrationThe study was registered in the Iranian Registry of Clinical Trials on 2018 -07-13 (IRCT20150310021420N4 at www.irct.ir, registered while recruiting).

Different doses of recombinant FSH and determining parameters of oxidative stress.

This study aimed to examine if there is a connection between recombinant FSH dose and OS parameters in serum and the impact on IVF outcome. This study consisted of 101 participants that went through IVF procedures. Parameter that were tested are SOD, SH groups and MDA. Serum samples were drawn before stimulation and on the last day of ovarian stimulation. Two groups were formed according to the dose of gonadotropins (rFSH). In both groups there were no significant differences in live-birth rate and miscarriage. In both groups mean serum MDA and SH-groups were significantly higher after ovarian stimulation, but mean serum SOD was significantly lower when compared to values before stimulation. There were less patients without OS before stimulation. Our results suggest that there is a difference in serum concentration in groups SOD, SH groups and MDA at the beginning and at the end ovarian stimulation. On the other hand, dose of rFSH is not related with change of parameters for oxidative stress, quality of oocytes, embryos, fertilization, pregnancies, and miscarriage rate. Patients without oxidative stress before the IVF procedure needed lower doses of gonadotropins during stimulation.

P-662 Embryo ploidy rates following PPOS or GnRH antagonist protocol. A prospective study with repeated ovarian stimulation

Abstract Study question Is there any difference in embryo ploidy rates following progesterone primed ovarian stimulation (PPOS) using micronized progesterone or GnRH antagonist protocol? Summary answer Pituitary downregulation with a PPOS protocol with micronized progesterone results in a comparable number of euploid blastocysts with a GnRH antagonist protocol. What is known already Although the GnRH antagonist is consider by most the gold standard protocol for controlling the LH surge during ovarian stimulation for IVF/ICSI, progesterone-primed ovarian stimulation protocols (PPOS) are being increasing used in freeze-all protocols. Still, despite the promising results of PPOS protocols, an early randomized trial reported potentially lower live births in recipients of oocytes resulting following downregulation with medroxyprogesterone acetate as compared with a GnRH antagonist protocol. The scope of the current prospective study was to investigate whether PPOS with micronized progesterone results in equivalent blastocyst euploidy rates as compared with a GnRH antagonist protocol. Study design, size, duration In this prospective study, performed between September 2019 to January 2022, 44 women underwent two consecutive ovarian stimulation (OS) protocols within a period of 6 months in a GnRH antagonist protocol or in a PPOS protocol with oral micronized progesterone. Participants/materials, setting, methods Overall, 44 women underwent two OS cycles with identical fixed dose of rFSH (225 or 300 IU) in both cycles. Downregulation in the 1st cycles was performed with the use of a flexible GnRH antagonist protocol and consecutively, after a washout period of one month, control of LH surge was performed with oral micronized progesterone from stimulation day 1. After the completion of both cycles, all generated blastocysts underwent genetic analysis for aneuploidy screening (PGT-A). Main results and the role of chance Comparisons between cycles did not reveal differences between the duration of OS neither in the gonadotrophins dose. Hormonal profile on the day of trigger revealed statistically significant differences between cycles in all the tested hormones except for FSH: with significantly higher serum E2 levels, more elevated LH levels and higher Progesterone levels in PPOS cycles as compared with antagonist cycles, respectively. PPOS protocol resulted in a significantly higher number of oocytes (10.27± 5.84 versus 12.68± 8.09), (DBM -2.4 [95% CI -4.1; -0.73]), MIIs (7.34±4.15 versus 9.09± 6.12), (DBM -1.8 [95% CI -3.1; -0.43]), and 2PNs (5.66±3.35 versus 7.14± 4.99), (DBM -1.5 [95% CI -2.6.1; -0.32]) as compared with the GnRH antagonist protocol. Nevertheless, no differences were observed regarding the mean number of blastocysts (2.84±2.12 versus 2.91±2.11), (DBM -0.07 [95% CI -0.67; 0.53]) and the mean number of biopsied blastocysts (2.86±2.15 versus 2.93±2.16), (DBM -0.07 [95% CI -0.70; 0.56]). Finally, no difference was observed for the primary outcome, with a mean number of euploid embryos of 0.86 ± 0.90 vs. 1.00 ± 1.12 for the comparison of PPOS vs. GnRh antagonist. Limitations, reasons for caution The study was powered to detect differences in the mean number of euploid embryos and not in terms of pregnancy outcomes. Additionally, per protocol there was no randomisation, the first cycle was always a GnRH antagonist cycle and the 2nd a PPOS with one month of washout period in between. Wider implications of the findings The current study provides evidence that the PPOS may result in equivalent blastocyst euploidy rates compared with antagonist protocol. This may imply that in case of a freeze-all protocol, clinicians may safely consider PPOS to control the LH surge and patients could benefit from the advantages (cost and oral administration). Trial registration number NCT04108039

O-134 Pretreatment with luteal estradiol for programming antagonist cycles compared to no pretreatment in advanced age women treated with corifollitropin alfa: a randomized controlled trial

Abstract Study question Does luteal estradiol pretreatment improve the number of oocytes retrieved in women aged 38 to 43 years treated in antagonist protocol with corifollitropin alfa? Summary answer Programming antagonist cycles with luteal estradiol did not increase the number of retrieved oocytes in advanced age women, however it was effective for programming. What is known already Previous studies showed that programming IVF cycles with estradiol pretreatment is a valuable tool for the organization of IVF centers and patients. However, variable effects of estradiol pretreatment were observed on the number of retrieved oocytes: no difference in normal responders, improved yield in oocyte donors and conflicting results in poor responders but no improvement in those fulfilling the bologna criteria. In advanced age women, recruitable follicles tends to decrease in number and to be more heterogeneous in size but it remains unclear if estradiol pretreatment could yield more oocytes through its negative feed-back effect on FSH intercycle rise. Study design, size, duration This non-blinded randomized controlled trial was conducted between 2016 and 2022. Participants were 324 women aged 38 to 43 years who requested IVF treatment. The primary endpoint was the comparison of the total number of retrieved oocytes between pretreated (P) and no pretreated (NP) patients. Secondary endpoints were duration and total dosage of recombinant FSH, cancellation rate, total number of metaphase II oocytes and obtained embryos, fresh transfer live birth rate (LBR) and cumulative LBR. Participants/materials, setting, methods This multicentric study (6 centers) enrolled women with regular cycles, weight &amp;gt;50 kg and BMI &amp;lt;32, IVF rank 1-2. According to computerized randomization, micronized estradiol 2mg twice a day was started on day (D) 20-24 continued until Wednesday beyond the onset of menses followed by administration of corifollitropin alfa on Friday i.e. stimulation (S)1 or from D1-3 of a natural cycle in unpretreated patients. GnRH antagonist was started at S6 and additional rFSH at S8. Main results and the role of chance Basal characteristics were similar in estradiol P (n = 147; number (mean [SD]) of treatment days 9.8[2.6]) and NP patients (n = 144), both displaying however a wide range of AMH levels (0.3 to 11.6 ng/ml) for advanced age patients. The cancellation rate showed a trend to be lower in P group (11 vs 19 %, p = 0.11). The mean number of retrieved oocytes was lower in P compared to NP patients (8.4[6.1] vs 9.1[6], respectively). The treatment difference was -0.7[-2.1 ;0.8] p = 0.03 one tailed test . No significant differences were observed in the number of MII oocytes (7[5.5] vs 7.3[5.2], p = 0.72) and the number of obtained embryos (5[4.6] vs 5.2[4.2], p = 0.60) between the two groups. Oocytes retrieval was more often on working days in P patients (91.9 vs 74.2 %, p &amp;lt; 0.001). However, the duration of stimulation was significantly longer (11.7[1.7] vs 10.8[1.8] days, p &amp;lt; 0.001) and the extra rFSH dosage in addition to corifollitropin alfa was significantly higher (1040[548] vs 778[504] IU, p &amp;lt; 0.001) in P than NP patients. The LBR after fresh transfer in P patients (16.2%[16/99]) was not significantly different from that in NP patients (18.5%[17/92]), p = 0.82, nor the cumulative LBR per patients 19.1 % vs 26.4 %, p = 0.16 respectively. Limitations, reasons for caution Our stimulated women older than 38 years obtained a wide range of collected oocytes (0 to 29) suggesting very different stages of ovarian aging in both groups. Estradiol pretreatment is more susceptible to increase oocyte yield at the stage of ovarian aging characterized by asynchrony of a reduced follicular cohort Wider implications of the findings Programming antagonist cycles with luteal estradiol pretreatment is effective in advanced age women to better schedule oocyte retrievals on working days. However, the potential benefit on the number of collected oocytes remains to be demonstrated in a more selected population displaying the characteristics of Poseïdon groups 3 and 4. Trial registration number ClinicalTrials.gov NCT02884245

Using an Interpretable Machine Learning Model to Predict Corifollitropin Alfa Protocol

Background: To demonstrate an interpretable machine learning (ML) model for a clinical prediction of corifollitropin alfa protocol. Methods: The retrospective study involved 1,221 cycles from 1,180 patients undergoing corifollitropin alfa protocol with oocyte retrieval events from a single in vitro fertilization (IVF) center. The ML models were assigned to the following tasks, which are the dosage of corifollitropin alfa, trigger type, the dosage of recombinant FSH (rFSH), the dosage of recombinant LH (rLH), the duration between the follow-up visit (FUV), and oocyte retrieval. Interpretable SHapley Additive exPlanations (SHAP) were selected to analyze the input features. The ranking of the prediction powers from each input feature reveals the level of contribution to the model. Result(s): Two series of interpretable ML models were developed to predict classification tasks and regression tasks. The areas under ROC (AUC) for predicting the dosage of corifollitropin alfa and trigger type were 0.933 ([Formula: see text] CI 0.907–0.958) and 0.891 ([Formula: see text] CI 0.864–0.918), while accuracies were 0.944 and 0.904. The mean absolute errors (MAEs) that predict the dosage of rFSH, the dosage of rLH, and the duration between the FUV and oocyte retrieval were 97.08 IU (rFSH), 105.61 IU (rLH), and 0.45 days. Conclusions: The study demonstrates a set of interpretable ML models predicting tasks involved in corifollitropin alfa protocol. The potential for the clinical application is to provide consistency in corifollitropin protocol adjustments.

The Ala307Thr polymorphism of the follicle-stimulating hormone receptor (FSHR) gene is associated with the dose of recombinant FSH received during IVF/ICSI treatment.

Follicle-stimulating hormone (FSH) is essential for folliculogenesis, acting through the follicle-stimulating hormone receptor (FSHR) that is present on the membrane of granulosa cells. Polymorphisms in the FSHR gene may lead to an altered pattern of receptor expression on the cell surface or to changes in affinity for FSH. The aim of this prospective study was to detect any association between the follicle-stimulating hormone receptor (FSHR) gene Ala307Thr polymorphism (rs6165) and ovarian reserve, ovarian response or clinical results in IVF/ICSI treatment. This prospective cohort study included 450 women who underwent IVF/ICSI cycles. DNA was extracted from peripheral blood, and the Ala307Thr FSHR polymorphism (rs6165) was genotyped using the TaqMan SNP genotyping assay. Participants were divided into three groups according to their Ala307Thr FSHR genotype: Thr/Thr (n:141), Thr/Ala (n=213) and Ala/Ala (n=96). The results were tested for associations with age, anti-Mullerian hormone (AMH) levels, antral follicle count (AFC), total dose of r-FSH, follicle size, number of retrieved oocytes, and clinical outcome of IVF/ICSI cycles. The statistical analyses were performed using Fisher's exact test and the Kruskal‒Wallis test. An association between the genotype of the FSHR (Ala307Thr) polymorphism and the dose of r-FSH was observed. Patients with the Ala/Ala genotype received a higher r-FSH dose than patients with the Ala/Thr (p=0.0002) and Thr/Thr (p=0.02) genotypes. No other correlation was observed. The Ala/Ala genotype was associated with the use of higher doses of recombinant FSH (r-FSH), suggesting that homozygosis of this allelic variant (Ala) provides lower sensitivity to r-FSH.

A real-world study of ART in France (REOLA) comparing a biosimilar rFSH against the originator according to rFSH starting dose

BackgroundSince the first launch of a biosimilar recombinant follicle stimulating hormone (rFSH), Bemfola®, in Europe in 2014, it has been possible to study in routine clinical care throughout France the effectiveness of a biosimilar rFSH including according to different rFSH starting doses. MethodsREOLA was a non-interventional, retrospective, real world study using anonymized data from 17 Assisted Reproductive Technology (ART) centres’ data management systems across France including 2,319 ART ovarian stimulation cycles with Bemfola® and 4,287 ART ovarian stimulation cycles with Gonal-f®. For both products, four populations were studied according to starting dose of rFSH: < 150 IU, 150 - 224 IU, 225 - 299 IU and ≥ 300 IU. The primary endpoint was the cumulative live birth rate (cLBR) per commenced ART ovarian stimulation cycle including all subsequent fresh and frozen-thawed embryo transfers starting during a follow up period of at least 1 year following oocyte retrieval. ResultsA direct relationship of increasing rFSH starting dose with increasing age, increasing basal FSH, decreasing AMH and increasing body mass index was noted. No clinically relevant differences were seen in all outcomes reported, including the cLBR, between Bemfola® and Gonal-f®, but for both drugs, an association was seen with increasing rFSH starting dose and decreasing cLBR. ConclusionsThe REOLA study demonstrates that the cLBR with Bemfola® is very similar to Gonal-f® across all patient subpopulations. The cLBR is inversely related to the rFSH starting dose irrespective of the drug used, and the REOLA study provides reassurance of the clinical effectiveness of a biosimilar rFSH used in a real world setting.

Comparison of blastocyst euploidy rates following luteal versus follicular phase stimulation in a GnRH antagonist protocol: a prospective study with repeated ovarian stimulation cycles.

Is there any difference in the mean number of euploid embryos following luteal phase start (LS) and follicular phase start (FS) of ovarian stimulation? The mean number of euploid blastocysts is equivalent independent of whether the inseminated oocytes are derived from FS or LS. Starting ovarian stimulation at any time of the cycle ('random-start') is commonly used for emergency fertility preservation in cancer patients. A few retrospective studies have been published evaluating LS in women undergoing ovarian stimulation in the context of IVF, but there is a lack of robust data on the comparative efficacy of LS versus FS.Although 'random start' is commonly used in cancer survivors, few retrospective and uncontrolled studies have been published evaluating luteal phase stimulation in women undergoing ovarian stimulation in the context of IVF. Owing to this evident lack of robust data on the efficacy of LS, guidelines typically recommend the LS approach only for medical reasons and not in the context of IVF. This is a prospective, equivalence study, with repeated stimulation cycles, conducted between May 2018 and December 2021. Overall, 44 oocyte donors underwent two identical consecutive ovarian stimulation cycles, one initiated in the FS and the other in the LS. The primary outcome of the study was to evaluate whether FS and LS in the same patient would result in equivalent numbers of euploid embryos following fertilization of oocytes with the same sperm sample. Overall, 44 oocyte donors underwent two consecutive ovarian stimulation protocols with 150 μg corifollitropin alpha followed by 200 IU recombinant FSH (rFSH) in a fixed GnRH antagonist protocol. The only difference between the two cycles was the day of initiation of ovarian stimulation, which was in the early follicular phase (FS) in one cycle, and in the luteal phase (LS) in the other. Forty-four oocyte recipients participated in the study receiving a mean of six metaphase II (MII) oocytes from each stimulation cycle (FS and LS). All MIIs were inseminated with the corresponding recipient's partner sperm (which had been previously frozen) or donor sperm, in order to safeguard the use of the same sample for either the FS or LS. Following fertilization and blastocyst culture, all generated embryos underwent genetic analysis for aneuploidy screening (preimplantation genetic testing for aneuploidy). FS resulted in a significantly shorter duration of ovarian stimulation (difference between means (DBM) -1.05 (95% CI -1.89; -0.20)) and a lower total additional dose of daily rFSH was needed (DBM -196.02 (95% CI -319.92; -72.12)) compared with LS. The donors' hormonal profile on the day of trigger was comparable between the two stimulation cycles, as well as the mean number of oocytes (23.70 ± 10.79 versus 23.70 ± 8.81) (DBM 0.00 (95% CI -3.03; 3.03)) and MII oocytes (20.27 ± 9.60 versus 20.73 ± 8.65) (DBM -0.45 (95% CI -2.82; 1.91)) between FS and LS cycles, respectively. Following fertilization, the overall blastocyst formation rate was 60.70% with a euploid rate of 57.1%. Comparisons between the two stimulation cycles did not reveal any significance differences in terms of fertilization rates (71.9% versus 71.4%), blastocyst formation rates (59.4% versus 62%) and embryo euploidy rates (56.9 versus 57.3%) for the comparison of FS versus LS, respectively. The mean number of euploid blastocysts was equivalent between the FS (1.59 ± 1.30) and the LS (1.61 ± 1.17), (DBM -0.02 (90%CI -0.48; 0.44)). The study was performed in young, potentially fertile oocyte donors who are patients with high blastocyst euploidy rates. Although results may be extrapolated to young infertile women with good ovarian reserve, caution is needed prior to generalizing the results to infertile women of older age. The current study provides evidence that initiation of ovarian stimulation in the luteal phase in young potentially fertile women may result in a comparable number of oocytes and comparable blastocyst euploidy rates compared with follicular phase stimulation. This may imply that in case of a freeze-all protocol in young patients with good ovarian reserve, clinicians may safely consider initiation of ovarian stimulation during the luteal phase. This research was supported by an unrestricted grant from MSD/Organon. N.P.P. has received Research grants and honoraria for lectures from: Merck Serono, MSD/Organon, Ferring Pharmaceuticals, Besins Intenational, Roche Diagnostics, IBSA, Theramex, Gedeon Richter. F.M., E.C., M.R. and S.G. declared no conflict of interests. The study was registered at Clinical Trials Gov (NCT03555942).

P-477 Ovarian stimulation and PGT-A outcomes with random start letrozole protocol for fertility preservation in breast cancer patients

Abstract Study question Data on random start letrozole ovarian stimulation (RSL) in breast cancer patients is scant. We studied RSL outcomes from a single-center with fertility preservation expertise. Summary answer RSL appears to result in high oocyte and embryo/blastocyst yield, with an age-appropriate aneuploidy rate. What is known already With increasing utility of neoadjuvant chemotherapy regimens in breast cancer, young women have limited time to undergo fertility preservation (FP) by oocyte or embryo cryopreservation. Random start stimulation protocols were developed to complete an ovarian stimulation cycle in the shortest time possible before cancer patients started chemotherapy. In addition, letrozole protocols were developed to mitigate the potential risks of high estrogen levels during ovarian stimulation in breast cancer patients. Even though both random start and letrozole supplemented ovarian stimulation have been studied individually, there is limited data on combining both strategies in random start letrozole (RSL) protocols. Study design, size, duration We reviewed our records from 2014 to 2021. We included the first FP cycle of all breast cancer patients undergoing RSL ovarian stimulation for oocyte or embryo cryopreservation. Cancer stages were grouped as 0-4, 0 being ductal carcinoma in situ (DCIS). Twenty-eight cycles met the criteria. Main outcome measures were number of oocytes retrieved and embryos cryopreserved. Secondary outcome measures were fertilization, blastocyst, and euploidy rates for the embryo cryopreservation group. Participants/materials, setting, methods Ovarian stimulation began on cycle days 3-26 with letrozole 5 mg/day initiated simultaneously with rFSH. The rFSH dose was based on age and antral follicle counts (AFC). Oocyte maturation was triggered with leuprolide acetate, rhCG or both 35h prior to retrieval. Main results and the role of chance The mean age was 33.5±4.9 years (range 26-40) and BMI 22.3±2.9(18.4-30.3). The mean breast cancer stage at FP was 1.7±0.9 (0-4), with 42.3% being stage-I disease. 77% of cases were estrogen receptor positive. AMH was 2.7±2.4 (0.16-7.9) and AFC 18.5±8.9(4-42). Ovarian stimulation began on cycle-day 13.7±7.3(2-26). 15(54%) cycles were started in the follicular and 13(46%) in the luteal-phase. The mean total rFSH dose was 3880±1615.4 IU(1275-8325) with a stimulation length of 11.8±2.4 days(8-19). Mean oocytes retrieved were 16.4±6.9(1-36) with a maturity rate of 71.4±17.9%(35.7-94.4). From 13 patients, 10.4±4.7(1-22) oocytes were cryopreserved. Among the 15 patients proceeding to embryo cryopreservation, the fertilization rate was 96.9±6.8%(80-100), resulting in 7.1±4.4(1-15) embryos. 24 embryos were cryopreserved on day-3 and 82 as blastocysts. PGT-A was performed in 72% of the cases, with euploidy rate of 37.7±24.4%(0-63.6). There was no difference in oocyte or embryo yield between follicular and luteal start patients. Likewise, fertilization rates and euploidy rates were similar. The comparative results are summarized in Table 1. To date, two patients conceived utilizing cryopreserved embryos. Both pregnancies resulted in healthy term babies. Limitations, reasons for caution Though our report is one of the largest series of RSL, it can be strengthened with longer follow up that includes pregnancy outcomes in a greater proportion of patients. Wider implications of the findings This is the first detailed report of the RSL protocol utility with PGT-A results in a breast-cancer population. RSL appears to result in high oocyte-yield, fertilization, and blastocyst formation rates, with age-appropriate euploidy. Additionally, the outcomes appear to be similar between patients undergoing follicular and luteal phase start RSL. Trial registration number not applicable

P-591 Cellular heterogeneity in the preovulatory follicle correlates with response to stimulation with recombinant FSH and affects somatic cell gene expression levels

Abstract Study question What is the difference in the composition and gene expression of pre-ovulatory follicular somatic cells between normo- and hyporesponder patients to rFSH stimulation? Summary answer Gene expression changes in the somatic cells of the preovulatory follicles of hyporesponder patients is partly attributable to distinct cell populations. What is known already Some women undergoing IVF respond suboptimally to the preceding ovarian stimulation without an indication of poor ovarian insufficiency nor advanced age. The gene expression levels in the somatic cells of their pre-ovulatory follicles have not been determined. However, the somatic cells play key roles in the transfer of gonadotrophin signals and in steroidogenesis. In the preovulatory follicle, being &amp;gt;20 mm in size, the somatic cells are exposed to various molecular gradients that drive their differentiation. Due to advancements in sequencing technologies the importance of somatic cell sub-populations in the etiologies of ovarian infertility can now be investigated by bioinformatic methods. Study design, size, duration Consecutive IVF patients &amp;lt;41 years of age were recruited at Nova Vita Clinic, Estonia, undergoing the antagonist stimulation protocol with rFSH administration. Ovarian puncture was performed for all patients after 36 hours of hCG stimulaton. rFSH dose &amp;gt;200 IU per one retrieved oocyte was considered as an indication for hyporesponse. Ten normo- and 9 hyporesponder patients were enrolled. Participants/materials, setting, methods Cells from the follicular fluid devoid of the cumulus oocyte complex were collected by centrifugation. RNA-seq was performed on the pooled cells of all participants. The study groups were compared by differential gene expression analysis by DESeq2. Single-cell RNA-seq was performed for &amp;gt;6000 individual cells of 3 normoresponder patients and cellular sub-populations were determined with Seurat package. CIBERTSORTx software was used to deconvolute the proportion of cell types from bulk RNA-seq data of all participants. Main results and the role of chance Alterations in the gene expression of ovarian somatic cells have been previously attributed to female age. We demonstrate that 407 genes are differentially expressed (FDR&amp;lt;0.05) between normo- and hyporesponder IVF patients after age adjustment. These genes were enriched into pathways of extracellular matrix reorganization (31 genes, FDR&amp;lt;0.005), post-translational protein phosphorylation (16 genes, FDR&amp;lt;0.006) and regulation of insulin-like growth factor transport and uptake by insulin-like growth factor binding proteins (16 genes, FDR&amp;lt;0.019). In addition, the sequencing of &amp;gt; 24 000 individual cells from 3 normoresponder patients revealed the somatic cell types of the preovulatory follicle in high resolution indicating the presence of immune cell populations (N = 4), epithelial cells, theca, cumulus cells and mural granulosa cell sub-populations (N = 7). Bioinformatic cell type deconvolution demonstrated significant differences between proportions of distinct cell sub-populations between normo- and hyporesponder patients and that several differentially expressed genes between the study groups can be attributed to a specific mural granulosa sub-population. Limitations, reasons for caution The number of study participants is small due to the high cost of the study methods. Wider implications of the findings The study demonstrates that hyporesponse to stimulation is associated with age-unrelated disturbances in gene expression of preovulatory somatic cells and that different somatic cell populations may have important underlying functions in the ovarian etiologies of infertility. Trial registration number not applicable

P-660 Polymorphisms in FSHR gene do not affect late follicular phase steroidogenic response in predicted normoresponders. Secondary analysis of a prospective multicenter cohort study

Abstract Study question Does the presence of FSHR SNPs influence late follicular phase progesterone and estradiol serum levels in predicted normoresponders treated with rFSH? Summary answer The presence of FSHR SNPs (rs6165, rs6166, rs1394205) had no statistically significant impact on late follicular phase serum progesterone and estradiol levels. What is known already Previous studies have shown that late follicular phase serum progesterone and estradiol levels are significantly correlated with the magnitude of ovarian response. Several authors have proposed that individual variability in the response to ovarian stimulation could be explained by variants in FSHR. So far, the literature is scarce on the influence of this genetic variability on late follicular phase steroidogenic response. Our aim is to determine whether genetic variants in the FSHR gene could modulate late follicular phase serum progesterone and estradiol levels. Study design, size, duration We performed a secondary analysis of a multicenter multinational prospective study including 366 patients from Vietnam, Belgium and Spain (166 from Europe and 200 from Asia), conducted from 11/2016-06/2019. All patients underwent ovarian stimulation followed by oocyte retrieval in an antagonist protocol with a fixed daily dose of 150IU rFSH. All patients had a serum progesterone and estradiol measurement on the day of trigger and were genotyped for 3 FSHR SNPs (rs6165, rs6166, rs1394205). Participants/materials, setting, methods Patients included were predicted normal responder women &amp;lt;38 years old undergoing their first or second ovarian stimulation cycle. The prevalence of late follicular phase elevated serum progesterone (EP), as well as mean serum progesterone and estradiol levels on the day of trigger were compared between the different FSHR SNPs genotypes. EP was defined as &amp;gt; 1.5 ng/ml. Main results and the role of chance The overall prevalence of EP was 15.8% (n = 58). No significant difference was found in the prevalence of EP in Caucasian and Asian patients (17.5% vs. 14.5%). Genetic model analysis revealed a similar prevalence of EP in co-dominant, dominant and recessive models for variants FSHR rs6166, rs6165 and rs1394205. Also, no statistically significant difference was found in the mean serum progesterone levels in the three genetic models. Likewise, FSHR SNPs genotypes had no statistically significant impact in the mean late follicular phase serum estradiol levels Limitations, reasons for caution This study is a post-hoc analysis of a multicenter multinational prospective cohort study. The results must be interpreted with caution considering the sample size of the EP group. The fact that a fixed daily dose of 150 IU rFSH was used in this population precludes the generalization of the results. Wider implications of the findings Based on our results, FSHR SNPs rs6165, rs6166, rs1394205 do not influence late follicular phase serum progesterone nor estradiol levels in predicted normal responders. These findings add to the controversy in the literature regarding the impact of individual genetic susceptibility in response to ovarian stimulation in this population. Trial registration number not-applicable