Identification of common genetic polymorphisms associated with down-regulated gonadotropin levels in an exome-wide association study.

OBJECTIVE: To investigate whether common genetic polymorphisms are associated with gonadotropin levels after down-regulation with daily gonadotropin-releasing hormone agonist and whether the polymorphisms of candidate variants influence the ovarian response to exogenous gonadotropins. DESIGN: Genetic association study. SETTING: University-affiliated in vitro fertilization center. PATIENTS: Subjects enrolled in an exploratory exome-wide association study (n = 862), a replication exome-wide association study (n = 86), and a classifier validation study (n = 148) were recruited from September 2016 to October 2018, September 2019 to September 2020, and January 2021 to December 2021, respectively. The included patients were aged ≤40 years and had a basal follicle-stimulating hormone (FSH) ≤12 IU/L. INTERVENTIONS: All participants received a luteal phase down-regulation long protocol. Genome DNA was extracted from the peripheral blood leukocytes. For the exploratory and replication cohorts, exome sequencing was conducted on a HiSeq 2500 sequencing platform. The multiplex polymerase chain reaction amplification technique and next-generation sequencing also were performed in the exploratory and replication cohorts. For the samples of the validation cohort, Sanger sequencing was performed. MAIN OUTCOME MEASURES: The primary endpoint was the gonadotropin levels after down-regulation, and the secondary endpoints were hormone levels and follicle diameters during stimulation, the total dose of FSH, duration of FSH stimulation, number of oocytes retrieved, and clinical pregnancy rate. RESULTS: In the exploratory cohort, we identified that FSHB rs6169 (P=2.71 × 10-24) and its single-nucleotide polymorphisms in high linkage disequilibrium were associated with the down-regulated FSH level. The same locus was confirmed in the replication cohort. Women carrying the C allele of FSHB rs6169 exhibited higher average estradiol level during stimulation (P=6.82 × 10-5), shorter duration of stimulation, and less amount of exogenous FSH (Pduration=0.0002; Pdose=0.0024). In the independent validation set, adding rs6169 genotypes into the prediction model for FSH level after down-regulation enhanced the area under the curve from 0.560 to 0.712 in a logistic regression model, and increased prediction accuracy by 41.05% when a support vector machine classifier was applied. CONCLUSION: The C allele of FSHB rs6169 is a susceptibility site for the relatively high level of FSH after down-regulation, which may be associated with increased ovarian FSH sensitivity.

Effect of the initiation of progesterone supplementation in in vitro fertilization-embryo transfer outcomes: a prospective randomized controlled trial.

OBJECTIVE: To analyze the influence of the start point of luteal support on clinical pregnancy rate, implantation rate, and live birth rate of in vitro fertilization and embryo transfer (IVF-ET) cycles. DESIGN: Single-center prospective randomized controlled trial. SETTING: University-affiliated IVF unit. PATIENT(S): Women ≤35 years of age with day 3 FSH levels <15 mIU/mL, who were undergoing their first IVF-ET cycles and received ovarian stimulation with the use of a GnRH agonist long protocol. INTERVENTION(S): The patients were randomized on the day of hCG trigger to receive luteal phase support either 1 day after oocyte retrieval (group A) or on the day of oocyte retrieval (group B). MAIN OUTCOME MEASURE(S): Clinical pregnancy rate, implantation rate, miscarriage rate in the first trimester of pregnancy, and live birth rate per embryo transfer cycle. RESULT(S): Two hundred thirty-three patients were enrolled in this study: 117 were assigned to group A and 116 to group B. The clinical pregnancy rate (group A vs. group B: 55.3% vs. 51.5%), implantation rate (38.4% vs. 38.0%), and miscarriage rate (7.7% vs. 7.5%) were similar between the two groups. The live birth rate also did not significantly differ between the two groups (45.7% vs. 46.6%). CONCLUSION(S): Our study indicated that the initiation of progesterone supplementation 1 day after oocyte retrieval did not decrease the clinical pregnancy rate, implantation rate, or live birth rate in women undergoing IVF-ET cycles with the use of the GnRH agonist long protocol. CLINICAL TRIAL REGISTRATION NUMBER: ChiCTR-IPR-14005293.

Self-control study on reduced-dose depot versus daily administration of gonadotrophin-releasing hormone agonists for pituitary desensitization in in vitro fertilization cycles.

AIM: Our aim was to analyze the effect of reducing the dose of depot gonadotrophin-regulating hormone-agonist (GnRH-a) to 1.0 mg on pituitary desensitization and clinical outcome of in vitro fertilization and embryo transfer cycles. MATERIAL AND METHODS: This retrospective self-control study was conducted on 143 patients who underwent repeated long-protocol treatment from 1 January 2011 to 31 May 2012 at our hospital. Of the 143 patients, 64 received reduced-dose depot (1.0 mg diphereline depot) GnRH-a for the first cycle and short-acting GnRH-a (0.05 mg diphereline) for the second cycle, while 79 patients received short-acting GnRH-a for the first cycle and reduced-dose depot GnRH-a for the second cycle. RESULTS: The serum follicle-stimulating hormone, luteinizing hormone and estradiol levels on the day of gonadotrophin initiation were significantly higher in the short-acting group compared with the long-acting group. Both number of days of gonadotrophin stimulation and gonadotrophin doses were significantly higher in the short-acting group. On the day of human chorionic gonadotrophin administration, the serum estradiol level was significantly higher while the progesterone level was significantly lower in the short-acting group. There were no significant differences with regard to the number of retrieved oocytes, fertilization rate, number of transferred embryos, clinical pregnancy rate, implantation rate, and early pregnancy loss rate between the two groups. However, the oocyte maturation rate was significantly higher in the long-acting group. CONCLUSION: Reduced-dose depot GnRH-a can be successfully used for pituitary desensitization in in vitro fertilization and embryo transfer. Deeper downregulation with reduced-dose depot GnRH-a indicates that the optimal dose of GnRH-a warrants future study.

Preliminary investigation of the impact of anticentromere antibody on oocyte maturation and embryo cleavage.

OBJECTIVE: To explore whether anticentromere antibody (ACA) is the most significant antibody among antinuclear antibodies (ANA), which adversely affect oocyte maturation, embryo cleavage, and pregnancy outcome in women undergoing an intracytoplasmic sperm injection program. DESIGN: Retrospective, nested case-control study. SETTING: Center for reproductive medicine, university hospital. PATIENT(S): A total of 187 women receiving the first intracytoplasmic sperm injection cycle were enrolled in this study, including 20 women with positive ACA and ANA (ACA[+]/ANA[+] group), 51 women with negative ACA and positive ANA(ACA[-]/ANA[+] group), and 116 patients with negative ACA and ANA (ACA[-]/ANA[-] group). Patients in the three groups were age-matched. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Percentages of germinal vesicle, metaphase I, and metaphase II oocytes, embryo cleavage rate, number of high-quality embryos, and rates of pregnancy and implantation. RESULT(S): The metaphase I oocyte percentage was markedly higher and the metaphase II oocyte percentage and the normal cleavage rate were significantly lower in the ACA[+]/ANA[+] group as compared with the ACA[-]/ANA[+] group. Furthermore, statistically significant differences were found in rates of pregnancy and implantation among the three groups. However, no significant difference was found between any two groups owing to the small sample size, except for a significantly lower implantation rate being found in the ACA[+]/ANA[+] group when compared with the ACA[-]/ANA[-] group. CONCLUSION(S): Our data suggest that ACA may be the essential marker for defective oocytes or embryos in infertile women with any type of ANA.

Mechanically expanding the zona pellucida of human frozen thawed embryos: a new method of assisted hatching.

OBJECTIVE: To determine whether a new assisted hatching (AH) method increases the implantation and clinical pregnancy rates of frozen-thawed day-3 (D3) embryos. DESIGN: Prospective study. SETTING: A university hospital in vitro fertilization (IVF) program. PATIENT(S): Patients who had their first IVF/intracytoplasmic sperm injection (ICSI) cycles between June 1, 2006, and December 31, 2008, with fresh IVF-embryo transfer failures or without fresh embryo transfer. INTERVENTION(S): The couples were randomized into thawed embryo transfer after AH versus no AH. In the AH group, the zona pellucida (ZP) of D3 frozen-thawed embryos was expanded by injected hydrostatic pressure after thawing. In the control group, embryos were pierced by ICSI needles without expanding the ZP. MAIN OUTCOME MEASURE(S): Clinical pregnancy and implantation rates. RESULT(S): The morphologic features of the blastomeres were carefully monitored and recorded. In the AH group, 244 embryos were thawed, and 178 (73.0%) survived; in the control group, 259 embryos were thawed, and 190 (73.4%) survived. Despite the transfer of a similar number of embryos, the AH group resulted in statistically significantly higher implantation and clinical pregnancy rates compared with the no AH group. CONCLUSION(S): Mechanically expanding the ZP of frozen-thawed D3 embryos with injected hydrostatic pressure after thawing increases the implantation rate compared with control embryos.