Reproductive Medicine Center, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, People's Republic of China; Guangdong Provincial Key Laboratory of Reproductive Medicine, Guangzhou, Guangdong, China.
Reproductive Medicine Center, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, People's Republic of China; Guangdong Provincial Key Laboratory of Reproductive Medicine, Guangzhou, Guangdong, China; Reproductive Medicine Center, The Affiliated Shenzhen Maternity and Child Healthcare Hospital of the South Medical University, Shenzhen, Guangdong, China.
Fertil Steril. 2023 Sep;120(3 Pt 2):671-681. doi: 10.1016/j.fertnstert.2023.03.031. Epub 2023 Mar 29.
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.
Genetic association study.
University-affiliated in vitro fertilization center.
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.
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.
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.
In the exploratory cohort, we identified that FSHB rs6169 (P=2.71 × 10) 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), shorter duration of stimulation, and less amount of exogenous FSH (P=0.0002; P=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.
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.
研究常见遗传多态性是否与每日促性腺激素释放激素激动剂下调后的促性腺激素水平有关，以及候选变异的多态性是否影响外源性促性腺激素对卵巢的反应。
遗传关联研究。
大学附属体外受精中心。
2016 年 9 月至 2018 年 10 月、2019 年 9 月至 2020 年 9 月和 2021 年 1 月至 2021 年 12 月分别招募了参加探索性外显子组全关联研究（n=862）、复制外显子组全关联研究（n=86）和分类器验证研究（n=148）的受试者。纳入的患者年龄≤40 岁，基础卵泡刺激素（FSH）≤12IU/L。
所有参与者均接受黄体期下调长方案。从外周血白细胞中提取基因组 DNA。对于探索性和复制队列，在外显子组测序平台上进行 HiSeq 2500 测序。在探索性和复制队列中也进行了多重聚合酶链反应扩增技术和下一代测序。对于验证队列的样本，进行 Sanger 测序。
主要终点是下调后的促性腺激素水平，次要终点是刺激期间的激素水平和卵泡直径、FSH 的总剂量、FSH 刺激持续时间、获卵数和临床妊娠率。
在探索性队列中，我们发现 FSHB rs6169（P=2.71×10）及其高度连锁不平衡的单核苷酸多态性与下调的 FSH 水平有关。在复制队列中也证实了相同的基因座。携带 FSHB rs6169 C 等位基因的女性在刺激期间表现出更高的平均雌二醇水平（P=6.82×10）、刺激持续时间更短、外源性 FSH 用量更少（P=0.0002；P=0.0024）。在独立的验证集中，将 rs6169 基因型添加到下调后 FSH 水平的预测模型中，Logistic 回归模型的曲线下面积从 0.560 提高到 0.712，当应用支持向量机分类器时，预测准确性提高了 41.05%。
FSHB rs6169 的 C 等位基因是下调后 FSH 水平相对较高的易感位点，可能与卵巢 FSH 敏感性增加有关。
