Research Laboratory on Human Reproduction, Faculté de Médecine, Université Libre de Bruxelles, Brussels, Belgium Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculté de Médecine, Université Libre de Bruxelles, Brussels, Belgium Fertility Clinic, Department of Gynecology and Obstetrics, Erasme Hospital, Université Libre de Bruxelles, Brussels, Belgium
Fertility Clinic, Department of Gynecology and Obstetrics, Erasme Hospital, Université Libre de Bruxelles, Brussels, Belgium.
Hum Reprod. 2015 May;30(5):1196-202. doi: 10.1093/humrep/dev042. Epub 2015 Mar 6.
Could anti-Müllerian hormone (AMH) mutations be implicated in the development of idiopathic premature ovarian insufficiency (POI)?
Three rare or unknown missense variants of the AMH gene were identified in a cohort of 55 POI patients; all three variants showed a drastically reduced in vitro bioactivity.
Genetic factors are implicated in 5-15% of cases of POI. However, only a few genes have been shown to be involved in its development. AMH inhibits the recruitment of primordial follicles in the ovary and defective or absent AMH leads to premature depletion of the primordial follicle pool in AMH null mice.
STUDY DESIGN, SIZE, DURATION: The whole coding sequence and the exon-intron junction of the AMH gene was sequenced in a cohort of 55 POI patients recruited over a period of 8 years. The studied variants were also sequenced in 197 ethnically matched controls.
PARTICIPANTS/MATERIALS, SETTING, METHODS: POI was defined as amenorrhea of more than 4 months with increased FSH before the age of 40. Patients with POI resulting from radio- or chemotherapy, surgery, chromosomal anomalies or FMR1 gene pre-mutation were excluded from the study. Recombinant human wild-type (wt) and mutated AMH proteins were produced in HEK293 T cells. KK-1 cells transfected with the AMH receptor type 2 (AMHR2) and a BMP responsive element coupled to a luciferase reporter vector were stimulated with different concentrations of wt AMH and the three tested variants.
The whole coding sequence of the AMH gene could be performed and analyzed for 50 POI patients: 16 variants were found, including 6 missense variants from which 1 was unknown (R444H) and 2 were very rare (G264R and D288E). The variant D288E was also found in one of the patient's mother who also underwent POI at 32 years old. The stimulation of the AMHR2 assessed by the luciferase activity was drastically reduced for the three variants when compared with the wt AMH.
LIMITATIONS, REASONS FOR CAUTION: The study is limited by a relatively small number of patients in the POI cohort.
This is the first time that the bioactivity of AMH variants related to POI patients is tested in vitro. The functional study showed a drastic reduction of the protein activity for the three variants, supporting their contribution to the development of the ovarian insufficiency. The familial segregation further supports the implication of AMH in the development of POI.
STUDY FUNDING/COMPETING INTERESTS: The study was performed thanks to funding from the 'Fondation Erasme'. No conflicts of interest are declared.
抗缪勒管激素（AMH）突变是否与特发性卵巢早衰（POI）的发生有关？
在 55 名 POI 患者的队列中发现了 AMH 基因的三个罕见或未知的错义变体；所有三种变体的体外生物活性都明显降低。
遗传因素在 5-15%的 POI 病例中起作用。然而，只有少数基因被证明与它的发展有关。AMH 抑制卵巢中原始卵泡的募集，AMH 缺陷或缺失导致 AMH 缺失小鼠中原始卵泡池的过早耗竭。
研究设计、大小、持续时间：在 8 年的时间里，对一组 55 名 POI 患者进行了 AMH 基因的整个编码序列和外显子-内含子连接测序。在 197 名具有相同种族的对照中也对研究变异进行了测序。
参与者/材料、设置、方法：POI 被定义为 40 岁前 FSH 升高的 4 个月以上的闭经。排除因放射或化学疗法、手术、染色体异常或 FMR1 基因前突变引起的 POI 患者。在 HEK293 T 细胞中产生重组人野生型（wt）和突变 AMH 蛋白。用不同浓度的 wt AMH 和三种测试变体刺激转染 AMH 受体 2（AMHR2）和与 BMP 反应元件偶联的荧光素酶报告载体的 KK-1 细胞。
对 50 名 POI 患者进行了 AMH 基因的整个编码序列的检测和分析：发现了 16 个变异，包括 6 个错义变异，其中 1 个是未知的（R444H），2 个是非常罕见的（G264R 和 D288E）。D288E 变异也在一名患者的母亲中发现，该患者的母亲也在 32 岁时经历了 POI。与 wt AMH 相比，三种变体的 AMHR2 刺激的荧光素酶活性明显降低。
局限性、谨慎的原因：该研究受到 POI 队列中患者数量相对较少的限制。
这是第一次在体外测试与 POI 患者相关的 AMH 变体的生物活性。功能研究表明，三种变体的蛋白活性明显降低，支持它们对卵巢功能不全的发展的贡献。家族性分离进一步支持了 AMH 在 POI 发展中的作用。
研究资金/利益冲突：该研究得到了“Erasme 基金会”的资助。没有利益冲突声明。
