Cadenas J, Poulsen L C, Nikiforov D, Grøndahl M L, Kumar A, Bahnu K, Englund A L M, Malm J, Marko-Varga G, Pla I, Sanchez A, Pors S E, Andersen C Yding
Laboratory of Reproductive Biology, Juliane Marie Centre for Women, Children and Reproduction, Copenhagen University Hospital, Rigshospitalet, Denmark.
Zealand Fertility Clinic, Zealand University Hospital, Køge, Denmark.
Hum Reprod. 2023 Apr 3;38(4):686-700. doi: 10.1093/humrep/dead024.
Which substances and signal transduction pathways are potentially active downstream to the effect of FSH and LH in the regulation of human oocyte maturation in vivo?
The regulation of human oocyte maturation appears to be a multifactorial process in which several different signal transduction pathways are active.
Many studies in animal species have provided insight into the mechanisms that govern the final maturation of oocytes. Currently, these studies have identified several different mechanisms downstream to the effects of FSH and LH. Some of the identified mechanisms include the regulation of cAMP/cGMP levels in oocytes involving C-type natriuretic peptide (CNP), effects of epidermal growth factor (EGF)-related peptides such as amphiregulin (AREG) and/or epiregulin (EREG), effect of TGF-β family members including growth differentiation factor 9 (GDF9) and morphogenetic protein 15 (BMP15), activins/inhibins, follicular fluid meiosis activating sterol (FF-MAS), the growth factor midkine (MDK), and several others. However, to what extent these pathways and mechanisms are active in humans in vivo is unknown.
STUDY DESIGN, SIZE, DURATION: This prospective cohort study included 50 women undergoing fertility treatment in a standard antagonist protocol at a university hospital affiliated fertility clinic in 2016-2018.
PARTICIPANTS/MATERIALS, SETTING, METHODS: We evaluated the substances and signalling pathways potentially affecting human oocyte maturation in follicular fluid (FF) and granulosa cells (GCs) collected at five time points during the final maturation of follicles. Using ELISA measurement and proteomic profiling of FF and whole genome gene expression in GC, the following substances and their signal transduction pathways were collectively evaluated: CNP, the EGF family, inhibin-A, inhibin-B, activins, FF-MAS, MDK, GDF9, and BMP15.
All the evaluated substances and signal transduction pathways are potentially active in the regulation of human oocyte maturation in vivo except for GDF9/BMP15 signalling. In particular, AREG, inhibins, and MDK were significantly upregulated during the first 12-17 h after initiating the final maturation of follicles and were measured at significantly higher concentrations than previously reported. Additionally, the genes regulating FF-MAS synthesis and metabolism were significantly controlled in favour of accumulation during the first 12-17 h. In contrast, concentrations of CNP were low and did not change during the process of final maturation of follicles, and concentrations of GDF9 and BMP15 were much lower than reported in small antral follicles, suggesting a less pronounced influence from these substances.
None.
LIMITATIONS, REASONS FOR CAUTION: Although GC and cumulus cells have many similar features, it is a limitation of the current study that information for the corresponding cumulus cells is not available. However, we seldom recovered a cumulus-oocyte complex during the follicle aspiration from 0 to 32 h.
Delineating the mechanisms governing the regulation of human oocyte maturation in vivo advances the possibility of developing a platform for IVM that, as for most other mammalian species, results in healthy offspring with good efficacy. Mimicking the intrafollicular conditions during oocyte maturation in vivo in small culture droplets during IVM may enhance oocyte nuclear and cytoplasmic maturation. The primary outlook for such a method is, in the context of fertility preservation, to augment the chances of achieving biological children after a cancer treatment by subjecting oocytes from small antral follicles to IVM. Provided that aspiration of oocytes from small antral follicles in vivo can be developed with good efficacy, IVM may be applied to infertile patients on a larger scale and can provide a cheap alternative to conventional IVF treatment with ovarian stimulation. Successful IVM has the potential to change current established techniques for infertility treatment.
STUDY FUNDING/COMPETING INTEREST(S): This research was supported by the University Hospital of Copenhagen, Rigshospitalet, the Independent Research Fund Denmark (grant number 0134-00448), and the Interregional EU-sponsored ReproUnion network. There are no conflicts of interest to be declared.
在体内调节人类卵母细胞成熟过程中，FSH和LH作用的下游哪些物质和信号转导途径可能是活跃的？
人类卵母细胞成熟的调节似乎是一个多因素过程，其中几种不同的信号转导途径是活跃的。
许多针对动物物种的研究深入探讨了控制卵母细胞最终成熟的机制。目前，这些研究已确定了FSH和LH作用下游的几种不同机制。一些已确定的机制包括涉及C型利钠肽（CNP）的卵母细胞中cAMP/cGMP水平的调节、表皮生长因子（EGF）相关肽如双调蛋白（AREG）和/或表皮调节素（EREG）的作用、转化生长因子-β家族成员包括生长分化因子9（GDF9）和形态发生蛋白15（BMP15）的作用、激活素/抑制素、卵泡液减数分裂激活甾醇（FF-MAS）、生长因子中期因子（MDK）以及其他一些物质的作用。然而，这些途径和机制在人类体内的活跃程度尚不清楚。
研究设计、规模、持续时间：这项前瞻性队列研究纳入了2016 - 2018年在一所大学附属医院附属生育诊所接受标准拮抗剂方案生育治疗的50名女性。
参与者/材料、设置、方法：我们评估了在卵泡最终成熟过程中的五个时间点收集的卵泡液（FF）和颗粒细胞（GCs）中可能影响人类卵母细胞成熟的物质和信号通路。通过ELISA测量以及FF的蛋白质组分析和GC中的全基因组基因表达，对以下物质及其信号转导途径进行了综合评估：CNP、EGF家族、抑制素-A、抑制素-B、激活素、FF-MAS、MDK、GDF9和BMP15。
除GDF9/BMP15信号通路外，所有评估的物质和信号转导途径在体内调节人类卵母细胞成熟过程中都可能是活跃的。特别是，AREG、抑制素和MDK在卵泡开始最终成熟后的最初12 - 17小时内显著上调，且测量浓度显著高于先前报道。此外，在最初12 - 17小时内，调节FF-MAS合成和代谢的基因受到显著调控，有利于其积累。相比之下，CNP浓度较低，在卵泡最终成熟过程中没有变化，GDF9和BMP15的浓度远低于小窦卵泡中的报道浓度，表明这些物质的影响不太明显。
无。
局限性、谨慎原因：尽管颗粒细胞和卵丘细胞有许多相似特征，但目前研究的一个局限性是没有相应卵丘细胞的信息。然而，在0至32小时的卵泡抽吸过程中，我们很少回收卵丘 - 卵母细胞复合体。
描绘体内调节人类卵母细胞成熟的机制增加了开发体外成熟（IVM）平台的可能性，与大多数其他哺乳动物物种一样，该平台能产生健康后代且效果良好。在IVM期间，在小培养滴中模拟卵母细胞体内成熟过程中的卵泡内环境可能会增强卵母细胞核和细胞质的成熟。在生育力保存的背景下，这种方法的主要前景是通过对来自小窦卵泡的卵母细胞进行IVM，增加癌症治疗后获得亲生孩子的机会。如果能够高效地在体内从小窦卵泡中抽吸卵母细胞，IVM可能会更广泛地应用于不孕患者，并可为传统的卵巢刺激体外受精治疗提供一种廉价的替代方法。成功的IVM有可能改变当前既定的不孕治疗技术。
研究资金/利益冲突：本研究得到了哥本哈根大学医院、里格霍斯医院、丹麦独立研究基金（资助编号0134 - 00448）以及欧盟资助的跨地区ReproUnion网络的支持。不存在需要申报的利益冲突。
