Deregulation of the endometrial stromal cell secretome precedes embryo implantation failure
Deregulation of the endometrial stromal cell secretome precedes embryo implantation failure
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DOI：
10.1093/molehr/gax023
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Lee, Yie Hou
中科院分区：
文献类型：
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作者：
Durairaj, Ruban Rex Peter;Aberkane, Asma;Lee, Yie Hou
ART 后的着床失败是否与子宫内膜基质细胞 (EnSC) 的蜕膜反应受到干扰有关？ ART 之前这一过渡途径的扰动与着床失败相关。尽管异常子宫内膜微环境在 IVF 失败中的作用仍不清楚，但着床是 ART 中的限速步骤。在成功或失败的 ART 周期之前分离的原代 EnSC 的蜕膜反应的时间变化的体外特征。对胚胎对未分化和蜕膜化 EnSC 分泌信号的反应进行了分析。该研究的主要临床结果是胚胎移植后 14 天尿妊娠试验呈阳性。从 IVF 治疗前获得的子宫内膜活检中分离出初级 EnSC 并冷冻保存。然后将来自 10 名怀孕患者和 10 名非怀孕患者的 EnSC 解冻，在培养物中扩增，进行克隆形成分析，并进行 2 或 8 天的蜕膜化。使用实时定量 PCR 分析蜕膜标记基因 [催乳素 (PRL)、胰岛素样生长因子结合蛋白 1 (IGFBP1) 和 11 β-羟基类固醇脱氢酶 (HSD11B1)] 的转录水平，并通过多重悬浮珠免疫测定法测量 45 种细胞因子、趋化因子和生长因子的时间分泌组变化。 EnSC分泌蛋白组对人类囊胚发育的影响进行了形态学评分；妊娠组和非妊娠组的 EnSC 培养物的克隆形成性和蜕膜标记基因的诱导具有可比性 (P > 0.05)。对 23 个分泌因子的分析表明，成功植入与蜕膜化 EnSC 中协调的分泌蛋白组变化相关，这种变化在分化第 2 天最为明显：相对于未分化（第 0 天）EnSC，蜕膜化第 2 天有 17 个差异分泌蛋白（P < 0.05）；相对于第 2 天，第 8 天有 11 个差异分泌蛋白（P < 0.05）；与第 0 天相比，第 8 天有 8 个差异分泌的蛋白质 (P < 0.05)。相比之下，植入失败与分泌体反应紊乱有关。与蜕膜化 EnSC 相比，在未分化 EnSC 条件培养基中培养 24 小时时，囊胚发育受到损害。对胚胎微滴的分析表明，人类囊胚对分化早期（第 2 天）而非晚期（第 8 天）产生的可溶性蜕膜因子产生分泌性细胞因子反应。怀孕组和非怀孕组的胚胎对蜕膜化 EnSC 分泌因子的反应相当（P > 0.05）。不适用。虽然本研究使用原代 EnSC 和人类胚胎，但由于研究的相关性和样本量有限，在将结果外推到体内情况时需要谨慎。我们的发现提出了 ART 之前进行子宫内膜分析可以最大限度地降低治疗失败风险的前景。这项工作得到了来自生殖健康生物医学研究单位，由考文垂和沃里克郡 NHS 信托大学医院和沃里克医学院、诺丁汉大学和 Nurture Fertility 以及新加坡国家医学研究理事会 (NMRC/BNIG14NOV023)、“Instituut voor Innovatie Door Wetenschap en Technologie”（IWT，佛兰德斯，比利时）、“Fonds voor”联合倡议Wetenschappelijk Onderzoek”（FWO，佛兰德斯，比利时）和“Wetenschappelijk Fonds Willy Gepts”（WFWG，UZ 布鲁塞尔）。作者声明不存在利益冲突。
Is implantation failure following ART associated with a perturbed decidual response in endometrial stromal cells (EnSCs)?Dynamic changes in the secretome of decidualizing EnSCs underpin the transition of a hostile to a supportive endometrial microenvironment for embryo implantation; perturbation in this transitional pathway prior to ART is associated with implantation failure.Implantation is the rate-limiting step in ART, although the contribution of an aberrant endometrial microenvironment in IVF failure remains ill defined.In vitro characterization of the temporal changes in the decidual response of primary EnSCs isolated prior to a successful or failed ART cycle. An analysis of embryo responses to secreted cues from undifferentiated and decidualizing EnSCs was performed. The primary clinical outcome of the study was a positive urinary pregnancy test 14 days after embryo transfer.Primary EnSCs were isolated from endometrial biopsies obtained prior to IVF treatment and cryopreserved. EnSCs from 10 pregnant and 10 non-pregnant patients were then thawed, expanded in culture, subjected to clonogenic assays, and decidualized for either 2 or 8 days. Transcript levels of decidual marker gene [prolactin (PRL), insulin-like growth factor binding protein 1 (IGFBP1) and 11 beta-hydroxysteroid dehydrogenase (HSD11B1)] were analysed using real-time quantitative PCR and temporal secretome changes of 45 cytokines, chemokines and growth factors were measured by multiplex suspension bead immunoassay. The impact of the EnSC secretome on human blastocyst development was scored morphologically; and embryo secretions in response to EnSC cues analyzed by multiplex suspension bead immunoassay.Clonogenicity and induction of decidual marker genes were comparable between EnSC cultures from pregnant and non-pregnant group groups (P > 0.05). Analysis of 23 secreted factors revealed that successful implantation was associated with co-ordinated secretome changes in decidualizing EnSCs, which were most pronounced on Day 2 of differentiation: 17 differentially secreted proteins on Day 2 of decidualization relative to undifferentiated (Day 0) EnSCs (P < 0.05); 11 differentially secreted proteins on Day 8 relative to Day 2 (P < 0.05); and eight differentially secreted proteins on Day 8 relative to Day 0 (P < 0.05). By contrast, failed implantation was associated with a disordered secretome response. Blastocyst development was compromised when cultured for 24 h in medium conditioned by undifferentiated EnSCs when compared to decidualizing EnSCs. Analysis of the embryo microdroplets revealed that human blastocysts mount a secretory cytokine response to soluble decidual factors produced during the early (Day 2) but not late phase (Day 8) of differentiation. The embryo responses to secreted factors from decidualizing EnSCs were comparable between the pregnant and non-pregnant group (P > 0.05).Not applicable.Although this study uses primary EnSCs and human embryos, caution is warranted when extrapolating the results to the in vivo situation because of the correlative nature of the study and limited sample size.Our finding raises the prospect that endometrial analysis prior to ART could minimize the risk of treatment failure.This work was supported by funds from the Biomedical Research Unit in Reproductive Health, a joint initiative of the University Hospitals Coventry & Warwickshire NHS Trust and Warwick Medical School, the University of Nottingham and Nurture Fertility, and the National Medical Research Council, Singapore (NMRC/BNIG14NOV023), the "Instituut voor Innovatie door Wetenschap en Technologie" (IWT, Flanders, Belgium), the "Fonds voor Wetenschappelijk Onderzoek" (FWO, Flanders, Belgium) and the "Wetenschappelijk Fonds Willy Gepts" (WFWG, UZ Brussel). The authors have declared that no conflict of interest exists.