Decreased PECAM1-mediated TGF-β1 expression in the mid-secretory endometrium in women with recurrent implantation failure
Decreased PECAM1-mediated TGF-β1 expression in the mid-secretory endometrium in women with recurrent implantation failure
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反复着床失败的女性中分泌期子宫内膜中 PECAM1 介导的 TGF-β 1 表达降低
DOI：
10.1093/humrep/dey022
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发表时间：
2018-05-01
影响因子：
6.1
通讯作者：
Zhang, Aijun
中科院分区：
文献类型：
--
作者：
Guo, Feng;Si, Chenchen;Zhang, Aijun
研究问题：复发性着床失败 (RIF) 是否与着床窗口期子宫内膜中血小板和内皮细胞粘附分子 I (PECAM1) 和转化生长因子 β 1 (TGF-β 1) 表达降低有关？总结答案：本研究表明，患有 RIF 的女性，分泌中期子宫内膜中 PECAM1 和 TGF-β 1 的表达显着降低，这可能是胚胎植入的原因失败。已知信息：RIF 已成为阻碍 IVF-胚胎移植 (IVF-ET) 妊娠率提高的瓶颈问题。 RIF 的病因很复杂，可能涉及多种生长因子、代谢物和炎症细胞因子的失调。目前，RIF的确切发病机制尚未阐明。 研究设计、规模、持续时间：这是一项前瞻性病例对照研究。子宫内膜组织样本取自2014年1月至2016年12月接受IVF的两组女性（RIF组，22名接受≥3次ET的女性，其中总共≥4个优质胚胎，但未妊娠；对照组，18名在第一个治疗周期受孕的女性）。同时，从 18 名因输卵管因素继发不孕的妇女中采集了月经周期早期增殖期、晚期增殖期和分泌中期期的样本（每组 n = 6）。用于分离原代人子宫内膜上皮细胞和基质细胞（HEEC 和 HESC）的样本是于 2017 年 12 月从 6 名继发于输卵管因素的不孕女性身上收集的。 参与者/材料、背景、方法：我们使用综合全基因组表达微阵列分析来研究基因表达，包括差异表达基因筛选、主成分分析和功能富集分析。 RTqPCR、蛋白质印迹、免疫组织化学、免疫荧光共定位分析和短发夹 RNA (shRNA) 质粒转染 Ishikawa 细胞系、HEEC 和 HESC 用于研究 PECAM I 和 TGF-β 1 的表达。 主要结果和机会的作用：全基因组表达谱的综合数据挖掘确定细胞粘附是 RIF 的关键调节因子。通过数据库检索和文献综述，筛选出一些可能参与胚胎着床的新型细胞粘附相关基因，包括PECAM I、细胞间粘附分子2（ICAM2）、整合素亚基β2（ITGB2）、选择素P（SELP）和TEK受体酪氨酸激酶（TEK）。在这些靶标中，RIF组中PECAM1的mRNA和蛋白水平显着低于对照组。继发性不孕女性月经周期中,PECAM1蛋白表达量在增殖早期最低,在增殖晚期略有升高,在分泌中期表达最高。而子宫内膜容受性标志物HOXA10的表达水平在增殖早期保持低水平，在增殖后期升高，在分泌中期保持高水平。此外，RIF 组中由 PECAM1 介导的 TGF-β 1 也显着降低。使用基于 shRNA 的方法，我们证明 PECAM1 的缺失显着降低了 Ishikawa 细胞以及原代 HEEC 和 HESC 中 TGF-β 1 的表达。这些结果表明，PECAM1 和 TGF-β 1 可能在调节子宫内膜容受性中发挥关键作用。 注意的局限性：虽然我们已经表明，PECAM1 和 TGF-β 1 在患有 RIF 的女性中下调，但这些因素对子宫内膜容受性影响的分子机制仍不清楚。 研究结果的更广泛意义：我们的研究结果提供了对 PECAM1 和 TGF-β 1 的贡献的深入了解。 TGF-β1 调节着床，可用于开发 RIF 的潜在治疗方法。
STUDY QUESTION: Is recurrent implantation failure (RIF) associated with decreased expression of platelet and endothelial cell adhesion molecule I (PECAM1) and transforming growth factor beta 1 (TGF-beta 1) in the endometrium during the implantation window?SUMMARY ANSWER: The present study demonstrates that the expression of PECAM1 and TGF-beta 1 is significantly decreased in the mid-secretory endometrium in women with RIF, which may account for embryo implantation failure.WHAT IS KNOWN ALREADY: RIF has become a bottleneck issue that hampers the improvement of pregnancy rates in IVF-embryo transfer (IVF-ET). The causes of RIF are complex and may involve the dysregulation of various growth factors, metabolites, and inflammatory cytokines. At present, the precise pathogenesis of RIF has not been elucidated.STUDY DESIGN, SIZE, DURATION: This was a prospective case-control study. Endometrial tissue samples were obtained from January 2014 to December 2016 from two groups of women who had undergone IVF (RIF group, 22 women who underwent >= 3 ETs including a total of >= 4 good-quality embryos without pregnancy, control group, 18 women who conceived in their first treatment cycle). At the same time, samples were obtained from 18 women with infertility secondary to tubal factor in the early proliferative, late proliferative and mid-secretory phases of the menstrual cycle (n = 6 per group). Samples used for isolation of primary human endometrial epithelial cells and stromal cells (HEECs and HESCs) were collected in December 2017 from six women with infertility secondary to tubal factor.PARTICIPANTS/MATERIALS, SETTING, METHODS: We investigated gene expression using integrative whole genome expression microarray analysis, including differentially expressed gene screening, principal component analysis, and functional enrichment analysis. RTqPCR, western blotting, immunohistochemistry, immunofluorescence co-localization analysis and short hairpin RNA (shRNA) plasmid transfection in Ishikawa cell line, HEECs and HESCs were used to investigate the expression of PECAM I and TGF-beta 1.MAIN RESULTS AND THE ROLE OF CHANCE: Integrative data mining of whole-genome expression profiles identified cell adhesion as a key regulator in RIF. Database retrieval and literature review screened several novel cell adhesion-related genes that might participate in embryo implantation, which include PECAM I, intercellular adhesion molecule 2 (ICAM2), integrin subunit beta 2 (ITGB2), selectin P (SELP) and TEK receptor tyrosine kinase (TEK). Among these targets, the mRNA and protein levels of PECAM1 were significantly lower in the RIF group than those in the control group. During the menstrual cycles of women with secondary infertility, the protein expression level of PECAM1 was the lowest in early proliferative phase, slighdy increased in late proliferative phase and was the highest in mid-secretory phase. While the expression level of HOXA10, an endometrial receptivity marker, kept at a low level in early proliferative phase and increased in late proliferative phase, then maintained at a high level in the mid-secretory phase. Furthermore, TGF-beta 1, mediated by PECAM1, was also decreased significantly in the RIF group. Using shRNA-based approach, we demonstrated that the depletion of PECAM1 significantly decreased the expression of TGF-beta 1 in Ishikawa cells, as well as in primary HEECs and HESCs. These results indicated that PECAM1 and TGF-beta 1 might play a pivotal role in modulating endometrial receptivity.LIMITATIONS REASONS FOR CAUTION: Although we have shown that PECAM1 and TGF-beta 1 were down-regulated in the women with RIF, the molecular mechanism of the effect of the factors on the endometrial receptivity remain unclear.WIDER IMPLICATIONS OF THE FINDINGS: Our findings provide insight into the contribution of PECAM1 and TGF-beta 1 in regulating implantation, which could be used to develop potential therapeutic methods for RIF.