Bouvier Sylvie, Paulmyer-Lacroix Odile, Molinari Nicolas, Bertaud Alexandrine, Paci Marine, Leroyer Aurélie, Robert Stéphane, Dignat George Françoise, Blot-Chabaud Marcel, Bardin Nathalie
Aix Marseille Univ, Inserm U1076, Marseille, France.
Assisted Reproductive Center, Laboratory of Reproduction, CHU La Conception, AP-HM, Marseille and Laboratory of Histology-Embryology/Biology of Reproduction, Aix-Marseille University, Marseille, France.
PLoS One. 2017 Mar 14;12(3):e0173724. doi: 10.1371/journal.pone.0173724. eCollection 2017.
Although progress was made in in vitro fertilization (IVF) techniques, the majority of embryos transferred fail to implant. Morphology embryo scoring is the standard procedure for most of IVF centres for choosing the best embryo, but remains limited since even the embryos classified as "top quality" may not implant. As it has been shown that i) CD146 is involved in embryo implantation and ii) membrane form is shed to generate soluble CD146 (sCD146), we propose that sCD146 in embryo supernatants may constitute a new biomarker of embryo selection. Immunocytochemical staining showed expression of CD146 in early embryo stages and sCD146 was detected by ELISA and Western-blot in embryo supernatants from D2. We retrospectively studied 126 couples who underwent IVF attempt. The embryo culture medium from each transferred embryo (n = 222) was collected for measurement of sCD146 by ELISA. Significantly higher sCD146 concentrations were present in embryo supernatants that did not implant (n = 185) as compared to those that successfully implanted (n = 37) (1310 +/- 1152 pg.mL-1 vs. 845+/- 1173 pg.mL-1, p = 0.024). Sensitivity analysis performed on single embryo transfers (n = 71) confirmed this association (p = 0.0054). The computed ROC curve established that the optimal sCD146 concentration for embryo implantation is under 1164 pg.mL-1 (sensitivity: 76%, specificity: 48%, PPV: 25% and NPV: 92%). Over this sCD146 threshold, the implantation rate was significantly lower (9% with sCD146 levels >1164 pg.ml-1 vs. 22% with sCD146 levels ≤ 1164 pg.mL-1, p = 0.01). Among the embryos preselected by morphologic scoring, sCD146 determination could allow a better selection of the embryo(s), thus improving the success of elective single embryo transfer. This study establishes the proof of concept for the use of sCD146 as a biomarker for IVF by excluding the embryo with the highest sCD146 level. A multicentre prospective study will now be necessary to further establish its use in clinical practice.
尽管体外受精(IVF)技术取得了进展,但大多数移植的胚胎未能着床。形态学胚胎评分是大多数IVF中心选择最佳胚胎的标准程序,但仍然存在局限性,因为即使被归类为“优质”的胚胎也可能无法着床。由于已经表明:i)CD146参与胚胎着床;ii)膜形式的CD146脱落会产生可溶性CD146(sCD146),我们提出胚胎上清液中的sCD146可能构成一种新的胚胎选择生物标志物。免疫细胞化学染色显示CD146在早期胚胎阶段表达,并且通过ELISA和蛋白质印迹法在第2天的胚胎上清液中检测到sCD146。我们回顾性研究了126对接受IVF尝试的夫妇。收集每个移植胚胎(n = 222)的胚胎培养基,通过ELISA测量sCD146。与成功着床的胚胎(n = 37)相比,未着床的胚胎(n = 185)的胚胎上清液中sCD146浓度显著更高(1310±1152 pg/mL vs. 845±1173 pg/mL,p = 0.024)。对单胚胎移植(n = 71)进行的敏感性分析证实了这种关联(p = 0.0054)。计算得出的ROC曲线确定,胚胎着床的最佳sCD146浓度低于1164 pg/mL(敏感性:76%,特异性:48%,阳性预测值:25%,阴性预测值:92%)。超过这个sCD146阈值,着床率显著降低(sCD146水平>1164 pg/mL时为9%,而sCD146水平≤1164 pg/mL时为22%,p = 0.01)。在通过形态学评分预选的胚胎中,sCD146的测定可以更好地选择胚胎,从而提高选择性单胚胎移植的成功率。本研究通过排除sCD146水平最高的胚胎,确立了将sCD146用作IVF生物标志物的概念验证。现在有必要进行一项多中心前瞻性研究,以进一步确定其在临床实践中的应用。