Vlismas Antonis, Bletsa Ritsa, Mavrogianni Despina, Mamali Georgina, Pergamali Maria, Dinopoulou Vasiliki, Partsinevelos George, Drakakis Peter, Loutradis Dimitris, Kiessling Ann A
1 1st Obstetrics and Gynecology Department of University of Athens, "Alexandra" Maternity Hospital , Athens, Greece .
2 Bedford Research Foundation , Bedford, Massachusetts.
Stem Cells Dev. 2016 Jan 15;25(2):160-77. doi: 10.1089/scd.2015.0284. Epub 2015 Dec 1.
Previous microarray analyses of RNAs from 8-cell (8C) human embryos revealed a lack of cell cycle checkpoints and overexpression of core circadian oscillators and cell cycle drivers relative to pluripotent human stem cells [human embryonic stem cells/induced pluripotent stem (hES/iPS)] and fibroblasts, suggesting growth factor independence during early cleavage stages. To explore this possibility, we queried our combined microarray database for expression of 487 growth factors and receptors. Fifty-one gene elements were overdetected on the 8C arrays relative to hES/iPS cells, including 14 detected at least 80-fold higher, which annotated to multiple pathways: six cytokine family (CSF1R, IL2RG, IL3RA, IL4, IL17B, IL23R), four transforming growth factor beta (TGFB) family (BMP6, BMP15, GDF9, ENG), one fibroblast growth factor (FGF) family [FGF14(FH4)], one epidermal growth factor member (GAB1), plus CD36, and CLEC10A. 8C-specific gene elements were enriched (73%) for reported circadian-controlled genes in mouse tissues. High-level detection of CSF1R, ENG, IL23R, and IL3RA specifically on the 8C arrays suggests the embryo plays an active role in blocking immune rejection and is poised for trophectoderm development; robust detection of NRG1, GAB1, -2, GRB7, and FGF14(FHF4) indicates novel roles in early development in addition to their known roles in later development. Forty-four gene elements were underdetected on the 8C arrays, including 11 at least 80-fold under the pluripotent cells: two cytokines (IFITM1, TNFRSF8), five TGFBs (BMP7, LEFTY1, LEFTY2, TDGF1, TDGF3), two FGFs (FGF2, FGF receptor 1), plus ING5, and WNT6. The microarray detection patterns suggest that hES/iPS cells exhibit suppressed circadian competence, underexpression of early differentiation markers, and more robust expression of generic pluripotency genes, in keeping with an artificial state of continual uncommitted cell division. In contrast, gene expression patterns of the 8C embryo suggest that it is an independent circadian rhythm-competent equivalence group poised to signal its environment, defend against maternal immune rejection, and begin the rapid commitment events of early embryogenesis.
先前对8细胞(8C)期人类胚胎RNA的微阵列分析显示,相对于多能人类干细胞[人类胚胎干细胞/诱导多能干细胞(hES/iPS)]和成纤维细胞,8C期胚胎缺乏细胞周期检查点,核心昼夜节律振荡器和细胞周期驱动因子过表达,这表明在早期卵裂阶段胚胎生长因子非依赖性。为了探究这种可能性,我们在综合微阵列数据库中查询了487种生长因子和受体的表达情况。相对于hES/iPS细胞,8C期胚胎微阵列上有51个基因元件检测值过高,其中14个检测值至少高出80倍,这些基因注释到多个信号通路:六个细胞因子家族(CSF1R、IL2RG、IL3RA、IL4、IL17B、IL23R),四个转化生长因子β(TGFB)家族(BMP6、BMP15、GDF9、ENG),一个成纤维细胞生长因子(FGF)家族[FGF14(FH4)],一个表皮生长因子成员(GAB1),加上CD36和CLEC10A。8C期胚胎特异性基因元件在小鼠组织中报道的昼夜节律控制基因中富集(73%)。8C期胚胎微阵列上CSF1R、ENG、IL23R和IL3RA的高水平检测表明,胚胎在阻断免疫排斥中发挥积极作用,并为滋养外胚层发育做好准备;NRG1、GAB1、-2、GRB7和FGF14(FHF4)的强检测信号表明,它们在早期发育中除了具有已知的后期发育作用外,还具有新的作用。8C期胚胎微阵列上有44个基因元件检测值过低,其中11个相对于多能细胞检测值至少低80倍:两个细胞因子(IFITM1、TNFRSF8),五个TGFB(BMP7、LEFTY1、LEFTY2、TDGF1、TDGF3),两个FGF(FGF2、FGF受体1),加上ING5和WNT6。微阵列检测模式表明,hES/iPS细胞表现出昼夜节律能力受抑制、早期分化标志物表达不足以及通用多能性基因表达更强,这与持续未分化细胞分裂的人工状态一致。相比之下,8C期胚胎的基因表达模式表明,它是一个具有独立昼夜节律能力的等价组,准备向其环境发出信号、抵御母体免疫排斥,并开始早期胚胎发育的快速定向分化事件。