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Low Focal Adhesion Signaling Promotes Ground State Pluripotency of Mouse Embryonic Stem Cells
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-09-13 00:00:00 , DOI: 10.1021/acs.jproteome.7b00322 Sara Taleahmad , Mehdi Mirzaei , Azam Samadian , Seyedeh-Nafiseh Hassani , Paul A. Haynes , Ghasem Hosseini Salekdeh 1 , Hossein Baharvand 2
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-09-13 00:00:00 , DOI: 10.1021/acs.jproteome.7b00322 Sara Taleahmad , Mehdi Mirzaei , Azam Samadian , Seyedeh-Nafiseh Hassani , Paul A. Haynes , Ghasem Hosseini Salekdeh 1 , Hossein Baharvand 2
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Mouse embryonic stem cells (mESCs) can be maintained in a pluripotent state when cultured with 2 inhibitors (2i) of extracellular signal-regulated kinase (MEK) and glycogen synthase kinase-3 (GSK3), and Royan 2 inhibitors (R2i) of FGF4 and TGFβ. The molecular mechanisms that control ESC self-renewal and pluripotency are more important for translating stem cell technologies to clinical applications. In this study, we used the shotgun proteomics technique to compare the proteome of the ground state condition (R2i- and 2i-grown cells) to that of serum. Out of 1749 proteins identified, 171 proteins were differentially expressed (p < 0.05) in the 2i, R2i, and serum samples. Gene ontology (GO) analysis of differentially abundant proteins showed that the focal adhesion signaling pathway significantly down-regulated under ground state conditions. mESCs had highly adhesive attachment under the serum condition, whereas in the 2i and R2i culture conditions, a loss of adhesion was observed and the cells were rounded and grew in compact colonies on gelatin. Quantitative RT-PCR showed reduced expression of the integrins family in the 2i and R2i conditions. The serum culture had more prominent phosphorylation of focal adhesion kinase (FAK) compared to 2i and R2i cultures. Activity of the extracellular signal-regulated kinase (ERK)1/2 decreased in the 2i and R2i cultures compared to serum. Activation of integrins by Mn2+ in the 2i and R2i cultures resulted in reduced Nanog and increased the expression of lineage marker genes. In this study, we demonstrated that reduced focal adhesion enabled mESCs to be maintained in an undifferentiated and pluripotent state.
中文翻译：
低局灶性粘附信号促进小鼠胚胎干细胞的基态全能。
与2种细胞外信号调节激酶（MEK）和糖原合酶激酶-3（GSK3）的抑制剂（2i）和FGF4的Royan 2抑制剂（R2i）培养时，小鼠胚胎干细胞（mESCs）可以保持多能状态。和TGFβ。控制ESC自我更新和多能性的分子机制对于将干细胞技术转化为临床应用更为重要。在这项研究中，我们使用the弹枪蛋白质组学技术将基态条件（R2i和2i生长的细胞）的蛋白质组与血清的蛋白质组进行了比较。在鉴定出的1749种蛋白质中，有171种蛋白质被差异表达（p<0.05）在2i，R2i和血清样本中。基因本体论（GO）分析差异丰富的蛋白质表明，在基态条件下，黏着斑信号通路显着下调。mESCs在血清条件下具有高度粘附性，而在2i和R2i培养条件下，观察到粘附力丧失，细胞被圆化并在明胶上的紧凑菌落中生长。定量RT-PCR显示在2i和R2i条件下整合素家族的表达降低。与2i和R2i培养相比，血清培养物的粘着斑激酶（FAK）磷酸化程度更高。与血清相比，2i和R2i培养物中细胞外信号调节激酶（ERK）1/2的活性降低。Mn 2+激活整合素在2i和R2i培养中，Nanog减少，谱系标记基因的表达增加。在这项研究中，我们证明减少的粘着斑使mESCs保持在未分化和多能的状态。
更新日期：2017-09-14
中文翻译：
低局灶性粘附信号促进小鼠胚胎干细胞的基态全能。
与2种细胞外信号调节激酶（MEK）和糖原合酶激酶-3（GSK3）的抑制剂（2i）和FGF4的Royan 2抑制剂（R2i）培养时，小鼠胚胎干细胞（mESCs）可以保持多能状态。和TGFβ。控制ESC自我更新和多能性的分子机制对于将干细胞技术转化为临床应用更为重要。在这项研究中，我们使用the弹枪蛋白质组学技术将基态条件（R2i和2i生长的细胞）的蛋白质组与血清的蛋白质组进行了比较。在鉴定出的1749种蛋白质中，有171种蛋白质被差异表达（p<0.05）在2i，R2i和血清样本中。基因本体论（GO）分析差异丰富的蛋白质表明，在基态条件下，黏着斑信号通路显着下调。mESCs在血清条件下具有高度粘附性，而在2i和R2i培养条件下，观察到粘附力丧失，细胞被圆化并在明胶上的紧凑菌落中生长。定量RT-PCR显示在2i和R2i条件下整合素家族的表达降低。与2i和R2i培养相比，血清培养物的粘着斑激酶（FAK）磷酸化程度更高。与血清相比，2i和R2i培养物中细胞外信号调节激酶（ERK）1/2的活性降低。Mn 2+激活整合素在2i和R2i培养中，Nanog减少，谱系标记基因的表达增加。在这项研究中，我们证明减少的粘着斑使mESCs保持在未分化和多能的状态。
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