Cardioinductive network guiding stem cell differentiation revealed by proteomic cartography of tumor necrosis factor α-primed endodermal secretome
Cardioinductive network guiding stem cell differentiation revealed by proteomic cartography of tumor necrosis factor α-primed endodermal secretome
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DOI：
10.1634/stemcells.2007-0599
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发表时间：
2008-02-01
期刊：
影响因子：
5.2
通讯作者：
Terzic, Andre
中科院分区：
文献类型：
--
作者：
Arrell, D. Kent;Niederlaender, Nicolas J.;Terzic, Andre
在发育中的胚胎中，来自腹侧内胚层的指导性指导确保了前外侧中胚层内的心脏程序诱导。然而，内胚层引导的心脏发生尚未在蛋白质组水平上得到解决。在这里，通过用重编程细胞因子肿瘤坏死因子α（TNF α）对内胚层进行心脏生成性启动，通过比较蛋白质组学描绘了胚胎干细胞心脏分化的候选效应子。差异二维凝胶电泳图谱显示，在TNF α引发的内胚层分泌组中，超过75%的蛋白质种类增加> 1.5倍。通过线性离子阱四极杆（LTQ）串联质谱（MS/MS）进行蛋白质斑点鉴定，并在多维色谱后通过鸟枪LTQ-傅立叶变换MS/MS进行验证，从153个斑点分配中绘制了99个独特的蛋白质。一个确定的一组48分泌组蛋白推导出迭代生物信息学筛选使用算法检测分泌的典型和非典型指数。蛋白质-蛋白质相互作用分析，结合各自的表达水平的变化，揭示了一个非随机的TNF α为中心的分泌组网络与无标度的层次结构。心血管发育是分解的TNF α锚定网络的主要发育功能。通过在胚胎干细胞上直接应用TNF α引发的分泌组，增强心脏定型和肌节生成，验证了衍生的心脏诱导网络的功能协同性。相反，抑制初级网络枢纽否定了TNF α引发的前心原性作用。因此，蛋白质组制图建立了一个系统生物学框架内胚层分泌网络指导干细胞心脏。
In the developing embryo, instructive guidance from the ventral endoderm secures cardiac program induction within the anterolateral mesoderm. Endoderm-guided cardiogenesis, however, has yet to be resolved at the proteome level. Here, through cardiopoietic priming of the endoderm with the reprogramming cytokine tumor necrosis factor alpha (TNF alpha), candidate effectors of embryonic stem cell cardiac differentiation were delineated by comparative proteomics. Differential two-dimensional gel electrophoretic mapping revealed that more than 75% of protein species increased > 1.5-fold in the TNF alpha-primed versus unprimed endodermal secretome. Protein spot identification by linear ion trap quadrupole (LTQ) tandem mass spectrometry (MS/MS) and validation by shotgun LTQ-Fourier transform MS/MS following multidimensional chromatography mapped 99 unique proteins from 153 spot assignments. A definitive set of 48 secretome proteins was deduced by iterative bioinformatic screening using algorithms for detection of canonical and noncanonical indices of secretion. Protein-protein interaction analysis, in conjunction with respective expression level changes, revealed a nonstochastic TNF alpha-centric secretome network with a scale-free hierarchical architecture. Cardiovascular development was the primary developmental function of the resolved TNF alpha-anchored network. Functional cooperativity of the derived cardioinductive network was validated through direct application of the TNF alpha-primed secretome on embryonic stem cells, potentiating cardiac commitment and sarcomerogenesis. Conversely, inhibition of primary network hubs negated the procardiogenic effects of TNF alpha priming. Thus, proteomic cartography establishes a systems biology framework for the endodermal secretome network guiding stem cell cardiopoiesis.