Quretec Ltd. Tartu, Estonia ; Faculty of Mathematics and Computer Science, Institute of Computer Science, University of Tartu Tartu, Estonia ; Faculty of Science and Technology, Institute of Molecular and Cellular Biology, University of Tartu Tartu, Estonia.
Front Physiol. 2013 Oct 28;4:303. doi: 10.3389/fphys.2013.00303. eCollection 2013.
Pluripotency in human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) is regulated by three transcription factors-OCT3/4, SOX2, and NANOG. To fully exploit the therapeutic potential of these cells it is essential to have a good mechanistic understanding of the maintenance of self-renewal and pluripotency. In this study, we demonstrate a powerful systems biology approach in which we first expand literature-based network encompassing the core regulators of pluripotency by assessing the behavior of genes targeted by perturbation experiments. We focused our attention on highly regulated genes encoding cell surface and secreted proteins as these can be more easily manipulated by the use of inhibitors or recombinant proteins. Qualitative modeling based on combining boolean networks and in silico perturbation experiments were employed to identify novel pluripotency-regulating genes. We validated Interleukin-11 (IL-11) and demonstrate that this cytokine is a novel pluripotency-associated factor capable of supporting self-renewal in the absence of exogenously added bFGF in culture. To date, the various protocols for hESCs maintenance require supplementation with bFGF to activate the Activin/Nodal branch of the TGFβ signaling pathway. Additional evidence supporting our findings is that IL-11 belongs to the same protein family as LIF, which is known to be necessary for maintaining pluripotency in mouse but not in human ESCs. These cytokines operate through the same gp130 receptor which interacts with Janus kinases. Our finding might explain why mESCs are in a more naïve cell state compared to hESCs and how to convert primed hESCs back to the naïve state. Taken together, our integrative modeling approach has identified novel genes as putative candidates to be incorporated into the expansion of the current gene regulatory network responsible for inducing and maintaining pluripotency.
人胚胎干细胞(hESCs)和诱导多能干细胞(iPSCs)的多能性由三个转录因子-OCT3/4、SOX2 和 NANOG 调控。为了充分利用这些细胞的治疗潜力,必须深入了解维持自我更新和多能性的机制。在这项研究中,我们展示了一种强大的系统生物学方法,首先通过评估受扰动实验靶向基因的行为,扩展包含多能性核心调控因子的文献基础网络。我们将注意力集中在高度调控的基因上,这些基因编码细胞表面和分泌蛋白,因为这些蛋白可以更容易地通过使用抑制剂或重组蛋白进行操作。基于布尔网络和计算机扰动实验的定性建模被用于识别新的多能性调控基因。我们验证了白细胞介素 11(IL-11),并证明这种细胞因子是一种新的多能性相关因子,能够在没有外源性添加 bFGF 的情况下支持自我更新。迄今为止,各种 hESCs 维持方案都需要补充 bFGF 以激活 TGFβ 信号通路中的激活素/ nodal 分支。支持我们发现的额外证据是,IL-11 属于与 LIF 相同的蛋白家族,已知 LIF 对于维持小鼠多能性是必需的,但对于人类 ESCs 则不是。这些细胞因子通过相同的 gp130 受体发挥作用,该受体与 Janus 激酶相互作用。我们的发现可以解释为什么 mESCs 比 hESCs 处于更原始的细胞状态,以及如何将已分化的 hESCs 转化回原始状态。总之,我们的整合建模方法已经确定了新的基因作为潜在的候选基因,以纳入负责诱导和维持多能性的当前基因调控网络的扩展。