Early role for IL-6 signalling during generation of induced pluripotent stem cells revealed by heterokaryon RNA-Seq.
Early role for IL-6 signalling during generation of induced pluripotent stem cells revealed by heterokaryon RNA-Seq.
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DOI：
10.1038/ncb2835
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发表时间：
2013-10
影响因子：
21.3
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中科院分区：
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对体细胞重新编程以诱导多能干细胞(IPS)的分子洞察将有助于再生医学，但由于其异质性，很难在iPS中阐明，因为经历重新编程的细胞相对较少(0.1-1%；参考文献，)。为了识别早期起作用的调控因子，我们利用了非分裂异核体(小鼠胚胎干细胞与人成纤维细胞融合)，在这种异核体中，向多能性方向重新编程是有效和快速的，使识别一开始所需的瞬时调控因子成为可能。我们使用双物种转录组范围的RNA-SEQ来量化人类体细胞核在重编程过程中的转录变化，以实现异核体的多能性。在异核体重编程过程中，在胚胎干细胞中检测不到的细胞因子白介素6(IL6)被诱导了50倍。在iPS重编程开始时，4天的培养用IL6取代了稳定转导的致癌c-Myc，因此只需要转导Oct4、Klf4和Sox2。IL6还激活了另一个JAK/STAT靶点，丝氨酸/苏氨酸激酶基因Pim1，这是IL6介导的iPS频率增加两倍的原因。相反，另一种被诱导的gp130配体Lif未能增加iPS频率或激活c-MycorPim1，从而揭示了两个Jak/Stat诱导子在iPS产生中的不同作用。这些发现证明了异核双物种全局RNA-seq在识别重编程的早期作用调节因子方面的能力，例如，稳定转导的转录因子的外在替换，如强大的癌基因c-Myc。
Molecular insights into somatic cell reprogramming to induced pluripotent stem cells (iPS) would aid regenerative medicine, but are difficult to elucidate in iPS because of their heterogeneity, as relatively few cells undergo reprogramming (0.1–1%; refs , ). To identify early acting regulators, we capitalized on non-dividing heterokaryons (mouse embryonic stem cells fused to human fibroblasts), in which reprogramming towards pluripotency is efficient and rapid, enabling the identification of transient regulators required at the onset. We used bi-species transcriptome-wide RNA-seq to quantify transcriptional changes in the human somatic nucleus during reprogramming towards pluripotency in heterokaryons. During heterokaryon reprogramming, the cytokine interleukin 6 (IL6), which is not detectable at significant levels in embryonic stem cells, was induced 50-fold. A 4-day culture with IL6 at the onset of iPS reprogramming replaced stably transduced oncogenic c-Myc such that transduction of only Oct4, Klf4 and Sox2 was required. IL6 also activated another Jak/Stat target, the serine/threonine kinase genePim1, which accounted for the IL6-mediated twofold increase in iPS frequency. In contrast, LIF, another induced GP130 ligand, failed to increase iPS frequency or activatec-MycorPim1, thereby revealing a differential role for the two Jak/Stat inducers in iPS generation. These findings demonstrate the power of heterokaryon bi-species global RNA-seq to identify early acting regulators of reprogramming, for example, extrinsic replacements for stably transduced transcription factors such as the potent oncogene c-Myc.