Analysis of alternative signaling pathways of endoderm induction of human embryonic stem cells identifies context specific differences.
Analysis of alternative signaling pathways of endoderm induction of human embryonic stem cells identifies context specific differences.
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人胚胎干细胞内胚层诱导的替代信号通路分析确定了环境特异性差异
DOI：
10.1186/1752-0509-6-154
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发表时间：
2012-12-15
影响因子：
--
通讯作者：
Banerjee I
中科院分区：
文献类型：
--
作者：
Mathew S;Jaramillo M;Zhang X;Zhang LA;Soto-Gutiérrez A;Banerjee I
背景人类胚胎干细胞（human embryonic stem cells，hESCs）的谱系特异性分化主要由特异性生长因子和细胞外基质分子介导。生长因子启动控制基因转录和细胞命运特化的信号级联。人们对诱导hESC进入内胚层命运非常感兴趣，内胚层命运作为通向更多功能细胞类型（如胰腺细胞）的途径。过去十年的研究已经建立了几种从hESC获得内胚层的强大途径，具有进一步成熟的能力。然而，根据我们的经验，这些内胚层衍生物的功能成熟，特别是胰腺谱系，很大程度上取决于内胚层诱导的特定途径。因此，了解介导这种诱导的潜在机制以及如何将其转化为进一步成熟将是很有趣的。在这项工作中，我们分析了调节相互作用介导的不同途径的内胚层induction. ResultsshESCs诱导使用激活素A和4种不同的生长因子（FGF 2（F），BMP 4（B），PI 3 KI（P），WNT 3A（W））及其组合，导致在15个总的实验条件。在分化结束时，通过qRT-PCR分析每种条件下的12种相关内胚层相关转录因子（TF）。作为第一种方法，我们使用层次聚类来确定哪些生长因子组合有利于不同基因的上调。在下一步中，我们使用双聚类算法鉴定了共调节转录因子的集合。实验数据的高度可变性通过将双聚类公式与自举重新采样相结合来确定共调节转录因子的鲁棒网络来解决。我们的结果表明，在高活化素下，FGF 2以及WNT 3A处理有利于从早期内胚层向晚期内胚层的转变。然而，晚期内胚层标记物的诱导是相对有利的WNT 3A下highactivin.ConclusionsUse的FGF 2，WNT 3A或PI 3 K抑制高激活素A可能服务以及在定形内胚层诱导，然后通过WNT 3A特异性信号直接定形内胚层到晚期内胚层谱系。其他组合，虽然对于内胚层诱导仍然可行，但在我们的实验中对于胰腺内胚层特化似乎不太有希望。
BackgroundLineage specific differentiation of human embryonic stem cells (hESCs) is largely mediated by specific growth factors and extracellular matrix molecules. Growth factors initiate a cascade of signals which control gene transcription and cell fate specification. There is a lot of interest in inducing hESCs to an endoderm fate which serves as a pathway towards more functional cell types like the pancreatic cells. Research over the past decade has established several robust pathways for deriving endoderm from hESCs, with the capability of further maturation. However, in our experience, the functional maturity of these endoderm derivatives, specifically to pancreatic lineage, largely depends on specific pathway of endoderm induction. Hence it will be of interest to understand the underlying mechanism mediating such induction and how it is translated to further maturation. In this work we analyze the regulatory interactions mediating different pathways of endoderm induction by identifying co-regulated transcription factors.ResultshESCs were induced towards endoderm using activin A and 4 different growth factors (FGF2 (F), BMP4 (B), PI3KI (P), and WNT3A (W)) and their combinations thereof, resulting in15total experimental conditions. At the end of differentiation each condition was analyzed by qRT-PCR for12relevant endoderm related transcription factors (TFs). As a first approach, we used hierarchical clustering to identify which growth factor combinations favor up-regulation of different genes. In the next step we identified sets of co-regulated transcription factors using a biclustering algorithm. The high variability of experimental data was addressed by integrating the biclustering formulation with bootstrap re-sampling to identify robust networks of co-regulated transcription factors. Our results show that the transition from early to late endoderm is favored by FGF2 as well as WNT3A treatments under high activin. However, induction of late endoderm markers is relatively favored by WNT3A under high activin.ConclusionsUse of FGF2, WNT3A or PI3K inhibition with high activin A may serve well in definitive endoderm induction followed by WNT3A specific signaling to direct the definitive endoderm into late endodermal lineages. Other combinations, though still feasible for endoderm induction, appear less promising for pancreatic endoderm specification in our experiments.
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影响因子：
5.2
作者：
McLean, Amanda B.;D'Amour, Kevin A.;Dalton, Stephen
通讯作者：
Dalton, Stephen
影响因子：
7.7
作者：
Mfopou JK;Chen B;Sui L;Sermon K;Bouwens L
通讯作者：
Bouwens L
影响因子：
4.6
作者：
Poulain, Morgane;Furthauer, Maximilian;Lepage, Thierry
通讯作者：
Lepage, Thierry
影响因子：
4.6
作者：
Nostro, M. Cristina;Sarangi, Farida;Keller, Gordon
通讯作者：
Keller, Gordon
DOI：
10.1109/tcbb.2004.2
发表时间：
2004-01-01
影响因子：
4.5
作者：
Madeira, SC;Oliveira, AL
通讯作者：
Oliveira, AL