BMP and TGF-β pathway mediators are critical upstream regulators of Wnt signaling during midbrain dopamine differentiation in human pluripotent stem cells.
BMP and TGF-β pathway mediators are critical upstream regulators of Wnt signaling during midbrain dopamine differentiation in human pluripotent stem cells.
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DOI：
10.1016/j.ydbio.2013.01.012
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发表时间：
2013-04-01
影响因子：
2.7
通讯作者：
Iacovitti, Lorraine
中科院分区：
文献类型：
--
作者：
Cai, Jingli;Schleidt, Stephanie;Pelta-Heller, Joshua;Hutchings, Danielle;Cannarsa, Gregory;Iacovitti, Lorraine
尽管许多实验室目前在方案中使用BMP（Dorsomorphin/DM）和TGF-β（SB 431542/SB）信号通路的小分子抑制剂来从hES和hiPS细胞产生中脑多巴胺（mDA）神经元，但直到现在，这些物质还没有被认为在mDA分化过程中发挥作用。我们在这里报告，组成型BMP（pSMADs 1，5，8）信号的瞬时抑制，无论是单独或与TGF-β抑制（pSMADs 2，3）的组合，是至关重要的在mDA祖细胞的Wnt 1-Lmx 1a信号的上游调控。我们推测DM或DM/SB介导这些效应的机制涉及SMAD相互作用蛋白1（SIP 1）的上调，这导致干细胞中Wnt拮抗剂分泌型卷曲相关蛋白1（Sfrp 1）的更大抑制。因此，SIP 1敲低逆转DM/SB对mDA分化的诱导作用，而Sfrp 1敲低/抑制模拟DM/SB。然而，SMAD抑制培养物中Wnt 1-Lmx 1a水平的升高伴随着SHH-Foxa 2水平的相互下调，导致产生少量共表达Foxa 2的TH+神经元。然而，如果外源性SHH/FGF 8与SMAD抑制剂一起沿着添加，则这两个重要途径将达到平衡，从而促进真正的（Lmx 1a + Foxa 2 +TH+）mDA神经元分化，同时在干细胞培养物中抑制替代细胞命运。这些数据表明，BMP和TGF-β信号传导的激活剂/抑制剂在人多能干细胞中的mDA分化过程中发挥关键的上游调节作用。
Although many laboratories currently use small molecule inhibitors of the BMP (Dorsomorphin/DM) and TGF-β (SB431542/SB) signaling pathways in protocols to generate midbrain dopamine (mDA) neurons from hES and hiPS cells, until now, these substances have not been thought to play a role in the mDA differentiation process. We report here that the transient inhibition of constitutive BMP (pSMADs 1, 5, 8) signaling, either alone or in combination with TGF-β inhibition (pSMADs 2, 3), is critically important in the upstream regulation of Wnt1-Lmx1a signaling in mDA progenitors. We postulate that the mechanism via which DM or DM/SB mediates these effects involves the up-regulation in SMAD-interacting protein 1 (SIP1), which results in greater repression of the Wnt antagonist, secreted frizzled related protein 1 (Sfrp1) in stem cells. Accordingly, knockdown of SIP1 reverses the inductive effects of DM/SB on mDA differentiation while Sfrp1 knockdown/inhibition mimics DM/SB. The rise in Wnt1-Lmx1a levels in SMAD-inhibited cultures is, however, accompanied by a reciprocal down-regulation in SHH-Foxa2 levels leading to the generation of few TH+ neurons that co-express Foxa2. If however, exogenous SHH/FGF8 is added along with SMAD inhibitors, equilibrium in these two important pathways is achieved such that authentic (Lmx1a+Foxa2+TH+) mDA neuron differentiation is promoted while alternate cell fates are suppressed in stem cell cultures. These data indicate that activators/inhibitors of BMP and TGF-β signaling play a critical upstream regulatory role in the mDA differentiation process in human pluripotent stem cells.
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DOI：
10.1073/pnas.0902396106
发表时间：
2009-05-05
影响因子：
11.1
作者：
Friling, Stina;Andersson, Elisabet;Ericson, Johan
通讯作者：
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影响因子：
4
作者：
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发表时间：
2011-04-01
影响因子：
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作者：
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通讯作者：
Knaus, Petra
影响因子：
2.9
作者：
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通讯作者：
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影响因子：
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作者：
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