BMP/SMAD Pathway Promotes Neurogenesis of Midbrain Dopaminergic Neurons In Vivo and in Human Induced Pluripotent and Neural Stem Cells
BMP/SMAD Pathway Promotes Neurogenesis of Midbrain Dopaminergic Neurons In Vivo and in Human Induced Pluripotent and Neural Stem Cells
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DOI：
10.1523/jneurosci.1540-17.2018
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发表时间：
2018-02-14
影响因子：
5.3
通讯作者：
Brodski, Claude
中科院分区：
文献类型：
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作者：
Jovanovic, Vukasin M.;Salti, Ahmad;Brodski, Claude
中脑多巴胺能（mDA）神经元在体内的胚胎形成为从干细胞体外分化mDA神经元提供了关键的指导，所述干细胞目前正在开发用于帕金森病细胞替代疗法。骨形态发生蛋白（BMP）/SMAD抑制通常用于干细胞分化方案的早期步骤，包括用于mDA神经元的产生。然而，BMP/SMAD通路在哺乳动物mDA神经元的体内规格的功能几乎是未知的。在这里，我们报告说，BMP 5/7缺陷小鼠（Bmp 5（-/-）; Bmp 7（-/-））缺乏mDA神经元，由于减少在mDA祖结构域的神经发生。作为解释Bmp 5（-/-）; Bmp 7（-/-）突变体中这些改变的分子机制，我们已经鉴定了BMP/SMAD靶基因MSX 1/2（msh同源框1/2）和SHH（音刺猬）的表达变化。小鼠体内神经干细胞（Smad 1（内斯））中SMAD 1的连接蛋白失活通过阻止细胞周期退出而阻碍祖细胞向mDA神经元的分化，尤其是阻止TH（+）SOX 6（+）（酪氨酸羟化酶，SRY-框6）和TH（+）GIRK 2（+）（钾电压门控通道亚家族-J成员-6）黑质神经元的分化。BMP 5/7在体外将人诱导多能干细胞和诱导神经干细胞向mDA神经元的分化增强了高达三倍。总之，我们已经确定了BMP/SMAD信号作为一种新的关键途径，协调哺乳动物体内mDA神经发生的重要步骤，平衡祖细胞增殖和分化。此外，我们证明了骨形态发生蛋白的潜力，以提高干细胞衍生的mDA神经元在体外的生成，强调顺序BMP/SMAD抑制和激活在这个过程中的重要性。
The embryonic formation of midbrain dopaminergic (mDA) neurons in vivo provides critical guidelines for the in vitro differentiation of mDA neurons from stem cells, which are currently being developed for Parkinson's disease cell replacement therapy. Bone morphogenetic protein (BMP)/SMAD inhibition is routinely used during early steps of stem cell differentiation protocols, including for the generation of mDA neurons. However, the function of the BMP/SMAD pathway for in vivo specification of mammalian mDA neurons is virtually unknown. Here, we report that BMP5/7-deficient mice (Bmp5(-/-); Bmp7(-/-)) lack mDA neurons due to reduced neurogenesis in the mDA progenitor domain. As molecular mechanisms accounting for these alterations in Bmp5(-/-); Bmp7(-/-) mutants, we have identified expression changes of the BMP/SMAD target genes MSX1/2 (msh homeobox 1/2) and SHH (sonic hedgehog). Conditionally inactivating SMAD1 in neural stem cells of mice in vivo (Smad1(Nes)) hampered the differentiation of progenitor cells into mDA neurons by preventing cell cycle exit, especially of TH(+)SOX6(+) (tyrosine hydroxylase, SRY-box 6) and TH(+)GIRK2(+) (potassium voltage-gated channel subfamily-J member-6) substantia nigra neurons. BMP5/7 robustly increased the in vitro differentiation of human induced pluripotent stem cells and induced neural stem cells to mDA neurons by up to threefold. In conclusion, we have identified BMP/SMAD signaling as a novel critical pathway orchestrating essential steps of mammalian mDA neurogenesis in vivo that balances progenitor proliferation and differentiation. Moreover, we demonstrate the potential of BMPs to improve the generation of stem-cell-derived mDA neurons in vitro, highlighting the importance of sequential BMP/SMAD inhibition and activation in this process.