Division of Regenerative Medicine, Institute of Cellular and System Medicine, and Center for Neuropsychiatric Research, National Health Research Institutes, Miaoli, Taiwan, Republic of China; Graduate Program of Biotechnology in Medicine, Institute of Biotechnology and Department of Life Science, Institute of Molecular Medicine, and Department of Medical Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China; Department of Psychiatry, Chang Gung Memorial Hospital at Linkou and Chang Gung University, Gueishan, Taoyuan, Taiwan, Republic of China; Department of Pharmacology, Tzu Chi University, Hualien, Taiwan, Republic of China; Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan, Republic of China.
Division of Regenerative Medicine, Institute of Cellular and System Medicine, and Center for Neuropsychiatric Research, National Health Research Institutes, Miaoli, Taiwan, Republic of China; Graduate Program of Biotechnology in Medicine, Institute of Biotechnology and Department of Life Science, Institute of Molecular Medicine, and Department of Medical Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China; Department of Psychiatry, Chang Gung Memorial Hospital at Linkou and Chang Gung University, Gueishan, Taoyuan, Taiwan, Republic of China; Department of Pharmacology, Tzu Chi University, Hualien, Taiwan, Republic of China; Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan, Republic of China
Stem Cells Transl Med. 2014 Jun;3(6):713-22. doi: 10.5966/sctm.2013-0111. Epub 2014 Apr 15.
Recent advances in somatic cell reprogramming have highlighted the plasticity of the somatic epigenome, particularly through demonstrations of direct lineage reprogramming of adult mouse and human fibroblasts to induced pluripotent stem cells (iPSCs) and induced neurons (iNs) under defined conditions. However, human cells appear to be less plastic and have a higher epigenetic hurdle for reprogramming to both iPSCs and iNs. Here, we show that SH2B adaptor protein 1β (SH2B1) can enhance neurite outgrowth of iNs reprogrammed from human fibroblasts as early as day 14, when combined with miR124 and transcription factors BRN2 and MYT1L (IBM) under defined conditions. These SH2B1-enhanced iNs (S-IBM) showed canonical neuronal morphology, and expressed multiple neuronal markers, such as TuJ1, NeuN, and synapsin, and functional proteins for neurotransmitter release, such as GABA, vGluT2, and tyrosine hydroxylase. Importantly, SH2B1 accelerated mature process of functional neurons and exhibited action potentials as early as day 14; without SH2B1, the IBM iNs do not exhibit action potentials until day 21. Our data demonstrate that SH2B1 can enhance neurite outgrowth and accelerate the maturation of human iNs under defined conditions. This approach will facilitate the application of iNs in regenerative medicine and in vitro disease modeling.
体细胞重编程的最新进展强调了体细胞核重编程的可塑性，特别是通过在特定条件下直接将成年小鼠和人成纤维细胞重编程为诱导多能干细胞（iPSCs）和诱导神经元（iNs）的实验，证明了这一点。然而，人类细胞的可塑性似乎较低，并且在重编程为 iPSCs 和 iNs 时，它们的表观遗传障碍更高。在这里，我们表明 SH2B 衔接蛋白 1β（SH2B1）可以增强从人成纤维细胞重编程而来的 iNs 的突起生长，早在第 14 天，当与 miR124 和转录因子 BRN2 和 MYT1L（IBM）在特定条件下组合使用时，就可以增强突起生长。这些 SH2B1 增强的 iNs（S-IBM）表现出典型的神经元形态，并表达多种神经元标记物，如 TuJ1、NeuN 和突触素，以及用于神经递质释放的功能性蛋白质，如 GABA、vGluT2 和酪氨酸羟化酶。重要的是，SH2B1 加速了功能性神经元的成熟过程，早在第 14 天就表现出动作电位；没有 SH2B1，IBM iNs 直到第 21 天才表现出动作电位。我们的数据表明，SH2B1 可以在特定条件下增强突起生长并加速人 iNs 的成熟。这种方法将促进 iNs 在再生医学和体外疾病建模中的应用。
