Scalable Production of iPSC-Derived Human Neurons to Identify Tau-Lowering Compounds by High-Content Screening.
Scalable Production of iPSC-Derived Human Neurons to Identify Tau-Lowering Compounds by High-Content Screening.
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DOI：
10.1016/j.stemcr.2017.08.019
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发表时间：
2017-10-10
影响因子：
5.9
通讯作者：
Gan L
中科院分区：
文献类型：
--
作者：
Wang C;Ward ME;Chen R;Liu K;Tracy TE;Chen X;Xie M;Sohn PD;Ludwig C;Meyer-Franke A;Karch CM;Ding S;Gan L
降低总tau水平是阿尔茨海默病和其他tau蛋白病的有吸引力的治疗策略。来源于人诱导多能干细胞（iPSC）的神经元的高通量筛选是鉴定tau靶向治疗剂的有力工具。然而，这种筛选受到异质性神经元生产、高成本和低产量以及多步分化程序的阻碍。我们设计了一个等基因iPSC系，该系含有可诱导的neurogenin 2转基因，这是一种快速将iPSC转化为神经元的转录因子，整合在AAVS1基因座上。使用简化的两步方案，我们将这些iPSC分化为具有最小孔间变异性的皮质神经元。我们开发了一种稳健的高含量筛选试验，以鉴定LOPAC中的tau降低化合物，并将肾上腺素能受体激动剂鉴定为一类降低内源性人tau的化合物。这些技术使人类神经元能够用于高通量筛选治疗神经退行性疾病的药物。可诱导的，同基因的，一种简单且可扩展的两步方案确保了最小的分化变异性。LOPAC AR激动剂中的降低化合物被鉴定为一类降低人tau的化合物Gan及其同事开发了一种简单且可扩展的技术，通过工程化产生大量同质的神经元能皮质神经元将神经生成素2表达盒连接到iPSC的AAVS1基因座。他们开发了一种高含量的筛选试验，并将肾上腺素能受体激动剂鉴定为一类降低内源性人类tau蛋白的化合物，这是阿尔茨海默病的关键致病因素。
Lowering total tau levels is an attractive therapeutic strategy for Alzheimer's disease and other tauopathies. High-throughput screening in neurons derived from human induced pluripotent stem cells (iPSCs) is a powerful tool to identify tau-targeted therapeutics. However, such screens have been hampered by heterogeneous neuronal production, high cost and low yield, and multi-step differentiation procedures. We engineered an isogenic iPSC line that harbors an inducible neurogenin 2 transgene, a transcription factor that rapidly converts iPSCs to neurons, integrated at the AAVS1 locus. Using a simplified two-step protocol, we differentiated these iPSCs into cortical glutamatergic neurons with minimal well-to-well variability. We developed a robust high-content screening assay to identify tau-lowering compounds in LOPAC and identified adrenergic receptors agonists as a class of compounds that reduce endogenous human tau. These techniques enable the use of human neurons for high-throughput screening of drugs to treat neurodegenerative disease. Inducible, isogenic, and integrated Ngn2 iPSCs were differentiated to pure neurons A simple and scalable two-step protocol ensures minimal differentiation variability An HCS assay was established to screen for tau-lowering compounds in LOPAC AR agonists were identified as a class of compounds that reduce human tau Gan and colleagues developed a simple and scalable technology to generate a large quantity of homogeneous glutamatergic cortical neurons by engineering a neurogenin 2-expressing cassette to the AAVS1 locus of iPSCs. They developed a high-content screening assay and identified adrenergic receptor agonists as a class of compounds that lower endogenous human tau, a key pathogenic factor in Alzheimer's disease.
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影响因子：
5.3
作者：
DeVos, Sarah L.;Goncharoff, Dustin K.;Miller, Timothy M.
通讯作者：
Miller, Timothy M.
DOI：
10.2174/1386207043328256
发表时间：
2004-12-01
影响因子：
1.8
作者：
Albrecht, H;Zbinden, P;Imbimbo, BP
通讯作者：
Imbimbo, BP
影响因子：
3.4
作者：
Harrill, Joshua A.;Robinette, Brian L.;Mundy, William R.
通讯作者：
Mundy, William R.
影响因子：
3.5
作者：
Baker, M;Litvan, I;Hutton, M
通讯作者：
Hutton, M
影响因子：
5.9
作者：
Chanda, Soham;Ang, Cheen Euong;Davila, Jonathan;Pak, ChangHui;Mall, Moritz;Lee, Qian Yi;Ahlenius, Henrik;Jung, Seung Woo;Suedhof, Thomas C.;Wernig, Marius
通讯作者：
Wernig, Marius