High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy.
High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy.
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DOI：
10.1038/s41598-021-96227-5
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发表时间：
2021-08-23
影响因子：
4.6
通讯作者：
Haggarty SJ
中科院分区：
文献类型：
--
作者：
Cheng C;Reis SA;Adams ET;Fass DM;Angus SP;Stuhlmiller TJ;Richardson J;Olafson H;Wang ET;Patnaik D;Beauchamp RL;Feldman DA;Silva MC;Sur M;Johnson GL;Ramesh V;Miller BL;Temple S;Kosik KS;Dickerson BC;Haggarty SJ
MAPT（微管相关蛋白tau）突变导致额颞叶痴呆（FTD）。MAPT突变与异常的tau磷酸化水平和错误折叠的tau蛋白的积累相关，所述错误折叠的tau蛋白可以在神经元之间传播，最终导致细胞死亡（tau蛋白病）。最近，p.A152T tau变体被确定为FTD、阿尔茨海默病和突触核蛋白病的风险因素。在这里，我们使用来自携带这种p.A152T变体的患者的诱导多能干细胞（iPSC）来创建一个强大的功能性细胞测定系统，用于探测病理生理学tau积累和磷酸化。使用稳定转导的iPSC衍生的神经祖细胞，其被工程化以使前神经转录因子Neurogenin 2（Ngn 2）的诱导型表达成为可能，我们以可扩展的、高通量筛选兼容的形式产生了疾病相关的皮质样神经元。利用自动共聚焦显微镜，和先进的图像处理管道优化形态复杂的人类神经元培养物的分析，我们报告定量，亚细胞定位特定的影响，多种激酶抑制剂对tau蛋白，包括临床研究中没有以前报道的影响tau蛋白磷酸化。这些结果证明了使用Tau病变的患者iPSC衍生的离体模型作为遗传上准确的疾病相关系统来探测Tau生物化学并支持发现Tau病变的新型治疗剂的潜力。
Mutations in MAPT (microtubule-associated protein tau) cause frontotemporal dementia (FTD). MAPT mutations are associated with abnormal tau phosphorylation levels and accumulation of misfolded tau protein that can propagate between neurons ultimately leading to cell death (tauopathy). Recently, a p.A152T tau variant was identified as a risk factor for FTD, Alzheimer's disease, and synucleinopathies. Here we used induced pluripotent stem cells (iPSC) from a patient carrying this p.A152T variant to create a robust, functional cellular assay system for probing pathophysiological tau accumulation and phosphorylation. Using stably transduced iPSC-derived neural progenitor cells engineered to enable inducible expression of the pro-neural transcription factor Neurogenin 2 (Ngn2), we generated disease-relevant, cortical-like glutamatergic neurons in a scalable, high-throughput screening compatible format. Utilizing automated confocal microscopy, and an advanced image-processing pipeline optimized for analysis of morphologically complex human neuronal cultures, we report quantitative, subcellular localization-specific effects of multiple kinase inhibitors on tau, including ones under clinical investigation not previously reported to affect tau phosphorylation. These results demonstrate the potential for using patient iPSC-derived ex vivo models of tauopathy as genetically accurate, disease-relevant systems to probe tau biochemistry and support the discovery of novel therapeutics for tauopathies.
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影响因子：
11.2
作者：
Brighton HE;Angus SP;Bo T;Roques J;Tagliatela AC;Darr DB;Karagoz K;Sciaky N;Gatza ML;Sharpless NE;Johnson GL;Bear JE
通讯作者：
Bear JE
影响因子：
8.8
作者：
Almeida S;Zhang Z;Coppola G;Mao W;Futai K;Karydas A;Geschwind MD;Tartaglia MC;Gao F;Gianni D;Sena-Esteves M;Geschwind DH;Miller BL;Farese RV Jr;Gao FB
通讯作者：
Gao FB
影响因子：
--
作者：
Cheng, Chialin;Fass, Daniel M;Haggarty, Stephen J
通讯作者：
Haggarty, Stephen J
影响因子：
2.7
作者：
Chartier M;Chénard T;Barker J;Najmanovich R
通讯作者：
Najmanovich R
影响因子：
12.7
作者：
Guo T;Noble W;Hanger DP
通讯作者：
Hanger DP