Department of Pathology, University of Washington, Seattle, Washington.
Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, Washington.
J Neuropathol Exp Neurol. 2018 May 1;77(5):353-360. doi: 10.1093/jnen/nly013.
Patient-specific stem cell technology from skin and other biopsy sources has transformed in vitro models of neurodegenerative disease, permitting interrogation of the effects of complex human genetics on neurotoxicity. However, the neuropathologic changes that underlie cognitive and behavioral phenotypes can only be determined at autopsy. To better correlate the biology of derived neurons with age-related and neurodegenerative changes, we generated leptomeningeal cell lines from well-characterized research subjects that have undergone comprehensive postmortem neuropathologic examinations. In a series of proof of principle experiments, we reprogrammed autopsy leptomeningeal cell lines to human-induced pluripotent stem cells (hiPSCs) and differentiated these into neurons. We show that leptomeningeal-derived hiPSC lines can be generated from fresh and frozen leptomeninges, are pluripotent, and retain the karyotype of the starting cell population. Additionally, neurons differentiated from these hiPSCs are functional and produce measurable Alzheimer disease-relevant analytes (Aβ and Tau). Finally, we used direct conversion protocols to transdifferentiate leptomeningeal cells to neurons. These resources allow the generation of in vitro models to test mechanistic hypotheses as well as diagnostic and therapeutic strategies in association with neuropathology, clinical and cognitive data, and biomarker studies, aiding in the study of late-onset Alzheimer disease and other age-related neurodegenerative diseases.
从皮肤和其他活检来源的患者特异性干细胞技术已经改变了神经退行性疾病的体外模型，允许研究复杂的人类遗传学对神经毒性的影响。然而，只有在尸检时才能确定导致认知和行为表型的神经病理学变化。为了更好地将衍生神经元的生物学与年龄相关和神经退行性变化相关联，我们从经过全面尸检神经病理学检查的特征明确的研究对象中生成软脑膜细胞系。在一系列原理验证实验中，我们将尸检软脑膜细胞系重新编程为人类诱导多能干细胞（hiPSC），并将这些细胞分化为神经元。我们表明，软脑膜衍生的 hiPSC 系可以从新鲜和冷冻的软脑膜中生成，具有多能性，并保留起始细胞群体的核型。此外，这些 hiPSC 分化而来的神经元具有功能并且可以产生可测量的阿尔茨海默病相关分析物（Aβ和 Tau）。最后，我们使用直接转化方案将软脑膜细胞转化为神经元。这些资源允许生成体外模型，以与神经病理学、临床和认知数据以及生物标志物研究一起测试机制假设以及诊断和治疗策略，从而有助于研究迟发性阿尔茨海默病和其他与年龄相关的神经退行性疾病。
