Endogenous recapitulation of Alzheimer's disease neuropathology through human 3D direct neuronal reprogramming.
Endogenous recapitulation of Alzheimer's disease neuropathology through human 3D direct neuronal reprogramming.
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通过人类 3D 直接神经元重编程内源性重现阿尔茨海默病神经病理学。
DOI：
10.1101/2023.05.24.542155
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Mennerick,Stev
中科院分区：
文献类型：
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作者：
Sun,Zhao;Kwon,Ji-Sun;Ren,Yudong;Chen,Shawei;Cates,Kitra;Lu,Xinguo;Walker,CourtneyK;Karahan,Hande;Sviben,Sanja;Fitzpatrick,JamesAJ;Valdez,Clarissa;Houlden,Henry;Karch,CelesteM;Bateman,RandallJ;Sato,Chihiro;Mennerick,Stev
阿尔茨海默病（AD）是一种主要影响老年个体的神经退行性疾病，其特征在于标志性神经元病理学，包括细胞外淀粉样蛋白-β（Aβ）斑块沉积、细胞内tau缠结和神经元死亡。然而，在患者源性神经元中重现这些年龄相关的神经元病理仍然是一个重大挑战，特别是对于迟发性AD（LOAD），最常见的疾病形式。在这里，我们应用高效率microRNA介导的直接神经元重编程的成纤维细胞从AD患者产生皮质神经元在三维（3D）基质胶和自组装的神经元球体。我们的研究结果表明，从常染色体显性AD（ADAD）和LOAD患者中重编程的神经元和球状体表现出与神经元相关的AD样表型，包括细胞外Aβ沉积、营养不良性神经突过度磷酸化、K63-泛素阳性、种子活性tau蛋白和培养中的自发神经元死亡。此外，在Aβ存款形成之前，在LOAD患者源性神经元和球状体中使用β-或γ-分泌酶抑制剂治疗显著降低了Aβ沉积以及tau蛋白病和神经变性。然而，在细胞已经形成Aβ沉积物后进行相同的处理仅具有轻微的影响。此外，通过用逆转录酶抑制剂拉米夫定治疗LOAD神经元和球状体来抑制年龄相关的逆转录转座因子（RTEs）的合成，减轻了AD神经病理学。总体而言，我们的结果表明，AD患者成纤维细胞在3D环境中的直接神经元重编程可以捕获年龄相关的神经病理学，并反映Aβ积累，tau失调和神经元死亡之间的相互作用。此外，基于miRNA的3D神经元转换提供了一种人类相关的AD模型，可用于鉴定可潜在改善AD相关病理和神经变性的化合物。
Alzheimer’s disease (AD) is a neurodegenerative disorder that primarily affects elderly individuals, and is characterized by hallmark neuronal pathologies including extracellular amyloid-β (Aβ) plaque deposition, intracellular tau tangles, and neuronal death. However, recapitulating these age-associated neuronal pathologies in patient-derived neurons has remained a significant challenge, especially for late-onset AD (LOAD), the most common form of the disorder. Here, we applied the high efficiency microRNA-mediated direct neuronal reprogramming of fibroblasts from AD patients to generate cortical neurons in three-dimensional (3D) Matrigel and self-assembled neuronal spheroids. Our findings indicate that neurons and spheroids reprogrammed from both autosomal dominant AD (ADAD) and LOAD patients exhibited AD-like phenotypes linked to neurons, including extracellular Aβ deposition, dystrophic neurites with hyperphosphorylated, K63-ubiquitin-positive, seed-competent tau, and spontaneous neuronal death in culture. Moreover, treatment with β- or γ-secretase inhibitors in LOAD patient-derived neurons and spheroids before Aβ deposit formation significantly lowered Aβ deposition, as well as tauopathy and neurodegeneration. However, the same treatment after the cells already formed Aβ deposits only had a mild effect. Additionally, inhibiting the synthesis of age-associated retrotransposable elements (RTEs) by treating LOAD neurons and spheroids with the reverse transcriptase inhibitor, lamivudine, alleviated AD neuropathology. Overall, our results demonstrate that direct neuronal reprogramming of AD patient fibroblasts in a 3D environment can capture age-related neuropathology and reflect the interplay between Aβ accumulation, tau dysregulation, and neuronal death. Moreover, miRNA-based 3D neuronal conversion provides a human-relevant AD model that can be used to identify compounds that can potentially ameliorate AD-associated pathologies and neurodegeneration.