Modeling Sporadic Progressive Supranuclear Palsy in 3D Midbrain Organoids: Recapitulating Disease Features for In Vitro Diagnosis and Drug Discovery

IF 8.1 1区医学 Q1 CLINICAL NEUROLOGY

Annals of Neurology Pub Date : 2025-01-28 DOI:10.1002/ana.27172

Elvira Immacolata Parrotta PhD, Valeria Lucchino PhD, Clara Zannino PhD, Desirèe Valente PhD, Stefania Scalise PhD, Davide Bressan PhD, Giorgia Lucia Benedetto PhD, Maria Roberta Iazzetta PhD, Mariagrazia Talarico PhD, Monica Gagliardi PhD, Francesco Conforti PhD, Silvia Di Agostino PhD, Alessandro Fiorenzano PhD, Aldo Quattrone MD, Giovanni Cuda MD, Andrea Quattrone MD, PhD

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引用次数: 0

Abstract

Objective

Progressive Supranuclear Palsy (PSP) is a severe neurodegenerative disease characterized by tangles of hyperphosphorylated tau protein and tufted astrocytes. Developing treatments for PSP is challenging due to the lack of disease models reproducing its key pathological features. This study aimed to model sporadic PSP-Richardson's syndrome (PSP-RS) using multi-donor midbrain organoids (MOs).

Methods

The MOs were generated by pooling induced pluripotent stem cells (iPSCs) from 4 patients with sporadic probable PSP-RS and compared them with MOs from 3 healthy control (HC) subjects. We performed comprehensive analyses of MOs over 120 days to assess neuronal death, reactive gliosis, and the accumulation of 4R-tau and hyperphosphorylated tau forms (pThr231, pSer396, pThr181, and pSer202/pThr205 [AT8]) using immunofluorescence microscopy and Western blot. On day 90, immunohistochemical analysis using pSer396 and AT8 antibodies was conducted to assess disease pathology.

Results

PSP-derived MOs showed progressive size reduction compared with HC-derived MOs, linked to upregulated apoptosis-related mRNA markers. Dopaminergic neuron degeneration was marked by decreased tyrosine hydroxylase (TH) and increased neurofilament light chain (NfL). Immunofluorescence and Western blot revealed accumulation of all investigated tau forms with a peak at 90 days, along with a significant rise in GFAP-positive cells in PSP-derived MOs. Immunochemistry confirmed typical PSP histological alterations, such as neurofibrillary tangles and tufted-shaped astrocytes, absent in HC-derived organoids.

Interpretation

We developed a robust in vitro PSP model reproducing the key molecular and histologic features of the disease. This result holds promise for advancing basic and clinical research in PSP, paving the way for in vitro molecular diagnosis and identification of novel therapeutic targets. ANN NEUROL 2025;97:845–859

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在三维中脑类器官中模拟散发性进行性核上性麻痹：概括疾病特征用于体外诊断和药物发现。

目的：进行性核上性麻痹（PSP）是一种严重的神经退行性疾病，其特征是过度磷酸化的tau蛋白缠结和簇状星形胶质细胞。由于缺乏重现其关键病理特征的疾病模型，开发治疗PSP的方法具有挑战性。本研究旨在利用多供体中脑类器官（MOs）建立散发性psp -理查德森综合征（PSP-RS）模型。方法：收集4例散发性疑似PSP-RS患者的诱导多能干细胞（iPSCs），并与3例健康对照（HC）患者的诱导多能干细胞进行比较。我们使用免疫荧光显微镜和Western blot技术对120天内的MOs进行了全面分析，以评估神经元死亡、反应性胶质瘤以及4R-tau和过度磷酸化tau形式（pThr231、pSer396、pThr181和pSer202/pThr205 [AT8]）的积累。第90天，使用pSer396和AT8抗体进行免疫组织化学分析以评估疾病病理。结果：与hc衍生的MOs相比，psp衍生的MOs显示出进行性大小减小，这与凋亡相关mRNA标记上调有关。多巴胺能神经元变性表现为酪氨酸羟化酶（TH）降低，神经丝轻链（NfL）增加。免疫荧光和Western blot显示，所有研究的tau形式的积累在第90天达到峰值，同时在psp衍生的MOs中gfap阳性细胞显著增加。免疫化学证实了典型的PSP组织学改变，如神经原纤维缠结和簇状星形胶质细胞，在hc来源的类器官中不存在。解释：我们开发了一个强大的体外PSP模型，再现了该疾病的关键分子和组织学特征。这一结果有望推进PSP的基础和临床研究，为体外分子诊断和新治疗靶点的鉴定铺平道路。Ann neurol 2025。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Annals of Neurology 医学-临床神经学

CiteScore

18.00

自引率

1.80%

发文量

270

审稿时长

3-8 weeks

期刊介绍： Annals of Neurology publishes original articles with potential for high impact in understanding the pathogenesis, clinical and laboratory features, diagnosis, treatment, outcomes and science underlying diseases of the human nervous system. Articles should ideally be of broad interest to the academic neurological community rather than solely to subspecialists in a particular field. Studies involving experimental model system, including those in cell and organ cultures and animals, of direct translational relevance to the understanding of neurological disease are also encouraged.