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

• Published in: Annals of neurology Vol. 97; no. 5; pp. 845 - 859
• Main Authors: Parrotta, Elvira Immacolata, Lucchino, Valeria, Zannino, Clara, Valente, Desirèe, Scalise, Stefania, Bressan, Davide, Benedetto, Giorgia Lucia, Iazzetta, Maria Roberta, Talarico, Mariagrazia, Gagliardi, Monica, Conforti, Francesco, Di Agostino, Silvia, Fiorenzano, Alessandro, Quattrone, Aldo, Cuda, Giovanni, Quattrone, Andrea
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.05.2025; Wiley Subscription Services, Inc
• Subjects: Accumulation; Aged; Apoptosis; Astrocytes; Basic Medicine; Degeneration; Diagnosis; Dopamine receptors; Drug Discovery - methods; Female; Glial fibrillary acidic protein; Gliosis; Humans; Immunofluorescence; Induced Pluripotent Stem Cells - pathology; Inhibitory postsynaptic potentials; Male; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Mesencephalon; Mesencephalon - metabolism; Mesencephalon - pathology; Middle Aged; mRNA; Neurodegeneration; Neurodegenerative diseases; Neurofibrillary tangles; Neurosciences; Neurovetenskaper; Organoids; Organoids - metabolism; Organoids - pathology; Pluripotency; Progressive supranuclear palsy; Stem cells; Supranuclear Palsy, Progressive - diagnosis; Supranuclear Palsy, Progressive - drug therapy; Supranuclear Palsy, Progressive - metabolism; Supranuclear Palsy, Progressive - pathology; Tau protein; tau Proteins - metabolism; Therapeutic targets; Tyrosine; Tyrosine 3-monooxygenase
• ISSN: 0364-5134; 1531-8249; 1531-8249
• DOI: 10.1002/ana.27172

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