Hebisch, Matthias Alexander Sebastian: Establishment of an inducible human pluripotent stem cell-based 3D model of Alzheimer's disease. - Bonn, 2020. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
Online-Ausgabe in bonndoc: https://nbn-resolving.org/urn:nbn:de:hbz:5-60401
urn: https://nbn-resolving.org/urn:nbn:de:hbz:5-60401,
author = {{Matthias Alexander Sebastian Hebisch}},
title = {Establishment of an inducible human pluripotent stem cell-based 3D model of Alzheimer's disease},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2020,
month = nov,

note = {Alzheimer’s disease (AD) is the most prevalent neurodegenerative disease in the aged population. The vast majority of cases are sporadic, however, various mutations may cause early-onset, familial AD (FAD) variants. AD is characterized by deposition of amyloid β (Aβ) plaques in the extracellular space and hyperphosphorylated tau (p-tau) within neurons. This project aimed to implement a human stem cell-based model in a 3D setting in order to more authentically model extracellular Aβ accumulation and its downstream pathogenic effects. To maximize production of aggregation-prone Aβ42, the safe-harbor locus AAVS1 of human induced pluripotent stem cells (iPSCs) was modified to conditionally overexpress FAD mutations in the genes encoding the amyloid precursor protein (APPSwe/Lon) and Presenilin-1 (PS-1ΔE9). Transgenic iPSCs were differentiated into long-term self-renewing neuroepithelial stem (lt-NES) cells, and embedded in a 200-300 µm thick Geltrex matrix for long-term differentiation in 3D. Induction with doxycycline caused 40-fold increased Aβ42 secretion in 2D cultures, whereas Aβ was efficiently entrapped in the gel matrix in 3D cultures. After 8 weeks, induced 3D cultures displayed Thioflavin T-positive Aβ deposits that strongly resemble amyloid plaques in size, structure and specific autofluorescence. After 16 weeks, induced 3D cultures contained TBS-insoluble Aβ. P-tau was present in induced 3D cultures from week 6, and fibril-specific tau epitopes could be detected after 16 weeks. As expected, p-tau pathology did not develop in the presence of γ-secretase inhibitors. Cellular energy metabolism was impaired in induced cultures as p-tau-positive neurons displayed hallmarks of mitochondrial fragmentation, alongside a reduction in essential respiratory chain complexes and lowered respiratory capacity. Induced cultures also showed neuronal degeneration compared to uninduced and γ-secretase-inhibited controls. It follows that 3D matrix cultures of lt-NES neurons represent a valid model to study intra- and extracellular AD pathology in an authentic human system.},
url = {http://hdl.handle.net/20.500.11811/8780}

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