Papandreou, Apostolos;
(2020)
Investigation of Disease Mechanisms and Screening for Treatments in Betapropeller Protein-Associated Neurodegeneration (BPAN).
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
INTRODUCTION: BPAN is a monogenic form of neurodegeneration, associated with brain iron accumulation and early-onset parkinsonism. It is caused by mutations in WDR45, an X-linked gene with role in early autophagy. The mechanisms linking autophagy, iron load and neurodegeneration are poorly understood, while there are no effective treatments for BPAN. I aimed to address these issues by developing a patient-derived, induced pluripotent stem cell (iPSC)-based dopaminergic neuronal cell model of the disease. METHODS: 3 paediatric patient-derived iPSC lines, 2 age-matched controls and 2 ‘isogenic controls’ (generated via CRISPR/Cas9-mediated mutation correction in 2 BPAN lines) were differentiated into dopaminergic neurons using a 65-day protocol. High content imaging assays were developed for a drug screen, looking for small molecules that reverse patient specific cellular phenotypes. RESULTS: At Day 65 there is absence of WDR45 protein in patient lines. RNA sequencing reveals differential gene expression in 190 genes when comparing patients to controls. When plated on multi-well plates at low density, BPAN ventral midbrain progenitors (Day 11 of differentiation) exhibit defective autophagy, with fewer LC3 puncta per cell forming compared to controls. This assay was used to perform a drug screen using the FDA-approved Prestwick library (1,280 compounds) and a series of novel autophagy activators. Some compound hits significantly enhance autophagy in all tested lines and show promise for further development. CONCLUSION: I have used a patient-derived dopaminergic model of BPAN for disease modelling and also as a platform for high content imaging-based drug screening. My work contributes towards identifying novel, effective treatments for BPAN, and increases our understanding of the link between autophagy and neurodegeneration.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Investigation of Disease Mechanisms and Screening for Treatments in Betapropeller Protein-Associated Neurodegeneration (BPAN) |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2020. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Neurosciences Dept |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10100057 |
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