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The functional relevance of BIN1: implications for endocytosis in Alzheimer’s disease

Burt, Anna 2017. The functional relevance of BIN1: implications for endocytosis in Alzheimer’s disease. PhD Thesis, Cardiff University.
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Abstract

Alzheimer’s disease is a complex neurodegenerative disorder, characterised by progressive neurological and cognitive impairment. Genome-wide association studies have shown the significant association of several endocytosis-related genes with Alzheimer’s disease, including BIN1. With multiple tissue-specific isoforms, BIN1 functions are wide-ranging and its involvement in mediating Alzheimer’s disease risk is not fully understood. The initial aim of this thesis was to develop a neuronal cell model from induced pluripotent stem cells (iPSCs) derived from peripheral blood mononuclear cells (PBMCs) for the study of endocytosis in Alzheimer’s disease. The development of PBMC-derived iPSC models is a novel field and this thesis demonstrates that PBMC-derived iPSCs can be differentiated into cortical neurons through dual SMAD inhibition. This protocol was carried out for the first time in the current lab and has therefore established these iPSC protocols for future use. Secondly, this thesis aimed to further develop knowledge on the role of BIN1 in clathrinmediated endocytosis (CME) in a non-neuronal context. The hCMEC/D3 human brain microvascular endothelial cell line was used as an in vitro model of the blood-brain barrier(BBB) to investigate the effect of siRNA-mediated BIN1 depletion. Evidence is presented suggesting BIN1 is involved in the amyloidogenic processing of APP by an increase in β-CTF upon BIN1 depletion, in the absence of changes in levels of APP, Aβ and soluble APP or putative α- β- and γ- secretases. This was not suggested to be due to altered CME as transferrin uptake and recycling remained unaffected. While caveolin-2 levels increased with BIN1 depletion, lactosylceramide internalisation, a measure of caveolar endocytosis, was not affected, suggesting BIN1 was not crucial for this mechanism in hCMEC/D3 cells. Impaired degradative clearance of β-CTF was also hypothesised, however an absence of gross changes in lysosome morphology suggested that BIN1 was not central to normal lysosome function. This thesis provides novel insight into the role of BIN1 in the amyloidogenic processing of APP and endocytosis at the BBB, furthering understanding on the functional relevance of AD risk mediated by BIN1.

Item Type: Thesis (PhD)
Status: Unpublished
Schools: Medicine
Date of First Compliant Deposit: 1 March 2018
Last Modified: 19 Oct 2019 02:39
URI: http://orca.cf.ac.uk/id/eprint/109392

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