Investigating how Late Onset Alzheimer’s Disease risk genes APOE4 and BIN1 impact neuronal function and interactions with microglia

Nowak, Karolina 2025. Investigating how Late Onset Alzheimer’s Disease risk genes APOE4 and BIN1 impact neuronal function and interactions with microglia. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Late onset Alzheimer’s disease (LOAD) is a complex and multifactorial progressive neurodegenerative disease with no disease-modifying therapies currently available. Genetics is one of the major factors influencing the risk of developing LOAD, with more than 75 risk genes been identified from genome wide association studies (GWAS), including APOE4 and BIN1, the two most significantly associated loci, respectively. Within the GERAD cohort, around one-third of people in the AD group have both APOE4 and a risk SNP in BIN1, but research to date has largely focused on individual risk genes in isolation, meaning that combined LOAD risk is still not well understood. BIN1 functions in endocytosis, a cellular pathway for internalizing, trafficking and recycling molecules. APOE is the major central nervous system lipoprotein and a key endocytic cargo, and the APOE4 isoform has been shown to cause endocytic defects including enlargement of neuronal early endosomes. Several other LOAD risk loci are endocytic genes, and maintained endocytosis has been linked with disease resilience, highlighting importance of endocytosis in LOAD. In this project, CRISPR-Cas9 was used to generate novel isogenic iPSC lines with APOE33 or APOE44 and a BIN1 knockout, to model their combined LOAD risk. The iPSCs were differentiated to cortical neurons which were functionally characterized to explore LOAD-associated phenotypes of the endo-lysosomal and autophagy pathways, using fixed and live high-content imaging, immunoblotting and Aβ profiling. Cortical neurons were also co-cultured with microglia differentiated from the APOE/BIN1 iPSCs to investigate how interactions between these two cell types were affected. The data did not show differences in neuronal endocytosis but suggested subtle lysosomal and autophagy effects with trends towards increased autophagy flux in neurons with a BIN1 knockout. However, there were no clear effects from combined presence of APOE44 and BIN1. Co-culture of neurons with microglia also showed an increase in network activity.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Schools > Medicine
Date of First Compliant Deposit:	6 March 2026
Last Modified:	06 Mar 2026 17:12
URI:	https://orca.cardiff.ac.uk/id/eprint/185570

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