To identify the molecular changes in signalling pathways triggered by altered apoE4 expression, we used lentiviral transduction to overexpress human apoE4 in 18-21 DIV primary hippocampal neurons. The molecular consequences of altered apoE4 expression in these neurons were the mapped by proteomic analysis. In silico investigations highlighted changes in proteins associated with mitochondrial dysfunction, sirtuin signalling and oxidative phosphorylation. Interestingly, by combining the data generated here with proteomic data from human patient AD APOE4 “spared” (BA17) and “affected” (BA41/42) synaptic isolates it is possible to identify physiological correlates in synaptic signalling pathways such as oxidative phosphorylation, sirtuin signalling, glutamate receptor signalling and calcium signalling. Thus, in combination with patient derived post mortem analyses, this viral apoE4 overexpression neuronal culture system could assist with delineation of apoE4 triggered molecular perturbations from chronic neurodegenerative processes present in end stage AD brain regions.

As explained above (using our standard workflows) the mass spec data output are provided in the files below –
The BiolayoutclustersforApoE4.xlsx files contains the Biolayout clusters identified by expression profile from the cell culture proteomic dataset.
The ApoE4Comparisonmolecules.xlsx file contains tabs of a number of signalling cascades containing proteins exhibiting log ratio fold changes between transduced cells and human tissue samples and include:
Sample details – including TMT tag information
LC-MS data filtered by number of unique peptides (2 or more only)

Data Citation

Wishart, Thomas. (2020). lentiviral mediated apoE4 transduction in primary neuronal culture, [dataset]. University of Edinburgh. Roslin Institute. https://doi.org/10.7488/ds/2919
Date made available1 Mar 2021
PublisherEdinburgh DataShare

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