Implementation of Multi-Electrode Arrays for Functional, Pre-Clinical Modeling of Alzheimer's Disease
Negri, Joseph M.
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CitationNegri, Joseph M. 2019. Implementation of Multi-Electrode Arrays for Functional, Pre-Clinical Modeling of Alzheimer's Disease. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences.
AbstractAlzheimer’s disease (AD) is the most common form of age-related dementia, affecting millions of patients globally. Unfortunately, the development of therapies for AD is lagging with no newly approved treatments in the last 15 years. This dearth of new therapies requires a reconsideration of the pre-clinical models of AD, in order to facilitate the discovery of efficacious treatments. Ideally such models would allow for monitoring the functional behavior of neurons in conditions approximating the AD brain. One potential instrument platform for such models are multi-electrode arrays (MEA), as these systems allow for prolonged, non-destructive recordings of spontaneously firing neurons in vitro. Towards this goal of developing more representative, pre-clinical models of AD; methods of experimental design and analysis for assessing neuronal firing using MEAs were established. This system was used to examine changes in neuronal activity following treatment with soluble human brain extract from a panel of 19 individuals comprised of: AD patients, unaffected persons not exhibiting AD pathology (low pathology not cognitively impaired, LP-NCI), and individuals not cognitively impaired who exhibited AD-related pathology post mortem (high pathology not cognitively impaired, HP-NCI). While the ‘amyloid hypothesis’ of AD posits that hydrophobic oligomers of the peptide amyloid-β (Aβ) lead to synaptotoxicity and neurodegeneration, experiments were unable to demonstrate that changes in spontaneous firing elicited by these brain extracts were directly mediated by Aβ. To determine the molecular etiology of these functional changes in an unbiased manner, label-free proteomic analysis was performed on a larger panel of 43 brain extracts of AD, LP-NCI, and HP-NCI individuals. The proteomic profiles of those brain extracts tested in the MEA-based neuronal firing assay were compared with a similar cohort of brain extracts tested in a morphological neurite integrity assay. This analysis revealed several candidates related to protein homeostasis and microtubule remodeling whose expression correlated with changes in neuronal structure and activity. The techniques and data herein provide a system for modeling the function of neurons and insights into proteins that may be mediating changes in cellular behavior within the AD brain.
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