The Effects of Everolimus and Lonafarnib on Hyperphosphorylated Tau and Mtor in Rat Hippocampal Astrocytes

Date of Award

6-2020

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Harold Komisky, PhD

Second Advisor

Richard White, PhD, FAHA

Third Advisor

Dennis Peffley, JD, PhD

Abstract

Dementia is a progressive syndrome that causes memory loss and affects an individual’s cognitive function and communication skills. Alzheimer’s Disease (AD) is an irreversible neurodegenerative disease and it is the most common form of Dementia. Currently, there are not any cures for AD. Here we decided to look into mechanisms within astrocytes that could slow down the progression of AD substantially. Astrocytes are glial cells that are important in maintaining neuronal homeostasis which is why they are quickly becoming the subjects in AD research. Our research targeted one of the key characteristics of AD, the accumulation of hyperphosphorylated Tau (pTau). Our other protein of interest was the mammalian target of Rapamycin (mTOR). mTOR is a protein kinase that is essential for various cellular processes, but it is also thought to increase the amount of hyperphosphorylated Tau when upregulated. We specifically studied how the drugs Lonafarnib and Everolimus effected mTOR and pTau in astrocytes. To conduct the research, we used Cell Counting Kits and Enzyme-Linked Immunosorbent Assays (ELISA). We also used western blotting techniques to analyze phosphorylated Tau and mTOR protein expression. The assays done with Everolimus were limited due to the pandemic. The CCK assay with Lonafarnib exhibited an increase in cell viability in the treatment groups in comparison to the controls. The ELISA kits with Lonafarnib showed an intracellular decrease in pTau with increasing concentrations of Lonafarnib, but no correlations were observed between Lonafarnib and intracellular mTOR levels. In conclusion, this preliminary data suggested that Lonafarnib may have a positive effect on astrocyte cell viability, but the mechanism of action remains unknown.

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