Date of Award

7-29-2013

Document Type

Thesis

Publisher

Santa Clara University

First Advisor

Zhiwen Zhang

Abstract

G-protein coupled receptors (GPCRs) are currently the largest class of membrane receptors and are targeted by a majority of the modern drug therapeutics. In addition, they partake in many physiological and pathological processes such as inflammation, growth, and hormone responses. Most importantly, GPCRs are targets of many disease-specific pathways such as Alzheimers, hypertension, leukemia, and depression. As a result, there is an immense interest in studying GPCRs as this area provides further knowledge into the specific disease pathways and allows the discovery of novel therapeutics. In order to have a better understanding of pathways, scientists have studied GPCR activation. The prostanoid receptors are of great interest because they coordinate a vast range of physiological processes such as regulating cardiovascular pathways, modulating neuronal activity and controlling immune responses. We collaborated with Multispan Inc., a biotech company focusing exclusively on drug discovery research targeting GPCRs, to monitor the agonist-induced internalization of GPCRs from the prostanoid family through live cell flow cytometry. From our experimental results, we have observed over 50 percent internalization for the EP1 and EP4 receptors within 60 minutes after being activated by iloprost and PGE2 respectively. Our initial experimental results have also shown over 40 percent internalization for the TP receptor within 120 minutes after exposure to PGD2. Overall, we have been able to utilize Multispan’s proprietary cell lines to overexpress a few of the prostanoid receptors. These assays are powerful tools for the discovery of novel therapeutics, as they enable the testing against libraries and screening from a few thousand to a few million compounds.

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