Year of Publication

2008

Degree Name

Doctor of Philosophy (PhD)

Document Type

Dissertation

College

Pharmacy

Department

Pharmaceutical Sciences

First Advisor

Dr. Kyung-Bo Kim

Abstract

The immunoproteasome, known to play an important role in MHC class I antigen processing and presentation, have been linked to neurodegenerative diseases and hematological cancers. However, the pathophysiological functions of the immunoproteasome in these diseases are still not very well established. This can be attributed mainly to the lack of appropriate molecular probes that selectively target the immunoproteasome catalytic subunits. Herein, we report the development of a small molecular inhibitor (AM) that selectively targets the major catalytic subunit, LMP2, of the immunoproteasome. We show that the compound covalently modifies the LMP2 subunit with high specificity in human prostate cancer cell. AM was also shown to selectively inhibit the chymotrypsin-like activity of LMP2 subunit. More importantly, the anti-proliferative activity of AM is more pronounced in prostate cancer cells that highly express LMP2 without inducing toxicity in normal cells. These results implicate an important role of LMP2 in regulating cell growth of malignant tumors that highly express LMP2.

Subsequently, the modes of action of AM were investigated. Prostate cancer cells that highly express LMP were shown to induce G2/M cell cycle arrest and apoptosis via PARP cleavage when treated with the compound. Similar to epoxomicin, the treatment of AM induced the accumulation of poly-ubiquitination in prostate cancer cells, which indicates the inhibition of proteolysis. However, unlike epoxomicin, the treatment of AM did not appear to inhibit the activation of inflammation. In conclusion, these results suggest that the LMP2 inhibitor, AM, may induce cytotoxicity prostate cancer cells that highly express LMP2 catalytic subunit in similar modes of action as epoxomicin but it does not involve the inflammatory pathway.

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