The essential roles of the ubiquitin-proteasome system (UPS) in many cellular processes are now well recognized. However, many questions still remain unanswered. Intriguing yet not fully understood are the functions of the immunoproteasome, an alternative form of the constitutive proteasome. It has been shown that the roles of the immunoproteasome in cells are not just limited to the generation of antigenic peptides, but far more multifaceted. In this regard, recent studies have shown that catalytic subunits of the immunoproteasome are implicated in various types of diseases including cancer. Despite these therapeutic implications, the immunoproteasome remains largely untapped for cancer drug discovery. A better understanding of immunoproteasome functions would be crucial in developing the optimum LMP2 inhibitor(s). In our continuing efforts to better understand the role of the immunoproteasome in cells, we have recently developed a small molecule probe ('UK-101') that specifically targets LMP2, a major catalytic subunit of the immunoproteasome. Further, we found that LMP2 is highly expressed in several types of primary tumor tissues and cancer cell lines. Interestingly, cancer cells that highly express LMP2 are markedly more sensitive to UK-101 than those expressing low level of LMP2. Consistent with these in vitro results, UK-101 inhibits tumor growth in a mouse xenograft model of human prostate tumor that highly expresses LMP2, but not that of tumor that expresses low level of LMP2. These observations further support a potential role of LMP2 in cancer cell growth and survival. The goals of this application are to determine the roles of the immunoproteasome in prostate cancer and to investigate therapeutic potentials of the immunoproteasome-targeting approaches in cancer therapy. Towards this goal, we propose to use innovative chemical probes that selectively target and visualize individual catalytic subunits of the immunoproteasome. In order to accomplish this goal, four specific aims are proposed: 1) To validate the immunoproteasome as the major pharmacological target of UK-101, 2) To determine dynamic cellular distribution of the catalytically active immunoproteasome, and 3) To determine the unique functions of the immunoproteasome compared to the constitutive proteasome and 4) To develop an optimum LMP2 inhibitor(s) by generating a structurally diversified small molecule library. The proposed work integrates both chemical and biological approaches. Successful completion of the project will significantly expand our knowledge on the role of the immunoproteasome. In addition, the study will yield critical information in developing immunoproteasome-specific inhibitors as a potential therapeutic agent. Finally, small molecule probes that will be used in this application will provide valuable research tools for the ubiquitin-proteasome biology community.

Public Health Relevance

Small molecule probes are valuable tools in deciphering protein functions. We have developed a novel cell permeable small molecule that selectively targets the immunoproteasome. Using this small molecule and other molecular tools, we will investigate the roles of the immunoproteasome in cancer cells and therapeutic potentials of the immunoproteasome-targeting approaches in cancer therapy.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01CA128903-05
Application #
8657838
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Fu, Yali
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Kentucky
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
City
Lexington
State
KY
Country
United States
Zip Code
40506
Miller, Zachary; Kim, Keun-Sik; Lee, Do-Min et al. (2015) Proteasome inhibitors with pyrazole scaffolds from structure-based virtual screening. J Med Chem 58:2036-41
Ao, Lin; Reichel, Derek; Hu, Di et al. (2015) Polymer micelle formulations of proteasome inhibitor carfilzomib for improved metabolic stability and anticancer efficacy in human multiple myeloma and lung cancer cell lines. J Pharmacol Exp Ther 355:168-73
Cornish Carmony, Kimberly; Sharma, Lalit Kumar; Lee, Do-Min et al. (2015) Elucidating the catalytic subunit composition of distinct proteasome subtypes: a crosslinking approach employing bifunctional activity-based probes. Chembiochem 16:284-92
Kasam, Vinod; Lee, Na-Ra; Kim, Kyung-Bo et al. (2014) Selective immunoproteasome inhibitors with non-peptide scaffolds identified from structure-based virtual screening. Bioorg Med Chem Lett 24:3614-7
Miller, Zachary; Lee, Wooin; Kim, Kyung Bo (2014) The immunoproteasome as a therapeutic target for hematological malignancies. Curr Cancer Drug Targets 14:537-48
Park, Ji Eun; Wu, Ying; Carmony, Kimberly Cornish et al. (2014) A FRET-based approach for identification of proteasome catalytic subunit composition. Mol Biosyst 10:196-200
Park, Ji Eun; Ao, Lin; Miller, Zachary et al. (2013) PSMB9 codon 60 polymorphisms have no impact on the activity of the immunoproteasome catalytic subunit B1i expressed in multiple types of solid cancer. PLoS One 8:e73732
Miller, Zachary; Ao, Lin; Kim, Kyung Bo et al. (2013) Inhibitors of the immunoproteasome: current status and future directions. Curr Pharm Des 19:4140-51
Carmony, Kimberly Cornish; Kim, Kyung Bo (2013) Activity-based imaging probes of the proteasome. Cell Biochem Biophys 67:91-101
Carmony, Kimberly Cornish; Lee, Do-Min; Wu, Ying et al. (2012) A bright approach to the immunoproteasome: development of LMP2/β1i-specific imaging probes. Bioorg Med Chem 20:607-13

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