Examining the Anti-Proliferative Properties of DUB Inhibitors in Primary Mesothelioma Cells

Maggie Cook, Biomedical EngineeringMentor: Dr. Christopher Plaisier, Assistant Professor

School of Biological Health Systems Engineering

Three different cell lines of two differentprimary MPM subtypes were tested:

INTRODUCTION

We are currently performing this drug screeninganalysis on other MPM cell lines from differentpatients. We are also in the middle of assessingother drug types, including those that influenceubiquitination levels of proteins. Ultimately, theIC50 values of all drugs and combinations will becompared with Cisplatin and Pemetrexed, twodrugs commonly used in chemotherapy, to seeif any of the drugs have a synergistic effect.

Ubiquitin is a key protein in many biologicalprocesses and typically marks proteins fordegradation. Deubiquitinating (DUB) enzymesremove the ubiquitin mark and can affectdownstream biological processes by changingthe ubiquitination of a specific set of proteins.They are frequently overexpressed in cancers1.DUB inhibitors block the deubiquitylation ofproteins leading to an excess of ubiquitinylatedproteins. Prior research has noted the efficacyof DUB inhibitors in immortalized cell lines2.Here we demonstrate that the DUB inhibitorPR-619 is significantly anti-proliferative forprimary malignant pleural mesothelioma cells,regardless of cancer subtype (epithelioid andsarcomatoid).

RESEARCH METHODS

RESULTS FUTURE DIRECTIONS

These figures show how increasing levels of PR-619 lead to adecrease in cell counts after 72 hours of treatment. The IC50values are also calculated for each sample in micromolars.

ACKNOWLEDGEMENTS

REFERENCES• M3T (Epithelioid)• M12T (Epithelioid)

• M24 (Sarcomatoid)

Each cell line was screened with PR-619 andcompared to a standard curve in order tocalculate the IC50 values, which represent theconcentration of drug required to inhibit abiological function to 50% of its typical capacity.

M3TIC50:6 µM

M12TIC50:7.4 µM

M24IC50:5.2 µM

NEW SKILLS

Previously in the Plaisier lab I conductedprimarily computational research. This semesterI was trained in and used primarily experimentalresearch techniques including culturing MPMcell lines, designing experiments, makingprotocols, calculating concentrations, andperforming experiments where I collected andinterpreted data.

Thanks to Sierra Wilfred for her training and teaching and toFURI for assistance in funding this research project.

[1] Morrow, J. K., Lin, H. K., Sun, S. C., & Zhang, S. (2015). Targeting ubiquitination for cancertherapies. Future medicinal chemistry, 7(17), 2333–2350. https://doi.org/10.4155/fmc.15.148[2] Kuo KL, Liu SH, Lin WC, et al. The Deubiquitinating Enzyme Inhibitor PR-619 Enhances theCytotoxicity of Cisplatin via the Suppression of Anti-Apoptotic Bcl-2 Protein: In Vitro and InVivo Study. Cells. 2019;8(10):1268. Published 2019 Oct 17. doi:10.3390/cells8101268

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PR-619, µM PR-619, µM

PR-619, µM PR-619, µM

PR-619, µM PR-619, µM