Examining the Anti-Proliferative Properties of DUB ...
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Examining the Anti-Proliferative Properties of DUB Inhibitors in Primary Mesothelioma Cells Maggie Cook, Biomedical Engineering Mentor: Dr. Christopher Plaisier, Assistant Professor School of Biological Health Systems Engineering Three different cell lines of two different primary MPM subtypes were tested: INTRODUCTION We are currently performing this drug screening analysis on other MPM cell lines from different patients. We are also in the middle of assessing other drug types, including those that influence ubiquitination levels of proteins. Ultimately, the IC50 values of all drugs and combinations will be compared with Cisplatin and Pemetrexed, two drugs commonly used in chemotherapy, to see if any of the drugs have a synergistic effect. Ubiquitin is a key protein in many biological processes and typically marks proteins for degradation. Deubiquitinating (DUB) enzymes remove the ubiquitin mark and can affect downstream biological processes by changing the ubiquitination of a specific set of proteins. They are frequently overexpressed in cancers 1 . DUB inhibitors block the deubiquitylation of proteins leading to an excess of ubiquitinylated proteins. Prior research has noted the efficacy of DUB inhibitors in immortalized cell lines 2 . Here we demonstrate that the DUB inhibitor PR-619 is significantly anti-proliferative for primary malignant pleural mesothelioma cells, regardless of cancer subtype (epithelioid and sarcomatoid). RESEARCH METHODS RESULTS FUTURE DIRECTIONS These figures show how increasing levels of PR-619 lead to a decrease in cell counts after 72 hours of treatment. The IC50 values are also calculated for each sample in micromolars. ACKNOWLEDGEMENTS REFERENCES • M3T (Epithelioid) • M12T (Epithelioid) • M24 (Sarcomatoid) Each cell line was screened with PR-619 and compared to a standard curve in order to calculate the IC50 values, which represent the concentration of drug required to inhibit a biological function to 50% of its typical capacity. M3T IC50: 6 μM M12T IC50: 7.4 μM M24 IC50: 5.2 μM NEW SKILLS Previously in the Plaisier lab I conducted primarily computational research. This semester I was trained in and used primarily experimental research techniques including culturing MPM cell lines, designing experiments, making protocols, calculating concentrations, and performing experiments where I collected and interpreted data. Thanks to Sierra Wilfred for her training and teaching and to FURI for assistance in funding this research project. [1] Morrow, J. K., Lin, H. K., Sun, S. C., & Zhang, S. (2015). Targeting ubiquitination for cancer therapies. 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 the Cytotoxicity of Cisplatin via the Suppression of Anti-Apoptotic Bcl-2 Protein: In Vitro and In Vivo Study. Cells. 2019;8(10):1268. Published 2019 Oct 17. doi:10.3390/cells8101268 Cells PR-619, μM PR-619, μM PR-619, μM PR-619, μM PR-619, μM PR-619, μM
Transcript of Examining the Anti-Proliferative Properties of DUB ...
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
