TY - JOUR
T1 - Kinome-level screening identifies inhibition of polo-like kinase-1 (PLK1) as a target for enhancing non-viral transgene expression
AU - Christensen, Matthew D.
AU - Elmer, Jacob J.
AU - Eaton, Seron
AU - Gonzalez-Malerva, Laura
AU - LaBaer, Joshua
AU - Rege, Kaushal
N1 - Funding Information:
Funding from the National Institute of General Medical Sciences (NIGMS), National Institutes of Health (Grant # 1R01GM093229-01A1 ) to KR is gratefully acknowledged. Entinostat used in this study was generously provided by Syndax Pharmaceuticals, Inc. through the Cancer Therapeutics Evaluation Program (CTEP) and the National Cancer Institute, NIH. We would also like to thank Dr. Tong Fu for assistance with training and operation of the FACSCalibur flow cytometer, and Dr. Thrimoorthy Potta and Taraka Sai Pavan Grandhi in the Rege group at ASU for several helpful discussions and generous technical help.
Publisher Copyright:
© 2015 Elsevier B.V. All rights reserved.
PY - 2015/4/28
Y1 - 2015/4/28
N2 - Human cells contain hundreds of kinase enzymes that regulate several cellular processes, which likely include transgene delivery and expression. We identified several kinases that influence gene delivery and/or expression by performing a kinome-level screen in which, we identified small-molecule kinase inhibitors that significantly enhanced non-viral (polymer-mediated) transgene (luciferase) expression in cancer cells. The strongest enhancement was observed with several small-molecule inhibitors of Polo-like Kinase 1 (PLK 1) (e.g., HMN-214 and BI 2536), which enhanced luciferase expression up to 30-fold by arresting cells in the G2/M phase of the cell cycle and influencing intracellular trafficking of plasmid DNA. Knockdown of PLK 1 using an shRNA-expressing lentivirus further confirmed the enhancement of polymer-mediated transgene expression. In addition, pairwise and three-way combinations of PLK1 inhibitors with the histone deacetylase-1 (HDAC-1) inhibitor Entinostat and the JAK/STAT inhibitor AG-490 enhanced luciferase expression to levels significantly higher than individual drug treatments acting alone. These findings indicate that inhibition of specific intracellular kinases (e.g., PLK1) can significantly enhance non-viral transgene expression for applications in biotechnology and medicine.
AB - Human cells contain hundreds of kinase enzymes that regulate several cellular processes, which likely include transgene delivery and expression. We identified several kinases that influence gene delivery and/or expression by performing a kinome-level screen in which, we identified small-molecule kinase inhibitors that significantly enhanced non-viral (polymer-mediated) transgene (luciferase) expression in cancer cells. The strongest enhancement was observed with several small-molecule inhibitors of Polo-like Kinase 1 (PLK 1) (e.g., HMN-214 and BI 2536), which enhanced luciferase expression up to 30-fold by arresting cells in the G2/M phase of the cell cycle and influencing intracellular trafficking of plasmid DNA. Knockdown of PLK 1 using an shRNA-expressing lentivirus further confirmed the enhancement of polymer-mediated transgene expression. In addition, pairwise and three-way combinations of PLK1 inhibitors with the histone deacetylase-1 (HDAC-1) inhibitor Entinostat and the JAK/STAT inhibitor AG-490 enhanced luciferase expression to levels significantly higher than individual drug treatments acting alone. These findings indicate that inhibition of specific intracellular kinases (e.g., PLK1) can significantly enhance non-viral transgene expression for applications in biotechnology and medicine.
KW - BI 2536
KW - Cell cycle
KW - HMN-214
KW - Kinases
KW - Polymer Gene Delivery
KW - Transient protein expression
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U2 - 10.1016/j.jconrel.2015.01.036
DO - 10.1016/j.jconrel.2015.01.036
M3 - Article
C2 - 25681050
AN - SCOPUS:84924171979
VL - 204
SP - 20
EP - 29
JO - Journal of Controlled Release
JF - Journal of Controlled Release
SN - 0168-3659
ER -