Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 20-29 |
| Number of pages | 10 |
| Journal | Journal of Controlled Release |
| Volume | 204 |
| DOIs | |
| State | Published - Apr 28 2015 |
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Keywords
- BI 2536
- Cell cycle
- HMN-214
- Kinases
- Polymer Gene Delivery
- Transient protein expression
ASJC Scopus subject areas
- Pharmaceutical Science
Cite this
Kinome-level screening identifies inhibition of polo-like kinase-1 (PLK1) as a target for enhancing non-viral transgene expression. / Christensen, Matthew D.; Elmer, Jacob J.; Eaton, Seron; Gonzalez-Malerva, Laura; LaBaer, Joshua; Rege, Kaushal.
In: Journal of Controlled Release, Vol. 204, 28.04.2015, p. 20-29.Research output: Contribution to journal › Article
}
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
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
UR - http://www.scopus.com/inward/record.url?scp=84924171979&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84924171979&partnerID=8YFLogxK
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 -