Kinome-level screening identifies inhibition of polo-like kinase-1 (PLK1) as a target for enhancing non-viral transgene expression

  • J Control Release. 2015 Apr 28;204:20-9. doi: 10.1016/j.jconrel.2015.01.036.
Matthew D Christensen  1 ,  Jacob J Elmer  1 ,  Seron Eaton  2 ,  Laura Gonzalez-Malerva  2 ,  Joshua LaBaer  2 ,  Kaushal Rege  3
Affiliations
  • 1. Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, USA.
  • 2. The Virginia G. Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, AZ, USA.
  • 3. Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, USA. Electronic address: [email protected].
PMID: 25681050 DOI: 10.1016/j.jconrel.2015.01.036
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.

Keywords
BI 2536; Cell cycle; HMN-214; Kinases; Polymer Gene Delivery; Transient protein expression.
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