title: VEGF inhibitor (Iressa) arrests histone deacetylase expression: single-cell cotransfection imaging cytometry for multi-target-multi-drug analysis.
abstract: Multi-target-multi-drug approaches are needed to accelerate the process of drug discovery screening and to design efficient therapeutic strategies against diseases that involve alterations in multiple cellular targets. Herein we report single-cell cotransfection imaging cytometry to quantitatively screen drug-induced off-target effects. Vascular endothelial growth factor (VEGF) and histone deacetylase (HDAC) genes amplified from the genomic DNA were cloned in fluorescently tagged gene constructs (RFP-HDAC/YFP-VEGF). These gene constructs were cotransfected in HEK-293 cells to explore the possibility of off-target effects of 4-phenylbutyrate and Iressa on the expression of VEGF and HDAC through single-cell imaging cytometry. Iressa (10 µM) treatment at the time of cotransfection or 48 h after cotransfection of RFP-HDAC/YFP-VEGF plasmids in HEK-293 cells resulted in off-target effects on HDAC expression. These results suggest possible applications of Iressa in the treatment of diseases in which expression of both HDAC and VEGF should be inhibited. 4-Phenylbutyrate (2.0 mM) did not show any off-target effects on VEGF expression. The developed quantitative multicolor live single-cell cotransfection imaging can be employed to select better drug combinations for faster screening and greater accuracy in multi-target-multi-drug analysis by increasing the on-target/desired off-target effects and eliminating the undesirable off-target effects.
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Yu Kyung
authors.initials: YK
authors.lastname: Tak
authors.forename: Pravin K
authors.initials: PK
authors.lastname: Naoghare
authors.forename: Eunyoung
authors.initials: E
authors.lastname: Han
authors.forename: Joon Myong
authors.initials: JM
authors.lastname: Song
mesh_terms: D055808:Drug Discovery / Q000379:methods*; D000077156:Gefitinib; D057809:HEK293 Cells; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000235:genetics; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D010654:Phenylbutyrates / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology*; D014162:Transfection / Q000379:methods; D042442:Vascular Endothelial Growth Factors / Q000037:antagonists & inhibitors / Q000235:genetics
chemical_list: D056572:Histone Deacetylase Inhibitors; D010654:Phenylbutyrates; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D042442:Vascular Endothelial Growth Factors; C075773:4-phenylbutyric acid; D006655:Histone Deacetylases; D000077156:Gefitinib