title: High-throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies.
abstract: Nonviral gene delivery methods are advantageous over viral vectors in terms of safety, cost, and flexibility in design and application, but suffer from lower gene transfer efficiency. In addition to modifications to nucleic acid design and nonviral carriers, new tools are sought to enhance transfection. Priming is the pharmacological modulation of transfection efficiency and transgene expression, and has demonstrated transfection increase in several compounds, for example, chloroquine and glucocorticoids. To develop a library of transfection priming compounds, a high-throughput screen was performed of the NIH Clinical Collection (NCC) to identify clinical compounds that prime polyethylenimine (PEI) transfection. HEK293T cells were treated with priming compounds, then transfected with enhanced green fluorescent protein (EGFP)-encoding plasmid by PEI. After 48-hr culture, primed and transfected cells were assayed for transfection, cell proliferation, and cell viability by fluorescence measurement of EGFP reporter, Hoechst 33342 nuclei stain, and resazurin metabolic assay. From the microscope image analysis and microplate measurements, transfection fold-changes were determined, and compounds resulting in statistically significant transfection fold-change were identified. NCC compounds were clustered using PubChem fingerprint similarity by Tanimoto coefficients in ChemmineTools. Fold-changes for each compound were linked to drug clusters, from which drug classes that prime transfection were identified. Among the identified drugs classes that primed transfection increases were antioxidants, GABAA receptor modulators, and glucocorticoids. Resveratrol and piceid, stilbenoid antioxidants found in grapes, and zolpidem, a GABAA modulator, increased transfection nearly three-fold. Literature indicate interaction of the identified transfection priming drug clusters with mitochondria, which may modulate mitochondrial dysfunction known to be associated with PEI transfection.
authors.affiliation: Dept. of Biological Systems Engineering University of Nebraska-Lincoln Lincoln NE 68583; Center for Nanohybrid Functional Materials University of Nebraska-Lincoln, Lincoln NE 68588; Dept. of Electrical and Computer Engineering University of Nebraska-Lincoln Lincoln NE 68588.
authors.forename: Albert
authors.initials: A
authors.lastname: Nguyen
authors.affiliation: Dept. of Biological Systems Engineering University of Nebraska-Lincoln Lincoln NE 68583.
authors.forename: Jared
authors.initials: J
authors.lastname: Beyersdorf
authors.affiliation: Bioinformatics Core Research Facility University of Nebraska-Lincoln Lincoln NE 68588; School of Biological Sciences University of Nebraska-Lincoln, Lincoln NE 68588.
authors.forename: Jean-Jack
authors.initials: JJ
authors.lastname: Riethoven
authors.affiliation: Dept. of Biological Systems Engineering University of Nebraska-Lincoln Lincoln NE 68583; Center for Nanohybrid Functional Materials University of Nebraska-Lincoln, Lincoln NE 68588.
authors.forename: Angela K
authors.initials: AK
authors.lastname: Pannier
keywords: NIH Clinical Collection; high‐throughput screen; mitochondrial dysfunction; nonviral gene delivery; polyethylenimine toxicity; priming transfection