title: Proteomic profiling of salivary gland after nonviral gene transfer mediated by conventional plasmids and minicircles.
abstract: In this study, we compared gene transfer efficiency and host response to ultrasound-assisted, nonviral gene transfer with a conventional plasmid and a minicircle vector in the submandibular salivary glands of mice. Initially, we looked at gene transfer efficiency with equimolar amounts of the plasmid and minicircle vectors, corroborating an earlier report showing that minicircle is more efficient in the context of a physical method of gene transfer. We then sought to characterize the physiological response of the salivary gland to exogenous gene transfer using global proteomic profiling. Somewhat surprisingly, we found that sonoporation alone, without a gene transfer vector present, had virtually no effect on the salivary gland proteome. However, when a plasmid vector was used, we observed profound perturbations of the salivary gland proteome that compared in magnitude to that seen in a previous report after high doses of AAV. Finally, we found that gene transfer with a minicircle induces only minor proteomic alterations that were similar to sonoporation alone. Using mass spectrometry, we assigned protein IDs to 218 gel spots that differed between plasmid and minicircle. Bioinformatic analysis of these proteins demonstrated convergence on 68 known protein interaction pathways, most notably those associated with innate immunity, cellular stress, and morphogenesis.
authors.affiliation: Gene Therapy Program, Allegheny Health Network, Pittsburgh, Pennsylvania, USA.
authors.forename: Ramaz
authors.initials: R
authors.lastname: Geguchadze
authors.affiliation: Gene Therapy Program, Allegheny Health Network, Pittsburgh, Pennsylvania, USA.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Gene Therapy Program, Allegheny Health Network, Pittsburgh, Pennsylvania, USA.
authors.forename: Lee
authors.initials: L
authors.lastname: Zourelias
authors.affiliation: National Institute of Dental and Craniofacial Research, Bethesda, Maryland, USA.
authors.forename: Paola
authors.initials: P
authors.lastname: Perez-Riveros
authors.affiliation: Department of Oral Pathology, Medicine and Radiology, Indiana University School of Dentistry, Indianapolis, Indiana, USA.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Edwards
authors.affiliation: Gene Therapy Program, Allegheny Health Network, Pittsburgh, Pennsylvania, USA.
authors.forename: Laurie
authors.initials: L
authors.lastname: Machen
authors.affiliation: Gene Therapy Program, Allegheny Health Network, Pittsburgh, Pennsylvania, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Passineau