title: Cell phenotype specific kinetics of expression of intratracheally injected manganese superoxide dismutase-plasmid/liposomes (MnSOD-PL) during lung radioprotective gene therapy.
abstract: Intratracheal (IT) injection of manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) complexes prior to whole lung irradiation of C57BL/6J mice provides significant protection from acute and chronic irradiation damage. We determined the duration of increased MnSOD biochemical activity and differential expression of a hemagglutinin (HA) epitope-tagged MnSOD transgene. HA-MnSOD-PL was IT injected at doses of 0-1000 microg, and mice were killed 1,2,3 or 4 days later. Other groups of mice were irradiated to 20 Gy to the pulmonary cavity 24 h after injection and killed at the same time points as non-irradiated mice. Both non-irradiated and irradiated groups of mice showed increased MnSOD biochemical activity with plasmid dose that plateaued at 100 microg of MnSOD plasmid DNA. In control mice, MnSOD biochemical activity decreased at 2, 3 or 4 days after injection. In irradiated mice, MnSOD biochemical activity decreased at day 2 but increased on days 3 and 4. HA-MnSOD expression decreased in broncheoalveolar macrophages and alveolar type-II cells 3 days after injection in non-irradiated and irradiated mice, but remained elevated in endothelial and epithelial cells past 4 days. The data provide a rationale for every second-day administration of intrapulmonary MnSOD-PL in clinical trials of radioprotective gene therapy. This should be sufficient to provide radioprotection during radiation treatments.
authors.affiliation: Department of Radiation Oncology, University of Pittsburgh Cancer Institute, 200 Lothrop Street, Pittsburgh, PA, USA.
authors.forename: M W
authors.initials: MW
authors.lastname: Epperly
authors.forename: H L
authors.initials: HL
authors.lastname: Guo
authors.forename: M
authors.initials: M
authors.lastname: Jefferson
authors.forename: S
authors.initials: S
authors.lastname: Nie
authors.forename: J
authors.initials: J
authors.lastname: Gretton
authors.forename: M
authors.initials: M
authors.lastname: Bernarding
authors.forename: D
authors.initials: D
authors.lastname: Bar-Sagi
authors.forename: H
authors.initials: H
authors.lastname: Archer
authors.forename: J S
authors.initials: JS
authors.lastname: Greenberger
mesh_terms: D000818:Animals; D004730:Endothelium, Vascular / Q000201:enzymology; D004847:Epithelial Cells / Q000201:enzymology; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D006388:Hemagglutinins / Q000235:genetics; D007267:Injections; D008081:Liposomes; D008168:Lung / Q000201:enzymology* / Q000528:radiation effects*; D016676:Macrophages, Alveolar / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D011650:Pulmonary Alveoli / Q000201:enzymology; D011833:Radiation Injuries, Experimental / Q000201:enzymology / Q000517:prevention & control*; D011837:Radiation-Protective Agents; D013482:Superoxide Dismutase / Q000235:genetics*; D013997:Time Factors
chemical_list: D006388:Hemagglutinins; D008081:Liposomes; D011837:Radiation-Protective Agents; D013482:Superoxide Dismutase