title: Electroporation-mediated delivery of the FER gene in the resolution of trauma-related fatal pneumonia.
abstract: Injured patients with lung contusion (LC) are at risk of developing bacterial pneumonia (PNA) followed by sepsis and death. A recent genome-wide association study (GWAS) showed FER gene expression positively correlating with survival rates among individuals with above conditions. We sought to determine whether electroporation (EP)-mediated delivery of FER gene could indeed improve survival, in a lethal model of combined LC and PNA. C57BL/6 mice sustained unilateral LC, which preceded a 500 Klebsiella colony forming unit (CFU) inoculation by 6 h. In-between these insults, human FER plasmid (pFER) was introduced into the lungs followed by eight EP pulses applied externally (10 ms at 200 V cm-1). Control groups included EP of empty vector (pcDNA3) or Na+/K+-ATPase genes (pPump) and no treatment (LC+PNA). We recorded survival, histology, lung mechanics, bronchial alveolar lavage (BAL) fluid, FER and inflammatory gene expression and bacteriology. The data show that 7-day survival was significantly improved by pFER compared with control groups. pFER increased BAL monocytes and activated antibacterial response genes (nitric oxide synthase (NOS), Fizz). pFER treatment showed decreased lung and blood Klebsiella counts reaching, in some cases, complete sterilization. In conclusion, FER gene delivery promoted survival in LC+PNA mice via recruitment of activated immune cells, improving efficiency of bacterial clearance within contused lung.
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: V A
authors.initials: VA
authors.lastname: Dolgachev
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: R
authors.initials: R
authors.lastname: Goldberg
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: M V
authors.initials: MV
authors.lastname: Suresh
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: B
authors.initials: B
authors.lastname: Thomas
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: N
authors.initials: N
authors.lastname: Talarico
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: Hemmila
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: K
authors.initials: K
authors.lastname: Raghavendran
authors.affiliation: Division of Acute Care Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
authors.forename: D
authors.identifier: 0000-0003-3716-962X
authors.initials: D
authors.lastname: Machado-Aranda
mesh_terms: D000818:Animals; D058491:Bacterial Load; D003288:Contusions / Q000150:complications*; D018274:Electroporation; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D007709:Klebsiella / Q000472:pathogenicity; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018410:Pneumonia, Bacterial / Q000209:etiology / Q000382:microbiology / Q000628:therapy*; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism
chemical_list: C052602:proto-oncogene protein c-fes-fps; D011505:Protein-Tyrosine Kinases; D000254:Sodium-Potassium-Exchanging ATPase