In vivo imaging of gene transfer to the respiratory tract

Griesenbach, Uta; Meng, Cuixiang; Farley, Raymond; Cheng, Seng H.; Scheule, Ronald K.; Davies, M. H.; Wolstenholme-Hogg, Paul; Hove, Willem ten; Hoeven, Paul van der; Sinn, Patrick L.; McCray, Paul B.; Geddes, Duncan M.; Hasegawa, Mamoru; Frankel, Gad; Wiles, Siouxsie; Alton, Eric W.F.W.

doi:10.1016/j.biomaterials.2007.11.017

Cited by 13 publications

(4 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, early successes in the lung were reported by Gambhir and colleagues (Hildebrandt et al, 2003) while examining tumor targeted PEI/DNA complexes. More recently BLI was used to study pulmonary expression conferred by Genzyme Lipid 67 DNA complexes (Griesenbach et al, 2008). Animals that received 80 μg of these complexes exhibited expression in the lung sufficient for detection by BLI.…”

Section: Discussionmentioning

confidence: 99%

Bioluminescent imaging of reporter gene expression in the lungs of wildtype and model mice following the administration of PEG‐stabilized DNA nanoparticles

Ziady

Kotlarchyk

Bryant

et al. 2010

Microscopy Res & Technique

View full text Add to dashboard Cite

DNA nanoparticles (DNPs) formed by compacting DNA with polyethyleneglycolylated poly-L-lysine are a nonviral vector shown to be safe and efficacious in animals and humans. To extend our capabilities of assessing the efficacy and duration of expression achieved by DNPs, we tested the utility of bioluminescent imaging (BLI) of transgene expression in wildtype and cystic fibrosis (CF) mouse models. We tested the effect of route of administration, mouse coat color, anesthesia, dose, and promoter sequence on the level and duration of expression. Furthermore, we investigated the correlation between imaging and direct analysis of luciferase expression in lung homogenates. We found that intratracheal instillation, and the use of deep and prolonged anesthesia with avertin produced significantly higher expression compared with intranasal administration, and the use of lighter anesthesia with isoflurane. Although similar expression was observed for both dark and light coat animals, imaging signal intensity was attenuated in mice with dark fur. Furthermore, good correlation between imaging and direct homogenate analysis was observed for single dose (r = 0.96), and dose response studies in wildtype (r = 0.82) and CF mice (r = 0.87). Finally, we used imaging to track gene expression over a 56-day time course. We found that the human ubiquitin B promoter gives stable transgene expression up to 49 days following nanoparticle administration, while expression with the cytomegalovirus promoter diminished after 2 days and returned to background levels by day 14. Taken together, our results demonstrate that BLI is an effective and useful modality for measuring gene expression conferred by DNPs in the lung.

show abstract

Section: Discussionmentioning

confidence: 99%

Bioluminescent imaging of reporter gene expression in the lungs of wildtype and model mice following the administration of PEG‐stabilized DNA nanoparticles

Ziady

Kotlarchyk

Bryant

et al. 2010

Microscopy Res & Technique

View full text Add to dashboard Cite

show abstract

“…We have previously shown that FLuc expression after non-viral gene transfer to the lung can be detected using bioluminescence in vivo imaging (BLI) [43] and have also shown that photon emission is enhanced if the substrate luciferin is delivered topically by nasal instillation rather than intraperitoneal administration, likely due to higher substrate concentrations reaching the lung [40]. Here, we assessed GLuc expression combined with topical administration of coelenterazine for BLI in the lung and showed that although GLuc expression can be detected in the lung, topical administration of the substrate leads to significant background photon emission in untransfected mice.…”

Section: Discussionmentioning

confidence: 99%

Secreted Gaussia luciferase as a sensitive reporter gene for in vivo and ex vivo studies of airway gene transfer

et al. 2011

Self Cite

View full text Add to dashboard Cite

“…Analogous systems have previously been reported for DNA and positive-strand RNA viruses [27] but have been elusive for negative-strand RNA viruses, largely due to virus attenuation [28] or genetic instability resulting from reporter gene insertion [29]. We considered SeV an ideal candidate for non-invasive imaging because (i) foreign-gene expression by paramyxovirus vectors is usually genetically stable [30], (ii) in vivo imaging of a non-replicating SeV in intact mice has been successfully demonstrated [31] and (iii) the match of SeV and the murine host allows pathogenesis studies [11]. The reporter virus rSeV-luc(M-F*) described here was found to express high levels of luciferase yet replicate and promote disease in mice similar to wild-type (WT) virus.…”

Section: Introductionmentioning

confidence: 99%

Illumination of Parainfluenza Virus Infection and Transmission in Living Animals Reveals a Tissue-Specific Dichotomy

et al. 2011

View full text Add to dashboard Cite

The parainfluenza viruses (PIVs) are highly contagious respiratory paramyxoviruses and a leading cause of lower respiratory tract (LRT) disease. Since no vaccines or antivirals exist, non-pharmaceutical interventions are the only means of control for these pathogens. Here we used bioluminescence imaging to visualize the spatial and temporal progression of murine PIV1 (Sendai virus) infection in living mice after intranasal inoculation or exposure by contact. A non-attenuated luciferase reporter virus (rSeV-luc(M-F*)) that expressed high levels of luciferase yet was phenotypically similar to wild-type Sendai virus in vitro and in vivo was generated to allow visualization. After direct intranasal inoculation, we unexpectedly observed that the upper respiratory tract (URT) and trachea supported robust infection under conditions that result in little infection or pathology in the lungs including a low inoculum of virus, an attenuated virus, and strains of mice genetically resistant to lung infection. The high permissivity of the URT and trachea to infection resulted in 100% transmission to naïve contact recipients, even after low-dose (70 PFU) inoculation of genetically resistant BALB/c donor mice. The timing of transmission was consistent with the timing of high viral titers in the URT and trachea of donor animals but was independent of the levels of infection in the lungs of donors. The data therefore reveals a disconnect between transmissibility, which is associated with infection in the URT, and pathogenesis, which arises from infection in the lungs and the immune response. Natural infection after transmission was universally robust in the URT and trachea yet limited in the lungs, inducing protective immunity without weight loss even in genetically susceptible 129/SvJ mice. Overall, these results reveal a dichotomy between PIV infection in the URT and trachea versus the lungs and define a new model for studies of pathogenesis, development of live virus vaccines, and testing of antiviral therapies.

show abstract

In vivo imaging of gene transfer to the respiratory tract

Cited by 13 publications

References 23 publications

Bioluminescent imaging of reporter gene expression in the lungs of wildtype and model mice following the administration of PEG‐stabilized DNA nanoparticles

Bioluminescent imaging of reporter gene expression in the lungs of wildtype and model mice following the administration of PEG‐stabilized DNA nanoparticles

Secreted Gaussia luciferase as a sensitive reporter gene for in vivo and ex vivo studies of airway gene transfer

Illumination of Parainfluenza Virus Infection and Transmission in Living Animals Reveals a Tissue-Specific Dichotomy

Contact Info

Product

Resources

About