Viscoelastic Gel Formulations Enhance Airway Epithelial Gene Transfer with Viral Vectors

Sinn, Patrick L.; Shah, Ankur J.; Donovan, Maureen D.; McCray, Paul B.

doi:10.1165/rcmb.2004-0410oc

Cited by 50 publications

(70 citation statements)

References 30 publications

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“…This dose was constant for each vector administration and for each protocol. Vector was formulated with 1% methylcellulose as previously described (40). An endotoxin assay revealed detectable levels of endotoxin (Ͻ100 endotoxin units) in the delivered volume of vehicle (data not shown).…”

Section: Methodsmentioning

confidence: 90%

Lentivirus Vector Can Be Readministered to Nasal Epithelia without Blocking Immune Responses

Sinn¹,

Arias²,

Brogden

et al. 2008

J Virol

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For many envisioned applications of lentivirus vectors as tools in respiratory biology and therapeutic gene delivery, the efficiency of gene transfer must be improved. We previously demonstrated stable, persistent (>11 months) in vivo expression following a single application of a feline immunodeficiency virus (FIV)-based lentivirus vector (GP64-FIV) to murine nasal epithelia. Here we investigate the efficacy of repeated administration of lentivirus vectors to the airways. Using quantitative bioluminescent imaging, we found that consecutive daily dosing achieved a linear increase in gene expression and greatly increased the number of epithelial cells targeted. Surprisingly, reporter gene expression also increased additively following each of seven doses of FIV delivered over consecutive weeks (1 dose/week), without the development of systemic or local neutralizing antibodies. This approach enhanced expression of both reporter and therapeutic transgenes. Transduction efficiency achieved following a single dose of FIV expressing mouse erythropoietin was insufficient to increase hematocrit, whereas seven consecutive daily doses significantly increased hematocrit. These unexpected results contrast strikingly with findings reported for adenovirus vectors. Prolonged gene expression has been observed in vivo following a single dose of virus vector; however, depending on the application, repeated administration of vector may be necessary to achieve stable, therapeutic gene expression.

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Section: Methodsmentioning

confidence: 90%

Lentivirus Vector Can Be Readministered to Nasal Epithelia without Blocking Immune Responses

Sinn¹,

Arias²,

Brogden

et al. 2008

J Virol

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“…Balb/c mice (strain code 01B05; Charles River Laboratories, Wilmington, MA) received FIV expressing firefly luciferase. Six to 8-week-old mice were transduced intranasally with FIV or adenovirus (Ad) vector formulated 1:1 with methylcellulose (50 ml total volume), as previously described (21). Animals were imaged at various time points, using the In Vivo Imaging Systems (IVIS) system (Caliper Life Sciences, Hopkinton, MA).…”

Section: Murine Studiesmentioning

confidence: 99%

“…mCC10, hBPIFA1, hCFTR, or hWDR65 candidate promoter elements driving Cre recombinase were packaged in a GP64-pseudotyped FIV vector. About 2.5 3 10 5 TU of each vector (maximum dose) were formulated with 1% methylcellulose (21) and delivered intranasally once a day for 3 days. Cre recombinase delivered to murine nasal epithelia excises the termination codon, which results in luciferase expression.…”

Section: Progenitor Populations Transducedmentioning

confidence: 99%

Transcriptional Targeting in the Airway Using Novel Gene Regulatory Elements

Burnight

Wang²,

McCray³

et al. 2012

Am J Respir Cell Mol Biol

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The delivery of cystic fibrosis transmembrane conductance regulator (CFTR) to airway epithelia is a goal of many gene therapy strategies to treat cystic fibrosis. Because the native regulatory elements of the CFTR are not well characterized, the development of vectors with heterologous promoters of varying strengths and specificity would aid in our selection of optimal reagents for the appropriate expression of the vector-delivered CFTR gene. Here we contrasted the performance of several novel gene-regulatory elements. Based on airway expression analysis, we selected putative regulatory elements from BPIFA1 and WDR65 to investigate. In addition, we selected a human CFTR promoter region (z 2 kb upstream of the human CFTR transcription start site) to study. Using feline immunodeficiency virus vectors containing the candidate elements driving firefly luciferase, we transduced murine nasal epithelia in vivo. Luciferase expression persisted for 30 weeks, which was the duration of the experiment. Furthermore, when the nasal epithelium was ablated using the detergent polidocanol, the mice showed a transient loss of luciferase expression that returned 2 weeks after administration, suggesting that our vectors transduced a progenitor cell population. Importantly, the hWDR65 element drove sufficient CFTR expression to correct the anion transport defect in CFTR-null epithelia. These results will guide the development of optimal vectors for sufficient, sustained CFTR expression in airway epithelia.Keywords: gene transfer; lung; cystic fibrosis Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene cause cystic fibrosis, a multiorgan disease that affects the sinuses, lung, sweat glands, intestines, liver, pancreas, and reproductive tracts (1-3). However, most of the morbidity and mortality associated with the disease results from chronic bacterial infections and inflammation in the lung. Developing gene transfer vectors for efficient, safe, and effective therapeutic expression in the appropriate cell types of the airways is an important consideration in the treatment of cystic fibrosis.CFTR mRNA is present in low copy numbers (1-2 copies per cell) in nasal, tracheal, and bronchial epithelia (4, 5). Previous work in Cftr-null murine models showed that achieving as little as 5% of normal CFTR concentrations resulted in a much larger correction of the chloride transport defect (50% of normal), suggesting that modest levels of transgene expression may confer a significant therapeutic benefit (6). Conversely, CFTR overexpression in airway epithelia may cause protein mislocalization to the basal surface and an overall net decrease in chloride current (7). The entire suite of regulatory elements for the CFTR locus is unknown, but several regions have been identified that span several hundred kilobases (8-12), a size too large to package efficiently in standard viral vectors. Thus, a need exists for vectors with appropriate transgene expression levels in the airway.To that end, we sought to develop vectors c...

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“…In addition, there are several intracellular obstacles, including clearance by uptake into endosomes, and the nuclear membrane that must be breached. 1 Efforts to overcome these barriers with the use of mucolytic agents, 2 viscoelastic gels, 3 or agents that disrupt tight junctions, thus allowing access to receptors present on the basolateral membrane, 4 have enhanced transfection efficiency to airway epithelium, but further improvements would always be welcome.…”

Section: Introductionmentioning

confidence: 99%

Use of ultrasound to enhance nonviral lung gene transfer in vivo

et al. 2007

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We have assessed if high-frequency ultrasound (US) can enhance nonviral gene transfer to the mouse lung. Cationic lipid GL67/pDNA, polyethylenimine (PEI)/pDNA and naked plasmid DNA (pDNA) were delivered via intranasal instillation, mixed with Optison microbubbles, and the animals were then exposed to 1 MHz US. Addition of Optison alone significantly reduced the transfection efficiency of all three gene transfer agents. US exposure did not increase GL67/ pDNA or PEI/pDNA gene transfer compared to Optisontreated animals. However, it increased naked pDNA transfection efficiency by approximately 15-fold compared to Optison-treated animals, suggesting that despite ultrasound being attenuated by air in the lung, sufficient energy penetrates the tissue to increase gene transfer. US-induced lung haemorrhage, assessed histologically, increased with prolonged US exposure. The left lung was more affected than the right and this was mirrored by a lesser increase in naked pDNA gene transfer, in the left lung. The positive effect of US was dependent on Optison, as in its absence US did not increase naked pDNA transfection efficiency. We have thus established proof of principle that US can increase nonviral gene transfer, in the air-filled murine lung.

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Viscoelastic Gel Formulations Enhance Airway Epithelial Gene Transfer with Viral Vectors

Cited by 50 publications

References 30 publications

Lentivirus Vector Can Be Readministered to Nasal Epithelia without Blocking Immune Responses

Lentivirus Vector Can Be Readministered to Nasal Epithelia without Blocking Immune Responses

Transcriptional Targeting in the Airway Using Novel Gene Regulatory Elements

Use of ultrasound to enhance nonviral lung gene transfer in vivo

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