Non–invasive liposome–mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice

Alton, Eric W.F.W.; Middleton, Peter G.; Caplen, Natasha J.; Smith, Stephen N.; Steel, D. M.; Munkonge, Felix M.; Jeffery, P. K.; Geddes, Duncan M.; Hart, SL; Williamson, Robert; Fasold, K. I.; Miller, Andrew D.; Dickinson, Paul; Stevenson, Barbara; McLachlan, Gerry; Dorin, Julia R.; Porteous, David J.

doi:10.1038/ng1093-135

Cited by 384 publications

(110 citation statements)

References 30 publications

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“…Host inflammatory or immune response to this ligand is not expected to occur. Recent results of the toxicity study of liposomes following their delivery to the lungs of human 32 and animals 33,34 showed minimal to no host inflammatory response. Therefore, we do not anticipate significant adverse immunological or inflammatory response to our transfection vector when applied in vivo.…”

Section: Discussionmentioning

confidence: 99%

Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells

Yanagihara

Cheng²,

Cheng³

2000

Cancer Gene Ther

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Poor transfection efficiency is the major drawback of lipofection. We showed previously that addition of transferrin (TF) to Lipofectin enhanced the expression of a reporter gene in HeLa cells by 120-fold and achieved close to 100% transfection efficiency. The purpose of this study was to determine whether TF and other ligands could improve the efficiency of lipofection in lung carcinoma cells. Confluent A549, Calu3, and H292 cells were transfected for 18 hours with a plasmid DNA (pCMVlacZ) using Lipofectin plus TF, insulin, or epidermal growth factor as the vector. The transfected cells were assessed for transfection efficiency by ␤-galactosidase activity (light units/g protein) and the percentage of blue cells following 5-bromo-4-chloro-3-indolyl ␤-Dgalactopyranoside staining. Lipofectin supplemented with epidermal growth factor yielded the largest enhancement of lipofection efficiency (Յ23-fold over that by Lipofectin alone) in all three cell lines. Insulin significantly enhanced the lipofection efficiency in A549 and Calu3 cells but not in H292 cells, whereas TF showed significant lipofection efficiency-enhancing effect in Calu3 and H292 cells but not in A549 cells. The transfection efficiency correlated well with the amounts of DNA delivered to the nucleus as well as the amounts of the receptor. These results indicate that the gene delivery strategy employing ligand-facilitated lipofection can achieve high transfection efficiency in human lung carcinoma cells. In addition, enhancement of the expression of the receptor may be a possible strategy for increasing the efficiency of gene targeting. Cancer Gene Therapy (2000) 7, 59 -65

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Section: Discussionmentioning

confidence: 99%

Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells

Yanagihara

Cheng²,

Cheng³

2000

Cancer Gene Ther

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“…Cationic liposomes such as DOTAP therefore have been used to enhance delivery of ASO to target cells (23,29,30). For example, the CFTR gene has been complexed with DOTAP and successfully administered to and expressed in the nasal epithelium of humans with cystic fibrosis (30).…”

Section: Discussionmentioning

confidence: 99%

Aerosolized Syk Antisense Suppresses Syk Expression, Mediator Release from Macrophages, and Pulmonary Inflammation

Stenton¹,

Kim

Nohara³

et al. 2000

The Journal of Immunology

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Syk protein tyrosine kinase (PTK) is involved in signaling in leukocytes. In macrophages, Fcγ-receptor cross-linking induces Syk PTK phosphorylation and activation, resulting in Syk-dependent events required for phagocytosis and mediator release. We hypothesized that Syk antisense oligodeoxynucleotides (ASO) delivered by aerosol to rat lungs in vivo would depress Syk PTK expression, mediator release from alveolar macrophages, and Syk-dependent pulmonary inflammation. RT-PCR and RT-in situ PCR demonstrated that aerosolized Syk ASO administration reduced Syk mRNA expression from alveolar macrophages compared with cells isolated from sham-treated rats. Western blot analysis confirmed that Syk PTK expression was reduced after Syk ASO treatment. Compared with sham-treated rats (scrambled oligodeoxynucleotide), Syk ASO treatment suppressed Fcγ-receptor-mediated nitric oxide (86.0 ± 8.3%) and TNF (73.1 ± 3.1%) production by alveolar macrophages stimulated with IgG-anti-IgG complexes. In contrast, Fcγ-receptor-induced IL-1β release was unaffected by Syk ASO treatment. Additionally, Syk ASO suppressed Ag-induced pulmonary inflammation, suggesting that Syk ASO may prove useful as an anti-inflammatory therapy in disorders such as asthma.

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“…Studies in cystic fibrosis transmembrane conductance regulator (Cftr) mutant mice are thought to be useful for testing whether a gene therapy vector can rescue the defects associated with lack of fully functional CFTR in the airway epithelium, such as loss of cyclic adenosine monophosphate-dependent chloride channel activity and/or regulation of epithelial sodium absorption via the sodium channel EnaC. [2][3][4] However, studies in the mouse alone are likely insufficient to predict clinical efficiency in CF patients because there are significant differences in disease pathology and airway physiology between mouse and human. Although large animal CF models continue to be developed [5][6][7] these are not yet widely available; thus, we have successfully used wild-type sheep to evaluate the safety and efficacy of lung-directed gene therapy in a clinically relevant way.…”

Section: Introductionmentioning

confidence: 99%

Pre-clinical evaluation of three non-viral gene transfer agents for cystic fibrosis after aerosol delivery to the ovine lung

et al. 2011

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We use both large and small animal models in our pre-clinical evaluation of gene transfer agents (GTAs) for cystic fibrosis (CF) gene therapy. Here, we report the use of a large animal model to assess three non-viral GTAs: 25 kDa-branched polyethyleneimine (PEI), the cationic liposome (GL67A) and compacted DNA nanoparticle formulated with polyethylene glycol-substituted lysine 30-mer. GTAs complexed with plasmids expressing human cystic fibrosis transmembrane conductance regulator (CFTR) complementary DNA were administered to the sheep lung (n¼8 per group) by aerosol. All GTAs gave evidence of gene transfer and expression 1 day after treatment. Vector-derived mRNA was expressed in lung tissues, including epithelial cell-enriched bronchial brushing samples, with median group values reaching 1-10% of endogenous CFTR mRNA levels. GL67A gave the highest levels of expression. Human CFTR protein was detected in small airway epithelial cells in some animals treated with GL67A (two out of eight) and PEI (one out of eight). Bronchoalveolar lavage neutrophilia, lung histology and elevated serum haptoglobin levels indicated that gene delivery was associated with mild local and systemic inflammation. Our conclusion was that GL67A was the best non-viral GTA currently available for aerosol delivery to the sheep lung, led to the selection of GL67A as our lead GTA for clinical trials in CF patients.

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Non–invasive liposome–mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice

Cited by 384 publications

References 30 publications

Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells

Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells

Aerosolized Syk Antisense Suppresses Syk Expression, Mediator Release from Macrophages, and Pulmonary Inflammation

Pre-clinical evaluation of three non-viral gene transfer agents for cystic fibrosis after aerosol delivery to the ovine lung

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