Mechanism of efficient transfection of the nasal airway epithelium by hypotonic shock

Lemoine, Jérôme; Farley, Raymond; Huang, Leaf

doi:10.1038/sj.gt.3302548

Cited by 24 publications

(14 citation statements)

References 38 publications

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“…Huang et al demonstrated that hypotonic shock enhanced uptake of plasmid DNA by nasal epithelium in vivo via the regulatory volume decrease (RVD) mechanism [317]. Specifically, they showed that sodium- and sucrose-based hypotonic vehicles provided enhanced transgene expression in the mouse nasal epithelium compared to isotonic vehicles with the greater effect observed with lower osmolality.…”

Section: Strategies To Overcome the Barriers To Inhaled Gene Therapymentioning

confidence: 99%

“…During this endocytic process, particulates, including inhaled gene vectors, in the vicinity of the apical membrane can be taken up by epithelial cells. The RVD effect has been shown to last up to 30 minutes [317, 319]. Potential safety concern resulting from hypotonic shock would need to be addressed before clinical implementation of this approach.…”

Section: Strategies To Overcome the Barriers To Inhaled Gene Therapymentioning

confidence: 99%

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Barriers to inhaled gene therapy of obstructive lung diseases: A review

Kim

Duncan

Hanes

et al. 2016

Journal of Controlled Release

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Knowledge of genetic origins of obstructive lung diseases has made inhaled gene therapy an attractive alternative to the current standards of care that are limited to managing disease symptoms. Initial lung gene therapy clinical trials occurred in the early 1990s following the discovery of the genetic defect responsible for cystic fibrosis (CF), a monogenic disorder. However, despite over two decades of intensive effort, gene therapy has yet to help patients with CF or any other obstructive lung disease. The slow progress is due in part to poor understanding of the biological barriers to inhaled gene therapy. Encouragingly, clinical trials have shown that inhaled gene therapy with various viral vectors and non-viral gene vectors is well tolerated by patients, and continued research has provided valuable lessons and resources that may lead to future success of this therapeutic strategy. In this review, we first introduce representative obstructive lung diseases and examine limitations of currently available therapeutic options. We then review key components for successful execution of inhaled gene therapy, including gene delivery systems, primary physiological barriers and strategies to overcome them, and advances in preclinical disease models with which the most promising systems may be identified for human clinical trials.

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Section: Strategies To Overcome the Barriers To Inhaled Gene Therapymentioning

confidence: 99%

Section: Strategies To Overcome the Barriers To Inhaled Gene Therapymentioning

confidence: 99%

Barriers to inhaled gene therapy of obstructive lung diseases: A review

Kim

Duncan

Hanes

et al. 2016

Journal of Controlled Release

View full text Add to dashboard Cite

show abstract

“…First, we applied hypotonicity mucosally in order to imitate mucosal osmotic changes encountered by simple epithelial cells in a physiologic or therapeutic setting (25)(26)(27). Mucosal hypotonicity of 18.4% elicited a robust and sustained rise in the I sc (DI sc ), with the current amplitudes and channel-inhibitor sensitivities being similar to those in DI sc activated by forskolin, a classic activator of CFTR ( Fig.…”

Section: Mucosal Hypotonicity Activated Apical Cftr In Calu-3 Epithelmentioning

confidence: 99%

Mechanosensitivity of wild‐type and G551D cystic fibrosis transmembrane conductance regulator (CFTR) controls regulatory volume decrease in simple epithelia

et al. 2015

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Mutations of cystic fibrosis transmembrane conductance regulator (CFTR), an epithelial ligand-gated anion channel, are associated with the lethal genetic disease cystic fibrosis. The CFTR G551D mutation impairs ATP hydrolysis and thereby makes CFTR refractory to cAMP stimulation. Both wild-type (WT) and G551D CFTR have been implicated in regulatory volume decrease (RVD), but the underlying mechanism remains incompletely understood. Here, we show that the channel activity of both WT and G551D CFTR is directly stimulated by mechanical perturbation induced by cell swelling at the single-channel, cellular, and tissue levels. Hypotonicity activated CFTR single channels in cell-attached membrane patches and WT-CFTR-mediated short-circuit current (I sc ) in Calu-3 cells, and this was independent of Ca 2+ and cAMP/PKA signaling. Genetic suppression and ablation but not G551D mutation of CFTR suppressed the hypotonicity-and stretch-induced I sc in Calu-3 cells and mouse duodena. Moreover, ablation but not G551D mutation of the CFTR gene inhibited the RVD of crypts isolated from mouse intestine; more importantly, CFTR-specific blockers markedly suppressed RVD in both WT-and G551D CFTR mice, demonstrating for the first time that the channel activity of both WT and G551D CFTR is required for epithelial RVD. Our findings uncover a previously unrecognized mechanism underlying CFTR involvement in epithelial RVD and suggest that the mechanosensitivity of G551D CFTR might underlie the mild phenotypes resulting from this

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“…Previous studies have suggested that hypotonic solutions can increase gene transfer efficacy. 36 Thus, we cannot exclude that the control animals (GL67/pDNA) showed values higher than if we had been able to use saline as the diluent for this group. To characterize the magnetic particles further, particle sizing was attempted but the polydispersity index indicated that all solutions were too heterogeneous in size to provide accurate data.…”

Section: Figurementioning

confidence: 99%