Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium

Maria D I Manunta, Robin J McAnulty, Aristides D Tagalakis, Stephen E Bottoms, Frederick Campbell, Helen C Hailes, Alethea B Tabor, Geoffrey J Laurent, Christopher O'Callaghan, Stephen L Hart

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

BACKGROUND: Gene therapy mediated by synthetic vectors may provide opportunities for new treatments for cystic fibrosis (CF) via aerosolisation. Vectors for CF must transfect the airway epithelium efficiently and not cause inflammation so they are suitable for repeated dosing. The inhaled aerosol should be deposited in the airways since the cystic fibrosis transmembrane conductance regulator gene (CFTR) is expressed predominantly in the epithelium of the submucosal glands and in the surface airway epithelium. The aim of this project was to develop an optimised aerosol delivery approach applicable to treatment of CF lung disease by gene therapy.

METHODOLOGY: The vector suspension investigated in this study comprises receptor-targeting peptides, cationic liposomes and plasmid DNA that self-assemble by electrostatic interactions to form a receptor-targeted nanocomplex (RTN) of approximately 150 nm with a cationic surface charge of +50 mV. The aerodynamic properties of aerosolised nanocomplexes produced with three different nebulisers were compared by determining aerosol deposition in the different stages of a Next Generation Pharmaceutical Impactor (NGI). We also investigated the yield of intact plasmid DNA by agarose gel electrophoresis and densitometry, and transfection efficacies in vitro and in vivo.

RESULTS: RTNs nebulised with the AeroEclipse II BAN were the most effective, compared to other nebulisers tested, for gene delivery both in vitro and in vivo. The biophysical properties of the nanocomplexes were unchanged after nebulisation while the deposition of RTNs suggested a range of aerosol aerodynamic sizes between 5.5 µm-1.4 µm cut off (NGI stages 3-6) compatible with deposition in the central and lower airways.

CONCLUSIONS: RTNs showed their ability at delivering genes via nebulisation, thus suggesting their potential applications for therapeutic interventions of cystic fibrosis and other respiratory disorders.

Original languageEnglish
Pages (from-to)e26768
JournalPLoS ONE
Volume6
Issue number10
DOIs
Publication statusPublished - 26 Oct 2011

Fingerprint

atomization
cystic fibrosis
Aerosols
gene transfer
Cystic Fibrosis
aerosols
epithelium
Epithelium
Genes
Gene therapy
receptors
Nebulizers and Vaporizers
atomizers
Genetic Therapy
gene therapy
aerodynamics
Aerodynamics
Plasmids
respiratory tract diseases
plasmids

Keywords

  • Cystic Fibrosis/therapy
  • Epithelium/metabolism
  • Gene Transfer Techniques
  • Genetic Therapy/methods
  • Molecular Targeted Therapy
  • Nanocomposites/administration & dosage
  • Nebulizers and Vaporizers
  • Plasmids/administration & dosage
  • Respiratory Mucosa/metabolism
  • Transfection

Cite this

Manunta, M. D. I., McAnulty, R. J., Tagalakis, A. D., Bottoms, S. E., Campbell, F., Hailes, H. C., ... Hart, S. L. (2011). Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium. PLoS ONE, 6(10), e26768. https://doi.org/10.1371/journal.pone.0026768
Manunta, Maria D I ; McAnulty, Robin J ; Tagalakis, Aristides D ; Bottoms, Stephen E ; Campbell, Frederick ; Hailes, Helen C ; Tabor, Alethea B ; Laurent, Geoffrey J ; O'Callaghan, Christopher ; Hart, Stephen L. / Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium. In: PLoS ONE. 2011 ; Vol. 6, No. 10. pp. e26768.
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abstract = "BACKGROUND: Gene therapy mediated by synthetic vectors may provide opportunities for new treatments for cystic fibrosis (CF) via aerosolisation. Vectors for CF must transfect the airway epithelium efficiently and not cause inflammation so they are suitable for repeated dosing. The inhaled aerosol should be deposited in the airways since the cystic fibrosis transmembrane conductance regulator gene (CFTR) is expressed predominantly in the epithelium of the submucosal glands and in the surface airway epithelium. The aim of this project was to develop an optimised aerosol delivery approach applicable to treatment of CF lung disease by gene therapy.METHODOLOGY: The vector suspension investigated in this study comprises receptor-targeting peptides, cationic liposomes and plasmid DNA that self-assemble by electrostatic interactions to form a receptor-targeted nanocomplex (RTN) of approximately 150 nm with a cationic surface charge of +50 mV. The aerodynamic properties of aerosolised nanocomplexes produced with three different nebulisers were compared by determining aerosol deposition in the different stages of a Next Generation Pharmaceutical Impactor (NGI). We also investigated the yield of intact plasmid DNA by agarose gel electrophoresis and densitometry, and transfection efficacies in vitro and in vivo.RESULTS: RTNs nebulised with the AeroEclipse II BAN were the most effective, compared to other nebulisers tested, for gene delivery both in vitro and in vivo. The biophysical properties of the nanocomplexes were unchanged after nebulisation while the deposition of RTNs suggested a range of aerosol aerodynamic sizes between 5.5 µm-1.4 µm cut off (NGI stages 3-6) compatible with deposition in the central and lower airways.CONCLUSIONS: RTNs showed their ability at delivering genes via nebulisation, thus suggesting their potential applications for therapeutic interventions of cystic fibrosis and other respiratory disorders.",
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Manunta, MDI, McAnulty, RJ, Tagalakis, AD, Bottoms, SE, Campbell, F, Hailes, HC, Tabor, AB, Laurent, GJ, O'Callaghan, C & Hart, SL 2011, 'Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium', PLoS ONE, vol. 6, no. 10, pp. e26768. https://doi.org/10.1371/journal.pone.0026768

Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium. / Manunta, Maria D I; McAnulty, Robin J; Tagalakis, Aristides D; Bottoms, Stephen E; Campbell, Frederick; Hailes, Helen C; Tabor, Alethea B; Laurent, Geoffrey J; O'Callaghan, Christopher; Hart, Stephen L.

In: PLoS ONE, Vol. 6, No. 10, 26.10.2011, p. e26768.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Nebulisation of receptor-targeted nanocomplexes for gene delivery to the airway epithelium

AU - Manunta, Maria D I

AU - McAnulty, Robin J

AU - Tagalakis, Aristides D

AU - Bottoms, Stephen E

AU - Campbell, Frederick

AU - Hailes, Helen C

AU - Tabor, Alethea B

AU - Laurent, Geoffrey J

AU - O'Callaghan, Christopher

AU - Hart, Stephen L

PY - 2011/10/26

Y1 - 2011/10/26

N2 - BACKGROUND: Gene therapy mediated by synthetic vectors may provide opportunities for new treatments for cystic fibrosis (CF) via aerosolisation. Vectors for CF must transfect the airway epithelium efficiently and not cause inflammation so they are suitable for repeated dosing. The inhaled aerosol should be deposited in the airways since the cystic fibrosis transmembrane conductance regulator gene (CFTR) is expressed predominantly in the epithelium of the submucosal glands and in the surface airway epithelium. The aim of this project was to develop an optimised aerosol delivery approach applicable to treatment of CF lung disease by gene therapy.METHODOLOGY: The vector suspension investigated in this study comprises receptor-targeting peptides, cationic liposomes and plasmid DNA that self-assemble by electrostatic interactions to form a receptor-targeted nanocomplex (RTN) of approximately 150 nm with a cationic surface charge of +50 mV. The aerodynamic properties of aerosolised nanocomplexes produced with three different nebulisers were compared by determining aerosol deposition in the different stages of a Next Generation Pharmaceutical Impactor (NGI). We also investigated the yield of intact plasmid DNA by agarose gel electrophoresis and densitometry, and transfection efficacies in vitro and in vivo.RESULTS: RTNs nebulised with the AeroEclipse II BAN were the most effective, compared to other nebulisers tested, for gene delivery both in vitro and in vivo. The biophysical properties of the nanocomplexes were unchanged after nebulisation while the deposition of RTNs suggested a range of aerosol aerodynamic sizes between 5.5 µm-1.4 µm cut off (NGI stages 3-6) compatible with deposition in the central and lower airways.CONCLUSIONS: RTNs showed their ability at delivering genes via nebulisation, thus suggesting their potential applications for therapeutic interventions of cystic fibrosis and other respiratory disorders.

AB - BACKGROUND: Gene therapy mediated by synthetic vectors may provide opportunities for new treatments for cystic fibrosis (CF) via aerosolisation. Vectors for CF must transfect the airway epithelium efficiently and not cause inflammation so they are suitable for repeated dosing. The inhaled aerosol should be deposited in the airways since the cystic fibrosis transmembrane conductance regulator gene (CFTR) is expressed predominantly in the epithelium of the submucosal glands and in the surface airway epithelium. The aim of this project was to develop an optimised aerosol delivery approach applicable to treatment of CF lung disease by gene therapy.METHODOLOGY: The vector suspension investigated in this study comprises receptor-targeting peptides, cationic liposomes and plasmid DNA that self-assemble by electrostatic interactions to form a receptor-targeted nanocomplex (RTN) of approximately 150 nm with a cationic surface charge of +50 mV. The aerodynamic properties of aerosolised nanocomplexes produced with three different nebulisers were compared by determining aerosol deposition in the different stages of a Next Generation Pharmaceutical Impactor (NGI). We also investigated the yield of intact plasmid DNA by agarose gel electrophoresis and densitometry, and transfection efficacies in vitro and in vivo.RESULTS: RTNs nebulised with the AeroEclipse II BAN were the most effective, compared to other nebulisers tested, for gene delivery both in vitro and in vivo. The biophysical properties of the nanocomplexes were unchanged after nebulisation while the deposition of RTNs suggested a range of aerosol aerodynamic sizes between 5.5 µm-1.4 µm cut off (NGI stages 3-6) compatible with deposition in the central and lower airways.CONCLUSIONS: RTNs showed their ability at delivering genes via nebulisation, thus suggesting their potential applications for therapeutic interventions of cystic fibrosis and other respiratory disorders.

KW - Cystic Fibrosis/therapy

KW - Epithelium/metabolism

KW - Gene Transfer Techniques

KW - Genetic Therapy/methods

KW - Molecular Targeted Therapy

KW - Nanocomposites/administration & dosage

KW - Nebulizers and Vaporizers

KW - Plasmids/administration & dosage

KW - Respiratory Mucosa/metabolism

KW - Transfection

U2 - 10.1371/journal.pone.0026768

DO - 10.1371/journal.pone.0026768

M3 - Article

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VL - 6

SP - e26768

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

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