Nebulisation of Receptor-Targeted Nanocomplexes for Gene Delivery to the Airway Epithelium 英文参考文献.docVIP

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Nebulisation of Receptor-Targeted Nanocomplexes for Gene Delivery to the Airway Epithelium 英文参考文献.doc

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Nebulisation of Receptor-Targeted Nanocomplexes for Gene Delivery to the Airway Epithelium 英文参考文献

NebulisationofReceptor-TargetedNanocomplexesfor GeneDeliverytotheAirwayEpithelium MariaD.I.Manunta1,RobinJ.McAnulty2,AristidesD.Tagalakis1,StephenE.Bottoms2 ,Frederick Campbell3,HelenC.Hailes3,AletheaB.Tabor3,GeoffreyJ.Laurent2,ChristopherO’Callaghan4, StephenL.Hart1* 1WolfsonCentreforGeneTherapyofChildhoodDisease,UCLInstituteofChildHealth,UniversityCollegeLondon,London,UnitedKingdom,2CentreforRespiratory Research, University College London, London, United Kingdom, 3Department of Chemistry, University College London, London, United Kingdom, 4Department of Infection,ImmunityandInflammation,UniversityofLeicester,LeicesterRoyalInfirmary,Leicester,UnitedKingdom Abstract Background:Genetherapymediatedbysyntheticvectorsmayprovideopportunitiesfornewtreatmentsforcysticfibrosis (CF)viaaerosolisation.VectorsforCFmusttransfecttheairwayepitheliumefficientlyandnotcauseinflammationsothey are suitable for repeated dosing. The inhaled aerosol should be deposited in the airways since the cystic fibrosis transmembraneconductanceregulatorgene(CFTR)isexpressedpredominantlyintheepitheliumofthesubmucosalglands and in the surface airway epithelium. The aim of this project was to develop an optimised aerosol delivery approach applicabletotreatmentofCFlungdiseasebygenetherapy. Methodology:Thevectorsuspensioninvestigatedinthisstudycomprisesreceptor-targetingpeptides,cationicliposomes and plasmid DNA that self-assemble by electrostatic interactions to form a receptor-targeted nanocomplex (RTN) of approximately150nmwithacationicsurfacechargeof+50mV.Theaerodynamicpropertiesofaerosolisednanocomplexes producedwiththreedifferentnebuliserswerecomparedbydeterminingaerosoldepositioninthedifferentstagesofaNext Generation Pharmaceutical Impactor (NGI). We also investigated the yield of intact plasmid DNA by agarose gel electrophoresisanddensitometry,andtransfectionefficaciesinvitroandinvivo. Results:RTNsnebulisedwiththeAeroEclipseIIBANwerethemosteffective,comparedtoothernebuliserstested,forgene deliv