Enhanced lung gene expression after aerosol delivery of concentrated pDNA/PEI complexes.
A major limitation of many self-assembling nonviral gene transfer formulations is that they are commonly prepared at relatively low component concentrations. While this typically has little impact on their use in cell culture, it can severely limit the progress of in vivo studies. In order to overcome this, we have developed a simple, scalable, pharmaceutically acceptable concentration method that has allowed us to increase the concentration of a commonly used pDNA/PEI formulation from 0.2 to >8 mg/ml plasmid DNA (pDNA). Crucially, the concentration method was found to have only minimal impact on the electrostatic properties or size of the pDNA/PEI particles. When delivered as an aerosol to the mouse lung, the concentrated pDNA/PEI formulations resulted in a 15-fold increase in lung reporter gene expression, with minimal impact in terms of inflammation or toxicity. Importantly, this performance advantage was replicated after aerosol administration to sheep lungs, with reporter gene expression being similarly approximately 15-fold higher than with the conventional pDNA/PEI formulation, and lung inflammation falling to background levels. These findings demonstrate that concentrated pDNA/PEI formulations offer increased aerosol gene transfer with decreased inflammatory sequelae, and represent a promising advance in the field of nonviral lung gene transfer. It seems likely that similar benefits might be achievable with alternative delivery routes and with other nonviral formulations.
Davies
Lee A
LA
Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK.
McLachlan
Gerry
G
Sumner-Jones
Stephanie G
SG
Ferguson
David
D
Baker
Alison
A
Tennant
Peter
P
Gordon
Catherine
C
Vrettou
Christina
C
Baker
Eilidh
E
Zhu
Jie
J
Alton
Eric W F W
EW
Collie
David D S
DD
Porteous
David J
DJ
Hyde
Stephen C
SC
Gill
Deborah R
DR
D000336:Aerosols; D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008168:Lung / Q000378:metabolism*; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D011094:Polyethyleneimine / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D012756:Sheep
D000336:Aerosols; D011094:Polyethyleneimine; D004247:DNA