Extracellular barriers to in Vivo PEI and PEGylated PEI polyplex-mediated gene delivery to the liver.
Polyplex-mediated gene therapy is a promising alternative to viral gene therapy. One challenge to these synthetic carriers is reduced transfection efficiencies in vivo compared to those achieved in vitro. Many of the intracellular barriers to gene delivery have been elucidated, but similar quantification of extracellular barriers to gene delivery remains a need. In this study, the unpackaging of polyplexes by serum proteins, soluble glycosaminoglycans, and an extracellular matrix extract was demonstrated by a YOYO-1 fluorescence quenching assay. Additionally, exposing polyplexes to serum or proteoglycans before in vitro transfection caused decreased cellular uptake of DNA. Lastly, PEI polyplexes and PEGylated PEI polyplexes were injected into the portal vein of mice, and the intrahepatic distributions of labeled DNA and polymer were assessed by confocal microscopy. PEI polyplexes delivered DNA to the liver, but extensive vector unpackaging was observed, with PEI primarily colocalized with the extracellular matrix. PEGylated polyplexes mediated less DNA delivery to the liver, possibly due to premature vector unpackaging in the blood. Through this work, both the blood and the extracellular matrix have been determined to be significant extracellular barriers to polyplex-mediated in vivo gene delivery to the liver.
Burke
Rob S
RS
Department of Bioengineering, University of Washington, 1705 NE Pacific Street, Seattle, WA 98195, USA.
Pun
Suzie H
SH
D016475:3T3 Cells; D000818:Animals; D002626:Chemistry, Pharmaceutical; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D004140:Dinitrophenols / Q000737:chemistry; D016503:Drug Delivery Systems; D005109:Extracellular Matrix / Q000378:metabolism / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism*; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011094:Polyethyleneimine / Q000031:analogs & derivatives* / Q000737:chemistry*; D045663:Quantum Dots; D014162:Transfection
D004140:Dinitrophenols; C521156:branched polyethyleneimine-block-polyethylene glycol; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D004247:DNA