Targeted delivery of complexes of biotin-PEG-polyethylenimine and NF-kappaB decoys to brain-derived endothelial cells in vitro.
OBJECTIVE
To evaluate the effect of re-directing the uptake mechanism of polyplexes containing oligodeoxynucleotide (ODN) decoys to nuclear factor kappa B (NF-kappaB) from absorptive-mediated to receptor-mediated endocytosis.


METHODS
Complexes of ODNs and a co-polymer of biotin-polyethylenglycol and polyethylenimine (BPP) were targeted to brain-derived endothelial cells with a conjugate of antibody 8D3 and streptavidin (8D3SA). Size and stability of ODN/BPP complexes was measured by dynamic light scattering. Cellular uptake was studied by confocal microscopy. Cell viability and pharmacological effects were investigated on murine bEnd5 cells stimulated with tumor necrosis factor.


RESULTS
ODN/BPP complexes showed sizes of 116+/-2.3 nm, which increased by 40 nm when coupled to 8D3SA, and were stable in physiological fluids. Targeted complexes were internalized intact into endosomal compartments. Treatment conditions, which yielded significant inhibitory effects on mRNA expression of VCAM-1, ICAM-1, IkappaBalpha and iNOS by bEnd5 cells, did not affect viability. At 0.5 microM, decoy ODN significantly inhibited monocyte adhesion to bEnd5 monolayers when delivered as 8D3SA-targeted complex, while higher concentrations of untargeted complex were ineffective.


CONCLUSIONS
The complex of NF-kappaB decoys and BPP, which can be targeted to transferrin receptors, is a promising drug candidate for neuroinflammatory diseases affecting the blood-brain barrier.
Bhattacharya
Raktima
R
Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center School of Pharmacy, 1300 Coulter Drive, Amarillo, Texas 79106, USA.
Osburg
Berit
B
Fischer
Dagmar
D
Bickel
Ulrich
U
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism* / Q000494:pharmacology; D001710:Biotin / Q000378:metabolism* / Q000633:toxicity; D001921:Brain / Q000098:blood supply*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002461:Cell Line, Transformed; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004337:Drug Carriers; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism* / Q000494:pharmacology; D010316:Particle Size; D011092:Polyethylene Glycols / Q000378:metabolism* / Q000633:toxicity; D011094:Polyethyleneimine / Q000378:metabolism* / Q000633:toxicity; D012333:RNA, Messenger / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism*; D013997:Time Factors; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D015815:Cell Adhesion Molecules; D004337:Drug Carriers; D016328:NF-kappa B; D009841:Oligonucleotides; D012333:RNA, Messenger; D011990:Receptors, Transferrin; D014409:Tumor Necrosis Factor-alpha; D011092:Polyethylene Glycols; D001710:Biotin; D011094:Polyethyleneimine