Inhibition of macrophage iNOS by selective targeting of antisense PNA.
Peptide nucleic acids (PNAs) are synthetic polynucleobases that bind to DNA and RNA with high affinity and specificity and with poor membrane permeability. Although PNAs have an enormous potential as antisense agents, the success of antisense PNA applications will require efficient cellular uptake. In this study, a unique antisense 14-mer anti-inducible nitric oxide synthase (iNOS) was encapsulated into erythrocytes (RBC) by hypotonic dialysis. RBC loaded with PNA (10.5 +/- 3.5 micromol/mL RBC) were targeted specifically to murine macrophages, taking advantage of an in vitro opsonization induced by ZnCl(2) and bis-sulfosuccynimidil-suberate (BS(3)). This in vitro opsonization enhanced the phagocytosis of loaded RBC and the delivery of PNA into macrophages (0.72 pmol/10(6) macrophages). The efficacy of this delivery system is demonstrated by decreases in NO production and iNOS protein expression inside the macrophage. Therefore, we can suggest this novel approach for biomedical application.
Chiarantini
Laura
L
Institute of Biochemistry Giorgio Fornaini Università degli Studi di Urbino, Italy. l.chiarantini@uniurb.it
Cerasi
Aurora
A
Fraternale
Alessandra
A
Andreoni
Francesca
F
Scarí
Sonia
S
Giovine
Marco
M
Clavarino
Emanuela
E
Magnani
Mauro
M
D000818:Animals; D016373:DNA, Antisense / Q000494:pharmacology*; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008264:Macrophages / Q000201:enzymology / Q000502:physiology; D051379:Mice; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors* / Q000097:blood / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D020135:Peptide Nucleic Acids / Q000494:pharmacology*; D010587:Phagocytosis; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism
D016373:DNA, Antisense; D020135:Peptide Nucleic Acids; D012333:RNA, Messenger; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse