TY - JOUR
T1 - Antisense peptide nucleic acidfunctionalized cationic nanocomplex for in vivo mRNA detection
AU - Shen, Yuefei
AU - Shrestha, Ritu
AU - Ibricevic, Aida
AU - Gunsten, Sean P.
AU - Welch, Michael J.
AU - Wooley, Karen L.
AU - Brody, Steven L.
AU - Taylor, John Stephen A.
AU - Liu, Yongjian
PY - 2013/6/6
Y1 - 2013/6/6
N2 - Acute lung injury (ALI) is a complex syndrome with many aetiologies, resulting in the upregulation of inflammatory mediators in the host, followed by dyspnoea, hypoxemia and pulmonary oedema. A central mediator is inducible nitric oxide synthase (iNOS) that drives the production of NO and continued inflammation. Thus, it is useful to have diagnostic and therapeutic agents for targeting iNOS expression. One general approach is to target the precursor iNOS mRNA with antisense nucleic acids. Peptide nucleic acids (PNAs) have many advantages that make them an ideal platform for development of antisense theranostic agents. Their membrane impermeability, however, limits biological applications. Here, we report the preparation of an iNOS imaging probe through electrostatic complexation between a radiolabelled antisense PNA-YR9. oligodeoxynucleotide (ODN) hybrid and a cationic shell-crosslinked knedel-like nanoparticle (cSCK). The Y (tyrosine) residue was used for 123I radiolabelling, whereas the R9 (arginine9) peptidewas included to facilitate cell exit of untargeted PNA.Complete binding of the antisense PNA-YR9. ODN hybrid to the cSCK was achieved at an 8: 1 cSCK amine to ODN phosphate (N/P) ratio by a gel retardation assay. The antisense PNA-YR9. ODN. cSCK nanocomplexes efficiently entered RAW264.7 cells, whereas the PNAYR9.ODN alone was not taken up. Low concentrations of 123I-labelled antisense PNA-YR9.ODN complexed with cSCK showed significantly higher retention of radioactivity when iNOS was induced in lipopolysaccharide{thorn} interferon-g-activated RAW264.7 cells when compared with a mismatched PNA.Moreover, statistically, greater retention of radioactivity from the antisense complex was also observed in vivo in an iNOS-induced mouse lung after intratracheal administration of the nanocomplexes. This study demonstrates the specificity and sensitivity by which the radiolabelled nanocomplexes can detect iNOS mRNA in vitro and in vivo and their potential for early diagnosis of ALI.
AB - Acute lung injury (ALI) is a complex syndrome with many aetiologies, resulting in the upregulation of inflammatory mediators in the host, followed by dyspnoea, hypoxemia and pulmonary oedema. A central mediator is inducible nitric oxide synthase (iNOS) that drives the production of NO and continued inflammation. Thus, it is useful to have diagnostic and therapeutic agents for targeting iNOS expression. One general approach is to target the precursor iNOS mRNA with antisense nucleic acids. Peptide nucleic acids (PNAs) have many advantages that make them an ideal platform for development of antisense theranostic agents. Their membrane impermeability, however, limits biological applications. Here, we report the preparation of an iNOS imaging probe through electrostatic complexation between a radiolabelled antisense PNA-YR9. oligodeoxynucleotide (ODN) hybrid and a cationic shell-crosslinked knedel-like nanoparticle (cSCK). The Y (tyrosine) residue was used for 123I radiolabelling, whereas the R9 (arginine9) peptidewas included to facilitate cell exit of untargeted PNA.Complete binding of the antisense PNA-YR9. ODN hybrid to the cSCK was achieved at an 8: 1 cSCK amine to ODN phosphate (N/P) ratio by a gel retardation assay. The antisense PNA-YR9. ODN. cSCK nanocomplexes efficiently entered RAW264.7 cells, whereas the PNAYR9.ODN alone was not taken up. Low concentrations of 123I-labelled antisense PNA-YR9.ODN complexed with cSCK showed significantly higher retention of radioactivity when iNOS was induced in lipopolysaccharide{thorn} interferon-g-activated RAW264.7 cells when compared with a mismatched PNA.Moreover, statistically, greater retention of radioactivity from the antisense complex was also observed in vivo in an iNOS-induced mouse lung after intratracheal administration of the nanocomplexes. This study demonstrates the specificity and sensitivity by which the radiolabelled nanocomplexes can detect iNOS mRNA in vitro and in vivo and their potential for early diagnosis of ALI.
KW - Acute lung injury
KW - Cationic nanoparticles
KW - Inducible nitric oxide synthase
KW - Peptide nucleic acid
KW - Radiolabelling
KW - Targeting
UR - http://www.scopus.com/inward/record.url?scp=84876528645&partnerID=8YFLogxK
U2 - 10.1098/rsfs.2012.0059
DO - 10.1098/rsfs.2012.0059
M3 - Article
C2 - 24427537
AN - SCOPUS:84876528645
SN - 2042-8898
VL - 3
JO - Interface Focus
JF - Interface Focus
IS - 3
ER -