Antisense peptide nucleic acidfunctionalized cationic nanocomplex for in vivo mRNA detection

Yuefei Shen, Ritu Shrestha, Aida Ibricevic, Sean P. Gunsten, Michael J. Welch, Karen L. Wooley, Steven L. Brody, John Stephen A. Taylor, Yongjian Liu

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

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.

Original languageEnglish
JournalInterface Focus
Volume3
Issue number3
DOIs
StatePublished - Jun 6 2013

Keywords

  • Acute lung injury
  • Cationic nanoparticles
  • Inducible nitric oxide synthase
  • Peptide nucleic acid
  • Radiolabelling
  • Targeting

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    Shen, Y., Shrestha, R., Ibricevic, A., Gunsten, S. P., Welch, M. J., Wooley, K. L., Brody, S. L., Taylor, J. S. A., & Liu, Y. (2013). Antisense peptide nucleic acidfunctionalized cationic nanocomplex for in vivo mRNA detection. Interface Focus, 3(3). https://doi.org/10.1098/rsfs.2012.0059