Bioavailability of antisense oligonucleotides in neuroblastoma cells: Comparison of efficacy among different types of molecules

Maria Valeria Corrias, Francesco Guarnaccia, Mirco Ponzoni

Research output: Contribution to journalArticle

Abstract

To evaluate the real effectiveness of various chemical modifications in enhancing the ability of antisense molecules to inhibit gene expression, the toxicity, stability, uptake, and intracellular localization of an identical sequence, synthetized either with a phosphodiester or a phosphorothioate backbone, with or without a cholesteryl moiety linked to the 3'-end, were compared in three different human neuroblastoma cell lines. The toxicity, assessed by inhibition of cell viability, greatly depend on the presence of the lipid moiety and to a less extent on the cell line used. At high doses all the antisenses caused a necrotic lysis of plasma membranes. Typical features of apoptotic cell death were never observed. The presence of the lipid moiety enhanced the uptake of antisense molecules while the phosphorothioate backbone, as expected, conferred higher stability. At late times, therefore, the combination of lipid conjugation and phosphorothioate backbone seems to be the most effective in obtaining a consistent antisense accumulation inside the cells. The presence of the cholesteryl moiety also caused a stronger association of the antisense to membraneous compartments, so that a quite different biodistribution occurred among the four antisenses tested. However, the actual amount of antisense molecules found inside NE cells was low in all the conditions tested. Only following cellular permeabilization a significant uptake was obtained, making the use of delivery system mandatory to achieve an efficient inhibition of highly expressed genes.

Original languageEnglish
Pages (from-to)171-180
Number of pages10
JournalJournal of Neuro-Oncology
Volume31
Issue number1-2
DOIs
Publication statusPublished - 1997

Fingerprint

Antisense Oligonucleotides
Neuroblastoma
Biological Availability
Lipids
Cell Line
Cell Survival
Cell Death
Cell Membrane
Gene Expression
Genes

Keywords

  • Antisense
  • Capped oligonucleotides
  • Intracellular distribution
  • Neuroblastoma
  • Oligonucleotides
  • Uptake

ASJC Scopus subject areas

  • Clinical Neurology
  • Cancer Research
  • Oncology
  • Neuroscience(all)

Cite this

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abstract = "To evaluate the real effectiveness of various chemical modifications in enhancing the ability of antisense molecules to inhibit gene expression, the toxicity, stability, uptake, and intracellular localization of an identical sequence, synthetized either with a phosphodiester or a phosphorothioate backbone, with or without a cholesteryl moiety linked to the 3'-end, were compared in three different human neuroblastoma cell lines. The toxicity, assessed by inhibition of cell viability, greatly depend on the presence of the lipid moiety and to a less extent on the cell line used. At high doses all the antisenses caused a necrotic lysis of plasma membranes. Typical features of apoptotic cell death were never observed. The presence of the lipid moiety enhanced the uptake of antisense molecules while the phosphorothioate backbone, as expected, conferred higher stability. At late times, therefore, the combination of lipid conjugation and phosphorothioate backbone seems to be the most effective in obtaining a consistent antisense accumulation inside the cells. The presence of the cholesteryl moiety also caused a stronger association of the antisense to membraneous compartments, so that a quite different biodistribution occurred among the four antisenses tested. However, the actual amount of antisense molecules found inside NE cells was low in all the conditions tested. Only following cellular permeabilization a significant uptake was obtained, making the use of delivery system mandatory to achieve an efficient inhibition of highly expressed genes.",
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AU - Ponzoni, Mirco

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