Pyridylthiourea-grafted polyethylenimine offers an effective assistance to siRNA-mediated gene silencing in vitro and in vivo

J Control Release. 2012 Feb 10;157(3):418-26. doi: 10.1016/j.jconrel.2011.10.007. Epub 2011 Oct 12.

Abstract

Success of synthetic interfering nucleic acids (siRNAs)-based therapy relies almost exclusively on effective, safe and preferably nanometric delivery systems which can be easily prepared, even at high concentrations. We prepared by chemical synthesis various self-assembling polymers to entrap siRNAs into stable polyplexes outside cells but with a disassembly potential upon sensing endosomal acidity. Our results revealed that pyridylthiourea-grafted polyethylenimine (πPΕΙ) followed the above-mentioned principles. It led to above 90% siRNA-mediated gene silencing in vitro on U87 cells at 10 nM siRNA concentration and did not have a hemolytic activity. Assembly of siRNA/πPΕΙ at high concentration was then studied and 4.5% glucose solution, pH 6.0, yielded stable colloidal solutions with sizes slightly below 100 nm for several hours. A single injection of these concentrated siRNA polyplexes into luciferase-expressing human glioblastoma tumors, which were subcutaneously xenografted into nude mice, led to a significant 30% siRNA-mediated luciferase gene silencing 4 days post-injection. Our results altogether substantiate the potential of self-assembling cationic polymers with a pH-sensitive disassembly switch for siRNA delivery in vitro and also in vivo experiments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Erythrocytes / drug effects
  • Erythrocytes / pathology
  • Gene Silencing*
  • Hemolysis / drug effects
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Male
  • Mice
  • Mice, Nude
  • Neoplasm Transplantation
  • Neoplasms / metabolism
  • Polyethyleneimine / administration & dosage*
  • Polyethyleneimine / chemistry
  • RNA, Small Interfering / administration & dosage*
  • RNA, Small Interfering / chemistry
  • Sheep
  • Thiourea / administration & dosage*
  • Thiourea / analogs & derivatives*
  • Thiourea / chemistry

Substances

  • RNA, Small Interfering
  • Polyethyleneimine
  • Thiourea