RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system

J Control Release. 2014 Jan 10:173:110-8. doi: 10.1016/j.jconrel.2013.10.003. Epub 2013 Oct 11.

Abstract

Angiogenesis is one of crucial processes associated with tumor growth and development, and consequently a prime target for cancer therapy. Although tumor endothelial cells (TECs) play a key role in pathological angiogenesis, investigating phenotypical changes in neovessels when a gene expression in TEC is suppressed is a difficult task. Small interfering RNA (siRNA) represents a potential agent due to its ability to silence a gene of interest. We previously developed a system for in vivo siRNA delivery to cancer cells that involves a liposomal-delivery system, a MEND that contains a unique pH-sensitive cationic lipid, YSK05 (YSK-MEND). In the present study, we report on the development of a system that permits the delivery of siRNA to TECs by combining the YSK-MEND and a ligand that is specific to TECs. Cyclo(Arg-Gly-Asp-D-Phe-Lys) (cRGD) is a well-known ligand to αVβ3 integrin, which is selectively expressed at high levels in TECs. We incorporated cRGD into the YSK-MEND (RGD-MEND) to achieve an efficient gene silencing in TECs. Quantitative RT-PCR and the 5' rapid amplification of cDNA ends PCR indicated that the intravenous injection of RGD-MEND at a dose of 4.0mg/kg induced a significant RNAi-mediated gene reduction in TEC but not in endothelial cells of other organs. Finally, we evaluated the therapeutic potency of the RGD-MEND encapsulating siRNA against vascular endothelial growth factor receptor 2. A substantial delay in tumor growth was observed after three sequential RGD-MEND injections on alternate days. In conclusion, the RGD-MEND represents a new approach for the characterization of TECs and for us in anti-angiogenic therapy.

Keywords: Active targeting; Anti-angiogenic therapy; Cyclic RGD; Liposome; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drug Delivery Systems
  • HEK293 Cells
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Lipids / chemistry
  • Liposomes / chemistry*
  • Liposomes / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Neoplasms / blood supply
  • Neoplasms / genetics
  • Neoplasms / pathology
  • Neoplasms / therapy*
  • Neovascularization, Pathologic / genetics
  • Neovascularization, Pathologic / pathology
  • Neovascularization, Pathologic / therapy*
  • Peptides, Cyclic / chemistry*
  • Peptides, Cyclic / metabolism
  • Piperidines / chemistry
  • Piperidines / metabolism
  • RNA Interference*
  • RNA, Small Interfering / administration & dosage*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / therapeutic use
  • Vascular Endothelial Growth Factor Receptor-2 / genetics*

Substances

  • Lipids
  • Liposomes
  • Peptides, Cyclic
  • Piperidines
  • RNA, Small Interfering
  • YSK05
  • cyclic arginine-glycine-aspartic acid peptide
  • Vascular Endothelial Growth Factor Receptor-2