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Exploring the HYDRAtion method for loading siRNA on liposomes: the interplay between stability and biological activity in human undiluted ascites fluid

Abstract

Delivery of small interfering RNA (siRNA) is recently gaining tremendous attention for the treatment of ovarian cancer. The present study investigated the potential of different liposomal formulations composed of (2,3-dioleoyloxy-propyl)-trimethylammonium (DOTAP) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) encapsulating siRNA (hydration method) for their ability to knockdown luciferase (Luc) activity in human ovarian cancer SKOV-3 cells. Fluorescence single particle tracking (fSPT) and fluorescence correlation spectroscopy (FCS) in human-undiluted ascites fluid obtained from a peritoneal carcinomatosis patient revealed that cationic hydra-lipoplexes (HYDRA-LPXs) and HYDRA-LPXs decorated with stable DSPE-PEG (DSPE HYDRA-LPXs) showed high stability during at least 24 h. HYDRA-LPXs decorated with sheddable C8 and C16 PEG-Ceramides (Cer HYDRA-LPXs) resulted in rapid and premature release of siRNA already in the first hours. Despite their role in preventing aggregation in vivo, liposomes decorated with stable PEG residues resulted in a poor transfection compared to the ones decorated with sheddable PEG residues in reduced serum conditions. Yet, the transfection efficiency of both Cer HYDRA-LPXs significantly decreased following 1 h of incubation in ascites fluid due to a drastic drop in the cellular uptake, while DSPE HYDRA-LPXs are still taken up by cells, but too stable to induce efficient gene silencing.

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Acknowledgments

GD is a doctoral fellow of the Flemish Government (Vlaamse overheid). WC is a senior clinical investigator of the Fund for Scientific Research—Flanders (FWO). We thank Dr. Pieter Baatsen from KU Leuven for his help with the characterization of the formulations. This work was supported by the Research Foundation-Flanders (research project G006714N). We thank Senne Corneils for his help with the experiments.

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Correspondence to Stefaan C. De Smedt or Katrien Remaut.

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Dakwar, G.R., Braeckmans, K., Ceelen, W. et al. Exploring the HYDRAtion method for loading siRNA on liposomes: the interplay between stability and biological activity in human undiluted ascites fluid. Drug Deliv. and Transl. Res. 7, 241–251 (2017). https://doi.org/10.1007/s13346-016-0329-4

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Keywords

  • siRNA delivery
  • siRNA encapsulation
  • Peritoneal metastasis
  • Sheddable PEG
  • Ascites