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J Control Release. 2018 Apr 28;276:50-58. doi: 10.1016/j.jconrel.2018.02.017. Epub 2018 Feb 20.

In vitro and in vivo delivery of siRNA via VIPER polymer system to lung cells.

Author information

1
Department of Oncology, Wayne State University School of Medicine, 4100 John R St, Detroit, MI 48201, United States.
2
Department of Bioengineering and Molecular Engineering and Sciences Institute, University of Washington, Seattle, WA, United States.
3
Department of Pharmacy, Pharmaceutical Technology and Biopharmacy, Ludwig-Maximilians- Universität München, 81337 Munich, Germany.
4
Department of Pharmaceutical Sciences, Wayne State University, 259 Mack Ave, Detroit, MI 48201, United States.
5
Department of Biology, Human Biology and Bioimaging, Ludwig-Maximilians- Universität München, 82152 Martinsried, Germany.
6
Department of Oncology, Wayne State University School of Medicine, 4100 John R St, Detroit, MI 48201, United States; Department of Pharmacy, Pharmaceutical Technology and Biopharmacy, Ludwig-Maximilians- Universität München, 81337 Munich, Germany; Department of Pharmaceutical Sciences, Wayne State University, 259 Mack Ave, Detroit, MI 48201, United States. Electronic address: olivia.merkel@lmu.de.

Abstract

The block copolymer VIPER (virus-inspired polymer for endosomal release) has been reported to be a promising novel delivery system of DNA plasmids both in vitro and in vivo. VIPER is comprised of a polycation segment for condensation of nucleic acids as well as a pH-sensitive segment that exposes the membrane lytic peptide melittin in acidic environments to facilitate endosomal escape. The objective of this study was to investigate VIPER/siRNA polyplex characteristics, and compare their in vitro and in vivo performance with commercially available transfection reagents and a control version of VIPER lacking melittin. VIPER/siRNA polyplexes were formulated and characterized at various charge ratios and shown to be efficiently internalized in cultured cells. Target mRNA knockdown was confirmed by both flow cytometry and qRT-PCR and the kinetics of knockdown was monitored by live cell spinning disk microscopy, revealing knockdown starting by 4 h post-delivery. Intratracheal instillation of VIPER particles formulated with sequence specific siRNA to the lung of mice resulted in a significantly more efficient knockdown of GAPDH compared to treatment with VIPER particles formulated with scrambled sequence siRNA. We also demonstrated using pH-sensitive labels that VIPER particles experience less acidic environments compared to control polyplexes. In summary, VIPER/siRNA polyplexes efficiently deliver siRNA in vivo resulting in robust gene silencing (>75% knockdown) within the lung.

KEYWORDS:

Endosomal escape; Melittin; Pulmonary delivery; pH-sensitive polymer; siRNA delivery

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