Self-assembled Micelle Interfering RNA for Effective and Safe Targeting of Dysregulated Genes in Pulmonary Fibrosis*

  1. Chun Geun Lee§3
  1. From the ‡Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea,
  2. the §Department of Molecular Microbiology and Immunology,
  3. ‖Department of Medicine, Alpert Medical School, Brown University, Providence, Rhode Island 02912,
  4. the Inhalation Toxicology Center, Korea Institute of Toxicology, Jeongeup Campus, Jeollabuk-do 580-185, Korea, and
  5. the **Division of Allergy and Clinical Immunology, Department of Internal Medicine, Seoul National University Hospital, Seoul 110-744, Korea
  1. ↵2 To whom correspondence may be addressed. Tel.: 82-63-570-8740; Fax: 82-63-570-8108; E-mail: khlee{at}kitox.re.kr.
  2. ↵3 To whom correspondence may be addressed. Tel.: 401-863-5932; Fax: 401-863-2925; E-mail: chun_lee{at}brown.edu.

  1. ↵1 These authors contributed equally to this work.

Abstract

The siRNA silencing approach has long been used as a method to regulate the expression of specific target genes in vitro and in vivo. However, the effectiveness of delivery and the nonspecific immune-stimulatory function of siRNA are the limiting factors for therapeutic applications of siRNAs. To overcome these limitations, we developed self-assembled micelle inhibitory RNA (SAMiRNA) nanoparticles made of individually biconjugated siRNAs with a hydrophilic polymer and lipid on their ends and characterized their stability, immune-stimulatory function, and in vivo silencing efficacy. SAMiRNAs form very stable nanoparticles with no significant degradation in size distribution and polydispersity index over 1 year. Overnight incubation of SAMiRNAs (3 μm) on murine peripheral blood mononuclear cells did not cause any significant elaboration of innate immune cytokines such as TNF-α, IL-12, or IL-6, whereas unmodified siRNAs or liposomes or liposome complexes significantly stimulated the expression of these cytokines. Last, the in vivo silencing efficacy of SAMiRNAs was evaluated by targeting amphiregulin and connective tissue growth factor in bleomycin or TGF-β transgenic animal models of pulmonary fibrosis. Intratracheal or intravenous delivery two or three times of amphiregulin or connective tissue growth factor SAMiRNAs significantly reduced the bleomycin- or TGF-β-stimulated collagen accumulation in the lung and substantially restored the lung function of TGF-β transgenic mice. This study demonstrates that SAMiRNA nanoparticle is a less toxic, stable siRNA silencing platform for efficient in vivo targeting of genes implicated in the pathogenesis of pulmonary fibrosis.

Footnotes

  • ↵* This work was supported by National Institutes of Health Grants RO1-HL115813 (to C. G. L.) and PO1-HL114501 (to J. A. E.) and Korea Drug Development Fund Grant 201312-11 funded by the Ministry of Science, ICT and Future Planning, Ministry of Trade, Industry, and Energy, and Ministry of Health and Welfare Republic of Korea (to P. O. Y. and C. G. L.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

  • ↵Graphic This article contains supplemental information and Figs. 1 and 2.

  • Received September 20, 2015.
  • Revision received January 23, 2016.
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  1. First Published on January 27, 2016 doi: 10.1074/jbc.M115.693671 The Journal of Biological Chemistry 291, 6433-6446.
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