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Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA

Nature Materials volume 8, pages 526–533 (2009)Cite this article

Abstract

Vaginal instillation of small-interfering RNA (siRNA) using liposomes has led to silencing of endogenous genes in the genital tract and protection against challenge from infectious disease. Although siRNA lipoplexes are easily formulated, several of the most effective transfection agents available commercially may be toxic to the mucosal epithelia and none are able to provide controlled or sustained release. Here, we demonstrate an alternative approach using nanoparticles composed entirely of FDA-approved materials. To render these materials effective for gene silencing, we developed novel approaches to load them with high amounts of siRNA. A single dose of siRNA-loaded nanoparticles to the mouse female reproductive tract caused efficient and sustained gene silencing. Knockdown of gene expression was observed proximal (in the vaginal lumen) and distal (in the uterine horns) to the site of topical delivery. In addition, nanoparticles penetrated deep into the epithelial tissue. This is the first report demonstrating that biodegradable polymer nanoparticles are effective delivery vehicles for siRNA to the vaginal mucosa.

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Figure 1: In vitro cytotoxicity and bioactivity of siRNA nanoparticles.
Figure 2: Fluorescent PLGA nanoparticles appear throughout the reproductive tract and penetrate deep within the vaginal tissue after topical administration.
Figure 3: Release of PLGA nanoparticles densely loaded with siRNA is sustained for several weeks.
Figure 4: Intravaginal delivery of siRNA using biodegradable nanoparticles causes gene silencing throughout the reproductive tract of transgenic GFP mice.
Figure 5: A single topical administration of PLGA nanoparticles loaded with siRNA causes sustained gene silencing throughout the reproductive tract.
Figure 6: Lipid delivery of siRNA may be inflammatory and cause epithelial disruption in the vaginal tissue.

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Acknowledgements

We are grateful to R. Flavell for use of the IVIS 200 imaging system. We are also grateful to A. Haberman and D. Gonzalez for assistance with the two-photon microscopy imaging. We thank T. Kyriakides, A. Iwasaki and S. Jay for helpful analysis and discussion of tissue sections used for immunohistochemistry. We also thank A. Sin for assistance with nanoparticle characterization and J. Blum for technical assistance with the animal experiments. Y. Cu, J. Saucier-Sawyer, A. Lin and J. Blum carried out blind scoring of the microscopy images for fluorescence. This work was supported by grants from the NIH to W.M.S. (RO1-EB0000487-18). K.A.W. acknowledges the fellowship support from the L’Oreal FWIS and a postdoctoral training fellowship (T32-HL007950-08).

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Authors and Affiliations

  1. Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA

    Kim A. Woodrow, Yen Cu, Jennifer K. Saucier-Sawyer, Monica J. Wood & W. Mark Saltzman

  2. Section of Comparative Medicine, Yale University, New Haven, Connecticut 06511, USA

    Carmen J. Booth

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Contributions

K.A.W. and W.M.S. designed the experiments. K.A.W. prepared and characterized the siRNA polymer nanoparticles. K.A.W., J.K.S.S. and M.J.W. carried out the in vitro cell culture experiments, and also collected and analysed the data. K.A.W. and J.K.S.S. carried out the in vivo animal experiments and collected the reproductive tissue, and K.A.W carried out the tissue sectioning, and the collection and analysis of microscopy images. K.A.W. and Y.C. prepared fluorescent nanoparticles and carried out biodistribution studies, and Y.C. collected and analysed data from these studies. K.A.W. carried out the immunohistochemical staining and C.J.B. carried out the histological examination of sectioned tissues.

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Correspondence to W. Mark Saltzman.

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Woodrow, K., Cu, Y., Booth, C. et al. Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA. Nature Mater 8, 526–533 (2009). https://doi.org/10.1038/nmat2444

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