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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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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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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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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DOI: https://doi.org/10.1038/nmat2444
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