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Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes

Abstract

To realize the therapeutic potential of RNA drugs, efficient, tissue-specific and nonimmunogenic delivery technologies must be developed. Here we show that exosomes—endogenous nano-vesicles that transport RNAs and proteins1,2—can deliver short interfering (si)RNA to the brain in mice. To reduce immunogenicity, we used self-derived dendritic cells for exosome production. Targeting was achieved by engineering the dendritic cells to express Lamp2b, an exosomal membrane protein, fused to the neuron-specific RVG peptide3. Purified exosomes were loaded with exogenous siRNA by electroporation. Intravenously injected RVG-targeted exosomes delivered GAPDH siRNA specifically to neurons, microglia, oligodendrocytes in the brain, resulting in a specific gene knockdown. Pre-exposure to RVG exosomes did not attenuate knockdown, and non-specific uptake in other tissues was not observed. The therapeutic potential of exosome-mediated siRNA delivery was demonstrated by the strong mRNA (60%) and protein (62%) knockdown of BACE1, a therapeutic target in Alzheimer's disease, in wild-type mice.

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Figure 1: Targeting peptide expressed with Lamp2b is expressed on the external surface of exosomes.
Figure 2: In vitro delivery of siRNA by targeted exosomes.
Figure 3: In vivo delivery of siRNA with targeted exosomes results in brain-specific gene knockdown.

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Acknowledgements

The authors would like to thank I. Sargent and his group, especially A. Brooks, for their assistance with the LM10-HS system, J. Morris for his help with the electron microscopy, D. Morrissey and his laboratory at Novartis, Basel, for supplying the RVG-9R peptide and the Wood laboratory members for critical reading of the manuscript. L.A.-E., C.B. and H.Y. are funded by the Muscular Dystrophy Ireland and the Muscular Dystrophy Campaign (UK). Y.S. is funded by the Agency for Science, Technology and Research (Singapore).

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L.A.-E., Y.S. and M.J.A.W. designed the experiments. L.A.-E. and Y.S. performed the experiments and analyzed the data except for intravenous injection, CFSE proliferation assay. H.Y. performed intravenous injections. C.B. assisted with dissection and harvesting of tissue. S.L. performed the CFSE proliferation assay. L.A.-E., Y.S. and M.J.A.W. wrote the manuscript.

Corresponding author

Correspondence to Matthew J A Wood.

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Supplementary Tables 1,2 and Supplementary Figs. 1–9 (PDF 1956 kb)

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Alvarez-Erviti, L., Seow, Y., Yin, H. et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat Biotechnol 29, 341–345 (2011). https://doi.org/10.1038/nbt.1807

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