Abstract
Dysfunctional endothelium contributes to more diseases than any other tissue in the body. Small interfering RNAs (siRNAs) can help in the study and treatment of endothelial cells in vivo by durably silencing multiple genes simultaneously, but efficient siRNA delivery has so far remained challenging. Here, we show that polymeric nanoparticles made of low-molecular-weight polyamines and lipids can deliver siRNA to endothelial cells with high efficiency, thereby facilitating the simultaneous silencing of multiple endothelial genes in vivo. Unlike lipid or lipid-like nanoparticles, this formulation does not significantly reduce gene expression in hepatocytes or immune cells even at the dosage necessary for endothelial gene silencing. These nanoparticles mediate the most durable non-liver silencing reported so far and facilitate the delivery of siRNAs that modify endothelial function in mouse models of vascular permeability, emphysema, primary tumour growth and metastasis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Change history
20 June 2014
In the version of this Article originally published online, the following authors' names were written incorrectly: Victor Koteliansky, Omar F. Khan and Kamaljeet Singh Sandhu. These have now been corrected in all versions of the Article.
References
Pober, J. S. & Sessa, W. C. Evolving functions of endothelial cells in inflammation. Nature Rev. Immunol. 7, 803–815 (2007).
Hagberg, C. E. et al. Targeting VEGF-B as a novel treatment for insulin resistance and type 2 diabetes. Nature 490, 426–430 (2012).
Kumar, V., Abbas, A., Fausto, N. & Aster, J. Robbins and Cotran Pathologic Basis of Disease 8th edn (Elsevier, 2009).
Kanasty, R., Dorkin, J. R., Vegas, A. & Anderson, D. Delivery materials for siRNA therapeutics. Nature Mater. 12, 967–977 (2013).
Love, K. T. et al. Lipid-like materials for low-dose, in vivo gene silencing. Proc. Natl Acad. Sci. USA 107, 1864–1869 (2010).
Semple, S. et al. Rational design of cationic lipids for siRNA delivery. Nature Biotechnol. 28, 172–176 (2010).
Whitehead, K. A., Langer, R. & Anderson, D. G. Knocking down barriers: advances in siRNA delivery. Nature Rev. Drug Discov. 8, 129–138 (2009).
Aleku, M. et al. Atu027, a liposomal small interfering RNA formulation targeting protein kinase N3, inhibits cancer progression. Cancer Res. 68, 9788–9798 (2008).
Aleku, M. et al. Intracellular localization of lipoplexed siRNA in vascular endothelial cells of different mouse tissues. Microvasc. Res. 76, 31–41 (2008).
Santel, A. et al. RNA interference in the mouse vascular endothelium by systemic administration of siRNA-lipoplexes for cancer therapy. Gene Ther. 13, 1360–1370 (2006).
Santel, A. et al. A novel siRNA-lipoplex technology for RNA interference in the mouse vascular endothelium. Gene Ther. 13, 1222–1234 (2006).
Polach, K. J. et al. Delivery of siRNA to the mouse lung via a functionalized lipopolyamine. Mol. Ther. 20, 91–100 (2012).
Kaufmann, J., Ahrens, K. & Santel, A. RNA interference for therapy in the vascular endothelium. Microvasc. Res. 80, 286–293 (2010).
Davis, M. E. et al. Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature 484, 1067–1070 (2010).
Rozema, D. B. et al. Dynamic polyconjugates for targeted in vivo delivery of siRNA to hepatocytes. Proc. Natl Acad. Sci. USA 104, 12982–12987 (2007).
Godbey, W. T., Wu, K. K. & Mikos, A. G. Poly(ethylenimine) and its role in gene delivery. J. Control. Rel. 60, 149–160 (1999).
Breunig, M., Lungwitz, U., Liebl, R. & Goepferich, A. Breaking up the correlation between efficacy and toxicity for nonviral gene delivery. Proc. Natl Acad. Sci. USA 104, 14454–14459 (2007).
Richards Grayson, A. C., Doody, A. M. & Putnam, D. Biophysical and structural characterization of polyethylenimine-mediated siRNA delivery in vitro. Pharm. Res. 8, 1868–1876 (2006).
Crawford, R. et al. Analysis of lipid nanoparticles by cryo-EM for characterizing siRNA delivery vehicles. Int. J. Pharm. 403, 237–244 (2011).
Jayaraman, M. et al. Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. Angew. Chem. Int. Ed. 51, 8529–8533 (2012).
Huang, H., Bhat, A., Woodnutt, G. & Lappe, R. Targeting the ANGPT-TIE2 pathway in malignancy. Nature Rev. Cancer 10, 575–585 (2010).
Novobrantseva, T. I. et al. Systemic RNAi-mediated gene silencing in nonhuman primate and rodent myeloid cells. Mol. Ther. Nucleic Acids 1, e4 (2012).
Kasahara, Y. et al. Inhibition of VEGF receptors causes lung cell apoptosis and emphysema. J. Clin. Invest. 106, 1311–1319 (2000).
Tuder, R. M. & Yun, J. H. Vascular endothelial growth factor of the lung: friend or foe. Curr. Opin. Pharmacol. 8, 255–260 (2008).
Thurston, G., Noguera-Troise, I. & Yancopoulos, G. D. The Delta paradox: DLL4 blockade leads to more tumour vessels but less tumour growth. Nature Rev. Cancer 7, 327–331 (2007).
Fischer, C., Mazzone, M., Jonckx, B. & Carmeliet, P. FLT1 and its ligands VEGFB and PlGF: drug targets for anti-angiogenic therapy? Nature Rev. Cancer 8, 942–956 (2008).
Lyden, D. et al. Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nature Med. 7, 1194–1201 (2001).
Kaplan, R. N. et al. VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche. Nature 438, 820–827 (2005).
Tammela, T. et al. VEGFR-3 controls tip to stalk conversion at vessel fusion sites by reinforcing Notch signalling. Nature Cell Biol. 13, 1202–1213 (2011).
Stevens, J. B. et al. Heterogeneity of cell death. Cytogenet. Genome Res. 139, 164–173 (2013).
Kuramoto, T. et al. Dll4-Fc, an inhibitor of Dll4-notch signaling, suppresses liver metastasis of small cell lung cancer cells through the downregulation of the NF-kappaB activity. Mol. Cancer Ther. 11, 2578–2587 (2012).
Garcia, A. & Kandel, J. J. Notch: a key regulator of tumor angiogenesis and metastasis. Histol. Histopathol. 27, 151–156 (2012).
Akinc, A. et al. Targeted delivery of RNAi therapeutics with endogenous and exogenous ligand-based mechanisms. Mol. Ther. 18, 1357–1364 (2010).
Monopoli, M. P., Aberg, C., Salvati, A. & Dawson, K. A. Biomolecular coronas provide the biological identity of nanosized materials. Nature Nanotech. 7, 779–786 (2012).
Chen, D. et al. Rapid discovery of potent siRNA-containing lipid nanoparticles enabled by controlled microfluidic formulation. J. Am. Chem. Soc. 134, 6948–6951 (2012).
Whitehead, K., Dahlman, J. E., Langer, R. S. & Anderson, D. G. Silencing or stimulation? siRNA delivery and the immune system. Ann. Rev. Chem. Biomol. Eng. 2, 77–96 (2011).
Panigrahy, D. et al. Epoxyeicosanoids stimulate multiorgan metastasis and tumor dormancy escape in mice. J. Clin. Invest. 122, 178–191 (2012).
Acknowledgements
The authors thank J. Cattie, T. O'Shea and T. Tammela. J.E.D. was supported by National Defense Science and Engineering (NDSEG), the National Science Foundation (NSF) and MIT Presidential Fellowships. D.P. was supported by R01 CA148663. M.W.K. was supported by the Stop and Shop Pediatric Brain Tumour Fund, as well as the Pediatric Brain Tumour Fund. H.S. was supported by the Deutsche Forschungsgemeinschaft (SA1668/2-1). Research was also supported by Alnylam and the Center for RNA Therapeutics and Biology.
Author information
Authors and Affiliations
Contributions
J.E.D., C.B., V.K., R.L. and D.G.A. conceived the experiments. J.E.D., C.B., O.F.K., A.T., S.J., T.E.S., Y.X., H.B.S., G.S., L.S., A.B., R.L.B., H.Y., T.R., Y.D., S.J., D.S., A.D., K.S.S., M.J.W., T.N., V.M.R., A.K.R.L.J., C.G.L., B.K., D.K.M., M.P., L.A., P.D., L.S., K.C., M.W.K., K.F., M.N., D.D., R.M.T., U.H.V.A., A.A., A.S. and D.P. performed experiments. J.E.D., C.B., V.K., R.L. and D.G.A. co-wrote the paper. All authors discussed the results and commented on the manuscript.
Corresponding author
Ethics declarations
Competing interests
R.L. is a shareholder and member of the Scientific Advisory Board of Alnylam. R.L and D.G.A have sponsored research grants from Alnylam. Alnylam also has a licence to certain intellectual property that was invented at the Massachusetts Institute of Technology by D.G.A. and R.L.
Supplementary information
Supplementary information
Supplementary Information (PDF 2132 kb)
Rights and permissions
About this article
Cite this article
Dahlman, J., Barnes, C., Khan, O. et al. In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight. Nature Nanotech 9, 648–655 (2014). https://doi.org/10.1038/nnano.2014.84
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nnano.2014.84