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A modular platform for targeted RNAi therapeutics

Nature Nanotechnology volume 13, pages 214–219 (2018)Cite this article

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Abstract

Previous studies have identified relevant genes and signalling pathways that are hampered in human disorders as potential candidates for therapeutics. Developing nucleic acid-based tools to manipulate gene expression, such as short interfering RNAs1,2,3 (siRNAs), opens up opportunities for personalized medicine. Yet, although major progress has been made in developing siRNA targeted delivery carriers, mainly by utilizing monoclonal antibodies (mAbs) for targeting4,5,6,7,8, their clinical translation has not occurred. This is in part because of the massive development and production requirements and the high batch-to-batch variability of current technologies, which rely on chemical conjugation. Here we present a self-assembled modular platform that enables the construction of a theoretically unlimited repertoire of siRNA targeted carriers. The self-assembly of the platform is based on a membrane-anchored lipoprotein that is incorporated into siRNA-loaded lipid nanoparticles that interact with the antibody crystallizable fragment (Fc) domain. We show that a simple switch of eight different mAbs redirects the specific uptake of siRNAs by diverse leukocyte subsets in vivo. The therapeutic potential of the platform is demonstrated in an inflammatory bowel disease model by targeting colon macrophages to reduce inflammatory symptoms, and in a Mantle Cell Lymphoma xenograft model by targeting cancer cells to induce cell death and improve survival. This modular delivery platform represents a milestone in the development of precision medicine.

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Fig. 1: ASSET platform design and construction.
Fig. 2: TsiLNP functionality in vitro and in vivo.
Fig. 3: In vivo targeting of TsiLNPs.
Fig. 4: TsiLNP-mediated therapeutic gene silencing in a murine colitis model.

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References

  1. Dahlman, J. E. et al. In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight. Nat. Nanotechnol. 9, 648–655 (2014).

    Article  Google Scholar 

  2. McNamara, J. O. II et al. Cell type-specific delivery of siRNAs with aptamer-siRNA chimeras. Nat. Biotechnol. 24, 1005–1015 (2006).

    Article  Google Scholar 

  3. Wittrup, A. & Lieberman, J. Knocking down disease: a progress report on siRNA therapeutics. Nat. Rev. Genet. 16, 543–552 (2015).

    Article  Google Scholar 

  4. Ramishetti, S. et al. Systemic gene silencing in primary T lymphocytes using targeted lipid nanoparticles. ACS Nano 9, 6706–6716 (2015).

    Article  Google Scholar 

  5. Weinstein, S. et al. Harnessing RNAi-based nanomedicines for therapeutic gene silencing in B-cell malignancies. Proc. Natl Acad. Sci. USA 113, E16–E22 (2016).

    Article  Google Scholar 

  6. Katakowski, J. A. et al. Delivery of siRNAs to dendritic cells using DEC205-targeted lipid nanoparticles to inhibit immune responses. Mol. Ther. 24, 146–155 (2016).

    Article  Google Scholar 

  7. Peer, D., Zhu, P., Carman, C. V., Lieberman, J. & Shimaoka, M. Selective gene silencing in activated leukocytes by targeting siRNAs to the integrin lymphocyte function-associated antigen-1. Proc. Natl Acad. Sci. USA 104, 4095–4100 (2007).

    Article  Google Scholar 

  8. Song, E. et al. Antibody mediated in vivo delivery of small interfering RNAs via cell-surface receptors. Nat. Biotechnol. 23, 709–717 (2005).

    Article  Google Scholar 

  9. Springer, T. A., Bhattacharya, A., Cardoza, J. T. & Sanchez-Madrid, F. Monoclonal antibodies specific for rat IgG1, IgG2a, and IgG2b subclasses, and kappa chain monotypic and allotypic determinants: reagents for use with rat monoclonal antibodies. Hybridoma 1, 257–273 (1982).

    Article  Google Scholar 

  10. Laukkanen, M. L., Teeri, T. T. & Keinanen, K. Lipid-tagged antibodies: bacterial expression and characterization of a lipoprotein-single-chain antibody fusion protein. Protein Eng. 6, 449–454 (1993).

    Article  Google Scholar 

  11. de Kruif, J., Storm, G., van Bloois, L. & Logtenberg, T. Biosynthetically lipid-modified human scFv fragments from phage display libraries as targeting molecules for immunoliposomes. FEBS Lett. 399, 232–236 (1996).

    Article  Google Scholar 

  12. Semple, S. C. et al. Rational design of cationic lipids for siRNA delivery. Nat. Biotechnol. 28, 172–176 (2010).

    Article  Google Scholar 

  13. Cohen, Z. R. et al. Localized RNAi therapeutics of chemoresistant grade IV glioma using hyaluronan-grafted lipid-based nanoparticles. ACS Nano. 9, 1581–1591 (2015).

    Article  Google Scholar 

  14. Tiisala, S., Paavonen, T. & Renkonen, R. Alpha E beta 7 and alpha 4 beta 7 integrins associated with intraepithelial and mucosal homing, are expressed on macrophages. Eur. J. Immunol. 25, 411–417 (1995).

    Article  Google Scholar 

  15. Peer, D., Park, E. J., Morishita, Y., Carman, C. V. & Shimaoka, M. Systemic leukocyte-directed siRNA delivery revealing cyclin D1 as an anti-inflammatory target. Science 319, 627–630 (2008).

    Article  Google Scholar 

  16. Zigmond, E. et al. Ly6C hi monocytes in the inflamed colon give rise to proinflammatory effector cells and migratory antigen-presenting cells. Immunity 37, 1076–1090 (2012).

    Article  Google Scholar 

  17. Grainger, J. R. et al. Inflammatory monocytes regulate pathologic responses to commensals during acute gastrointestinal infection. Nat. Med. 19, 713–721 (2013).

    Article  Google Scholar 

  18. Benhar, I. & Pastan, I. Cloning, expression and characterization of the Fv fragments of the anti-carbohydrate mAbs Bl and B5 as single-chain immunotoxins. Protein Eng. 7, 1509–1515 (1994).

    Article  Google Scholar 

  19. Vaks, L. & Benhar, I. Production of stabilized scFv antibody fragments in the E. coli bacterial cytoplasm. Methods Mol. Biol. 1060, 171–184 (2014).

    Article  Google Scholar 

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Acknowledgements

This work was supported in part by grants from the Dotan Hemato-oncology Center at Tel Aviv University, by The Leona M. and Harry B. Helmsley Nanotechnology Research Fund, by the Kenneth Rainin Foundation and by the ERC grant LeukoTheranostics (number 647410) awarded to D.P.

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Author notes

  1. These authors contributed equally: R. Kedmi and N. Veiga.

Authors and Affiliations

  1. Laboratory of Precision NanoMedicine, School of Molecular Cell Biology and Biotechnology, George S. Wise Faculty of Life Sciences, Department of Materials Sciences and Engineering, Iby and Aladar Fleischman Faculty of Engineering, Center for Nanoscience and Nanotechnology, Cancer Biology Research Center, Tel Aviv University, Tel Aviv, Israel

    Ranit Kedmi, Nuphar Veiga, Srinivas Ramishetti, Meir Goldsmith, Daniel Rosenblum, Niels Dammes, Inbal Hazan-Halevy, Shani Leviatan-Ben-Arye & Dan Peer

  2. School of Molecular Cell Biology and Biotechnology, Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel

    Limor Nahary & Itai Benhar

  3. Veterinary Service Center, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

    Michael Harlev

  4. Integrated DNA Technologies Inc., Coralville, IA, USA

    Mark Behlke

  5. Program in Cellular and Molecular Medicine, Boston Children’s Hospital, and Department of Pediatrics, Harvard Medical School, Boston, MA, USA

    Judy Lieberman

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  1. Ranit Kedmi
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Contributions

R.K., N.V. and D.P. conceived and designed the project. R.K., N.V. S.R., M.G., D.R. N.D., L.N., I.H.-H., S.L.-B.-A. and M.H. performed the experimental work. R.K., N.V., M.B., I.B. J.L. and D.P. analysed the data. R.K. N.V. and D.P. wrote the manuscript. All authors discussed the results.

Corresponding author

Correspondence to Dan Peer.

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Competing interests

M.B. is an employee of Integrated DNA Technologies, Inc. J.L. is on the Scientific Advisory Board of Alnylam Pharmaceuticals. D.P. declares financial interests in Quiet Therapeutics. The rest of the authors declare no competing financial interests.

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Kedmi, R., Veiga, N., Ramishetti, S. et al. A modular platform for targeted RNAi therapeutics. Nature Nanotech 13, 214–219 (2018). https://doi.org/10.1038/s41565-017-0043-5

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