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Biomimetic proteolipid vesicles for targeting inflamed tissues

Abstract

A multitude of micro- and nanoparticles have been developed to improve the delivery of systemically administered pharmaceuticals, which are subject to a number of biological barriers that limit their optimal biodistribution. Bioinspired drug-delivery carriers formulated by bottom-up or top-down strategies have emerged as an alternative approach to evade the mononuclear phagocytic system and facilitate transport across the endothelial vessel wall. Here, we describe a method that leverages the advantages of bottom-up and top-down strategies to incorporate proteins derived from the leukocyte plasma membrane into lipid nanoparticles. The resulting proteolipid vesicles—which we refer to as leukosomes—retained the versatility and physicochemical properties typical of liposomal formulations, preferentially targeted inflamed vasculature, enabled the selective and effective delivery of dexamethasone to inflamed tissues, and reduced phlogosis in a localized model of inflammation.

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Figure 1: Leukosome synthesis and formulation.
Figure 2: Characterization of leukosomes’ physicochemical features.
Figure 3: Analysis of the leukocyte membrane proteins transferred to the leukosome’s lipid bilayer.
Figure 4: Leukosomes retain drug loading and release properties similar to control liposomes.
Figure 5: Leukosomes preferentially adhere to inflamed vasculature in vivo and improve tissue healing by preserving its architecture and reducing neutrophil infiltration.
Figure 6: Immunogenicity and safety of leukosomes.

References

  1. Torchilin, V. P. Multifunctional, stimuli-sensitive nanoparticulate systems for drug delivery. Nature Rev. Drug Discov. 13, 813–827 (2014).

    CAS  Article  Google Scholar 

  2. Mitragotri, S., Burke, P. A. & Langer, R. Overcoming the challenges in administering biopharmaceuticals: formulation and delivery strategies. Nature Rev. Drug Discov. 13, 655–672 (2014).

    CAS  Article  Google Scholar 

  3. Tasciotti, E. et al. Mesoporous silicon particles as a multistage delivery system for imaging and therapeutic applications. Nature Nanotech. 3, 151–157 (2008).

    CAS  Article  Google Scholar 

  4. Parodi, A. et al. Bromelain surface modification increases the diffusion of silica nanoparticles in the tumor extracellular matrix. ACS Nano 8, 9874–9883 (2014).

    CAS  Article  Google Scholar 

  5. Mura, S., Nicolas, J. & Couvreur, P. Stimuli-responsive nanocarriers for drug delivery. Nature Mater. 12, 991–1003 (2013).

    CAS  Article  Google Scholar 

  6. Kudgus, R. A. et al. Tuning pharmacokinetics and biodistribution of a targeted drug delivery system through incorporation of a passive targeting component. Sci. Rep. 4, 5669 (2014).

    CAS  Article  Google Scholar 

  7. Luk, B. T. & Zhang, L. Cell membrane-camouflaged nanoparticles for drug delivery. J. Control. Rel. 220, 600–607 (2015).

    CAS  Article  Google Scholar 

  8. Hu, C.-M. J. et al. Erythrocyte membrane-camouflaged polymeric nanoparticles as a biomimetic delivery platform. Proc. Natl Acad. Sci. USA 108, 10980–10985 (2011).

    CAS  Article  Google Scholar 

  9. Hu, C.-M. J. et al. Nanoparticle biointerfacing by platelet membrane cloaking. Nature 526, 118–121 (2015).

    CAS  Article  Google Scholar 

  10. Parodi, A. Synthetic nanoparticles functionalized with biomimetic leukocyte membranes possess cell-like functions. Nature Nanotech. 8, 61–68 (2013).

    CAS  Article  Google Scholar 

  11. Hammer, D. A. et al. Leuko-polymersomes. Faraday Discuss 139, 129–141 (2008).

    CAS  Article  Google Scholar 

  12. Doshi, N. et al. Platelet mimetic particles for targeting thrombi in flowing blood. Adv. Mater. 24, 3864–3869 (2012).

    CAS  Article  Google Scholar 

  13. Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nature Biotechnol. 33, 941–951 (2015).

    CAS  Article  Google Scholar 

  14. Robbins, G. P. et al. Tunable leuko-polymersomes that adhere specifically to inflammatory markers. Langmuir 26, 14089–14096 (2010).

    CAS  Article  Google Scholar 

  15. Anselmo, A. C. et al. Platelet-like nanoparticles: mimicking shape, flexibility, and surface biology of platelets to target vascular injuries. ACS Nano 8, 11243–11253 (2014).

    CAS  Article  Google Scholar 

  16. Toledano Furman, N. E. et al. Reconstructed stem cell nanoghosts: a natural tumor targeting platform. Nano Lett. 13, 3248–3255 (2013).

    CAS  Article  Google Scholar 

  17. Yoo, J-W., Irvine, D. J., Discher, D. E. & Mitragotri, S. Bio-inspired, bioengineered and biomimetic drug delivery carriers. Nature Rev. Drug Discov. 10, 521–535 (2011).

    CAS  Article  Google Scholar 

  18. Alvarez-Lorenzo, C. & Concheiro, A. Bioinspired drug delivery systems. Curr. Opin. Biotechnol. 24, 1167–1173 (2013).

    CAS  Article  Google Scholar 

  19. Gutiérrez Millán, C., Colino Gandarillas, C. I., Sayalero Marinero, M. L. & Lanao, J. M. Cell-based drug-delivery platforms. Ther. Deliver. 3, 25–41 (2012).

    Article  Google Scholar 

  20. Millan, C. G., Marinero, M. A. L. S., Castaneda, A. Z. & Lanao, J. M. Drug, enzyme and peptide delivery using erythrocytes as carriers. J. Control. Release 95, 27–49 (2004).

    CAS  Article  Google Scholar 

  21. Bretscher, M. S. Asymmetrical lipid bilayer structure for biological membranes. Nature 236, 11–12 (1972).

    CAS  Article  Google Scholar 

  22. Demetzos, C. Differential scanning calorimetry (DSC): a tool to study the thermal behavior of lipid bilayers and liposomal stability. J. Liposome Res. 18, 159–173 (2008).

    CAS  Article  Google Scholar 

  23. Manconi, M. et al. Ex vivo skin delivery of diclofenac by transcutol containing liposomes and suggested mechanism of vesicle–skin interaction. Eur. J. Pharm. Biopharm. 78, 27–35 (2011).

    CAS  Article  Google Scholar 

  24. Mura, S., Manconi, M., Sinico, C., Valenti, D. & Fadda, A. M. Penetration enhancer-containing vesicles (PEVs) as carriers for cutaneous delivery of minoxidil. Intl. J. Pharm. 380, 72–79 (2009).

    CAS  Article  Google Scholar 

  25. Chow, T. S. Nanoscale surface roughness and particle adhesion on structured substrates. Nanotechnology 18, 115713 (2007).

    Article  Google Scholar 

  26. Schaap, I. A., Eghiaian, F., des Georges, A. & Veigel, C. Effect of envelope proteins on the mechanical properties of influenza virus. J. Biol. Chem. 287, 41078–41088 (2012).

    CAS  Article  Google Scholar 

  27. Mereghetti, P. et al. A Fourier transform infrared spectroscopy study of cell membrane domain modifications induced by docosahexaenoic acid. Biochim. Biophys. Acta 1840, 3115–3122 (2014).

    CAS  Article  Google Scholar 

  28. Lodish, H. et al. Molecular Cell Biology (W. H. Freeman & Co., 2000).

    Google Scholar 

  29. Benmerah, A., Scott, M., Poupon, V. & Marullo, S. Nuclear functions for plasma membrane associated proteins? Traffic 4, 503–511 (2003).

    CAS  Article  Google Scholar 

  30. Durr, E. et al. Direct proteomic mapping of the lung microvascular endothelial cell surface in vivo and in cell culture. Nature Biotechnol. 22, 985–992 (2004).

    CAS  Article  Google Scholar 

  31. Lund, R., Leth-Larsen, R., Jensen, O. N. & Ditzel, H. J. Efficient isolation and quantitative proteomic analysis of cancer cell plasma membrane proteins for identification of metastasis-associated cell surface markers. J. Proteome Res. 8, 3078–3090 (2009).

    CAS  Article  Google Scholar 

  32. Liu, X., Zhang, M., Go, V. L. W. & Hu, S. Membrane proteomic analysis of pancreatic cancer cells. J. Biomed. Sci. 17, 74 (2010).

    Article  Google Scholar 

  33. Corbo, C. et al. Proteomic profiling of a biomimetic drug delivery platform. Curr. Drug Targets 16, 1540–1547 (2015).

    CAS  Article  Google Scholar 

  34. Zarbock, A., Ley, K., McEver, R. P. & Hidalgo, A. Leukocyte ligands for endothelial selectins: specialized glycoconjugates that mediate rolling and signaling under flow. Blood 118, 6743–6751 (2011).

    CAS  Article  Google Scholar 

  35. Soto Pantoja, D. R., Kaur, S., Miller, T. W., Isenberg, J. S. & Roberts, D. D. Leukocyte surface antigen CD47. UCSD Mol. Pages 2, http://dx.doi.org/10.6072/H0.MP.A005186.01 (2013).

  36. Hu, C.-M. J. et al. ‘Marker-of-self’ functionalization of nanoscale particles through a top-down cellular membrane coating approach. Nanoscale 5, 2664–2668 (2013).

    CAS  Article  Google Scholar 

  37. Allen, T. Liposomal drug formulations. Drugs 56, 747–756 (1998).

    CAS  Article  Google Scholar 

  38. Cosco, D., Paolino, D., Cilurzo, F., Casale, F. & Fresta, M. Gemcitabine and tamoxifen-loaded liposomes as multidrug carriers for the treatment of breast cancer diseases. Intl. J. Pharm. 422, 229–237 (2012).

    CAS  Article  Google Scholar 

  39. Bernsdorff, C., Reszka, R. & Winter, R. Interaction of the anticancer agent Taxol TM (paclitaxel) with phospholipid bilayers. J. Biomed. Mater. Res. 46, 141–149 (1999).

    CAS  Article  Google Scholar 

  40. Franchimont, D., Kino, T., Galon, J., Meduri, G. U. & Chrousos, G. Glucocorticoids and inflammation revisited: the state of the art. Neuroimmunomodulation 10, 247–260 (2002).

    Article  Google Scholar 

  41. Gross, S. et al. Bioluminescence imaging of myeloperoxidase activity in vivo. Nature Med. 15, 455–461 (2009).

    CAS  Article  Google Scholar 

  42. Azzopardi, E. A., Ferguson, E. L. & Thomas, D. W. The enhanced permeability retention effect: a new paradigm for drug targeting in infection. J. Antimicrob. Chemother. 68, 257–274 (2013).

    CAS  Article  Google Scholar 

  43. Ishibashi, M. et al. Integrin LFA-1 regulates cell adhesion via transient clutch formation. Biochem. Biophys. Res. Commun. 464, 459–466 (2015).

    CAS  Article  Google Scholar 

  44. Arroyo, A. G. et al. Induction of tyrosine phosphorylation during ICAM-3 and LFA-1-mediated intercellular adhesion, and its regulation by the CD45 tyrosine phosphatase. J. Cell Biol. 126, 1277–1286 (1994).

    CAS  Article  Google Scholar 

  45. Sigal, A. et al. The LFA-1 integrin supports rolling adhesions on ICAM-1 under physiological shear flow in a permissive cellular environment. J. Immunol. 165, 442–452 (2000).

    CAS  Article  Google Scholar 

  46. Lorenz, H. M. et al. CD45 mAb induces cell adhesion in peripheral blood mononuclear cells via lymphocyte function-associated antigen-1 (LFA-1) and intercellular cell adhesion molecule 1 (ICAM-1). Cell. Immunol. 147, 110–128 (1993).

    CAS  Article  Google Scholar 

  47. Chen, X. et al. Inflamed leukocyte-mimetic nanoparticles for molecular imaging of inflammation. Biomaterials 32, 7651–7661 (2011).

    CAS  Article  Google Scholar 

  48. Sherman, M. B. et al. Removal of divalent cations induces structural transitions in red clover necrotic mosaic virus, revealing a potential mechanism for RNA release. J. Virol. 80, 10395–10406 (2006).

    CAS  Article  Google Scholar 

  49. Chiappini, C. et al. Biodegradable silicon nanoneedles delivering nucleic acids intracellularly induce localized in vivo neovascularization. Nature Mater. 14, 532–539 (2015).

    CAS  Article  Google Scholar 

  50. Copp, J. A. et al. Clearance of pathological antibodies using biomimetic nanoparticles. Proc. Natl Acad. Sci. USA 111, 13481–13486 (2014).

    CAS  Article  Google Scholar 

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Acknowledgements

The authors would like to gratefully acknowledge M. Ferrari for valuable and stimulating discussions about the study. The authors would like to thank J. You for his help in the animal procedures. The authors acknowledge support from the National Institute of Health (1R21CA173579-01A1 and 5U54CA143837 PSOC Pilot project), the Department of Defense (W81XWH-12-10414 BCRP Innovator Expansion), George J. and Angelina P. Kostas Charitable Foundation, Brown Foundation Inc., William Randolph Hearst Foundation, and The Regenerative Medicine Program Cullen Trust for Health Care to E.T.; R.M. was supported by grant RF-2010-2305526; C.C. and A.P. were supported by grant RF-2010-2318372 from Italian Ministry of Health. We thank Associazione Bianca Garavaglia, Via C. Cattaneo, 8, 21052 Busto Arsizio Varese, Italy and Project CREME ‘Campania Research in Experimental Medicine’ POR Campania FSE 2007/2013. We ackowledge D. A. Engler and the Proteomics Core, D. Haviland and the Flow Cytometry Core, A. L. Rivera and the Research Pathology Core at HMRI. We thank M. G. Landry and M. Evangelopoulos for graphical assistance with the creation of the schematics. The authors also acknowledge the Sealy Center for Structural Biology and Molecular Biophysics at the University of Texas Medical Branch at Galveston for providing research resources.

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Contributions

E.T. conceived the leukosome platform, wrote the paper and was the principal investigator of the major supporting grants. E.T. and R.M. designed the research project and defined the goals of the present study. R.M. developed and optimized the protocols for leukosome assembly, supervised all the experiments, and evaluated the therapeutic efficacy with contributions from J.O.M. and E.D.R.; C.C. performed all the proteomic experiments and the interpretation of the data on protein enrichment; J.O.M. and E.D.R. performed the intravital microscopy experiments and analysis; M.E. performed flow cytometry and optimized the in vitro flow systems. J.O.M. carried out bioluminescence imaging (BLI) analysis and revised the manuscript; F.T. performed FTIR and AFM analyses; S.M. performed DSC analysis; F.T., S.M., and A.D.V. performed H&E and immunofluorescence staining and optical and confocal laser microscopy imaging; M.B.S. performed Cryo-TEM and assisted with analysis; I.K.Y. performed cytokine and organ functionality analyses; P.Z. performed the immunological analysis of leukosomes and gave his expert advice about the study of the immunogenic response; N.E.T.F. performed dexamethasone loading and release experiments; X.W. performed the PCR analysis; A.P. assisted with the editing of the manuscript and mentored the authors during the development of the project.

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Correspondence to E. Tasciotti.

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Molinaro, R., Corbo, C., Martinez, J. et al. Biomimetic proteolipid vesicles for targeting inflamed tissues. Nature Mater 15, 1037–1046 (2016). https://doi.org/10.1038/nmat4644

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