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Biomimetic proteolipid vesicles for targeting inflamed tissues

Nature Materials volume 15, pages 1037–1046 (2016)Cite this article

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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.

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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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Authors and Affiliations

  1. Department of Regenerative Medicine, Houston Methodist Research Institute, Houston, Texas 77030, USA

    R. Molinaro, C. Corbo, J. O. Martinez, F. Taraballi, M. Evangelopoulos, S. Minardi, I. K. Yazdi, E. De Rosa, N. E. Toledano Furman, X. Wang, A. Parodi & E. Tasciotti

  2. CEINGE-Biotecnologie Avanzate s.c.a.r.l., Via G. Salvatore 486, 80145 Naples, Italy

    C. Corbo

  3. IRCCS SDN, Via Gianturco 113, 80143 Naples, Italy

    C. Corbo & A. Parodi

  4. Pain Therapy Service, Fondazione IRCCS Policlinico San Matteo, Pavia 27100, Italy

    F. Taraballi

  5. Department of Pathology and Genomic Medicine, Houston Methodist Research Institute, Houston, Texas 77030, USA

    P. Zhao

  6. Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, Texas 77555, USA

    M. B. Sherman

  7. Osteoncology and Rare Tumors Center, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), 47014 Meldola, Italy

    A. De Vita

  8. Department of Orthopedics, Houston Methodist Hospital, Houston, Texas 77030, USA

    E. Tasciotti

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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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