Skip to main content
SpringerLink
Log in

Polymersomes scalably fabricated via flash nanoprecipitation are non-toxic in non-human primates and associate with leukocytes in the spleen and kidney following intravenous administration

Nano Research volume 11, pages 5689–5703 (2018)Cite this article

Abstract

Vesicular nanocarrier formulations confer the ability to deliver hydrophobic and hydrophilic cargos simultaneously to cells of interest in vivo. While liposomal formulations reached the clinic long ago, younger technologies such as polymeric vesicles (polymersomes) have yet to make the transition to clinical approval and use, in part due to difficulties in ensuring their safe and scalable production. In this work, we demonstrate the scalable production of poly(ethylene glycol)-block-poly(propylene sulfide) (PEG-bl-PPS) polymersomes via flash nanoprecipitation, and further show the safe administration of these nanocarriers to mice and non-human primates. In mice, PEG-bl-PPS polymersomes were found to be well tolerated at up to 200 mg/(kg·week). Following the administration of a more relevant 20 mg/(kg·week) dosage in non-human primates, polymersomes were found to associate with numerous phagocytic immune cell populations, including a remarkable 68% of plasmacytoid dendritic cells and > 95% of macrophages in the spleen, while showing no toxicity or abnormalities in the liver, kidney, spleen, or blood. Despite the presence of a dense PEG corona, neither anti-PEG antibodies nor complement activation were detected. This work provides evidence of the translatability of PEG-bl-PPS polymersomes into the clinic for therapeutic applications in humans.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

References

  1. Bobo, D.; Robinson, K. J.; Islam, J.; Thurecht, K. J.; Corrie, S. R. Nanoparticle-based medicines: A review of FDA-approved materials and clinical trials to date. Pharm. Res. 2016, 33, 2373–2387.

    Article  Google Scholar 

  2. Torchilin, V. P. Multifunctional, stimuli-sensitive nanoparticulate systems for drug delivery. Nat. Rev. Drug. Discov. 2014, 13, 813–827.

    Article  Google Scholar 

  3. Singh, R.; Lillard, J. W. Nanoparticle-based targeted drug delivery. Exp. Mol. Pathol. 2009, 86, 215–223.

    Article  Google Scholar 

  4. Allen, S.; Osorio, O.; Liu, Y. G.; Scott, E. Facile assembly and loading of theranostic polymersomes via multi-impingement flash nanoprecipitation. J. Control. Release 2017, 262, 91–103.

    Article  Google Scholar 

  5. Dowling, D. J.; Scott, E. A.; Scheid, A.; Bergelson, I.; Joshi, S.; Pietrasanta, C.; Brightman, S.; Sanchez-Schmitz, G.; Van Haren, S. D.; Ninkovic, J. et al. Toll-like receptor 8 agonist nanoparticles mimic immunomodulating effects of the live BCG vaccine and enhance neonatal innate and adaptive immune responses. J. Allergy Clin. Immunol. 2017, 140, 1339–1350.

    Article  Google Scholar 

  6. Discher, D. E.; Eisenberg, A. Polymer vesicles. Science 2002, 297, 967–973.

    Article  Google Scholar 

  7. Cerritelli, S.; O'Neil, C. P.; Velluto, D.; Fontana, A.; Adrian, M.; Dubochet, J.; Hubbell, J. A. Aggregation behavior of poly(ethylene glycol-bl-propylene sulfide) Di-and triblock copolymers in aqueous solution. Langmuir 2009, 25, 11328–11335.

    Article  Google Scholar 

  8. Yi, S. J.; Allen, S. D.; Liu, Y. G.; Ouyang, B. Z.; Li, X. M.; Augsornworawat, P.; Thorp, E. B.; Scott, E. A. Tailoring nanostructure morphology for enhanced targeting of dendritic cells in atherosclerosis. ACS Nano 2016, 10, 11290–11303.

    Article  Google Scholar 

  9. Bobbala, S.; Allen, S. D.; Scott, E. A. Flash nanoprecipitation permits versatile assembly and loading of polymeric bicontinuous cubic nanospheres. Nanoscale 2018, 10, 5078–5088.

    Article  Google Scholar 

  10. Karabin, N. B.; Allen, S.; Kwon, H. K.; Bobbala, S.; Firlar, E.; Shokuhfar, T.; Shull, K. R.; Scott, E. A. Sustained micellar delivery via inducible transitions in nanostructure morphology. Nat. Commun. 2018, 9, 624.

    Article  Google Scholar 

  11. Napoli, A.; Valentini, M.; Tirelli, N.; Müller, M.; Hubbell, J. A. Oxidation-responsive polymeric vesicles. Nat. Mater. 2004, 3, 183–189.

    Article  Google Scholar 

  12. Vasdekis, A. E.; Scott, E. A.; O'Neil, C. P.; Psaltis, D.; Hubbell, J. A. Precision intracellular delivery based on optofluidic polymersome rupture. ACS Nano 2012, 6, 7850–7857.

    Article  Google Scholar 

  13. Scott, E. A.; Stano, A.; Gillard, M.; Maio-Liu, A. C.; Swartz, M. A.; Hubbell, J. A. Dendritic cell activation and T cell priming with adjuvant-and antigen-loaded oxidation-sensitive polymersomes. Biomaterials 2012, 33, 6211–6219.

    Article  Google Scholar 

  14. Stano, A.; Scott, E. A.; Dane, K. Y.; Swartz, M. A.; Hubbell, J. A. Tunable T cell immunity towards a protein antigen using polymersomes vs. solid-core nanoparticles. Biomaterials 2013, 34, 4339–4346.

    Article  Google Scholar 

  15. Eetezadi, S.; Ekdawi, S. N.; Allen, C. The challenges facing block copolymer micelles for cancer therapy: In vivo barriers and clinical translation. Adv. Drug. Deliver. Rev. 2015, 91, 7–22.

    Article  Google Scholar 

  16. Anselmo, A. C.; Prabhakarpandian, B.; Pant, K.; Mitragotri, S. Clinical and commercial translation of advanced polymeric nanoparticle systems: Opportunities and material challenges. Transl. Mater. Res. 2017, 4, 014001.

    Article  Google Scholar 

  17. O'Neil, C. P.; Suzuki, T.; Demurtas, D.; Finka, A.; Hubbell, J. A. A novel method for the encapsulation of biomolecules into polymersomes via direct hydration. Langmuir 2009, 25, 9025–9029.

    Article  Google Scholar 

  18. Rameez, S.; Bamba, I.; Palmer, A. F. Large scale production of vesicles by hollow fiber extrusion: A novel method for generating polymersome encapsulated hemoglobin dispersions. Langmuir 2010, 26, 5279–5285.

    Article  Google Scholar 

  19. Tang, C.; Amin, D.; Messersmith, P. B.; Anthony, J. E.; Prud'homme, R. K. Polymer directed self-assembly of pH-responsive antioxidant nanoparticles. Langmuir 2015, 31, 3612–3620.

    Article  Google Scholar 

  20. Tang C.; Amin D.; Messersmith P.B.; Anthony J. E.; Prud'homme R. K. Polymer directed self-assembly of pH-responsive antioxidant nanoparticles. Langmuir 2015, 31, 3612–3620.

    Article  Google Scholar 

  21. Johnson, B. K.; Prud'homme, R. K. Flash nano precipitation of organic actives and block copolymers using a confined impinging jets mixer. Aust. J. Chem. 2003, 56, 1021–1024.

    Article  Google Scholar 

  22. Han, J.; Zhu, Z. X.; Qian, H. T.; Wohl, A. R.; Beaman, C. J.; Hoye, T. R.; Macosko, C. W. A simple confined impingement jets mixer for flash nanoprecipitation. J. Pharm. Sci. 2012, 101, 4018–4023.

    Article  Google Scholar 

  23. Johnson, B. K.; Prud'homme, R. K. Chemical processing and micromixing in confined impinging jets. AIChE J 2003, 49, 2264–2282.

    Article  Google Scholar 

  24. Saad, W. S.; Prud'homme, R. K. Principles of nanoparticle formation by flash nanoprecipitation. Nano Today 2016, 11, 212–227.

    Article  Google Scholar 

  25. Liu, Y.; Cheng, C. Y.; Liu, Y.; Prud'homme, R. K.; Fox, R. O. Mixing in a multi-inlet vortex mixer (MIVM) for flash nano-precipitation. Chem. Eng. Sci. 2008, 63, 2829–2842.

    Article  Google Scholar 

  26. Maecker, H. T.; McCoy, J. P.; Nussenblatt, R. Standardizing immunophenotyping for the Human Immunology Project. Nat. Rev. Immunol. 2012, 12, 471.

    Article  Google Scholar 

  27. Guilliams, M.; Ginhoux, F.; Jakubzick, C.; Naik, S. H.; Onai, N.; Schraml, B. U.; Segura, E.; Tussiwand, R.; Yona, S. Dendritic cells, monocytes and macrophages: A unified nomenclature based on ontogeny. Nat. Rev. Immunol. 2014, 14, 571–578.

    Article  Google Scholar 

  28. Malyala, P.; Singh, M. Endotoxin limits in formulations for preclinical research. J. Pharm. Sci. 2008, 97, 2041–2044.

    Article  Google Scholar 

  29. Dane, K. Y.; Nembrini, C.; Tomei, A. A.; Eby, J. K.; O'Neil, C. P.; Velluto, D.; Swartz, M. A.; Inverardi, L.; Hubbell, J. A. Nano-sized drug-loaded micelles deliver payload to lymph node immune cells and prolong allograft survival. J. Control. Release 2011, 156, 154–160.

    Article  Google Scholar 

  30. Hoffman, W. P.; Ness, D. K.; van Lier, R. B. Analysis of rodent growth data in toxicology studies. Toxicol. Sci. 2002, 66, 313–319.

    Article  Google Scholar 

  31. Hall, A. P.; Elcombe, C. R.; Foster, J. R.; Harada, T.; Kaufmann, W.; Knippel, A.; Küttler, K.; Malarkey, D. E.; Maronpot, R. R.; Nishikawa, A. et al. Liver hypertrophy: A review of adaptive (adverse and non-adverse) changes—conclusions from the 3rd International ESTP Expert Workshop. Toxicol. Pathol. 2012, 40, 971–994.

    Article  Google Scholar 

  32. Lake, B. G.; Evans, J. G.; Gray, T. J. B.; Körösi, S. A.; North, C. J. Comparative studies on nafenopin-induced hepatic peroxisome proliferation in the rat, Syrian hamster, guinea pig, and marmoset. Toxicol. Appl. Pharmacol. 1989, 99, 148–160.

    Article  Google Scholar 

  33. Ray, K. Liver: Clearance of nanomaterials in the liver. Nat. Rev. Gastroenterol. Hepatol. 2016, 13, 560.

    Article  Google Scholar 

  34. Armstrong, J. K.; Hempel, G.; Koling, S.; Chan, L. S.; Fisher, T.; Meiselman, H. J.; Garratty, G. Antibody against poly(ethylene glycol) adversely affects PEG-asparaginase therapy in acute lymphoblastic leukemia patients. Cancer 2007, 110, 103–111.

    Article  Google Scholar 

  35. Richter, A. W.; Åkerblom, E. Polyethylene glycol reactive antibodies in man: Titer distribution in allergic patients treated with monomethoxy polyethylene glycol modified allergens or placebo, and in healthy blood donors. Int. Arch. Allergy Appl. Immunol. 1984, 74, 36–39.

    Article  Google Scholar 

  36. Yang, Q.; Lai, S. K. Anti-PEG immunity: Emergence, characteristics, and unaddressed questions. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 2015, 7, 655–677.

    Article  Google Scholar 

  37. Judge, A.; McClintock, K.; Phelps, J. R.; Maclachlan, I. Hypersensitivity and loss of disease site targeting caused by antibody responses to PEGylated liposomes. Mol. Ther. 2006, 13, 328–337.

    Article  Google Scholar 

  38. Wang, X. Y.; Ishida, T.; Kiwada, H. Anti-PEG IgM elicited by injection of liposomes is involved in the enhanced blood clearance of a subsequent dose of PEGylated liposomes. J. Control. Release 2007, 119, 236–244.

    Article  Google Scholar 

  39. Yang, W.; Liu, S. J.; Bai, T.; Keefe, A. J.; Zhang, L.; Ella-Menye, J. R.; Li, Y. T.; Jiang, S. Y. Poly(carboxybetaine) nanomaterials enable long circulation and prevent polymerspecific antibody production. Nano Today 2014, 9, 10–16.

    Article  Google Scholar 

  40. Szebeni, J.; Baranyi, L.; Savay, S.; Milosevits, J.; Bunger, R.; Laverman, P.; Metselaar, J. M.; Storm, G.; Chanan-Khan, A.; Liebes, L. et al. Role of complement activation in hypersensitivity reactions to doxil and hynic PEG liposomes: Experimental and clinical studies. J. Liposome Res. 2002, 12, 165–172.

    Article  Google Scholar 

  41. Szebeni, J.; Baranyi, L.; Savay, S.; Bodo, M.; Morse, D. S.; Basta, M.; Stahl, G. L.; Bünger, R.; Alving, C. R. Liposome-induced pulmonary hypertension: Properties and mechanism of a complement-mediated pseudoallergic reaction. Am. J. Physiol. Heart Circ. Physiol. 2000, 279, H1319–H1328.

    Article  Google Scholar 

  42. Park, J. K.; Utsumi, T.; Seo, Y. E.; Deng, Y.; Satoh, A.; Saltzman, W. M.; Iwakiri, Y. Cellular distribution of injected PLGA-nanoparticles in the liver. Nanomed.-Nanotechnol. Biol. Med. 2016, 12, 1365–1374.

    Article  Google Scholar 

  43. Huh, Y.; Smith, D. E.; Feng, M. R. Interspecies scaling and prediction of human clearance: Comparison of small-and macro-molecule drugs. Xenobiotica 2011, 41, 972–987.

    Article  Google Scholar 

  44. Vaure, C.; Liu, Y. Q. A comparative review of toll-like receptor 4 expression and functionality in different animal species. Front. Immunol. 2014, 5, 316.

    Article  Google Scholar 

  45. Chiarelli, P. A.; Revia, R. A.; Stephen, Z. R.; Wang, K.; Jeon, M.; Nelson, V.; Kievit, F. M.; Sham, J.; Ellenbogen, R. G.; Kiem, H. P. et al. Nanoparticle biokinetics in mice and nonhuman primates. ACS Nano 2017, 11, 9514–9524.

    Article  Google Scholar 

  46. Velásquez-Lopera, M. M.; Correa, L. A.; García, L. F. Human spleen contains different subsets of dendritic cells and regulatory T lymphocytes. Clin. Exp. Immunol. 2008, 154, 107–114.

    Article  Google Scholar 

  47. Du, F. F.; Liu, Y. G.; Scott, E. A. Immunotheranostic polymersomes modularly assembled from tetrablock and diblock copolymers with oxidation-responsive fluorescence. Cell. Mol. Bioeng. 2017, 10, 357–370.

    Article  Google Scholar 

Download references

Acknowledgements

We acknowledge staff and instrumentation support from the Structural Biology Facility at Northwestern University, the Robert H Lurie Comprehensive Cancer Center of Northwestern University and NCI CCSG P30 CA060553. The Gatan K2 direct electron detector was purchased with funds provided by the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust. SAXS experiments were performed at the DuPont-Northwestern-Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source (APS). DND-CAT is supported by Northwestern University, E.I. DuPont de Nemours & Co., and The Dow Chemical Company. This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02- 06CH11357. This work made use of the EPIC facility of Northwestern University’s NUANCE Center, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205); the MRSEC program (NSF DMR-1121262) at the Materials Research Center; the International Institute for Nanotechnology (IIN); the Keck Foundation; and the State of Illinois, through the IIN. This work made use of the IMSERC at Northwestern University, which has received support from the NSF (CHE-1048773); Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205); the State of Illinois and International Institute for Nanotechnology (IIN). This work was supported by the Northwestern University–Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI CA060553). Imaging work was performed at the Northwestern University Center for Advanced Molecular Imaging generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. The authors acknowledge Jonathan Remis (Structural Biology Facility, NU) for his contribution to cryoTEM image acquisition. We acknowledge Sierra M. Paxton and Courtney R. Burkett for their excellent technical assistance with the NHP study.

Author information

Authors and Affiliations

  1. Interdisciplinary Biological Sciences, Northwestern University, Evanston, IL, 60208, USA

    Sean D. Allen & Evan A. Scott

  2. Department of Biomedical Engineering, Northwestern University, Evanston, 60208, IL, USA

    Yu-Gang Liu, Sharan Bobbala & Evan A. Scott

  3. Saha Cardiovascular Research Center, University of Kentucky, Lexington, KY, 40506, USA

    Lei Cai, Peter I. Hecker & Ryan Temel

  4. Department of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, 40506, USA

    Peter I. Hecker & Ryan Temel

Authors
  1. Sean D. Allen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yu-Gang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sharan Bobbala
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lei Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter I. Hecker
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ryan Temel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Evan A. Scott
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Evan A. Scott.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Allen, S.D., Liu, YG., Bobbala, S. et al. Polymersomes scalably fabricated via flash nanoprecipitation are non-toxic in non-human primates and associate with leukocytes in the spleen and kidney following intravenous administration. Nano Res. 11, 5689–5703 (2018). https://doi.org/10.1007/s12274-018-2069-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12274-018-2069-x

Keywords

search

Navigation