Skip to main content
SpringerLink
Log in

Targeting, Endocytosis, and Lysosomal Delivery of Active Enzymes to Model Human Neurons by ICAM-1-Targeted Nanocarriers

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose

Delivery of therapeutics to neurons is paramount to treat neurological conditions, including many lysosomal storage disorders. However, key aspects of drug-carrier behavior in neurons are relatively unknown: the occurrence of non-canonical endocytic pathways (present in other cells); whether carriers that traverse the blood–brain barrier are, contrarily, retained within neurons; if neuron-surface receptors are accessible to bulky carriers compared to small ligands; or if there are differences regarding neuronal compartments (neuron body vs. neurites) pertaining said parameters. We have explored these questions using model polymer nanocarriers targeting intercellular adhesion molecule-1 (ICAM-1).

Methods

Differentiated human neuroblastoma cells were incubated with anti-ICAM-coated polystyrene nanocarriers and analyzed by fluorescence microscopy.

Results

ICAM-1 expression and nanocarrier binding was enhanced in altered (TNFα) vs. control conditions. While small ICAM-1 ligands (anti-ICAM) preferentially accessed the cell body, anti-ICAM nanocarriers bound with faster kinetics to neurites, yet reached similar saturation over time. Anti-ICAM nanocarriers were also endocytosed with faster kinetics and lower saturation levels in neurites. Non-classical cell adhesion molecule (CAM) endocytosis ruled uptake, and neurite-to-cell body transport was inferred. Nanocarriers trafficked to lysosomes, delivering active enzymes (dextranase) with substrate reduction in a lysosomal-storage disease model.

Conclusion

ICAM-1-targeting holds potential for intracellular delivery of therapeutics to neurons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

BBB:

Blood–brain barrier

Bmax:

Maximal binding

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

ICAM-1:

Intercellular adhesion molecule-1

IgG:

Immunoglobulin G

Imax:

Maximal internalization

LSD:

Lysosomal storage disorder

MDC:

Monodansylcadaverine

NC:

Nanocarrier

PBS:

Phosphate buffer saline

PDI:

Polydispersity index

Tmax:

Maximal transport to lysosomes.

References

  1. Barchet TM, Amiji MM. Challenges and opportunities in CNS delivery of therapeutics for neurodegenerative diseases. Expert Opin Drug Deliv. 2009;6(3):211–25.

    Article  CAS  PubMed  Google Scholar 

  2. Abbott NJ. Blood–brain barrier structure and function and the challenges for CNS drug delivery. J Inherit Metab Dis. 2013;36(3):437–49.

    Article  CAS  PubMed  Google Scholar 

  3. Dhuria SV, Hanson LR, Frey WH. 2nd. Intranasal delivery to the central nervous system: mechanisms and experimental considerations. J Pharm Sci. 2010;99(4):1654–73.

    CAS  PubMed  Google Scholar 

  4. Lakhal S, Wood MJ. Exosome nanotechnology: an emerging paradigm shift in drug delivery: exploitation of exosome nanovesicles for systemic in vivo delivery of RNAi heralds new horizons for drug delivery across biological barriers. BioEssays. 2011;33(10):737–41.

    Article  CAS  PubMed  Google Scholar 

  5. Neuwelt E, Abbott NJ, Abrey L, Banks WA, Blakley B, Davis T, et al. Strategies to advance translational research into brain barriers. Lancet Neurol. 2008;7(1):84–96.

    Article  CAS  PubMed  Google Scholar 

  6. Muro S. Strategies for delivery of therapeutics into the central nervous system for treatment of lysosomal storage disorders. Drug Deliv Transl Res. 2012;2(3):169–86.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Hoffman AS. The origins and evolution of “controlled” drug delivery systems. J Control Release. 2008;132(3):153–63.

    Article  CAS  PubMed  Google Scholar 

  8. Muro S. Challenges in design and characterization of ligand-targeted drug delivery systems. J Control Release. 2012;164(2):125–37.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Albertazzi L, Gherardini L, Brondi M, Sulis Sato S, Bifone A, Pizzorusso T, et al. In vivo distribution and toxicity of PAMAM dendrimers in the central nervous system depend on their surface chemistry. Mol Pharm. 2012;10(1):249–60.

    Article  PubMed  Google Scholar 

  10. Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A. 1995;92(1):7297–301.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. O’Mahony AM, Godinho BM, Ogier J, Devocelle M, Darcy R, Cryan JF, et al. Click-modified cyclodextrins as nonviral vectors for neuronal siRNA delivery. ACS Chem Neurosci. 2012;3(10):744–52.

    Article  PubMed Central  PubMed  Google Scholar 

  12. Boado RJ, Zhang Y, Wang Y, Pardridge WM. Engineering and expression of a chimeric transferrin receptor monoclonal antibody for blood–brain barrier delivery in the mouse. Biotechnol Bioeng. 2009;102(4):1251–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. LeBowitz JH, Grubb JH, Maga JA, Schmiel DH, Vogler C, Sly WS. Glycosylation-independent targeting enhances enzyme delivery to lysosomes and decreases storage in mucopolysaccharidosis type VII mice. Proc Natl Acad Sci U S A. 2004;101(9):3083–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Osborn MJ, McElmurry RT, Peacock B, Tolar J, Blazar BR. Targeting of the CNS in MPS-IH using a nonviral transferrin-alpha-L-iduronidase fusion gene product. Mol Ther. 2008;16(8):1459–66.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Budzinski KL, Sgro AE, Fujimoto BS, Gadd JC, Shuart NG, Gonen T, et al. Synaptosomes as a platform for loading nanoparticles into synaptic vesicles. ACS Chem Neurosci. 2011;2(5):236–41.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Deinhardt K, Berninghausen O, Willison HJ, Hopkins CR, Schiavo G. Tetanus toxin is internalized by a sequential clathrin-dependent mechanism initiated within lipid microdomains and independent of epsin1. J Cell Biol. 2006;174(3):459–71.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Francesconi A, Kumari R, Zukin RS. Regulation of group I metabotropic glutamate receptor trafficking and signaling by the caveolar/lipid raft pathway. J Neurosci. 2009;29(11):3590–602.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Rothlein R, Springer TA. The requirement for lymphocyte function-associated antigen 1 in homotypic leukocyte adhesion stimulated by phorbol ester. J Exp Med. 1986;163(5):1132–49.

    Article  CAS  PubMed  Google Scholar 

  19. Hsu J, Rappaport J, Muro S. Specific binding, uptake, and transport of ICAM-1-targeted nanocarriers across endothelial and subendothelial cell components of the blood–brain barrier. Pharm Res. 2014;31(7):1855–66.

    Article  CAS  PubMed  Google Scholar 

  20. Hsu J, Serrano D, Bhowmick T, Kumar K, Shen Y, Kuo YC, et al. Enhanced endothelial delivery and biochemical effects of alpha-galactosidase by ICAM-1-targeted nanocarriers for Fabry disease. J Control Release. 2011;149(3):323–31.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Papademetriou I, Garnacho C, Serrano D, Bhowmick T, Schuchman EH, Muro S. Comparative binding, endocytosis, and biodistribution of antibodies and antibody-coated carriers for targeted delivery of lysosomal enzymes to ICAM-1 versus transferrin receptor. J Inherit Metab Dis. 2013;36(3):467–77.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Ansar M, Serrano D, Papademetriou I, Bhowmick TK, Muro S. Biological functionalization of drug delivery carriers to bypass size restrictions of receptor-mediated endocytosis independently from receptor targeting. ACS Nano. 2013;7(12):10597–611.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Muro S, Garnacho C, Champion JA, Leferovich J, Gajewski C, Schuchman EH, et al. Control of endothelial targeting and intracellular delivery of therapeutic enzymes by modulating the size and shape of ICAM-1-targeted carriers. Mol Ther. 2008;16(8):1450–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Serrano D, Bhowmick T, Chadha R, Garnacho C, Muro S. Intercellular adhesion molecule 1 engagement modulates sphingomyelinase and ceramide, supporting uptake of drug carriers by the vascular endothelium. Arterioscler Thromb Vasc Biol. 2012;32(5):1178–85.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Muro S, Wiewrodt R, Thomas A, Koniaris L, Albelda SM, Muzykantov VR, et al. A novel endocytic pathway induced by clustering endothelial ICAM-1 or PECAM-1. J Cell Sci. 2003;116(Pt 8):1599–609.

    Article  CAS  PubMed  Google Scholar 

  26. Ghaffarian R, Bhowmick T, Muro S. Transport of nanocarriers across gastrointestinal epithelial cells by a new transcellular route induced by targeting ICAM-1. J Control Release. 2012;163(1):25–33.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Muro S. A DNA device that mediates selective endosomal escape and intracellular delivery of drugs and biologicals. Adv Funct Mater. 2014;24(19):2899–906.

    Article  CAS  PubMed  Google Scholar 

  28. Gimenez-Cassina A, Lim F, Diaz-Nido J. Differentiation of a human neuroblastoma into neuron-like cells increases their susceptibility to transduction by herpesviral vectors. J Neurosci Res. 2006;84(4):755–67.

    Article  CAS  PubMed  Google Scholar 

  29. Muro S, Cui X, Gajewski CM, Murciano J-C, Muzykantov VR, Koval M. Slow intracellular trafficking of catalase nanoparticles targeted to ICAM-1 protects endothelial cells from oxidative stress. Am J Phys Cell Physiol. 2003;285(5):C1339–47.

    Article  CAS  Google Scholar 

  30. Birdsall HH. Induction of ICAM-1 on human neural cells and mechanisms of neutrophil-mediated injury. Am J Pathol. 1991;139(6):1341–50.

    PubMed Central  CAS  PubMed  Google Scholar 

  31. Muro S, Dziubla T, Qiu W, Leferovich J, Cui X, Berk E. Muzykantov VR. Endothelial targeting of high-affinity multivalent polymer nanocarriers directed to intercellular adhesion molecule 1. J Pharmacol Exp Ther. 2006;317(3):1161–9.

    Article  CAS  PubMed  Google Scholar 

  32. Khalikova E, Susi P, Korpela T. Microbial dextran-hydrolyzing enzymes: fundamentals and applications. Microbiol Mol Biol Rev. 2005;69(2):306–25.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  33. Augustin R, Riley J, Moley KH. GLUT8 contains a [DE] XXXL [LI] sorting motif and localizes to a late endosomal/lysosomal compartment. Traffic. 2005;6:1196–212.

    Article  CAS  PubMed  Google Scholar 

  34. Garnacho C, Albelda SM, Muzykantov VR, Muro S. Differential intra-endothelial delivery of polymer nanocarriers targeted to distinct PECAM-1 epitopes. J Control Release. 2008;130(3):226–33.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Rappaport J, Garnacho C, Muro S. Clathrin-mediated endocytosis is impaired in type A-B Niemann-Pick disease model cells and can be restored by ICAM-1-mediated enzyme replacement. Mol Pharm. 2014;11(8):2887–95.

    Article  CAS  PubMed  Google Scholar 

  36. Blanpied TA, Scott DB, Ehlers MD. Dynamics and regulation of clathrin coats at specialized endocytic zones of dendrites and spines. Neuron. 2002;36(3):435–49.

    Article  CAS  PubMed  Google Scholar 

  37. Roberts VJ, Gorenstein C. Examination of the transient distribution of lysosomes in neurons of developing rat brains. Dev Neurosci. 1987;9(4):255–64.

    Article  CAS  PubMed  Google Scholar 

  38. Deinhardt K, Salinas S, Verastegui C, Watson R, Worth D, Hanrahan S, et al. Rab5 and Rab7 control endocytic sorting along the axonal retrograde transport pathway. Neuron. 2006;52(2):293–305.

    Article  CAS  PubMed  Google Scholar 

  39. Millecamps S, Julien JP. Axonal transport deficits and neurodegenerative diseases. Nat Rev Neurosci. 2013;14(3):161–76.

    Article  CAS  PubMed  Google Scholar 

  40. Torchilin VP. Recent approaches to intracellular delivery of drugs and DNA and organelle targeting. Annu Rev Biomed Eng. 2006;8:343–75.

    Article  CAS  PubMed  Google Scholar 

Download references

ACKNOWLEDGMENTS AND DISCLOSURES

The authors thank Dr. Estrella Rubio Solsona (Program in Rare and Genetic Diseases & IBV/CSIC Associated Unit, Centro de Investigación Príncipe Felipe, Valencia, Spain) for her guidance in propagating and differentiating SH-SY5Y cells. We also thank Rachel Manthe (Department of Bioengineering, University of Maryland, College Park, MD, USA) for help with grammar edits. This work was supported by NIH grant R01-HL09816 and NSF award CBET-1402756 (S.M.).

Author information

Authors and Affiliations

  1. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland, 20742, USA

    Janet Hsu & Silvia Muro

  2. Program in Rare and Genetic Diseases & IBV/CSIC Associated Unit Centro de Investigación Príncipe Felipe, Valencia, Spain

    Janet Hoenicka

  3. Centro de Investigación Biomédica en Red de Salud Mental (CIBERSAM) ISCIII, Valencia, Spain

    Janet Hoenicka

  4. Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, Maryland, 20742, USA

    Silvia Muro

Authors
  1. Janet Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Janet Hoenicka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Silvia Muro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Silvia Muro.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hsu, J., Hoenicka, J. & Muro, S. Targeting, Endocytosis, and Lysosomal Delivery of Active Enzymes to Model Human Neurons by ICAM-1-Targeted Nanocarriers. Pharm Res 32, 1264–1278 (2015). https://doi.org/10.1007/s11095-014-1531-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-014-1531-z

KEY WORDS

Search

Navigation