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Imaging the Cytosolic Drug Delivery Mechanism of HDL-Like Nanoparticles

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ABSTRACT

Purpose

Molecular therapeutics often require an effective nanoparticle-based delivery strategy to transport them to cytosolic organelles to be functional. Recently, a cytosolic delivery strategy based on the scavenger receptor class B type I (SR-BI) mediated pathway has shown great potential for the effective delivery of theranostics agents into the cytoplasm of cells without detrimental endosomal entrapment. This study elucidates this unique delivery mechanism for improving cytosolic drug delivery.

Methods

Multifluorophore-labeled HDL-mimicking peptide phospholipid scaffold (HPPS) nanoparticles were developed. Fluorescence imaging was utilized to examine HPPS transporting payloads into cells step by step through sequential inhibition studies.

Results

HPPS specifically recognizes and binds to SR-BI, then interacts with SR-BI, which results in direct transport of payload molecules into the cell cytoplasm without entire particles internalization. The cytosolic transport of payloads occurred through a temperature- and energy-independent pathway, and was also different from actin- and clathrin-mediated endocytosis. Furthermore, this transport was significantly inhibited by disruption of lipid rafts using filipin or methyl-β-cyclodextrin.

Conclusions

The cytosolic delivery of payloads by HPPS via SR-BI targeting is predominately mediated through a lipid rafts/caveolae-like pathway. This cytosolic delivery strategy can be utilized for transporting molecular therapeutics that require their action sites to be within cytosolic organelles to enhance therapeutic effect.

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REFERENCES

  1. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998;391(6669):806–11.

    Article  CAS  PubMed  Google Scholar 

  2. Vasir JK, Labhasetwar V. Biodegradable nanoparticles for cytosolic delivery of therapeutics. Adv Drug Deliv Rev. 2007;59(8):718–28.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Mathew E, Hardee GE, Bennett CF, Lee KD. Cytosolic delivery of antisense oligonucleotides by listeriolysin O-containing liposomes. Gene Ther. 2003;10(13):1105–15.

    Article  CAS  PubMed  Google Scholar 

  4. Lee SH, Choi SH, Kim SH, Park TG. Thermally sensitive cationic polymer nanocapsules for specific cytosolic delivery and efficient gene silencing of siRNA: swelling induced physical disruption of endosome by cold shock. J Control Release. 2008;125(1):25–32.

    Article  CAS  PubMed  Google Scholar 

  5. Yezhelyev MV, Qi L, O’Regan RM, Nie S, Gao X. Proton-sponge coated quantum dots for siRNA delivery and intracellular imaging. J Am Chem Soc. 2008;130(28):9006–12.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Raoof M, Corr SJ, Kaluarachchi WD, Massey KL, Briggs K, Zhu C, et al. Stability of antibody-conjugated gold nanoparticles in the endolysosomal nanoenvironment: implications for noninvasive radiofrequency-based cancer therapy. Nanomedicine. 2012;8(7):1096–105.

    Google Scholar 

  7. Berg K, Selbo PK, Prasmickaite L, Tjelle TE, Sandvig K, Moan J, et al. Photochemical internalization: a novel technology for delivery of macromolecules into cytosol. Cancer Res. 1999;59(6):1180–3.

    CAS  PubMed  Google Scholar 

  8. Hu Y, Litwin T, Nagaraja AR, Kwong B, Katz J, Watson N, et al. Cytosolic delivery of membrane-impermeable molecules in dendritic cells using pH-responsive core-shell nanoparticles. Nano Lett. 2007;7(10):3056–64.

    Article  CAS  PubMed  Google Scholar 

  9. Petros RA, DeSimone JM. Strategies in the design of nanoparticles for therapeutic applications. Nat Rev Drug Discov. 2010;9(8):615–27.

    Article  CAS  PubMed  Google Scholar 

  10. Madani F, Lindberg S, Langel U, Futaki S, Graslund A. Mechanisms of cellular uptake of cell-penetrating peptides. J Biophys. 2011;2011:414729.

    PubMed Central  PubMed  Google Scholar 

  11. Richard JP, Melikov K, Vives E, Ramos C, Verbeure B, Gait MJ, et al. Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake. J Biol Chem. 2003;278(1):585–90.

    Article  CAS  PubMed  Google Scholar 

  12. Kunisawa J, Masuda T, Katayama K, Yoshikawa T, Tsutsumi Y, Akashi M, et al. Fusogenic liposome delivers encapsulated nanoparticles for cytosolic controlled gene release. J Control Release. 2005;105(3):344–53.

    Article  CAS  PubMed  Google Scholar 

  13. Partlow KC, Lanza GM, Wickline SA. Exploiting lipid raft transport with membrane targeted nanoparticles: a strategy for cytosolic drug delivery. Biomaterials. 2008;29(23):3367–75.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Zhang Z, Cao W, Jin H, Lovell JF, Yang M, Ding L, et al. Biomimetic nanocarrier for direct cytosolic drug delivery. Angew Chem Int Ed Engl. 2009;48(48):9171–5.

    Article  CAS  PubMed  Google Scholar 

  15. Frias JC, Williams KJ, Fisher EA, Fayad ZA. Recombinant HDL-like nanoparticles: a specific contrast agent for MRI of atherosclerotic plaques. J Am Chem Soc. 2004;126(50):16316–7.

    Article  CAS  PubMed  Google Scholar 

  16. Cormode DP, Skajaa T, van Schooneveld MM, Koole R, Jarzyna P, Lobatto ME, et al. Nanocrystal core high-density lipoproteins: a multimodality contrast agent platform. Nano Lett. 2008;8(11):3715–23.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Lacko AG, Nair M, Paranjape S, Johnso S, McConathy WJ. High density lipoprotein complexes as delivery vehicles for anticancer drugs. Anticancer Res. 2002;22(4):2045–9.

    CAS  PubMed  Google Scholar 

  18. Yang M, Chen J, Cao W, Ding L, Ng KK, Jin H, et al. Attenuation of nontargeted cell-kill using a high-density lipoprotein-mimicking peptide–phospholipid nanoscaffold. Nanomedicine (Lond). 2011;6(4):631–41.

    Article  CAS  Google Scholar 

  19. Shahzad MM, Mangala LS, Han HD, Lu C, Bottsford-Miller J, Nishimura M, et al. Targeted delivery of small interfering RNA using reconstituted high-density lipoprotein nanoparticles. Neoplasia. 2011;13(4):309–19.

    CAS  PubMed Central  PubMed  Google Scholar 

  20. McMahon KM, Mutharasan RK, Tripathy S, Veliceasa D, Bobeica M, Shumaker DK, et al. Biomimetic high density lipoprotein nanoparticles for nucleic acid delivery. Nano Lett. 2011;11(3):1208–14.

    Article  CAS  PubMed  Google Scholar 

  21. Lin Q, Chen J, Jin H, Ng KK, Yang M, Cao W, et al. Efficient systemic delivery of siRNA by using high-density lipoprotein-mimicking peptide lipid nanoparticles. Nanomedicine (Lond). 2012 Jul 26.

  22. Acton S, Rigotti A, Landschulz KT, Xu S, Hobbs HH, Krieger M. Identification of scavenger receptor SR-BI as a high density lipoprotein receptor. Science. 1996;271(5248):518–20.

    Article  CAS  PubMed  Google Scholar 

  23. Kratzer I, Wernig K, Panzenboeck U, Bernhart E, Reicher H, Wronski R, et al. Apolipoprotein A-I coating of protamine-oligonucleotide nanoparticles increases particle uptake and transcytosis in an in vitro model of the blood–brain barrier. J Control Release. 2007;117(3):301–11.

    Article  CAS  PubMed  Google Scholar 

  24. Rhainds D, Bourgeois P, Bourret G, Huard K, Falstrault L, Brissette L. Localization and regulation of SR-BI in membrane rafts of HepG2 cells. J Cell Sci. 2004;117(Pt 15):3095–105.

    Article  CAS  PubMed  Google Scholar 

  25. Rodrigueza WV, Thuahnai ST, Temel RE, Lund-Katz S, Phillips MC, Williams DL. Mechanism of scavenger receptor class B type I-mediated selective uptake of cholesteryl esters from high density lipoprotein to adrenal cells. J Biol Chem. 1999;274(29):20344–50.

    Article  CAS  PubMed  Google Scholar 

  26. Yu M, Romer KA, Nieland TJ, Xu S, Saenz-Vash V, Penman M, et al. Exoplasmic cysteine Cys384 of the HDL receptor SR-BI is critical for its sensitivity to a small-molecule inhibitor and normal lipid transport activity. Proc Natl Acad Sci U S A. 2011;108(30):12243–8.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Simons K, Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol. 2000;1(1):31–9.

    Article  CAS  PubMed  Google Scholar 

  28. Nieland TJ, Penman M, Dori L, Krieger M, Kirchhausen T. Discovery of chemical inhibitors of the selective transfer of lipids mediated by the HDL receptor SR-BI. Proc Natl Acad Sci U S A. 2002;99(24):15422–7.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Conner SD, Schmid SL. Regulated portals of entry into the cell. Nature. 2003;422(6927):37–44.

    Article  CAS  PubMed  Google Scholar 

  30. Podbilewicz B, Mellman I. ATP and cytosol requirements for transferrin recycling in intact and disrupted MDCK cells. EMBO J. 1990;9(11):3477–87.

    CAS  PubMed Central  PubMed  Google Scholar 

  31. Schnitzer JE, Oh P, Pinney E, Allard J. Filipin-sensitive caveolae-mediated transport in endothelium: reduced transcytosis, scavenger endocytosis, and capillary permeability of select macromolecules. J Cell Biol. 1994;127(5):1217–32.

    Article  CAS  PubMed  Google Scholar 

  32. Masereel B, Pochet L, Laeckmann D. An overview of inhibitors of Na(+)/H(+) exchanger. Eur J Med Chem. 2003;38(6):547–54.

    Article  CAS  PubMed  Google Scholar 

  33. Krieger M. Charting the fate of the “good cholesterol”: identification and characterization of the high-density lipoprotein receptor SR-BI. Annu Rev Biochem. 1999;68:523–58.

    Article  CAS  PubMed  Google Scholar 

  34. Wang N, Lan D, Chen W, Matsuura F, Tall AR. ATP-binding cassette transporters G1 and G4 mediate cellular cholesterol efflux to high-density lipoproteins. Proc Natl Acad Sci U S A. 2004;101(26):9774–9.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Yang S, Damiano MG, Zhang H, Tripathy S, Luthi AJ, Rink JS, et al. Biomimetic, synthetic HDL nanostructures for lymphoma. Proc Natl Acad Sci U S A. 2013;110(7):2511–6.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. Cabral H, Matsumoto Y, Mizuno K, Chen Q, Murakami M, Kimura M, et al. Accumulation of sub-100 nm polymeric micelles in poorly permeable tumours depends on size. Nat Nanotechnol. 2011;6(12):815–23.

    Article  CAS  PubMed  Google Scholar 

  37. Nieland TJ, Ehrlich M, Krieger M, Kirchhausen T. Endocytosis is not required for the selective lipid uptake mediated by murine SR-BI. Biochim Biophys Acta. 2005;1734(1):44–51.

    Article  CAS  PubMed  Google Scholar 

  38. Pagler TA, Rhode S, Neuhofer A, Laggner H, Strobl W, Hinterndorfer C, et al. SR-BI-mediated high density lipoprotein (HDL) endocytosis leads to HDL resecretion facilitating cholesterol efflux. J Biol Chem. 2006;281(16):11193–204.

    Article  CAS  PubMed  Google Scholar 

  39. Wustner D, Mondal M, Huang A, Maxfield FR. Different transport routes for high density lipoprotein and its associated free sterol in polarized hepatic cells. J Lipid Res. 2004;45(3):427–37.

    Article  PubMed  Google Scholar 

  40. Graf GA, Connell PM, van der Westhuyzen DR, Smart EJ. The class B, type I scavenger receptor promotes the selective uptake of high density lipoprotein cholesterol ethers into caveolae. J Biol Chem. 1999;274(17):12043–8.

    Article  CAS  PubMed  Google Scholar 

  41. Peng Y, Akmentin W, Connelly MA, Lund-Katz S, Phillips MC, Williams DL. Scavenger receptor BI (SR-BI) clustered on microvillar extensions suggests that this plasma membrane domain is a way station for cholesterol trafficking between cells and high-density lipoprotein. Mol Biol Cell. 2004;15(1):384–96.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  42. Ahras M, Naing T, McPherson R. Scavenger receptor class B type I localizes to a late endosomal compartment. J Lipid Res. 2008;49(7):1569–76.

    Article  CAS  PubMed  Google Scholar 

  43. Koivusalo M, Welch C, Hayashi H, Scott CC, Kim M, Alexander T, et al. Amiloride inhibits macropinocytosis by lowering submembranous pH and preventing Rac1 and Cdc42 signaling. J Cell Biol. 2010;188(4):547–63.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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ACKNOWLEDGMENTS AND DISCLOSURES

This study was conducted with the support of the China-Canada Joint Health Research Initiative (NSFC-30911120489, CIHR CCI-102936), DLVR Therapeutics, Canadian Institutes of Health Research, Ontario Institute for Cancer Research, Natural Sciences and Engineering Research Council of Canada, Canada Foundation for Innovation, National Natural Science Foundation of China (Grant No. 81172153), 111 Project of China (B07038), and the Joey and Toby Tanenbaum/Brazilian Ball Chair in Prostate Cancer Research.

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Authors and Affiliations

  1. Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics Huazhong University of Science & Technology, Wuhan, 430074, China

    Qiaoya Lin & Zhihong Zhang

  2. Department of Medical Biophysics, University of Toronto, Toronto, Canada

    Qiaoya Lin & Gang Zheng

  3. Ontario Cancer Institute Campbell Family Cancer Research Institute and Techna Institute UHN, Toronto, Canada

    Qiaoya Lin, Juan Chen, Kenneth K. Ng, Weiguo Cao, Zhihong Zhang & Gang Zheng

  4. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada

    Kenneth K. Ng & Gang Zheng

  5. Department of Chemistry, Shanghai University, Shanghai, China

    Weiguo Cao

  6. University of Toronto, TMDT 5-363, 101 College Street, Toronto, Ontario, M5G 1L7, Canada

    Gang Zheng

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  1. Qiaoya Lin
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  2. Juan Chen
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  4. Weiguo Cao
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Correspondence to Zhihong Zhang or Gang Zheng.

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Lin, Q., Chen, J., Ng, K.K. et al. Imaging the Cytosolic Drug Delivery Mechanism of HDL-Like Nanoparticles. Pharm Res 31, 1438–1449 (2014). https://doi.org/10.1007/s11095-013-1046-z

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  • DOI: https://doi.org/10.1007/s11095-013-1046-z

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