Abstract
DNA-functionalized gold nanoparticles (AuNP–DNA) were hybridized with complementary di-N-acetyllactosamine-(di-LacNAc, [3Gal(β1-4)GlcNAc(β1-]2)-modified oligonucleotides to form glycol-functionalized particles, AuNP–DNA–di-LacNAc. While AuNP–DNA are known to be taken up by cells via scavenger receptors, glycol-functionalized particles have shown to be taken up via asialoglycoprotein receptors (ASGP-R). In this work, the interaction of these new particles with HepG2 cells was analyzed, which express scavenger receptors class B type I (SR-BI) and ASGP-R. To study the contribution of these receptors as potential mediators for cellular uptake, receptor-blocking experiments were performed with d-lactose, a ligand for ASGP-R, Fucoidan, a putative ligand for SR-BI, and a SR-BI blocking antibody. Labeling with Cy5-modified DNA ligands enabled us to monitor the particle uptake by confocal fluorescence microscopy and flow cytometry, in order to discriminate the two putative pathways by competitive binding studies. While SR-BI-antibody and d-lactose had no inhibiting effects on particle uptake Fucoidan led to a complete inhibition. Thus, a receptor-mediated uptake by the two receptors studied could not be proven and therefore other uptake mechanisms have to be considered.
References
Acuna GP, Bucher M, Stein IH, Steinhauer C, Kuzyk A, Holzmeister P, Schreiber R, Moroz A, Stefani FD, Liedl T, Simmel FC, Tinnefeld P (2012) Distance dependence of single-fluorophore quenching by gold nanoparticles studied on DNA origami. ACS Nano 6:3189–3195
Arnáiz B, Martínez-Ávila O, Falcon-Perez JM, Penadés S (2012) Cellular uptake of gold nanoparticles bearing HIV gp120 oligomannosides. Bioconjug Chem 23:814–825
Bergen JM, von Recum HA, Goodman TT, Massay AP, Pun SH (2006) Gold nanoparticles as a versatile platform for optimizing physicochemical parameters for targeted drug delivery. Macromol Biosci 6:506–516
Biessen EA, Vietsch H, Rump ET, Fluiter K, Kuiper J, Bijsterbosch MK, von Berkel TJC (1999) Targeted delivery of oligodeoxynucleotides to parenchymal liver cells in vivo. Biochem J 340:783–792
Dong P, Xie T, Zhou X, Hu W, Chen Y, Duan Y, Li X, Han J (2011) Induction of macrophage scavenger receptor type BI expression by tamoxifen and 4-hydroxytamoxifen. Atherosclerosis 218:435–442. doi:10.1016/j.atherosclerosis.2011.06.048
Drickamer K, Taylor ME (1993) Biology of animal lectins. Annu Rev Cell Biol 9:237–264
Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nat Phys Sci 241:20–22
Gallo J, Genicio N, Penadés S (2012) Uptake and intracellular fate of fluorescent-magnetic glyco-nanoparticles. Adv Healthcare Mater 1:302–307
Giljohann DA, Seferos DS, Patel PC, Millstone JE, Rosi NL, Mirkin CA (2007) Oligonucleotide loading determines cellular uptake of DNA-modified gold nanoparticles. Nano Lett 7:3818–3821
Giljohann DA, Seferos DS, Prigodich AE, Patel PC, Mirkin CA (2009) Gene regulation with polyvalent siRNA-nanoparticle conjugates. J Am Chem Soc 131:2072–2073
Giljohann DA, Seferos DS, Daniel WL, Massich MD, Patel PC, Mirkin CA (2010) Gold nanoparticles for biology and medicine. Angew Chem Int Ed Engl 49:3280–3294
Hangeland JJ, Flesher JE, Deamond SF, Lee YC, Ts’O PO, Frost JJ (1997) Tissue distribution and metabolism of the [32P]-labeled oligodeoxynucleoside methylphosphonate-neoglycopeptide conjugate, [YEE(ah-GalNAc)3]-SMCC-AET-pUmpT7, in the mouse. Antisense Nucleic Acid Drug Dev 7:141–149
He H, MacKinnon KM, Genovese KJ, Nerren JR, Swaggerty CL, Nisbet DJ, Kogut MH (2009) Chicken scavenger receptors and their ligand-induced cellular immune responses. Mol Immunol 46:2218–2225
Huang G, Diakur J, Xu Z, Wiebe LI (2008) Asialoglycoprotein receptor-targeted superparamagnetic iron oxide nanoparticles. Int J Pharm 360:197–203
Hurst SJ, Lytton-Jean AKR, Mirkin CA (2006) Maximizing DNA loading on a range of gold nanoparticle sizes. Anal Chem 78:8313–8318
Husemann J, Loike JD, Anankov R, Febbraio M, Silverstein SC (2002) Scavenger receptors in neurobiology and neuropathology: their role on microglia and other cells of the nervous system. Glia 40:195–205
Kanaras AG, Wang Z, Bates AD, Cosstick R, Brust M (2003) Towards multistep nanostructure synthesis: programmed enzymatic self-assembly of DNA/gold systems. Angew Chem Int Ed 42:191–194
Kikkeri R, Lepenies B, Adibekian A, Laurino P, Seeberger PH (2009) In vitro imaging and in vivo liver targeting with carbohydrate capped quantum dots. J Am Chem Soc 131:2110–2112
Kim CK, Kalluru RR, Singh JP, Fortner A, Griffin J, Darbha GK, Ray PC (2006) Gold-nanoparticle-based miniaturized laser-induced fluorescence probe for specific DNA hybridization detection: studies on size-dependent optical properties. Nanotechnology 17:3085–3093
Lazarides A, Schatz G (2000) DNA-linked metal nanosphere materials: structural basis for the optical properties. J Phys Chem B 104:460–467
Lee RT, Lin P, Lee YC (1984) New synthetic cluster ligands for galactose/N-acetylgalactosamine-specific lectin of mammalian liver. Biochemistry 23:4255–4261
Lee J-S, Han MS, Mirkin CA (2007) Colorimetric detection of mercuric ion (Hg2+) in aqueous media using DNA-functionalized gold nanoparticles. Angew Chem Int Ed Engl 46:4093–4096
Li B, Lu F, Wei X, Zhao R (2008) Fucoidan: structure and bioactivity. Molecules 13:1671–1695
Mirkin CA, Letsinger RL, Mucic RC, Storhoff JJ (1996) A DNA-based method for rationally assembling nanoparticles into macroscopic materials. Nature 382:607–609
Murphy RF, Powers S, Cantor CR (1984) Endosome pH measured in single cells by dual fluorescence flow cytometry: rapid acidification of insulin to pH 6. J Cell Biol 98:1757–1762
Nitin N, Javier DJ, Richards-Kortum R (2007) Oligonucleotide-coated metallic nanoparticles as a flexible platform for molecular imaging agents. Bioconjug Chem 18:2090–2096
Patel PC, Giljohann DA, Daniel WL, Zheng AE, Prigodich AE, Mirkin CA (2010) Scavenger receptors mediate cellular uptake of polyvalent oligonucleotide-functionalized gold nanoparticles. Bioconjug Chem 21:2250–2256
Pathak A, Vyas SP, Gupta KC (2008) Nano-vectors for efficient liver specific gene transfer. Int J Nanomed 3:31–49
Rech C, Rosencrantz RR, Křenek K, Pelantová H, Bojarová, Römer CE, Hanisch F-G, Křen V, Elling L (2011) Combinatorial one-pot synthesis of poly-N-acetyllactosamine oligosaccharides with leloir-glycosyltransferases. Adv Synth Catal 353:2492–2500
Rhainds D, Falstrault L, Tremblay C, Brissette L (1999) Uptake and fate of class B scavenger receptor ligands in HepG2 cells. Eur J Biochem 261:227–235
Rosi NL, Mirkin CA (2005) Nanostructures in biodiagnostics. Chem Rev 105:1547–1562
Rosi NL, Giljohann DA, Thaxton CS, Lytton-Jean AKR, Han MS, Mirkin CA (2006) Oligonucleotide-modified gold nanoparticles for intracellular gene regulation. Science 312:1027–1030
Rybak SL, Murphy RF (1998) Primary cell cultures from murine kidney and heart differ in endosomal pH. J Cell Physiol 176:216–222
Sauerzapfe B, Křenek K, Schmiedel J, Wakarchuk WW, Pelantová, Křen V, Elling L (2009) Chemo-enzymatic synthesis of poly-N-acetyllactosamine (poly-LacNAc) structures and their characterization for CGL2-galectin-mediated binding of ECM glycoproteins to biomaterial surfaces. Glycoconj J 26:141–159
Seferos DS, Prigodich AE, Giljohann DA, Patel PC, Mirkin CA (2009) Polyvalent DNA nanoparticle conjugates stabilize nucleic acids. Nano Lett 9:308–311
Song X, Fischer P, Chen X, Burton C, Wang J (2009) An apoA-I mimetic peptide facilitates off-loading cholesterol from HDL to liver cells through scavenger receptor BI. Int J Biol Sci 5:637–646
Spiess M (1990) The asialoglycoprotein receptor: a model for endocytic transport receptors. Biochemistry 29:10009–10018
Stockert RJ (1995) The asialoglycoprotein receptor: relationships between structure, function, and expression. Physiol Rev 75:591–609
Su G, Zhou H, Mu Q, Zhang Y, Li L, Jiao P, Jiang G, Yan B (2012) Effective surface charge density determines the electrostatic attraction between nanoparticles and cells. J Phys Chem C 116:4993–4998
Takae S, Akiyama Y, Otsuka H, Nakamura T, Nagasaki Y, Kataoka K (2005) Ligand density effect on biorecognition by PEGylated gold nanoparticles: regulated interaction of RCA120 lectin with lactose installed to the distal end of tethered PEG strands on gold surface. Biomacromolecules 6:818–824
Tian X, Pai J, Baek K-H, Ko S-K, Shin I (2011) Fluorophore-labeled, peptide-based glycoclusters: synthesis, binding properties for lectins, and detection of carbohydrate-binding proteins in cells. Chem Asian J 6:2107–2113
Turkevich J, Stevenson PC, Hillier J (1951) A study of the nucleation and growth processes in the synthesis of colloidal gold. Discuss Faraday Soc 11:55–75
Westerlind U, Westman J, Törnquist E, Smith CIE, Oscarson S, Lahmann M, Norberg T (2004) Ligands of the asialoglycoprotein receptor for targeted gene delivery, Part 1: synthesis of and binding studies with biotinylated cluster glycosides containing N-acetylgalactosamine. Glycoconj J 21:227–241
Witten KG (2012) Glyko-DNA-funktionalisierte goldnanopartikel: synthese und wechselwirkung mit lektinen und zellen. PhD Thesis RWTH, Aachen University
Witten KG, Bretschneider JC, Eckert T, Richtering W, Simon U (2008) Assembly of DNA-functionalized gold nanoparticles studied by UV/Vis-spectroscopy and dynamic light scattering. Phys Chem Chem Phys 10:1870–1875
Witten KG, Rech C, Eckert T, Charrak S, Richtering W, Elling L, Simon U (2011) Glyco-DNA–gold nanoparticles: lectin-mediated assembly and dual-stimuli response. Small 7:1954–1960
Yan H, Tram K (2007) Glycotargeting to improve cellular delivery efficiency of nucleic acids. Glycoconj J 24:107–123
Yang Y, Zhang Z, Chen L, Gu W, Li Y (2010) Galactosylated poly(2-(2-aminoethyoxy)ethoxy)phosphazene/DNA complex nanoparticles: in vitro and in vivo evaluation for gene delivery. Biomacromolecules 11:927–933
Acknowledgments
We thank Prof. E. Weinhold and Dr. S. Charrak (Institute of Organic Chemistry, RWTH Aachen University) for HPLC, Dr. W. Bettray (Institute of Organic Chemistry, RWTH Aachen University) for ESI–MS, Dr. Yu Pan (Biomedical Engineering, Biointerface Laboratory RWTH Aachen University) for Nanodrop analytics and the Immunohistochemistry, and Confocal Laser Scanning Microscopy Facility, a core facility of the IZKF within the Faculty of Medicine, RWTH Aachen University. We are grateful to Liangliang Hao for valuable comments on this manuscript. This research is a part of the project “ForSaTum”, co-funded by the European Union (European Regional Development Fund—Investing in your future) and the German federal state North Rhine-Westphalia (NRW). Furthermore, the work was financially supported by the Deutsche Forschungsgemeinschaft Graduate School “Biointerface” (No. 1035) and the Excellence Initiative of the German federal and state Governments (ERS Seed Fund Project).
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Witten, K.G., Ruff, J., Mohr, A. et al. Cellular uptake of fluorophore-labeled glyco-DNA–gold nanoparticles. J Nanopart Res 15, 1992 (2013). https://doi.org/10.1007/s11051-013-1992-8
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DOI: https://doi.org/10.1007/s11051-013-1992-8