The effective drug delivery systems for cancer treatment are currently on high demand. In this paper, biological behavior of the novel hybrid copolymers based on polysaccharide glycogen were characterized. The copolymers were modified by fluorescent dyes for flow cytometry, confocal microscopy, and in vivo fluorescence imaging. Moreover, the effect of oxazoline grafts on degradation rate was examined. Intracellular localization, cytotoxicity, and internalization route of the modified copolymers were examined on HepG2 cell line. Biodistribution of copolymers was addressed by in vivo fluorescence imaging in C57BL/6 mice. Our results indicate biocompatibility, biodegradability, and non-toxicity of the glycogen-based hybrid copolymers. Copolymers were endocyted into the cytoplasm, most probably via caveolae-mediated endocytosis. Higher content of oxazoline in polymers slowed down cellular uptake. No strong colocalization of the glycogen-based probe with lysosomes was observed; thus, it seems that the modified externally administered glycogen is degraded in the same way as an endogenous glycogen. In vivo experiment showed relatively fast biodistribution and biodegradation. In conclusion, this novel nanoprobe offers unique chemical and biological attributes for its use as a novel drug delivery system that might serve as an efficient carrier for cancer therapeutics with multimodal imaging properties.

Department of Physiology Faculty of Science Charles University Prague Viničná 7 128 44 Praha 2 Czech Republic

Institute for Clinical and Experimental Medicine Videnska 1958 9 140 21 Prague 4 Czech Republic

Institute of Biophysics and Informatics 1st Medicine Faculty Charles University Salmovská 1 12000 Prague 2 Czech Republic

Institute of Macromolecular Chemistry of the Academy of Sciences of the Czech Republic Heyrovsky Sq 2 162 06 Prague 6 Czech Republic

Institute of Physiology of the Academy of Sciences of the Czech Republic Videnska 1083 142 20 Prague 4 Czech Republic

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Cho K, Wang X, Nie S, Chen ZG, Shin DM. Therapeutic nanoparticles for drug delivery in cancer. Clin Cancer Res Off J Am Assoc Cancer Res. 2008;14:1310–6. DOI

Fang J, Nakamura H, Maeda H. The EPR effect: unique features of tumor blood vessels for drug delivery, factors involved, and limitations and augmentation of the effect. Adv Drug Deliv Rev. 2011;63:136–51. PubMed DOI

Taurin S, Nehoff H, Greish K. Anticancer nanomedicine and tumor vascular permeability; where is the missing link? J Control Release Off J Control Release Soc. 2012;164:265–75. DOI

Talelli M, Iman M, Rijcken CJF, van Nostrum CF, Hennink WE. Targeted core-crosslinked polymeric micelles with controlled release of covalently entrapped doxorubicin. J Control Release 2010;148:e121–e122.

Barar J, Omidi Y. Dysregulated pH in tumor microenvironment checkmates cancer therapy. BioImpacts BI. 2013;3:149–62. PubMed PMC

Omidi Y, Barar J. Targeting tumor microenvironment: crossing tumor interstitial fluid by multifunctional nanomedicines. BioImpacts BI. 2014;4:55–67. PubMed PMC

Tirone TA, Brunicardi FC. Overview of glucose regulation. World J Surg. 2001;25:461–7. PubMed DOI

Filippov SK, Sedlacek O, Bogomolova A, Vetrik M, Jirak D, Kovar J, et al. Glycogen as a biodegradable construction nanomaterial for in vivo use. Macromol Biosci. 2012;12:1731–8. PubMed DOI

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;6:815–23. PubMed DOI

Rybicka KK. Glycosomes—the organelles of glycogen metabolism. Tissue Cell. 1996;28:253–65. PubMed DOI

Izawa H, Nawaji M, Kaneko Y, Kadokawa J. Preparation of glycogen-based polysaccharide materials by phosphorylase-catalyzed chain elongation of glycogen. Macromol Biosci. 2009;9:1098–104. PubMed DOI

Duncan R, Gilbert HRP, Carbajo RJ, Vicent MJ. Polymer masked−unmasked protein therapy. 1. Bioresponsive dextrin−trypsin and −melanocyte stimulating hormone conjugates designed for α-amylase activation. Biomacromolecules. 2008;9:1146–54. PubMed DOI

Hreczuk-Hirst D, Chicco D, German L, Duncan R. Dextrins as potential carriers for drug targeting: tailored rates of dextrin degradation by introduction of pendant groups. Int J Pharm. 2001;230:57–66. PubMed DOI

Pospisilova A, Filippov SK, Bogomolova A, Turner S, Sedlacek O, Matushkin N, et al. Glycogen-graft-poly(2-alkyl-2-oxazolines)—the new versatile biopolymer-based thermoresponsive macromolecular toolbox. RSC Adv. 2014;4:61580–8. DOI

Aleksandrova LA, Kutuzova TM, Kraevskii AA, Kukhanova MK, Gottikh BP. Aminoacyl derivatives of nucleosides, nucleotides, and polynucleotides. Bull Acad Sci USSR Div Chem Sci. 1977;26:583–9. DOI

Chen RF. Dansyl labeled proteins: determination of extinction coefficient and number of bound residues with radioactive dansyl chloride. Anal Biochem. 1968;25:412–6. PubMed DOI

Hoechst Stains [Internet]. 2015. Available from: https://tools.thermofisher.com/content/sfs/manuals/mp21486.pdf

LysoTracker® and LysoSensor™ Probes [Internet]. 2015. Available from: https://tools.thermofisher.com/content/sfs/manuals/mp07525.pdf

Freshney RI. Culture of animal cells: a manual of basic technique. Wiley; 2000.

Lloyd JB, Mason RW, editors. Biology of the lysosome [Internet]. Boston, MA: Springer US; 1996 [cited 2015 Dec 4]. Available from: http://link.springer.com/10.1007/978-1-4615-5833-0

Fernández-Novell JM, Bellido D, Vilaró S, Guinovart JJ. Glucose induces the translocation of glycogen synthase to the cell cortex in rat hepatocytes. Biochem J. 1997;321:227–31. PubMed DOI PMC

García-Rocha M, Roca A, De La Iglesia N, Baba O, Fernández-Novell JM, Ferrer JC, et al. Intracellular distribution of glycogen synthase and glycogen in primary cultured rat hepatocytes. Biochem J. 2001;357:17–24. PubMed DOI PMC

Ros S, García-Rocha M, Domínguez J, Ferrer JC, Guinovart JJ. Control of liver glycogen synthase activity and intracellular distribution by phosphorylation. J Biol Chem. 2009;284:6370–8. PubMed DOI

Fernández-Novell JM, López-Iglesias C, Ferrer JC, Guinovart JJ. Zonal distribution of glycogen synthesis in isolated rat hepatocytes. FEBS Lett. 2002;531:222–8. PubMed DOI

Rewatkar PV, Parton RG, Parekh HS, Parat M-O. Are caveolae a cellular entry route for non-viral therapeutic delivery systems? Adv Drug Deliv Rev. 2015;91:92–108. PubMed DOI

Oh N, Park J-H. Endocytosis and exocytosis of nanoparticles in mammalian cells. Int J Nanomedicine. 2014;9:51–63. PubMed PMC

Chang Y, Lee GH, Kim T-J, Chae K-S. Toxicity of magnetic resonance imaging agents: small molecule and nanoparticle. Curr Top Med Chem. 2013;13:434–45. PubMed DOI

Zhou Z, Lu Z-R. Gadolinium-based contrast agents for magnetic resonance cancer imaging. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2013;5:1–18. PubMed DOI

Lansman JB. Blockade of current through single calcium channels by trivalent lanthanide cations. Effect of ionic radius on the rates of ion entry and exit. J Gen Physiol. 1990;95:679–96. PubMed DOI

Biagi BA, Enyeart JJ. Gadolinium blocks low- and high-threshold calcium currents in pituitary cells. Am J Physiol - Cell Physiol. 1990;259:C515–20. DOI

Glycogen as an advantageous polymer carrier in cancer theranostics: Straightforward in vivo evidence

Fluorine polymer probes for magnetic resonance imaging: quo vadis?