Glutathione-mediated biotransformation in the liver is a well-known detoxification process to eliminate small xenobiotics, but its impacts on nanoparticle retention, targeting and clearance are much less understood than liver macrophage uptake, even though both processes are involved in liver detoxification. By designing a thiol-activatable fluorescent gold nanoprobe that can bind to serum protein and be transported to the liver, we non-invasively imaged the biotransformation kinetics in vivo at high specificity and examined this process at the chemical level. Our results show that glutathione efflux from hepatocytes results in high local concentrations of both glutathione and cysteine in liver sinusoids, which transforms the nanoparticle surface chemistry, reduces its affinity to serum protein and significantly alters its blood retention, targeting and clearance. With this biotransformation, liver detoxification, a long-standing barrier in nanomedicine translation, can be turned into a bridge toward maximizing targeting and minimizing nanotoxicity.
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The data that support the plots within this paper and other findings of this study are available from the corresponding author upon reasonable request.
Tsoi, K. M. et al. Mechanism of hard-nanomaterial clearance by the liver. Nat. Mater. 15, 1212–1221 (2016).
Wilhelm, S. et al. Analysis of nanoparticle delivery to tumours. Nat. Rev. Mater. 1, 16014 (2016).
Fischer, H. C., Liu, L., Pang, K. S. & Chan, W. C. Pharmacokinetics of nanoscale quantum dots: in vivo distribution, sequestration and clearance in the rat. Adv. Funct. Mater. 16, 1299–1305 (2006).
Ye, L. et al. A pilot study in non-human primates shows no adverse response to intravenous injection of quantum dots. Nat. Nanotechnol. 7, 453–458 (2012).
Balasubramanian, S. K. et al. Biodistribution of gold nanoparticles and gene expression changes in the liver and spleen after intravenous administration in rats. Biomaterials 31, 2034–2042 (2010).
Gu, X. & Manautou, J. E. Molecular mechanisms underlying chemical liver injury. Exp. Rev. Mol. Med. 14, e4 (2012).
Braet, F. & Wisse, E. Structural and functional aspects of liver sinusoidal endothelial cell fenestrae: a review. Comp. Hepatol. 1, 1 (2002).
Kaplowitz, N., Aw, T. Y. & Ookhtens, M. The regulation of hepatic glutathione. Annu. Rev. Pharm. Toxicol. 25, 715–744 (1985).
Ballatori, N., Krance, S. M., Marchan, R. & Hammond, C. L. Plasma membrane glutathione transporters and their roles in cell physiology and pathophysiology. Mol. Aspects Med. 30, 13–28 (2009).
Dickinson, D. A. & Forman, H. J. Cellular glutathione and thiols metabolism. Biochem. Pharmacol. 64, 1019–1026 (2002).
Singhal, R. K., Anderson, M. E. & Meister, A. Glutathione, a first line of defense against cadmium toxicity. FASEB J. 1, 220–223 (1987).
Du, B. et al. Glomerular barrier behaves as an atomically precise bandpass filter in a sub-nanometre regime. Nat. Nanotechnol. 12, 1096–1102 (2017).
Shinohara, H. et al. Direct measurement of hepatic indocyanine green clearance with near-infrared spectroscopy: separate evaluation of uptake and removal. Hepatology 23, 137–144 (1996).
Sun, S. et al. Dimerization of organic dyes on luminescent gold nanoparticles for ratiometric pH sensing. Angew. Chem. Int. Ed. 128, 2467–2470 (2016).
Choi, H. S. et al. Renal clearance of quantum dots. Nat. Biotechnol. 25, 1165–1170 (2007).
Dreaden, E. C., Austin, L. A., Mackey, M. A. & El-Sayed, M. A. Size matters: gold nanoparticles in targeted cancer drug delivery. Ther. Deliv. 3, 457–478 (2012).
Hirn, S. et al. Particle size-dependent and surface charge-dependent biodistribution of gold nanoparticles after intravenous administration. Eur. J. Pharm. Biopharm. 77, 407–416 (2011).
Ookhtens, M., Hobdy, K., Corvasce, M., Aw, T. Y. & Kaplowitz, N. Sinusoidal efflux of glutathione in the perfused rat liver. Evidence for a carrier-mediated process. J. Clin. Invest. 75, 258–265 (1985).
Plummer, J. L., Smith, B. R., Sies, H. & Bend, J. R. Chemical depletion of glutathione in vivo. Methods Enzymol. 77, 50–59 (1981).
Winters, R. A., Zukowski, J., Ercal, N., Matthews, R. H. & Spitz, D. R. Analysis of glutathione, glutathione disulfide, cysteine, homocysteine and other biological thiols by high-performance liquid chromatography following derivatization by n-(1-pyrenyl) maleimide. Anal. Biochem. 227, 14–21 (1995).
Parmentier, C., Leroy, P., Wellman, M. & Nicolas, A. Determination of cellular thiols and glutathione-related enzyme activities: versatility of high-performance liquid chromatography–spectrofluorimetric detection. J. Chromatogr. B 719, 37–46 (1998).
Jocelyn, P. The standard redox potential of cysteine–cystine from the thiol–disulphide exchange reaction with glutathione and lipoic acid. FEBS J. 2, 327–331 (1967).
Wu, G., Fang, Y.-Z., Yang, S., Lupton, J. R. & Turner, N. D. Glutathione metabolism and its implications for health. J. Nutr. 134, 489–492 (2004).
Liu, J. et al. PEGylation and Zwitterionization: pros and cons in the renal clearance and tumor targeting of near-IR-emitting gold nanoparticles. Angew. Chem. Int. Ed. 125, 12804–12808 (2013).
Tate, S. S. & Meister, A. in The Biological Effects of Glutamic Acid and Its Derivatives (ed. Najjar, V. A.) 357–368 (Springer, 1981).
Paolicchi, A. et al. γ-Glutamyl transpeptidase catalyses the extracellular detoxification of cisplatin in a human cell line derived from the proximal convoluted tubule of the kidney. Eur. J. Cancer 39, 996–1003 (2003).
Hanigan, M. H. & Pitot, H. C. Gamma-glutamyl transpeptidase—its role in hepatocarcinogenesis. Carcinogenesis 6, 165–172 (1985).
Peng, C. et al. Targeting orthotopic gliomas with renal-clearable luminescent gold nanoparticles. Nano Res. 10, 1366–1376 (2017).
Wu, Z., Suhan, J. & Jin, R. One-pot synthesis of atomically monodisperse, thiol-functionalized Au25 nanoclusters. J. Mater. Chem. 19, 622–626 (2009).
Dhar, S., Daniel, W. L., Giljohann, D. A., Mirkin, C. A. & Lippard, S. J. Polyvalent oligonucleotide gold nanoparticle conjugates as delivery vehicles for platinum (IV) warheads. J. Am. Chem. Soc. 131, 14652–14653 (2009).
Weber, C. A., Duncan, C. A., Lyons, M. J. & Jenkinson, S. G. Depletion of tissue glutathione with diethyl maleate enhances hyperbaric oxygen toxicity. Am. J. Physiol. Lung Cell. Mol. Physiol. 258, L308–L312 (1990).
Adams, J., Lauterburg, B. & Mitchell, J. Plasma glutathione and glutathione disulfide in the rat: regulation and response to oxidative stress. J. Pharmacol. Exp. Ther. 227, 749–754 (1983).
Rahman, I., Kode, A. & Biswas, S. K. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat. Protoc. 1, 3159 (2006).
The authors acknowledge financial support from the National Institutes of Health (NIH; R01DK103363 and R01DK115986), the Cancer Prevention Research Institute of Texas (CPRIT; RP160866), the Welch Research Foundation (AT-1974-20180324) and the Cecil H. and Ida Green Professorship in System Biology (to J.Z.) from the University of Texas at Dallas. The authors also thank E. Hernandez and J.T. Hsieh from The University of Texas Southwestern Medical Center for tissue slide preparation.
The authors declare no competing interests.
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