Dexamethasone Conjugation to Biodegradable Avidin-Nucleic-Acid-Nano-Assemblies Promotes Selective Liver Targeting and Improves Therapeutic Efficacy in an Autoimmune Hepatitis Murine Model.

Steroids are the standard therapy for autoimmune hepatitis (AIH) but the long-lasting administration is hampered by severe side effects. Methods to improve the tropism of the drug toward the liver are therefore required. Among them, conjugation to nanoparticles represents one possible strategy. In this study, we exploited the natural liver tropism of Avidin-Nucleic-Acid-Nano-Assemblies (ANANAS) to carry dexamethasone selectively to the liver in an AIH animal model. An acid-labile biotin-hydrazone linker was developed for reversible dexamethasone loading onto ANANAS. The biodistribution, pharmacokinetics and efficacy of free and ANANAS-linked dexamethasone (ANANAS-Hz-Dex) in healthy and AIH mice were investigated upon intraperitoneal administration. In ANANAS-treated animals, the free drug was detected only in the liver. Super-resolution microscopy showed that nanoparticles segregate inside lysosomes of liver immunocompetent cells, mainly involved in AIH progression. In agreement with these observational results, chronic low-dose treatment with ANANAS-Hz-Dex reduced the expression of liver inflammation markers and, in contrast to the free drug, also the levels of circulating AIH-specific autoantibodies. These data suggest that the ANANAS carrier attenuates AIH-related liver damage without drug accumulation in off-site tissues. The safety and biodegradability of the ANANAS carrier make this formulation a promising tool for the treatment of autoimmune liver disorders.

[1]  P. Olinga,et al.  Targeting dexamethasone to Kupffer cells: Effects on liver inflammation and fibrosis in rats , 2001, Hepatology.

[2]  N. Realdon,et al.  Optimized avidin nucleic acid nanoassemblies by a tailored PEGylation strategy and their application as molecular amplifiers in detection. , 2010, Bioconjugate chemistry.

[3]  T. Park,et al.  Diverse Applications of Nanomedicine , 2017, ACS nano.

[4]  M. Manns,et al.  Breaking tolerance to the natural human liver autoantigen cytochrome P450 2D6 by virus infection , 2008, The Journal of experimental medicine.

[5]  Bangmao Wang,et al.  Altered function of monocytes/macrophages in patients with autoimmune hepatitis , 2016, Molecular medicine reports.

[6]  T. Luedde,et al.  Fluorescent cell-traceable dexamethasone-loaded liposomes for the treatment of inflammatory liver diseases. , 2015, Biomaterials.

[7]  M. Morbidelli,et al.  Biocompatible Polymer Nanoformulation To Improve the Release and Safety of a Drug Mimic Molecule Detectable via ICP-MS. , 2017, Molecular pharmaceutics.

[8]  K. Fu,et al.  Synthesis and Evaluation of a Well-defined HPMA Copolymer–Dexamethasone Conjugate for Effective Treatment of Rheumatoid Arthritis , 2008, Pharmaceutical Research.

[9]  K. Dawson,et al.  Elution of Labile Fluorescent Dye from Nanoparticles during Biological Use , 2011, PLoS ONE.

[10]  A. Buda,et al.  Discrimination between ulcerative colitis and Crohn's disease using phage display identified peptides and virus-mimicking synthetic nanoparticles. , 2017, Nanomedicine : nanotechnology, biology, and medicine.

[11]  M. Ammirati,et al.  Efficient Liver Targeting by Polyvalent Display of a Compact Ligand for the Asialoglycoprotein Receptor. , 2017, Journal of the American Chemical Society.

[12]  J. Neuberger,et al.  Transplantation in autoimmune liver diseases. , 2008, World journal of gastroenterology.

[13]  Pietro Invernizzi,et al.  Geoepidemiology of autoimmune liver diseases. , 2010, Journal of autoimmunity.

[14]  W J Jusko,et al.  Prednisolone Pharmacokinetics and Pharmacodynamics in Relation to Sex and Race , 2001, Journal of clinical pharmacology.

[15]  U. Christen,et al.  The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice , 2012, Journal of visualized experiments : JoVE.

[16]  M. Fernández-Rojo,et al.  Caveolin-1 Function in Liver Physiology and Disease. , 2016, Trends in molecular medicine.

[17]  M. Wilchek,et al.  Natural antibodies to avidin in human serum. , 1993, Immunology letters.

[18]  J. Pfeilschifter,et al.  Mechanism of autoimmune hepatic fibrogenesis induced by an adenovirus encoding the human liver autoantigen cytochrome P450 2D6. , 2013, Journal of autoimmunity.

[19]  N. Realdon,et al.  In vivo fate of avidin-nucleic acid nanoassemblies as multifunctional diagnostic tools. , 2014, ACS nano.

[20]  J. Pfeilschifter,et al.  Non-alcoholic fatty liver disease (NAFLD) potentiates autoimmune hepatitis in the CYP2D6 mouse model. , 2016, Journal of autoimmunity.

[21]  Ian D. McGilvray,et al.  Mechanism of hard nanomaterial clearance by the liver , 2016, Nature materials.

[22]  Giovanni Paganelli,et al.  Therapeutic use of avidin is not hampered by antiavidin antibodies in humans. , 2010, Cancer biotherapy & radiopharmaceuticals.

[23]  P. Invernizzi Liver auto-immunology: the paradox of autoimmunity in a tolerogenic organ. , 2013, Journal of autoimmunity.

[24]  A. Czaja,et al.  Decreased fibrosis during corticosteroid therapy of autoimmune hepatitis. , 2004, Journal of hepatology.

[25]  Aurelian Radu,et al.  DNA condensation by high‐affinity interaction with avidin , 2004, Journal of molecular recognition : JMR.

[26]  N. Realdon,et al.  Improvement and extension of anti-EGFR targeting in breast cancer therapy by integration with the Avidin-Nucleic-Acid-Nano-Assemblies , 2018, Nature Communications.

[27]  N. Realdon,et al.  Characterization of multifunctional nanosystems based on the avidin-nucleic acid interaction as signal enhancers in immuno-detection. , 2012, Analytical chemistry.

[28]  N. Realdon,et al.  A New ELISA Using the ANANAS Technology Showing High Sensitivity to diagnose the Bovine Rhinotracheitis from Individual Sera to Pooled Milk , 2016, PloS one.

[29]  K. Landfester,et al.  Imaging of Polymeric Nanoparticles: Hard Challenge for Soft Objects , 2016 .

[30]  J. Bostwick,et al.  Psychiatric adverse effects of corticosteroids. , 2006, Mayo Clinic proceedings.

[31]  Akshay Jain,et al.  The principles and applications of avidin‐based nanoparticles in drug delivery and diagnosis , 2017, Journal of controlled release : official journal of the Controlled Release Society.

[32]  M. Wauben,et al.  Complete remission of experimental arthritis by joint targeting of glucocorticoids with long-circulating liposomes. , 2003, Arthritis and rheumatism.

[33]  G. Sims,et al.  A method for the estimation of polyethylene glycol in plasma protein fractions. , 1980, Analytical biochemistry.

[34]  S. Duarte,et al.  Matrix metalloproteinases in liver injury, repair and fibrosis , 2015, Matrix biology : journal of the International Society for Matrix Biology.

[35]  Yechezkel Barenholz,et al.  Amphipathic weak acid glucocorticoid prodrugs remote-loaded into sterically stabilized nanoliposomes evaluated in arthritic rats and in a Beagle dog: a novel approach to treating autoimmune arthritis. , 2008, Arthritis and rheumatism.

[36]  G. Mieli-Vergani,et al.  Autoimmune hepatitis. , 2011, Journal of hepatology.

[37]  J. Pfeilschifter,et al.  Molecular mimicry rather than identity breaks T-cell tolerance in the CYP2D6 mouse model for human autoimmune hepatitis. , 2013, Journal of autoimmunity.