Highly Versatile Polyelectrolyte Complexes for Improving the Enzyme Replacement Therapy of Lysosomal Storage Disorders.

Lysosomal storage disorders are currently treated by enzyme replacement therapy (ERT) through the direct administration of the unprotected recombinant protein to the patients. Herein we present an ionically cross-linked polyelectrolyte complex (PEC) composed of trimethyl chitosan (TMC) and α-galactosidase A (GLA), the defective enzyme in Fabry disease, with the capability of directly targeting endothelial cells by incorporating peptide ligands containing the RGD sequence. We assessed the physicochemical properties, cytotoxicity, and hemocompatibility of RGD-targeted and untargeted PECs, the uptake by endothelial cells and the intracellular activity of PECs in cell culture models of Fabry disease. Moreover, we also explored the effect of different freeze-drying procedures in the overall activity of the PECs. Our results indicate that the use of integrin-binding RGD moiety within the PEC increases their uptake and the efficacy of the GLA enzyme, while the freeze-drying allows the activity of the therapeutic protein to remain intact. Overall, these results highlight the potential of TMC-based PECs as a highly versatile and feasible drug delivery system for improving the ERT of lysosomal storage disorders.

[1]  Ignacio Insua,et al.  Polyion complex (PIC) particles: Preparation and biomedical applications , 2016, European polymer journal.

[2]  J. Veciana,et al.  α‐Galactosidase‐A Loaded‐Nanoliposomes with Enhanced Enzymatic Activity and Intracellular Penetration , 2016, Advanced healthcare materials.

[3]  S. Schwartz,et al.  Multifunctionalized polyurethane-polyurea nanoparticles: hydrophobically driven self-stratification at the o/w interface modulates encapsulation stability. , 2015, Journal of materials chemistry. B.

[4]  D. Lockhart,et al.  Oral Migalastat HCl Leads to Greater Systemic Exposure and Tissue Levels of Active α-Galactosidase A in Fabry Patients when Co-Administered with Infused Agalsidase , 2015, PloS one.

[5]  C. Remuñán-López,et al.  Hybrid nanosystems based on natural polymers as protein carriers for respiratory delivery: Stability and toxicological evaluation. , 2015, Carbohydrate polymers.

[6]  J. Esko,et al.  GNeosomes: Highly Lysosomotropic Nanoassemblies for Lysosomal Delivery. , 2015, ACS nano.

[7]  Yoram Tekoah,et al.  Characterization of a chemically modified plant cell culture expressed human α-Galactosidase-A enzyme for treatment of Fabry disease. , 2015, Molecular genetics and metabolism.

[8]  Surajit Ghosh,et al.  Novel lysosome targeted molecular transporter built on a guanidinium-poly-(propylene imine) hybrid dendron for efficient delivery of doxorubicin into cancer cells. , 2015, Chemical communications.

[9]  Ahmed Jalal Khan Chowdhury,et al.  Impact of chitosan composites and chitosan nanoparticle composites on various drug delivery systems: A review , 2014, Journal of food and drug analysis.

[10]  Zhen Gu,et al.  Stimuli-responsive nanomaterials for therapeutic protein delivery. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[11]  K. Raghu,et al.  A lysosome-targeted drug delivery system based on sorbitol backbone towards efficient cancer therapy. , 2014, Organic & biomolecular chemistry.

[12]  C. Arús,et al.  Ex vivo assessment of polyol coated-iron oxide nanoparticles for MRI diagnosis applications: toxicological and MRI contrast enhancement effects , 2014, Journal of Nanoparticle Research.

[13]  V. Rotello,et al.  Direct delivery of functional proteins and enzymes to the cytosol using nanoparticle-stabilized nanocapsules. , 2013, ACS nano.

[14]  T. Kirkegaard Emerging therapies and therapeutic concepts for lysosomal storage diseases , 2013 .

[15]  Eleonore Fröhlich,et al.  The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles , 2012, International journal of nanomedicine.

[16]  V. Préat,et al.  RGD-based strategies to target alpha(v) beta(3) integrin in cancer therapy and diagnosis. , 2012, Molecular pharmaceutics.

[17]  A. Hajitou,et al.  Clathrin-mediated Endocytosis and Subsequent Endo-Lysosomal Trafficking of Adeno-associated Virus/Phage* , 2012, The Journal of Biological Chemistry.

[18]  Ruth Duncan,et al.  Endocytosis and intracellular trafficking as gateways for nanomedicine delivery: opportunities and challenges. , 2012, Molecular pharmaceutics.

[19]  XinRan Li,et al.  Lysosomal delivery of a lipophilic gemcitabine prodrug using novel acid-sensitive micelles improved its antitumor activity. , 2012, Bioconjugate chemistry.

[20]  Carlos A. Muniesa,et al.  Surface-modified silica nanoparticles for tumor-targeted delivery of camptothecin and its biological evaluation. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[21]  V. Torchilin,et al.  Screening and optimization of ligand conjugates for lysosomal targeting. , 2011, Bioconjugate chemistry.

[22]  E. Vázquez,et al.  Integrated approach to produce a recombinant, his‐tagged human α‐galactosidase a in mammalian cells , 2011, Biotechnology progress.

[23]  Kinam Park,et al.  Targeted drug delivery to tumors: myths, reality and possibility. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[24]  J. Kamps,et al.  Targeted siRNA delivery to diseased microvascular endothelial cells—Cellular and molecular concepts , 2011, IUBMB life.

[25]  M. Garcia-Parajo,et al.  pH-responsive polysaccharide-based polyelectrolyte complexes as nanocarriers for lysosomal delivery of therapeutic proteins. , 2011, Biomacromolecules.

[26]  Xiaoyuan Chen,et al.  Integrin Targeted Delivery of Chemotherapeutics , 2011, Theranostics.

[27]  F. Kiessling,et al.  Integrin Targeted Delivery of Radiotherapeutics , 2011, Theranostics.

[28]  Silvia Muro,et al.  Enhanced endothelial delivery and biochemical effects of α-galactosidase by ICAM-1-targeted nanocarriers for Fabry disease. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[29]  B. Evrard,et al.  Development of pH-responsive nanocarriers using trimethylchitosans and methacrylic acid copolymer for siRNA delivery. , 2010, Biomaterials.

[30]  T. Xia,et al.  Understanding biophysicochemical interactions at the nano-bio interface. , 2009, Nature materials.

[31]  W. Hennink,et al.  Influence of the degree of acetylation on the enzymatic degradation and in vitro biological properties of trimethylated chitosans. , 2009, Biomaterials.

[32]  V. Mourya,et al.  Trimethyl chitosan and its applications in drug delivery , 2009, Journal of materials science. Materials in medicine.

[33]  W. Hennink,et al.  Synthesis, characterization and in vitro biological properties of O-methyl free N,N,N-trimethylated chitosan. , 2008, Biomaterials.

[34]  Samir Mitragotri,et al.  Control of endothelial targeting and intracellular delivery of therapeutic enzymes by modulating the size and shape of ICAM-1-targeted carriers. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[35]  D. Bundle,et al.  A novel linker methodology for the synthesis of tailored conjugate vaccines composed of complex carbohydrate antigens and specific TH-cell peptide epitopes. , 2008, Chemistry.

[36]  R. Brady,et al.  Elevated globotriaosylsphingosine is a hallmark of Fabry disease , 2008, Proceedings of the National Academy of Sciences.

[37]  H. Uludaǧ,et al.  Conjugation of arginine-glycine-aspartic acid peptides to poly(ethylene oxide)-b-poly(epsilon-caprolactone) micelles for enhanced intracellular drug delivery to metastatic tumor cells. , 2007, Biomacromolecules.

[38]  H. Junginger,et al.  Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[39]  J. Shayman,et al.  An in vitro model of Fabry disease. , 2005, Journal of the American Society of Nephrology : JASN.

[40]  W. Mark Saltzman,et al.  Targeted for drug delivery , 2005 .

[41]  Anthony H. Futerman,et al.  The cell biology of lysosomal storage disorders , 2004, Nature Reviews Molecular Cell Biology.

[42]  D. Garboczi,et al.  The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. , 2004, Journal of molecular biology.

[43]  W. Sly,et al.  Glycosylation-independent targeting enhances enzyme delivery to lysosomes and decreases storage in mucopolysaccharidosis type VII mice. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[44]  R. Desnick,et al.  Enzyme replacement and enhancement therapies: lessons from lysosomal disorders , 2002, Nature Reviews Genetics.

[45]  S. W. Kim,et al.  An angiogenic, endothelial-cell-targeted polymeric gene carrier. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[46]  X. Dai,et al.  An improved synthesis of a selective αvβ3-integrin antagonist cyclo(-RGDfK-) , 2000 .

[47]  J. Brussee,et al.  Preparation and NMR characterization of highly substitutedN-trimethyl chitosan chloride , 1998 .

[48]  I. Pastan,et al.  alpha-Galactosidase A deficient mice: a model of Fabry disease. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[49]  R. Swerlick,et al.  HMEC-1: establishment of an immortalized human microvascular endothelial cell line. , 1992, The Journal of investigative dermatology.

[50]  Thomas J. Raub,et al.  Adsorptive endocytosis and membrane recycling by cultured primary bovine brain microvessel endothelial cell monolayers. , 1990, Journal of cell science.

[51]  R. Braun,et al.  Investigation of the bicinchoninic acid protein assay: identification of the groups responsible for color formation. , 1988, Analytical biochemistry.

[52]  J. Scheerer,et al.  Differential assay for lysosomal alpha-galactosidases in human tissues and its application to Fabry's disease. , 1981, Clinica chimica acta; international journal of clinical chemistry.

[53]  K. J. Dean,et al.  Studies on human liver alpha-galactosidases. I. Purification of alpha-galactosidase A and its enzymatic properties with glycolipid and oligosaccharide substrates. , 1979, The Journal of biological chemistry.

[54]  R. Desnick,et al.  Fabry's disease: enzymatic diagnosis of hemizygotes and heterozygotes. Alpha-galactosidase activities in plasma, serum, urine, and leukocytes. , 1973, The Journal of laboratory and clinical medicine.

[55]  J. Kint Fabry's Disease: Alpha-Galactosidase Deficiency , 1970, Science.

[56]  R. Brady,et al.  Enzymatic defect in Fabry's disease. Ceramidetrihexosidase deficiency. , 1967, The New England journal of medicine.

[57]  Eun Seong Lee,et al.  Poly(L-aspartic acid) nanogels for lysosome-selective antitumor drug delivery. , 2013, Colloids and surfaces. B, Biointerfaces.

[58]  L. Rajendran,et al.  Subcellular targeting strategies for drug design and delivery , 2010, Nature Reviews Drug Discovery.

[59]  Monty Liong,et al.  Cationic polystyrene nanosphere toxicity depends on cell-specific endocytic and mitochondrial injury pathways. , 2008, ACS nano.

[60]  V. Muzykantov,et al.  Lysosomal enzyme delivery by ICAM-1-targeted nanocarriers bypassing glycosylation- and clathrin-dependent endocytosis. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[61]  H. Sakuraba,et al.  Urinary excretion of the vitronectin receptor (integrin αVβ3) in patients with Fabry disease , 1999 .