Comparison between Nanoparticle Encapsulation and Surface Loading for Lysosomal Enzyme Replacement Therapy

Abstract

Poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) enhance the delivery of therapeutic enzymes for replacement therapy of lysosomal storage disorders. Previous studies examined NPs encapsulating or coated with enzymes, but these formulations have never been compared. We examined this using hyaluronidase (HAse), deficient in mucopolysaccharidosis IX, and acid sphingomyelinase (ASM), deficient in types A–B Niemann–Pick disease. Initial screening of size, PDI, ζ potential, and loading resulted in the selection of the Lactel II co-polymer vs. Lactel I or Resomer, and Pluronic F68 surfactant vs. PVA or DMAB. Enzyme input and addition of carrier protein were evaluated, rendering NPs having, e.g., 181 nm diameter, 0.15 PDI, −36 mV ζ potential, and 538 HAse molecules encapsulated per NP. Similar NPs were coated with enzyme, which reduced loading (e.g., 292 HAse molecules/NP). NPs were coated with targeting antibodies (> 122 molecules/NP), lyophilized for storage without alterations, and acceptably stable at physiological conditions. NPs were internalized, trafficked to lysosomes, released active enzyme at lysosomal conditions, and targeted both peripheral organs and the brain after i.v. administration in mice. While both formulations enhanced enzyme delivery compared to free enzyme, encapsulating NPs surpassed coated counterparts (18.4- vs. 4.3-fold enhancement in cells and 6.2- vs. 3-fold enhancement in brains), providing guidance for future applications.

Defect-free graphene enhances enzyme delivery to fibroblasts derived from patients with lysosomal storage disorders

“…15 At present, polymeric nanoparticles, owing to their biocompatibility and stability, are one of the most explored NMs in ERT. [16][17][18][19][20] Enzyme encapsulation or surface coating have been proposed so far. 20 However, this approach might involve complex chemistry due to surface modifications needed, in order to trigger cellular uptake and trafficking to the lysosomes.…”

“…[16][17][18][19][20] Enzyme encapsulation or surface coating have been proposed so far. 20 However, this approach might involve complex chemistry due to surface modifications needed, in order to trigger cellular uptake and trafficking to the lysosomes. [17][18][19] Thus, GBMs could provide a simpler option for both the loading and delivery of enzymes: high enzyme loading can be achieved through non-covalent interactions between the enzyme and the material, while subcellular localization within the lysosomes occurs through a natural intracellular trafficking route of GBMs.…”

Harnessing the Materials Chemistry of Mesoporous Silicon Nanoparticles to Prepare “Armor-Clad” Enzymes

“…These hosts serve to stabilize the proteins and optimize their function . Similarly, the incorporation of enzymes within nanomaterial-based hosts offers a strategy to improve performance, extend lifetime, and increase operating stability under conditions that would normally be detrimental to enzyme function. − Here broader considerations in terms of processability, biocompatibility, leaching, and direct delivery of enzymes into a targeted location are of significant importance for applications in biomedical and other health-related fields. − Extensive research effort has been expended on the development of enzyme host materials and immobilization strategies based on adsorption, covalent conjugation, pore infiltration and in situ synthesis (i.e., coprecipitation, biomineralization). − However, it remains a challenge to develop general approaches to prepare encapsulated enzymes that satisfy all of the above constraints.…”

Role of the Lactide:Glycolide Ratio in PLGA Nanoparticle Stability and Release under Lysosomal Conditions for Enzyme Replacement Therapy of Lysosomal Storage Disorders

“…For these reasons, several studies have explored PLGA NPs for lysosomal ERT, including formulations loaded with acid sphingomyelinase for treatment of Niemann–Pick disease types A or B, hyaluronidase (HAse) for MPS type IX, iduronate-2-sulfatase for MPS type II, palmitoyl-protein thioesterase-1 for neuronal ceroid lipofuscinosis 1, galactosylceramidase for Krabbe disease, and acid α-glucosidase for Pompe disease [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ]. Some of these PLGA NP formulations additionally displayed targeting elements for receptor-mediated transport across the BBB and/or inside target cells, such as intercellular adhesion molecule 1 (ICAM-1) antibodies, angiopep-2 oligopeptide, transferrin-binding peptide Tf2, or g7 glycopeptide [ 31 , 32 , 33 , 34 , 36 , 37 , 38 , 39 , 40 ]. All these studies have demonstrated enhanced delivery and/or effects of lysosomal enzymes in cell culture and/or animal models [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ].…”

“…Some of these PLGA NP formulations additionally displayed targeting elements for receptor-mediated transport across the BBB and/or inside target cells, such as intercellular adhesion molecule 1 (ICAM-1) antibodies, angiopep-2 oligopeptide, transferrin-binding peptide Tf2, or g7 glycopeptide [ 31 , 32 , 33 , 34 , 36 , 37 , 38 , 39 , 40 ]. All these studies have demonstrated enhanced delivery and/or effects of lysosomal enzymes in cell culture and/or animal models [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ]. However, their delivery efficacy and associated effects varied among different formulations [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ].…”