Use of Polylactide-Co-Glycolide-Nanoparticles for Lysosomal Delivery of a Therapeutic Enzyme in Glycogenosis Type II Fibroblasts.
Glycogenosis type II, or Pompe Disease, is a lysosomal storage disease caused by the deficiency of acid alpha-glucosidase (GAA), leading to glycogen accumulation in muscles. A recombinant human GAA (rhGAA, Myozyme®) is currently used for enzyme replacement therapy. Despite its efficacy in most of patients, some of them show a diminished response to the treatment with rapidly progressive clinical deterioration, due to immuno-mediated enzyme inactivation. To demonstrate that Nanoparticles (NPs) could be profitably exploited to carry macromolecules, PLGA NPs loaded with rhGAA (GAA-NPs) were prepared by double emulsion solvent evaporation. Their surface morphology, particle size, zeta-potential and biochemical activity were assessed. "Pulse and chase" experiments were made by administrating GAA-NPs on patients' fibroblasts. Biochemical activity tests showed a more efficient cellular uptake of rhGAA loaded to NPs and a more significant stability of the enzyme (up to 7 days) in vitro, if compared to the same amount of rhGAA free enzyme. This data allows to envision in vivo experiments, in significant animal models, to further characterize lysosomal enzyme loaded-NPs' efficacy and toxicity.
Brunella
B
Tancini
Giovanni
G
Tosi
Barbara
B
Bortot
Diego
D
Dolcetta
Alessandro
A
Magini
Eleonora
E
De Martino
Lorena
L
Urbanelli
Barbara
B
Ruozi
Flavio
F
Forni
Carla
C
Emiliani
Maria Angela
MA
Vandelli
Giovanni Maria
GM
Severini
D002478:Cells, Cultured; D016503:Drug Delivery Systems; D005347:Fibroblasts; D006009:Glycogen Storage Disease Type II; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry* / Q000493:pharmacokinetics; D008247:Lysosomes / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000493:pharmacokinetics; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012313:RNA / Q000737:chemistry* / Q000493:pharmacokinetics; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000493:pharmacokinetics
C039919:RNA, recombinant; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D012313:RNA; C509951:GAA protein, human; D000520:alpha-Glucosidases