Comparison of the effects of agalsidase alfa and agalsidase beta on cultured human Fabry fibroblasts and Fabry mice

Abstract

We compared two recombinant a-galactosidases developed for enzyme replacement therapy for Fabry disease, agalsidase alfa and agalsidase beta, as to specific a-galactosidase activity, stability in plasma, mannose 6-phosphate (M6P) residue content, and effects on cultured human Fabry fibroblasts and Fabry mice. The specific enzyme activities of agalsidase alfa and agalsidase beta were 1.70 and 3.24 mmol h À1 mg protein À1 , respectively, and there was no difference in stability in plasma between them. The M6P content of agalsidase beta (3.6 mol/mol protein) was higher than that of agalsidase alfa (1.3 mol/mol protein). The administration of both enzymes resulted in marked increases in a-galactosidase activity in cultured human Fabry fibroblasts, and Fabry mouse kidneys, heart, spleen and liver. However, the increase in enzyme activity in cultured fibroblasts, kidneys, heart and spleen was higher when agalsidase beta was used. An immunocytochemical analysis revealed that the incorporated recombinant enzyme degraded the globotriaosyl ceramide accumulated in cultured Fabry fibroblasts in a dose-dependent manner, with the effect being maintained for at least 7 days. Repeated administration of agalsidase beta apparently decreased the number of accumulated lamellar inclusion bodies in renal tubular cells of Fabry mice.

Establishment of immortalized Schwann cells from Fabry mice and their low uptake of recombinant α-galactosidase

“…A decrease in the number of inclusion bodies was not observed after 24 h in the culture medium containing 3.0 lg/mL agalsidase beta (data not shown), although intracellularly deposited globotriaosylceramide was apparently decreased in the case of cultured Fabry fibroblasts (F377), as described previously (Sakuraba et al 2006a(Sakuraba et al , 2006b). The administration of a high dose (10.0 lg/mL) of the enzyme apparently decreased the number of inclusion bodies in the cells (Fig.…”

“…Actin was used as an internal control. Lane 1, brain; lane 2, kidney; lane 3, liver completely sufficient to remove the storage materials from human Fabry fibroblasts (Sakuraba et al 2006a(Sakuraba et al , 2006b.…”

Reduction of Plasma Globotriaosylsphingosine Levels After Switching from Agalsidase Alfa to Agalsidase Beta as Enzyme Replacement Therapy for Fabry Disease

“…However, agalsidase beta contains more fully sialylated oligosaccharides and more mannose-6-phosphate than agalsidase alfa (Lee et al 2003;Togawa et al 2014); cellular uptake of recombinant agalsidases is mediated by mannose-6-phosphate receptors. Greater uptake of agalsidase beta than of agalsidase alfa has been suggested in vitro (in skin fibroblasts from patients with FD) (Keslová-Veselíková et al 2008;Lee et al 2003) and in vivo (in the kidney and heart after administration of the same dose of the two enzymes in a mouse model of FD) (Desnick and Schuchman 2012;Sakuraba et al 2006). Although not adequately powered, the results from a study in patients with FD suggest that the two agents have similar clinical effects when administered at the same dose of 0.2 mg/kg every other week (EOW) (Vedder et al 2007).…”

Nicotiana benthamiana α-galactosidase A1.1 can functionally complement human α-galactosidase A deficiency associated with Fabry disease

“…Mannose-6-phosphate moieties (Man-6-P) on the 3 N-linked glycans of α-Gal A mediate the transport of newly formed enzyme to lysosomes by Man-6-P receptors (MPRs). Alternative sorting via the mannose receptor pathway was suggested by Shen and colleagues (18,19). Dysfunction or absence of α-Gal A leads to Fabry disease (FD), an X-linked lysosomal disorder characterized by accumulation of glycosphingolipids with terminal galactosyl moieties in tissues and body fluids of FD patients (9,20).…”

“…Two therapeutic enzymes (agalsidase alpha, Replagal ® and agalsidase beta, Fabrazyme ® ) produced in mammalian cells are in use and a plant-produced enzyme is being developed (18,22). The efficacy of present ERT interventions is considered to be poor (23).…”