Use of a modified alpha-N-acetylgalactosaminidase in the development of enzyme replacement therapy for Fabry disease.
A modified alpha-N-acetylgalactosaminidase (NAGA) with alpha-galactosidase A (GLA)-like substrate specificity was designed on the basis of structural studies and was produced in Chinese hamster ovary cells. The enzyme acquired the ability to catalyze the degradation of 4-methylumbelliferyl-alpha-D-galactopyranoside. It retained the original NAGA's stability in plasma and N-glycans containing many mannose 6-phosphate (M6P) residues, which are advantageous for uptake by cells via M6P receptors. There was no immunological cross-reactivity between the modified NAGA and GLA, and the modified NAGA did not react to serum from a patient with Fabry disease recurrently treated with a recombinant GLA. The enzyme cleaved globotriaosylceramide (Gb3) accumulated in cultured fibroblasts from a patient with Fabry disease. Furthermore, like recombinant GLA proteins presently used for enzyme replacement therapy (ERT) for Fabry disease, the enzyme intravenously injected into Fabry model mice prevented Gb3 storage in the liver, kidneys, and heart and improved the pathological changes in these organs. Because this modified NAGA is hardly expected to cause an allergic reaction in Fabry disease patients, it is highly promising as a new and safe enzyme for ERT for Fabry disease.
Department of Clinical Genetics, Meiji Pharmaceutical University, Tokyo 204-8588, Japan.
Youichi
Y
Tajima
Ikuo
I
Kawashima
Takahiro
T
Tsukimura
Kanako
K
Sugawara
Mayuko
M
Kuroda
Toshihiro
T
Suzuki
Tadayasu
T
Togawa
Yasunori
Y
Chiba
Yoshifumi
Y
Jigami
Kazuki
K
Ohno
Tomoko
T
Fukushige
Takuro
T
Kanekura
Kohji
K
Itoh
Toya
T
Ohashi
Hitoshi
H
Sakuraba
D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002384:Catalysis; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004195:Disease Models, Animal; D004355:Drug Stability; D056947:Enzyme Replacement Therapy / Q000379:methods*; D000795:Fabry Disease / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects; D005456:Fluorescent Dyes / Q000378:metabolism; D005697:Galactosides / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D007150:Immunohistochemistry; D007668:Kidney / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D008099:Liver / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D008958:Models, Molecular; D008970:Molecular Weight; D009206:Myocardium / Q000473:pathology / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000627:therapeutic use; D012190:Retroviridae / Q000235:genetics; D014162:Transfection; D014281:Trihexosylceramides / Q000378:metabolism; D048809:alpha-N-Acetylgalactosaminidase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use*
D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D005697:Galactosides; D011994:Recombinant Proteins; D014281:Trihexosylceramides; D006923:Hymecromone; C026845:4-methylumbelliferyl-galactopyranoside; C018549:globotriaosylceramide; D048809:alpha-N-Acetylgalactosaminidase