title: Screening for pharmacological chaperones in Fabry disease.
abstract: As a prerequisite for clinical trials of pharmacological chaperone therapy (PCT) for Fabry disease, we developed a rapid screening assay for enhancement of endogenous alpha-galactosidase A (alpha-Gal A) in patient-derived cells. We used a T-cell based system to screen 11 mutations causing Fabry disease for enhanceability using 1-deoxygalactonojirimycin (DGJ). When patient-derived T-cells were grown in the presence of DGJ, alpha-Gal A activity increased to more than 50% of normal in several mutations but was unaffected in others. In addition to the mutation R301Q, reported previously, A97V, R112H, R112C, A143T, and L300P were enhanceable, but R356W, G132R, A143P, R220X, and 30delG were not. The level of alpha-Gal A activity achieved provides a basis for the therapeutic trial of DGJ in patients with similarly enhanceable enzyme. This assay method has general utility in other disorders in assessing the degree of enhancement of activity of mutated proteins by PCT.
authors.affiliation: Developmental and Metabolic Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 10 Center Drive, Building 10, Room 3D04, MSC 1260, Bethesda, MD 20892-1260, USA.
authors.forename: Sang-Hoon
authors.initials: SH
authors.lastname: Shin
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Murray
authors.forename: Stefanie
authors.initials: S
authors.lastname: Kluepfel-Stahl
authors.forename: Adele M
authors.initials: AM
authors.lastname: Cooney
authors.forename: Jane M
authors.initials: JM
authors.lastname: Quirk
authors.forename: Raphael
authors.initials: R
authors.lastname: Schiffmann
authors.forename: Roscoe O
authors.initials: RO
authors.lastname: Brady
authors.forename: Christine R
authors.initials: CR
authors.lastname: Kaneski
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002478:Cells, Cultured; D015195:Drug Design; D000795:Fabry Disease / Q000378:metabolism*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008297:Male; D018832:Molecular Chaperones / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D000519:alpha-Galactosidase / Q000378:metabolism*
chemical_list: D018832:Molecular Chaperones; D000519:alpha-Galactosidase