Akademik Veri Yönetim Sistemi
SSIEM Annual Symposium, Almanya, 30 Ağustos - 02 Eylül 2022, sa.1418955
Background: Gaucher disease is a lysosomal storage disease due to beta-glucocerebrosidase enzyme deficiency caused by mutations in the GBA gene. Enzyme replacement therapy seems to have no effect on neurological symptoms in chronic neuronopathic type due to the limitations in crossing the blood-brain barrier. In this study, beta-glucocerebrosidase recovery potential of four chemical chaperones were investigated for the first time in cultured fibroblasts obtained from a patient with chronic neuronopathic type of Gaucher Disease resulting from homozygous p.L483P mutation. Case Study / Methods: Skin fibroblasts were cultured in DMEM/%10 FBS at 37 degree C in %5 CO2. Phenylbutyrate, ursodeoxycholic acid, betaine and pyrimethamine which are already licensed for different indications but also known to act as chemical chaperones and can cross blood-brain barrier were selected for the experiment. The cells were incubated for 72 hours with phenylbutyrate at 0,1-0,5-2,5-5mM concentrations and with ursodeoxycholic acid at 5,10,25,50 μmol/L concentrations, betaine 10,25,50,100 mM concentrations and pyrimethamine 0,1-0,25-0,5-1 μg/ml concentrations. Glucocerebrosidase activity, glucosylsphingosine levels, gene expression, western blot and staining were performed in treated cells. Results: An increase in enzyme activities was demonstrated in fibroblasts treated with phenylbutyrate at a dose of 0,1 mM andwith ursodeoxycholic acid at doses of 5 and 25 μmol/L. In addition, reductions of up to 80% in glucosylsphingosine levels were detected with phenylbutyrate at a dose of 5 mM, and up to 75% with ursodeoxycholic acid at a dose of 10 μmol/L. Neuronal cell, animal, and clinical studies are planned. Conclusion / Discussion: It was for the first time shown that phenylbutyrate and ursodeoxycholic acid would act as chaperones to recover glucocerebrosidase activity in Gaucher Disease fibroblasts resulting from homozygous p.L483P mutation in the GBA gene and hold the promise for being novel treatments for chronic neuropathic type.