CHEMICAL CHAPERONE THERAPY FOR GAUCHER DISEASE: N-OCTYL-BETA-VALIENAMINE INCREASES THE CELLULAR ACTIVITY OF NOT ONLY F213I BUT ALSO N188S BETAGLUCOCEREBROSIDASES

Objectives: Chemical chaperone therapy is a novel therapeutic strategy for glycolipid storage disorders to stabilize and accelerate transport of mutant enzymes by using a low molecular chemical that mimics a carbohydrate and inhibits the enzyme activity in vitro. We reported a carbohydrate mimic N-octyl-beta-valienamine (NOV) up-regulated cellular enzyme activity of Gaucher disease fibroblasts with a F213I beta-Glc mutation (2004). We have screened whether NOV works as a chemical chaperone for other beta-Glc mutant forms.

Methods: We have screened cultured fibroblasts from patients with Gaucher disease, in which beta-Glc activity is activated in the presence of NOV. Fibroblasts are cultured for 4 days in the presence or absence of NOV and beta-Glc activity in cell lysates was determined.

Results: We found beta-Glc activity in fibroblasts from a patient with N188S/G193W was activated in the presence of NOV. A FLAG-tagged beta-Glc cNDA with either N188S or G193W mutation was cloned in an expression vector and the construct was transfected into COS cells. We found that the protein level and activity of N188S were increased in the presence of NOV. Conclusion: NOV works as a chemical chaperone to up-regulate the F188S mutant beta-Glc and may suggest a therapeutic value of this compound for Gaucher disease with F213I and N188S mutations.