Objective: The aim was to evaluate the effectiveness of the restoration of GCase activity in macrophages from GBA-PD patients using pharmacological chaperone ambroxol.

Background: Mutations in the GBA gene lead to a deficiency of glucocerebrosidase (GCase) enzymatic activity and to the development of Gaucher disease (GD) which belongs to lysosomal storage diseases. At the same time GBA mutations increase the risk of Parkinson’s disease (PD) in 7-8 times. It was discussed in recent years that pharmacological chaperones could potentially enhance GCase activity and therefore could be used for GD and GBA-PD treatment. One of the most promising molecule is ambroxol. This drug is currently undergoing first clinical trial targeting patients with GBA-associated PD (GBA-PD).

Method: Mononuclear fraction was isolated from whole blood of GBA-PD patients (with mutations in GBA gene: N370S (n=3) and with L444P (n=3)) and healthy controls (n=5). With subsequent differentiated into macrophages using RPMI supplemented with 10% bovine serum, 1% streptomycin-penicillin and 10 ng/ml M-CSF for 4 days, with daily media changes. GCase enzymatic activity and concertation of lysosphingolipids – hexosylsphingosine HexSph (GlcSph+GalSph) were measured by liquid chromatography with tandem-mass spectrometry (LC-MS/MS) in dry macrophage cell spots on filter paper with the concentration of 2×10^6 cells/ml. Macrophages were treated with final concentration concentrations of ambroxol hydrochloride 50 µM.

Results: We evaluated the effects of ambroxol treatment on GCase activity. In our study, we showed an increase of GCase activity in macrophages derived from GBA-PD patients with ambroxol 56,80 (29,35 – 115,14) mmol/l/h compared with cells without ambroxol 19,52 (8,30-27,67) (p<0.001). We also shown the decrease of HexSph concentration in macrophages from GBA-PD patients with presence of ambroxol 31,68 (19,73 – 37,92) ng/ml compared with cells without ambroxol 48,26 (32,15 – 86,66) ng/ml (p=0.015).

Conclusion: Ambroxol increases GCase activity and decrease lysoscphingolipids concentration in primary macrophages from GBA-PD patients. We propose that ambroxol should be further investigated as a potential drug for PD treatment. Macrophage culture is suitable for screening of new potential GCase pharmacological chaperones. The study was supported by RSF № 17-75-20159

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MDS Abstracts - https://www.mdsabstracts.org/abstract/ambroxol-treatment-of-primary-macrophages-derived-from-patients-with-gba-associated-parkinsons-disease/