Ambroxol increases glucocerebrosidase (GCase) activity and restores GCase translocation in primary patient-derived macrophages in Gaucher disease and Parkinsonism

Kopytova, Alena E.; Rychkov, Georgy; Николаев, Михаил; Baydakova, Galina; Cheblokov, Aleksandr; Senkevich, Konstantin; Bogdanova, D.; Bolshakova, O. I.; Miliukhina, Irina; Bezrukikh, Vadim; Salogub, Galina; Саранцева, С. В.; Usenko, Tatiana; Zakharova, Ekaterina; Anton, Emil; Pchelina, Sofya

doi:10.1016/j.parkreldis.2021.02.003

Cited by 36 publications

(36 citation statements)

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“…Mutant GCase is recognized in cells as a misfolded protein, and instead of being trafficked to the lysosomes, it gets re-translocated to the cytoplasm, where it is degraded via the ubiquitin-proteasome system [ 18 ]. Several studies and our results confirm that AMB increases GCase trafficking to the lysosome and rescues the misfolded enzyme from degradation in GD macrophages and fibroblasts [ 16 , 18 , 36 ]. However, little is known about the effects of EGT on the trafficking and activity of GCase, aside from the fact that EGT did not inhibit enzyme activity [ 37 ].…”

Section: Discussionsupporting

confidence: 84%

“…AMB demonstrated good tolerability while enhancing GCase activity and improving neurological manifestations [ 14 ]. AMB interacts with active and non-active sites of enzymes, explaining the mixed type of activation/inhibition and pH-dependent activity [ 11 , 15 , 16 ]. Ambroxol stabilizes GCase, and demonstrates inhibitory GCase activity at neutral pH and absence of inhibitory effect at the acidic pH of lysosomes [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

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Cellular and biochemical response to chaperone versus substrate reduction therapies in neuropathic Gaucher disease

et al. 2021

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Gaucher disease (GD) is caused by deficiency of the lysosomal membrane enzyme glucocerebrosidase (GCase) and the subsequent accumulation of its substrate, glucosylceramide (GC). Mostly missense mutations of the glucocerebrosidase gene (GBA) cause GCase misfolding and inhibition of proper lysosomal trafficking. The accumulated GC leads to lysosomal dysfunction and impairs the autophagy pathway. GD types 2 and 3 (GD2-3), or the neuronopathic forms, affect not only the Central Nervous System (CNS) but also have severe systemic involvement and progressive bone disease. Enzyme replacement therapy (ERT) successfully treats the hematologic manifestations; however, due to the lack of equal distribution of the recombinant enzyme in different organs, it has no direct impact on the nervous system and has minimal effect on bone involvement. Small molecules have the potential for better tissue distribution. Ambroxol (AMB) is a pharmacologic chaperone that partially recovers the mutated GCase activity and crosses the blood-brain barrier. Eliglustat (EGT) works by inhibiting UDP-glucosylceramide synthase, an enzyme that catalyzes GC biosynthesis, reducing GC influx load into the lysosome. Substrate reduction therapy (SRT) using EGT is associated with improvement in GD bone marrow burden score and bone mineral density parallel with the improvement in hematological parameters. We assessed the effects of EGT and AMB on GCase activity and autophagy-lysosomal pathway (ALP) in primary cell lines derived from patients with GD2-3 and compared to cell lines from healthy controls. We found that EGT, same as AMB, enhanced GCase activity in control cells and that an individualized response, that varied with GBA mutations, was observed in cells from patients with GD2-3. EGT and AMB enhanced the formation of lysosomal/late endosomal compartments and improved autophagy, independent of GBA mutations. Both AMB and EGT increased mitochondrial mass and density in GD2-3 fibroblasts, suggesting enhancement of mitochondrial function by activating the mitochondrial membrane potential. These results demonstrate that EGT and AMB, with different molecular mechanisms of action, enhance GCase activity and improve autophagy-lysosome dynamics and mitochondrial functions.

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Section: Discussionsupporting

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Cellular and biochemical response to chaperone versus substrate reduction therapies in neuropathic Gaucher disease

et al. 2021

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“…Monocyte-derived macrophages represent one of the most promising models for investigating molecular mechanisms of GCase dysfunction, as this cell type is vulnerable for disturbances in ceramide metabolism [16,17]. In particular, we and others demonstrated high potential of peripheral blood monocyte-derived macrophages to reflect individual sensitivity for drugs influencing GCase activity [18,19]. Here, we first generated the transcriptomic profiles for GBA-PD patients, asymptomatic GBA mutation carriers (GBA carriers), and controls in monocyte-derived macrophages, in order to investigate what variations in monocytederived macrophage transcriptomes can be attributed to the presence of GBA mutation and what can be viewed as a trigger of PD in GBA mutation carriers.…”

Section: Introductionmentioning

confidence: 76%

“…PBMCs were differentiated by the macrophage colony-stimulating factor (M-CSF) (10 ng/ml) (Sigma-Aldrich, Burlington, MA, USA) in RPMI 1640 medium (Gibco, Waltham, MA, USA) supplemented with 10% FCS (Gibco, Waltham, MA, USA) with harvesting after 5 days. Phenotypical maturation of monocyte-derived macrophages was confirmed by light microscopy and flow cytometry with specific antibodies to CD14+ and CD68+ (eBioscience, San Diego, CA, USA), as described earlier [18,21].…”

Section: Differentiation Of Human Monocytes To Macrophagesmentioning

confidence: 99%

Comparative Transcriptome Analysis in Monocyte-Derived Macrophages of Asymptomatic GBA Mutation Carriers and Patients with GBA-Associated Parkinson’s Disease

Usenko

Bezrukova

Basharova

et al. 2021

Genes

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Mutations of the GBA gene, encoding for lysosomal enzyme glucocerebrosidase (GCase), are the greatest genetic risk factor for Parkinson’s disease (PD) with frequency between 5% and 20% across the world. N370S and L444P are the two most common mutations in the GBA gene. PD carriers of severe mutation L444P in the GBA gene is characterized by the earlier age at onset compared to N370S. Not every carrier of GBA mutations develop PD during one’s lifetime. In the current study we aimed to find common gene expression signatures in PD associated with mutation in the GBA gene (GBA-PD) using RNA-seq. We compared transcriptome of monocyte-derived macrophages of 5 patients with GBA-PD (4 L444P/N, 1 N370S/N) and 4 asymptomatic GBA mutation carriers (GBA-carriers) (3 L444P/N, 1 N370S/N) and 4 controls. We also conducted comparative transcriptome analysis for L444P/N only GBA-PD patients and GBA-carriers. Revealed deregulated genes in GBA-PD independently of GBA mutations (L444P or N370S) were involved in immune response, neuronal function. We found upregulated pathway associated with zinc metabolism in L444P/N GBA-PD patients. The potential important role of DUSP1 in the pathogenesis of GBA-PD was suggested.

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“…Our team and other authors have shown that a primary culture of macrophages derived from the pe- ripheral blood monocytes of GBA-PD and GD patients can be used for personalized screening and assessment of the effectiveness of pharmacological chaperones [ 124 , 125 ]. Peripheral blood macrophages from GD and GBA-PD patients, which were cultured in the presence of ambroxol, demonstrated an increase in GCase activity and a decrease in the concentration of lysosphingolipids [ 124 , 125 , 126 ]. Recent data have demonstrated that the effects of ambroxol can depend on the type of GBA gene mutations.…”

Section: Potential Therapeutic Approaches For Gba-pdmentioning

confidence: 99%

Parkinson’s Disease Associated with GBA Gene Mutations: Molecular Aspects and Potential Treatment Approaches

et al. 2021

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Parkinsons disease (PD) is a multifactorial neurodegenerative disease. To date, genome-wide association studies have identified more than 70 loci associated with the risk of PD. Variants in the GBA gene encoding glucocerebrosidase are quite often found in PD patients in all populations across the world, which justifies intensive investigation of this gene. A number of biochemical features have been identified in patients with GBA-associated Parkinsons disease (GBA-PD). In particular, these include decreased activity of glucocerebrosidase and accumulation of the glucosylceramide substrate. These features were the basis for putting forward a hypothesis about treatment of GBA-PD using new strategies aimed at restoring glucocerebrosidase activity and reducing the substrate concentration. This paper discusses the molecular and genetic mechanisms of GBA-PD pathogenesis and potential approaches to the treatment of this form of the disease.

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Ambroxol increases glucocerebrosidase (GCase) activity and restores GCase translocation in primary patient-derived macrophages in Gaucher disease and Parkinsonism

Cited by 36 publications

References 53 publications

Cellular and biochemical response to chaperone versus substrate reduction therapies in neuropathic Gaucher disease

Cellular and biochemical response to chaperone versus substrate reduction therapies in neuropathic Gaucher disease

Comparative Transcriptome Analysis in Monocyte-Derived Macrophages of Asymptomatic GBA Mutation Carriers and Patients with GBA-Associated Parkinson’s Disease

Parkinson’s Disease Associated with GBA Gene Mutations: Molecular Aspects and Potential Treatment Approaches

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