Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells
Heterozygous mutations in GBA1 gene, encoding for lysosomal enzyme glucocerebrosidase (GCase), are a major risk factor for sporadic Parkinson's disease (PD). Defective GCase has been reported in fibroblasts of GBA1-mutant PD patients and pharmacological chaperone ambroxol has been shown to corr...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-10-01
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| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996115002260 |
| _version_ | 1818610517336391680 |
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| author | Giulia Ambrosi Cristina Ghezzi Roberta Zangaglia Giovanna Levandis Claudio Pacchetti Fabio Blandini |
| author_facet | Giulia Ambrosi Cristina Ghezzi Roberta Zangaglia Giovanna Levandis Claudio Pacchetti Fabio Blandini |
| author_sort | Giulia Ambrosi |
| collection | DOAJ |
| description | Heterozygous mutations in GBA1 gene, encoding for lysosomal enzyme glucocerebrosidase (GCase), are a major risk factor for sporadic Parkinson's disease (PD). Defective GCase has been reported in fibroblasts of GBA1-mutant PD patients and pharmacological chaperone ambroxol has been shown to correct such defect. To further explore this issue, we investigated GCase and elements supporting GCase function and trafficking in fibroblasts from sporadic PD patients — with or without heterozygous GBA1 mutations — and healthy subjects, in basal conditions and following in vitro exposure to ambroxol. We assessed protein levels of GCase, lysosomal integral membrane protein-2 (LIMP-2), which mediates GCase trafficking to lysosomes, GCase endogenous activator saposin (Sap) C and parkin, which is involved in degradation of defective GCase. We also measured activities of GCase and cathepsin D, which cleaves Sap C from precursor prosaposin. GCase activity was reduced in fibroblasts from GBA1-mutant patients and ambroxol corrected this defect. Ambroxol increased cathepsin D activity, GCase and Sap C protein levels in all groups, while LIMP-2 levels were increased only in GBA1-mutant PD fibroblasts. Parkin levels were slightly increased only in the PD group without GBA1 mutations and were not significantly modified by ambroxol. Our study confirms that GCase activity is deficient in fibroblasts of GBA1-mutant PD patients and that ambroxol corrects this defect. The drug increased Sap C and LIMP-2 protein levels, without interfering with parkin. These results confirm that chemical chaperone ambroxol modulates lysosomal markers, further highlighting targets that may be exploited for innovative PD therapeutic strategies. |
| first_indexed | 2024-12-16T15:15:41Z |
| format | Article |
| id | doaj.art-0c9263ad7acd43849db15d4f02295f00 |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-16T15:15:41Z |
| publishDate | 2015-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-0c9263ad7acd43849db15d4f02295f002022-12-21T22:26:47ZengElsevierNeurobiology of Disease1095-953X2015-10-0182235242Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cellsGiulia Ambrosi0Cristina Ghezzi1Roberta Zangaglia2Giovanna Levandis3Claudio Pacchetti4Fabio Blandini5Center for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyCenter for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyCenter for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyCenter for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyCenter for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyCorresponding author at: C. Mondino National Neurological Institute, Via Mondino, 2, 27100, Pavia, Italy.; Center for Research in Neurodegenerative Diseases, C. Mondino National Neurological Institute, Pavia, ItalyHeterozygous mutations in GBA1 gene, encoding for lysosomal enzyme glucocerebrosidase (GCase), are a major risk factor for sporadic Parkinson's disease (PD). Defective GCase has been reported in fibroblasts of GBA1-mutant PD patients and pharmacological chaperone ambroxol has been shown to correct such defect. To further explore this issue, we investigated GCase and elements supporting GCase function and trafficking in fibroblasts from sporadic PD patients — with or without heterozygous GBA1 mutations — and healthy subjects, in basal conditions and following in vitro exposure to ambroxol. We assessed protein levels of GCase, lysosomal integral membrane protein-2 (LIMP-2), which mediates GCase trafficking to lysosomes, GCase endogenous activator saposin (Sap) C and parkin, which is involved in degradation of defective GCase. We also measured activities of GCase and cathepsin D, which cleaves Sap C from precursor prosaposin. GCase activity was reduced in fibroblasts from GBA1-mutant patients and ambroxol corrected this defect. Ambroxol increased cathepsin D activity, GCase and Sap C protein levels in all groups, while LIMP-2 levels were increased only in GBA1-mutant PD fibroblasts. Parkin levels were slightly increased only in the PD group without GBA1 mutations and were not significantly modified by ambroxol. Our study confirms that GCase activity is deficient in fibroblasts of GBA1-mutant PD patients and that ambroxol corrects this defect. The drug increased Sap C and LIMP-2 protein levels, without interfering with parkin. These results confirm that chemical chaperone ambroxol modulates lysosomal markers, further highlighting targets that may be exploited for innovative PD therapeutic strategies.http://www.sciencedirect.com/science/article/pii/S0969996115002260GlucocerebrosidaseSaposin CLIMP-2Parkinson's diseaseLysosomesFibroblasts |
| spellingShingle | Giulia Ambrosi Cristina Ghezzi Roberta Zangaglia Giovanna Levandis Claudio Pacchetti Fabio Blandini Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells Neurobiology of Disease Glucocerebrosidase Saposin C LIMP-2 Parkinson's disease Lysosomes Fibroblasts |
| title | Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells |
| title_full | Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells |
| title_fullStr | Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells |
| title_full_unstemmed | Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells |
| title_short | Ambroxol-induced rescue of defective glucocerebrosidase is associated with increased LIMP-2 and saposin C levels in GBA1 mutant Parkinson's disease cells |
| title_sort | ambroxol induced rescue of defective glucocerebrosidase is associated with increased limp 2 and saposin c levels in gba1 mutant parkinson s disease cells |
| topic | Glucocerebrosidase Saposin C LIMP-2 Parkinson's disease Lysosomes Fibroblasts |
| url | http://www.sciencedirect.com/science/article/pii/S0969996115002260 |
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