Abstract
There is still no treatment for polyglutamine disorders, but clearance of mutant proteins might represent a potential therapeutic strategy. Autophagy, the major pathway for organelle and protein turnover, has been implicated in these diseases. To determine whether the autophagy/lysosome system contributes to the pathogenesis of spinocerebellar ataxia type 7 (SCA7), caused by expansion of a polyglutamine tract in the ataxin-7 protein, we looked for biochemical, histological and transcriptomic abnormalities in components of the autophagy/lysosome pathway in a knock-in mouse model of the disease, postmortem brain and peripheral blood mononuclear cells (PBMC) from patients. In the mouse model, mutant ataxin-7 accumulated in inclusions immunoreactive for the autophagy-associated proteins mTOR, beclin-1, p62 and ubiquitin. Atypical accumulations of the autophagosome/lysosome markers LC3, LAMP-1, LAMP2 and cathepsin-D were also found in the cerebellum of the SCA7 knock-in mice. In patients, abnormal accumulations of autophagy markers were detected in the cerebellum and cerebral cortex of patients, but not in the striatum that is spared in SCA7, suggesting that autophagy might be impaired by the selective accumulation of mutant ataxin-7. In vitro studies demonstrated that the autophagic flux was impaired in cells overexpressing full-length mutant ataxin-7. Interestingly, the expression of the early autophagy-associated gene ATG12 was increased in PBMC from SCA7 patients in correlation with disease severity. These results provide evidence that the autophagy/lysosome pathway is impaired in neurons undergoing degeneration in SCA7. Autophagy/lysosome-associated molecules might, therefore, be useful markers for monitoring the effects of potential therapeutic approaches using modulators of autophagy in SCA7 and other autophagy/lysosome-associated neurodegenerative disorders.
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Abbreviations
- SCA7:
-
Spinocerebellar ataxia type 7
- polyQ:
-
Polyglutamine
- ATXN7:
-
Ataxin-7
- KI:
-
Knock-in
- PBMC:
-
Peripheral blood mononuclear cells
- PC:
-
Purkinje cell
- ATG:
-
Autophagy-related protein
- LC3:
-
Microtubule-associated protein 1 light chain (MAP1) light chain 3
- Lamp-1:
-
Lysosomal-associated membrane protein 1
- Lamp-2:
-
Lysosomal associated membrane protein 2
- PML:
-
Promyelocytic leukemia protein
- APP:
-
Amyloid precursor protein
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Acknowledgments
This study was supported by grants from the French National Research Agency (ANR-07-MRAR-025-01 to A.S), the French Association against Myopathies (AFM, to AB and long-term fellowship to SA), the French association Connaitre les Syndrômes Cérébelleux (short-term fellowship to SA), and the French Foundation for Medical Research (FRM, to JCC and FC-D), as well as the “Investissements d’avenir” program ANR-10-IAIHU-06 (to the Brain and Spine Institute, Paris). We thank Prof. H. Zoghbi (Baylor College of Medicine, Houston, Texas, USA) for the SCA7 KI mice. We are grateful to Pr. C. Duyckaerts for brain samples and Drs. C. Marelli and C. Jauffret for blood samples from SCA7 patients. We would also like to thank the Cellular Imaging Platform of the Pitié Salpêtrière, especially Dr. A. Dauphin, for advice on confocal imaging, and J. Garrigue and W. Carpentier for technical assistance. The authors have no additional financial interests.
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F. Cormier-Dequaire and M. Marinello contributed equally.
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Alves, S., Cormier-Dequaire, F., Marinello, M. et al. The autophagy/lysosome pathway is impaired in SCA7 patients and SCA7 knock-in mice. Acta Neuropathol 128, 705–722 (2014). https://doi.org/10.1007/s00401-014-1289-8
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DOI: https://doi.org/10.1007/s00401-014-1289-8