Boertien, L.M. (2016) The effect of mutant SOD1 on cell viability. thesis, Medicine.
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Abstract
Mutations in the gene encoding for superoxide dismutase 1 (mSOD1) lead to protein aggregation and are associated with 20% of familial amyotrophic lateral sclerosis (ALS) cases. Multiple molecular chaperones are known to influence mSOD1 aggregation and cytotoxicity. Still, the relation between protein aggregation and cytotoxicity remains elusive. In the current project, human osteosarcoma cell lines stably expressing SOD1-WT or SOD1-A4V (U2OS-WT, U2OS-A4V) were exposed to endoplasmatic reticular stress, proteasomal inhibition and autophagy inhibition. Phenotypes of SOD1-A4V cytotoxicity were then correlated to levels of mSOD1 expression and aggregation. Additionally, overexpression of the molecular chaperones HSPA1A and HSPA1L was investigated for their potential to rescue the mSOD1 associated phenotype. Endoplasmatic reticular (ER) stress induced by Tunicamycin treatment was the only stressor that produced a reproducible phenotype characterised by a decrease in U2OS-A4V cell viability compared to U2OS-WT. No correlation with mSOD1 aggregation could be established. HSPA1A and HSPA1L overexpression both resulted in a marginal relative rescue of cell viability specific to U2OS-A4V under Tunicamycin treatment. Interpretation of the results was hampered by the marginal differences in cell viability, the low expression levels of SOD1-A4V and the expression of endogenous SOD1. An experimental endogenous SOD1 knockout model, with expression levels of the proteins of interest close to endogenous expression of SOD1, is proposed to substantiate the findings of this study. Further investigation of the relationship between ER stress and mSOD1 cytotoxicity can be centred around the in literature described interaction between mSOD1 and Derlin-1, through which mSOD1 is proposed to mediate its cytotoxicity.
| Item Type: | Thesis (Thesis) |
|---|---|
| Supervisor name: | Faculty Supervisor: and Bergink, S. (Steven) and Secondary Supervisor: and Serlidaki, D. (Despina) and Department of Medical Cell Biology UMCG |
| Faculty: | Medical Sciences |
| Date Deposited: | 25 Jun 2020 11:02 |
| Last Modified: | 25 Jun 2020 11:02 |
| URI: | https://umcg.studenttheses.ub.rug.nl/id/eprint/2195 |
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