Effects of Cellular Pathway Disturbances on Misfolded Superoxide Dismutase-1 in Fibroblasts Derived from ALS Patients

摘要: Mutations in superoxide dismutase-1 (SOD1) are a common known cause of amyotrophic lateral sclerosis (ALS). The neurotoxicity mutant SOD1s is most likely caused by misfolded molecular species, but disease pathogenesis still not understood. Proposed mechanisms include impaired mitochondrial function, induction endoplasmic reticulum stress, reduction the activities proteasome and autophagy, formation neurotoxic aggregates. Here we examined whether perturbations these cellular pathways turn influence levels SOD1 potentially amplifying neurotoxicity. For study used fibroblasts, which express at physiological under regulation native promoter. cells were derived from ALS patients expressing 9 different mutants widely variable characteristics, as well carrying GGGGCC-repeat-expansion C9orf72 non-disease controls. A specific ELISA was to quantify soluble, SOD1, aggregated analysed western blotting. Misfolded detected all lines. Levels found be much lower control non-SOD1 This enabled us validate patient fibroblasts for use subsequent perturbation studies. Mitochondrial inhibition, stress or autophagy inhibition did affect soluble cases, detergent-resistant aggregates detected. However, led uniformly large increases cell lines an increase aggregation some. Thus ubiquitin-proteasome pathway principal determinant both controls decline activity with aging could one factors behind mid-to late-life onset inherited ALS.