Contributions
Abstract: EP1508
Type: ePoster
Abstract Category: Pathology and pathogenesis of MS - 19 Neurodegeneration
Background: Multiple sclerosis (MS) is a chronic autoimmune condition of the central nervous system associated with demyelination and neurological impairment. Autophagy is a mechanism of intracellular lysosomal degradation of dysfunctional cytoplasmic components. It plays a critical role in the functioning of the immune system and failures in autophagic processes have been implicated in neurodegeneration. As such, dysregulation of autophagy may play a role in the pathophysiology of MS. Recent evidence suggests that Simvastatin (a HMG-CoA reductase inhibitor) induces autophagy in a variety of mammalian cells, and there is growing interest in the therapeutic potential of statins in the treatment of MS. Following promising results from a phase 2 trial, a phase 3 trial of Simvastatin treatment in secondary progressive MS is due to be conducted.
Objective: To evaluate the autophagy-modulating effects of Simvastatin in a variety of cell types, using flow cytometry.
Methods: LC3B immunofluorescence labelling was used to measure autophagy in Simvastatin-treated K562 (human erythroleukaemia cells) and Jurkat cells (human acute T cell leukaemia), as well as PC12 cells (rat neuroblastic cells). PC12 cells resemble neuronal cells when differentiated with nerve growth factor. Fluorescent labelling of organelles was used to analyse organelle-specific autophagic processes in mitochondria and lysosomes, and a Proteostat assay detected protein aggregation in PC12 cells. Two time points were analysed: 24 and 48 hours.
Results: Simvastatin induced autophagy in a dose-dependent manner in Jurkat (at 24h; p< 0.05) and K562 cells (at 48h; p< 0.05) treated at the following concentrations: 0µM, 0.1µM, 1µM and 10µM. The greatest upregulation of LC3B expression was seen at 1µM Simvastatin in both Jurkat (p=0.032) and K562 (p=0.0432) cells. Simvastatin-treatment also slightly upregulated mitochondrial mass in Jurkat and K562 cells; increased lysosome expression in K562 cells, and upregulated LC3B and Proteostat expression in PC12 cells at 48 hours.
Conclusion: These preliminary findings suggest that Simvastatin influences autophagy, and this could be one mechanism by which Simvastatin may have therapeutic potential in the treatment of neurodegenerative diseases, such as MS. Further investigation is merited.
Disclosure:
E James: Nothing to disclose
P Dua: Nothing to disclose
G Warnes: Nothing to disclose
UC Meier: Nothing to disclose
Abstract: EP1508
Type: ePoster
Abstract Category: Pathology and pathogenesis of MS - 19 Neurodegeneration
Background: Multiple sclerosis (MS) is a chronic autoimmune condition of the central nervous system associated with demyelination and neurological impairment. Autophagy is a mechanism of intracellular lysosomal degradation of dysfunctional cytoplasmic components. It plays a critical role in the functioning of the immune system and failures in autophagic processes have been implicated in neurodegeneration. As such, dysregulation of autophagy may play a role in the pathophysiology of MS. Recent evidence suggests that Simvastatin (a HMG-CoA reductase inhibitor) induces autophagy in a variety of mammalian cells, and there is growing interest in the therapeutic potential of statins in the treatment of MS. Following promising results from a phase 2 trial, a phase 3 trial of Simvastatin treatment in secondary progressive MS is due to be conducted.
Objective: To evaluate the autophagy-modulating effects of Simvastatin in a variety of cell types, using flow cytometry.
Methods: LC3B immunofluorescence labelling was used to measure autophagy in Simvastatin-treated K562 (human erythroleukaemia cells) and Jurkat cells (human acute T cell leukaemia), as well as PC12 cells (rat neuroblastic cells). PC12 cells resemble neuronal cells when differentiated with nerve growth factor. Fluorescent labelling of organelles was used to analyse organelle-specific autophagic processes in mitochondria and lysosomes, and a Proteostat assay detected protein aggregation in PC12 cells. Two time points were analysed: 24 and 48 hours.
Results: Simvastatin induced autophagy in a dose-dependent manner in Jurkat (at 24h; p< 0.05) and K562 cells (at 48h; p< 0.05) treated at the following concentrations: 0µM, 0.1µM, 1µM and 10µM. The greatest upregulation of LC3B expression was seen at 1µM Simvastatin in both Jurkat (p=0.032) and K562 (p=0.0432) cells. Simvastatin-treatment also slightly upregulated mitochondrial mass in Jurkat and K562 cells; increased lysosome expression in K562 cells, and upregulated LC3B and Proteostat expression in PC12 cells at 48 hours.
Conclusion: These preliminary findings suggest that Simvastatin influences autophagy, and this could be one mechanism by which Simvastatin may have therapeutic potential in the treatment of neurodegenerative diseases, such as MS. Further investigation is merited.
Disclosure:
E James: Nothing to disclose
P Dua: Nothing to disclose
G Warnes: Nothing to disclose
UC Meier: Nothing to disclose