Neuroserpin polymers cause oxidative stress in a neuronal model of the dementia FENIB
Autor: | Claudia Moriconi, Valerio Licursi, Giuseppe Lupo, Emanuela D'Acunto, Emanuele Cacci, Elena Miranda, Nicoletta Carucci, Noemi Antonella Guadagno, Rodolfo Negri, Antonella De Jaco, Paola S. Nisi |
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Jazyk: | angličtina |
Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Polymers Epilepsies Myoclonic Protein conformational disease MTT 3-(4 5-dimethylthiazol-2-yl)-2 5-diphenyltetrazolium bromide Mice FENIB familial encephalopathy with neuroserpin inclusion bodies GST glutathione transferase Cells Cultured ApoE apolipoprotein E Neurons Cerebral Cortex wild type WT GPx glutathione peroxidase Neurodegeneration HRP horseradish peroxidase Prdx6 peroxiredoxin 6 3. Good health Scara scavenger receptor class A Neurology Heredodegenerative Disorders Nervous System Intracellular Familial encephalopathy with neuroserpin inclusion bodies NS neuroserpin Cell Survival Neuroserpin Mice Transgenic Biology Serpin Article lcsh:RC321-571 Neural progenitor cells Oxidative stress Serpin polymers ER endoplasmic reticulum 03 medical and health sciences SOD superoxide dismutase medicine unfolded protein response UPR Animals Humans A1AT alpha1 antitrypsin lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry Serpins Endoplasmic reticulum Neuropeptides Wild type medicine.disease Molecular biology Oxidative Stress 030104 developmental biology NFκB nuclear factor κB Unfolded protein response Aldh aldehyde dehydrogenase Dementia |
Zdroj: | Neurobiology of Disease Neurobiology of Disease, Vol 103, Iss, Pp 32-44 (2017) |
ISSN: | 1095-953X 0969-9961 |
Popis: | The serpinopathies are human pathologies caused by mutations that promote polymerisation and intracellular deposition of proteins of the serpin superfamily, leading to a poorly understood cell toxicity. The dementia FENIB is caused by polymerisation of the neuronal serpin neuroserpin (NS) within the endoplasmic reticulum (ER) of neurons. With the aim of understanding the toxicity due to intracellular accumulation of neuroserpin polymers, we have generated transgenic neural progenitor cell (NPC) cultures from mouse foetal cerebral cortex, stably expressing the control protein GFP (green fluorescent protein), or human wild type, G392E or delta NS. We have characterised these cell lines in the proliferative state and after differentiation to neurons. Our results show that G392E NS formed polymers that were mostly retained within the ER, while wild type NS was correctly secreted as a monomeric protein into the culture medium. Delta NS was absent at steady state due to its rapid degradation, but it was easily detected upon proteasomal block. Looking at their intracellular distribution, wild type NS was found in partial co-localisation with ER and Golgi markers, while G392E NS was localised within the ER only. Furthermore, polymers of NS were detected by ELISA and immunofluorescence in neurons expressing the mutant but not the wild type protein. We used control GFP and G392E NPCs differentiated to neurons to investigate which cellular pathways were modulated by intracellular polymers by performing RNA sequencing. We identified 747 genes with a significant upregulation (623) or downregulation (124) in G392E NS-expressing cells, and we focused our attention on several genes involved in the defence against oxidative stress that were up-regulated in cells expressing G392E NS (Aldh1b1, Apoe, Gpx1, Gstm1, Prdx6, Scara3, Sod2). Inhibition of intracellular anti-oxidants by specific pharmacological reagents uncovered the damaging effects of NS polymers. Our results support a role for oxidative stress in the cellular toxicity underlying the neurodegenerative dementia FENIB. Graphical abstract Image 1 Highlights • A cell model system for FENIB was generated using mouse neural progenitor cells. • Cells expressing polymerogenic neuroserpin upregulated several anti-oxidant genes. • Inhibition of the anti-oxidant defences led to apoptosis of these cells. • Oxidative stress and lipid oxidation may contribute to neurodegeneration in FENIB. |
Databáze: | OpenAIRE |
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