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
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
Externí odkaz: