Header

UZH-Logo

Maintenance Infos

Evaluation of NADPH oxidases as drug targets in a mouse model of familial amyotrophic lateral sclerosis


Seredenina, Tamara; Nayernia, Zeynab; Sorce, Silvia; Maghzal, Ghassan J; Filippova, Aleksandra; Ling, Shuo-Chien; Basset, Olivier; Plastre, Olivier; Daali, Youssef; Rushing, Elisabeth J; Giordana, Maria T; Cleveland, Don W; Aguzzi, Adriano; Stocker, Roland; Krause, Karl-Heinz; Jaquet, Vincent (2016). Evaluation of NADPH oxidases as drug targets in a mouse model of familial amyotrophic lateral sclerosis. Free Radical Biology & Medicine, 97:95-108.

Abstract

Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease characterized by progressive loss of motor neurons, gliosis, neuroinflammation and oxidative stress. The aim of this study was to evaluate the involvement of NADPH oxidases (NOX) in the oxidative damage and progression of ALS neuropathology. We examined the pattern of NOX expression in spinal cords of patients and mouse models of ALS and analyzed the impact of genetic deletion of the NOX1 and 2 isoforms as well as pharmacological NOX inhibition in the SOD1G93A ALS mouse model.

A substantial (10–60 times) increase of NOX2 expression was detected in three etiologically different ALS mouse models while up-regulation of some other NOX isoforms was model-specific. In human spinal cord samples, high NOX2 expression was detected in microglia. In contrast to previous publications, survival of SOD1G93A mice was not modified upon breeding with constitutive NOX1 and NOX2 deficient mice. As genetic deficiency of a single NOX isoform is not necessarily predictive of a pharmacological intervention, we treated SOD1G93A mice with broad-spectrum NOX inhibitors perphenazine and thioridazine. Both compounds reached in vivo CNS concentrations compatible with NOX inhibition and thioridazine significantly decreased superoxide levels in the spinal cord of SOD1G93A mice in vivo. Yet, neither perphenazine nor thioridazine prolonged survival. Thioridazine, but not perphenazine, dampened the increase of microglia markers in SOD1G93A mice. Thioridazine induced an immediate and temporary enhancement of motor performance (rotarod) but its precise mode of action needs further investigation. Additional studies using specific NOX inhibitors will provide further evidence on the relevance of NOX as drug targets for ALS and other neurodegenerative disorders.

List of abbreviations
ALS, amyotrophic lateral sclerosis; BBB, blood brain barrier; 2-Cl-E+, 2-chloroethidium; CNS, central nervous system; DMSO, Dimethylsulfoxide; DPI, diphenyleneiodonium; DMEM, Dulbecco's modified Eagle Medium; DNA, deoxyribonucleic acid; DUOX, dual oxidase; E+, ethidium; EC50, half maximal efficient concentration; FBS, fetal bovine serum; FUS/TLS, Fused in Sarcoma/Translocated in Sarcoma; GM-CSF, granulocyte-macrophage colony-stimulating factor; HBSS, Hank's buffered salt solution; HE, hydroethidine; HOCl, hypochlorous acid; H2O2, hydrogen peroxide; 2-OH-E+, 2-hydroxyethidium; LC/MS/MS, Liquid chromatography-tandem mass spectrometry; MPO, myeloperoxidase; NADPH, nicotinamide adenine dinucleotide phosphate; NOX, NADPH oxidase; NOXO1, NADPH oxidase organiser type 1; NC, non-carrier; PBS, phosphate buffered saline; PMA, phorbol myristate acetate; qPCR, quantitative polymerase chain reaction; ROS, reactive oxygen species; O2∙−, superoxide radical anion; SOD, superoxide dismutase; TDP 43, TAR DNA binding protein 43; WST-1, 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt; WT, wild type

Abstract

Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease characterized by progressive loss of motor neurons, gliosis, neuroinflammation and oxidative stress. The aim of this study was to evaluate the involvement of NADPH oxidases (NOX) in the oxidative damage and progression of ALS neuropathology. We examined the pattern of NOX expression in spinal cords of patients and mouse models of ALS and analyzed the impact of genetic deletion of the NOX1 and 2 isoforms as well as pharmacological NOX inhibition in the SOD1G93A ALS mouse model.

A substantial (10–60 times) increase of NOX2 expression was detected in three etiologically different ALS mouse models while up-regulation of some other NOX isoforms was model-specific. In human spinal cord samples, high NOX2 expression was detected in microglia. In contrast to previous publications, survival of SOD1G93A mice was not modified upon breeding with constitutive NOX1 and NOX2 deficient mice. As genetic deficiency of a single NOX isoform is not necessarily predictive of a pharmacological intervention, we treated SOD1G93A mice with broad-spectrum NOX inhibitors perphenazine and thioridazine. Both compounds reached in vivo CNS concentrations compatible with NOX inhibition and thioridazine significantly decreased superoxide levels in the spinal cord of SOD1G93A mice in vivo. Yet, neither perphenazine nor thioridazine prolonged survival. Thioridazine, but not perphenazine, dampened the increase of microglia markers in SOD1G93A mice. Thioridazine induced an immediate and temporary enhancement of motor performance (rotarod) but its precise mode of action needs further investigation. Additional studies using specific NOX inhibitors will provide further evidence on the relevance of NOX as drug targets for ALS and other neurodegenerative disorders.

List of abbreviations
ALS, amyotrophic lateral sclerosis; BBB, blood brain barrier; 2-Cl-E+, 2-chloroethidium; CNS, central nervous system; DMSO, Dimethylsulfoxide; DPI, diphenyleneiodonium; DMEM, Dulbecco's modified Eagle Medium; DNA, deoxyribonucleic acid; DUOX, dual oxidase; E+, ethidium; EC50, half maximal efficient concentration; FBS, fetal bovine serum; FUS/TLS, Fused in Sarcoma/Translocated in Sarcoma; GM-CSF, granulocyte-macrophage colony-stimulating factor; HBSS, Hank's buffered salt solution; HE, hydroethidine; HOCl, hypochlorous acid; H2O2, hydrogen peroxide; 2-OH-E+, 2-hydroxyethidium; LC/MS/MS, Liquid chromatography-tandem mass spectrometry; MPO, myeloperoxidase; NADPH, nicotinamide adenine dinucleotide phosphate; NOX, NADPH oxidase; NOXO1, NADPH oxidase organiser type 1; NC, non-carrier; PBS, phosphate buffered saline; PMA, phorbol myristate acetate; qPCR, quantitative polymerase chain reaction; ROS, reactive oxygen species; O2∙−, superoxide radical anion; SOD, superoxide dismutase; TDP 43, TAR DNA binding protein 43; WST-1, 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt; WT, wild type

Statistics

Citations

5 citations in Web of Science®
4 citations in Scopus®
Google Scholar™

Altmetrics

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > University Hospital Zurich > Institute of Neuropathology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Uncontrolled Keywords:NADPH oxidase; NOX; Amyotrophic lateral sclerosis; Microglia; Phenothiazine; Perphenazine; Thioridazine; SOD1G93A mice
Date:2016
Deposited On:07 Jun 2016 08:49
Last Modified:08 Dec 2017 19:37
Publisher:Elsevier
ISSN:0891-5849
Publisher DOI:https://doi.org/10.1016/j.freeradbiomed.2016.05.016
PubMed ID:27212019

Download

Full text not available from this repository.
View at publisher