bims-noxint 2020-01-12 papers

bims-noxint

Biomed News

on NADPH oxidases in tumorigenesis

Issue of 2020–01–12
seven papers selected by
Laia Caja Puigsubira, Uppsala University

Exogenous 8-hydroxydeoxyguanosine ameliorates liver fibrosis through the inhibition of Rac1-NADPH oxidase signaling.
Cyba-deficient mice display an increase in hematopoietic stem cells and an overproduction of immunoglobulins.
Maximum reproductive lifespan correlates with CD33rSIGLEC gene number: Implications for NADPH oxidase-derived reactive oxygen species in aging.
Cardioprotective effects of α-cardiac actin on oxidative stress in a dilated cardiomyopathy mouse model.
KRIT1 loss-mediated upregulation of NOX1 in stromal cells promotes paracrine pro-angiogenic responses.
Enhancer of zeste homolog 2 modulates oxidative stress-mediated pyroptosis in vitro and in a mouse kidney ischemia-reperfusion injury model.
Prevention of Connexin43 remodeling protects against duchenne muscular dystrophy cardiomyopathy.

J Gastroenterol Hepatol. 2020 Jan 06.

Exogenous 8-hydroxydeoxyguanosine ameliorates liver fibrosis through the inhibition of Rac1-NADPH oxidase signaling.

Seung Kak Shin, Kyung-Ok Kim, Se-Hee Kim, Oh Sang Kwon, Cheol Soo Choi, Sung Hwan Jeong, Yun Soo Kim, Ju Hyun Kim, Myung-Hee Chung.

   BACKGROUND AND AIM: Exogenous 8-hydroxydeoxyguanosine (8-OHdG) was suggested as an inhibitor of Rac1 and NADPH oxidase (NOX). The aim of this study was to evaluate the effects of the exogenous 8-OHdG on hepatic fibrogenesis in vitro and in vivo model of liver fibrosis.
METHODS: Adult Sprague-Dawley rats were allocated to sham-operated rats (n=7), rats underwent bile duct ligation (BDL) (n=6), and BDL rats treated with 8-OHdG (60 mg/kg/day by gavage, n=6). All rats were sacrificed on day 21. Double immunofluorescence staining between either NOX1 or NOX2 and α-smooth muscle actin (SMA) in liver was performed. Hepatic fibrotic contents were assessed by hydroxyproline assay and quantified by Sirius red staining. In vitro, hepatic stellate cell (HSC) line LX-2 and HHSteC cells were stimulated by angiotensin-II (10μM). The ROS production was measured by confocal microsopy. The expressions of NOX1, NOX2, α-SMA, transforming growth factor (TGF)-β1, and collagen Iα were analyzed by quantitative real-time polymerase chain reaction or immunoblotting.
RESULTS: The 8-OHdG treatment in BDL rats reduced the NOX1 and NOX2 protein expression which overlapped with α-SMA compared to BDL rats. The 8-OHdG treatment in BDL rats significantly decreased the mRNA expression of NOX1, NOX2, α-SMA, TGF-β1, and collagen Iα, and fibrotic contents. Increases of reactive oxygen species (ROS) production, Rac1 activation, NOX1, NOX2, and fibronectin expression induced by angiotensin II in HSCs were attenuated by 8-OHdG.
CONCLUSIONS: Rac1 activation and NOX-derived ROS are implicated to liver fibrosis. The 8-OHdG ameliorates liver fibrosis through the inhibition of Rac1 activation and NOX-derived ROS.

Keywords:  8-hydroxydeoxyguanosine; NADPH oxidase; Rac1; liver fibrosis

DOI:  https://doi.org/10.1111/jgh.14979
Haematologica. 2020 Jan 09. pii: haematol.2019.233064. [Epub ahead of print]

Cyba-deficient mice display an increase in hematopoietic stem cells and an overproduction of immunoglobulins.

Rodrigo Prieto-Bermejo, Marta Romo-González, Alejandro Pérez-Fernández, Ignacio García-Tuñón, Manuel Sánchez-Martín, Ángel Hernández-Hernández.

  The regulation of protein function by reversible oxidation is increasingly recognized as a key mechanism for the control of cellular signaling, modulating crucial biological processes such as cell differentiation. In this scenario, NADPH oxidases must occupy a prominent position. Our results show that hematopoietic stem and progenitor cells express three p22phox-dependent NADPH oxidases members (NOX1, NOX2 and NOX4). By deleting the p22phox coding gene (Cyba), here we have analyzed the importance of this family of enzymes during in vivo hematopoiesis. Cyba-/- mice show a myeloid bias, and an enrichment of hematopoietic stem cell populations. By means of hematopoietic transplant experiments we have also tried to dissect the specific role of the NADPH oxidases. While the absence of NOX1 or NOX2 provides a higher level of reconstitution, a lack of NOX4 rendered the opposite result, suggesting a functional specificity among the different NADPH oxidases. Cyba-/- cells showed a hampered activation of AKT1 and a sharp decrease in STAT5 protein. This is in line with the diminished response to IL-7 shown by our results, which could explain the overproduction of immunoglobulins observed in Cyba-/- mice.

Keywords:  Cyba; Granulocytes, Monocytes, Macrophages; Hematopoiesis; Hematopoietic Stem Cell; NADPH oxidase

DOI:  https://doi.org/10.3324/haematol.2019.233064
FASEB J. 2019 Dec 24.

Maximum reproductive lifespan correlates with CD33rSIGLEC gene number: Implications for NADPH oxidase-derived reactive oxygen species in aging.

Naazneen Khan, Stuart K Kim, Pascal Gagneux, Laura L Dugan, Ajit Varki.

  Humans and orcas are among the very rare species that have a prolonged post-reproductive lifespan (PRLS), during which the aging process continues. Reactive oxygen species (ROS) derived from mitochondria and from the NADPH oxidase (NOX) enzymes of innate immune cells are known to contribute to aging, with the former thought to be dominant. CD33-related-Siglecs are immune receptors that recognize self-associated-molecular-patterns and modulate NOX-derived-ROS. We herewith demonstrate a strong correlation of lifespan with CD33rSIGLEC gene number in 26 species, independent of body weight or phylogeny. The correlation is stronger when considering total CD33rSIGLEC gene number rather than those encoding inhibitory and activating subsets, suggesting that lifetime balancing of ROS is important. Combining independent lines of evidence including the short half-life and spontaneous activation of neutrophils, we calculate that even without inter-current inflammation, a major source of lifetime ROS exposure may actually be neutrophil NOX-derived. However, genomes of human supercentenarians (>110 years) do not harbor a significantly higher number of functional CD33rSIGLEC genes. Instead, lifespan correlation with CD33rSIGLEC gene number was markedly strengthened by excluding the post-reproductive lifespan of humans and orcas (R2 = 0.83; P < .0001). Thus, CD33rSIGLEC modulation of ROS likely contributes to maximum reproductive lifespan, but other unknown mechanisms could be important to PRLS.

Keywords:   CD33rSIGLEC ; NADPH-oxidase; prolonged post-reproductive lifespan; reactive oxygen species

DOI:  https://doi.org/10.1096/fj.201902116R
FASEB J. 2019 Dec 29.

Cardioprotective effects of α-cardiac actin on oxidative stress in a dilated cardiomyopathy mouse model.

Aude Angelini, Mark-Alexander Gorey, Florent Dumont, Nathalie Mougenot, Maria Chatzifrangkeskou, Antoine Muchir, Zhenlin Li, Mathias Mericskay, Jean-Francois Decaux.

  The expression of α-cardiac actin, a major constituent of the cytoskeleton of cardiomyocytes, is dramatically decreased in a mouse model of dilated cardiomyopathy triggered by inducible cardiac-specific serum response factor (Srf) gene disruption that could mimic some forms of human dilated cardiomyopathy. To investigate the consequences of the maintenance of α-cardiac actin expression in this model, we developed a new transgenic mouse based on Cre/LoxP strategy, allowing together the induction of SRF loss and a compensatory expression of α-cardiac actin. Here, we report that maintenance of α-cardiac actin within cardiomyocytes temporally preserved cytoarchitecture from adverse cardiac remodeling through a positive impact on both structural and transcriptional levels. These protective effects were accompanied in vivo by the decrease of ROS generation and protein carbonylation and the downregulation of NADPH oxidases NOX2 and NOX4. We also show that ectopic expression of α-cardiac actin protects HEK293 cells against oxidative stress induced by H2 O2 . Oxidative stress plays an important role in the development of cardiac remodeling and contributes also to the pathogenesis of heart failure. Taken together, these findings indicate that α-cardiac actin could be involved in the regulation of oxidative stress that is a leading cause of adverse remodeling during dilated cardiomyopathy development.

Keywords:  cardiac remodeling; dilated cardiomyopathy; oxidative stress; α-cardiac actin

DOI:  https://doi.org/10.1096/fj.201902389R
Cell Signal. 2020 Jan 06. pii: S0898-6568(20)30004-8. [Epub ahead of print] 109527

KRIT1 loss-mediated upregulation of NOX1 in stromal cells promotes paracrine pro-angiogenic responses.

Federica Finetti, Irene Schiavo, Jasmine Ercoli, Enrica Boda, Saverio Francesco Retta, Lorenza Trabalzini.

  Cerebral cavernous malformation (CCM) is a cerebrovascular disorder of proven genetic origin characterized by abnormally dilated and leaky capillaries occurring mainly in the central nervous system, with a prevalence of 0.3-0.5% in the general population. Genetic studies have identified causative mutations in three genes, CCM1/KRIT1, CCM2 and CCM3, which are involved in the maintenance of vascular homeostasis. However, distinct studies in animal models have clearly shown that CCM gene mutations alone are not sufficient to cause CCM disease, but require additional contributing factors, including stochastic events of increased oxidative stress and inflammation. Consistently, previous studies have shown that up-regulation of NADPH oxidase-mediated production of reactive oxygen species (ROS) in KRIT1 deficient endothelium contributes to the loss of microvessel barrier function. In this study, we demonstrate that KRIT1 loss-of-function in stromal cells, such as fibroblasts, causes the up-regulation of NADPH oxidase isoform 1 (NOX1) and the activation of inflammatory pathways, which in turn promote an enhanced production of proangiogenic factors, including vascular endothelial growth factor (VEGF) and prostaglandin E2 (PGE2). Furthermore and importantly, we show that conditioned media from KRIT1 null fibroblasts induce proliferation, migration, matrix metalloproteinase 2 (MMP2) activation and VE-cadherin redistribution in wild type human endothelial cells. Taken together, our results demonstrate that KRIT1 loss-of-function in stromal cells affects the surrounding microenvironment through a NOX1-mediated induction and release of angiogenic factors that are able to promote paracrine proangiogenic responses in human endothelial cells, thus pointing to a novel role for endothelial cell-nonautonomous effects of KRIT1 mutations in CCM pathogenesis, and opening new perspectives for disease prevention and treatment.

Keywords:  CCM; COX-2; Cerebrovascular disease; KRIT1; NOX1; PGE2

DOI:  https://doi.org/10.1016/j.cellsig.2020.109527
FASEB J. 2020 Jan;34(1): 835-852

Enhancer of zeste homolog 2 modulates oxidative stress-mediated pyroptosis in vitro and in a mouse kidney ischemia-reperfusion injury model.

Hao Liu, Zhiyuan Chen, Xiaodong Weng, Hui Chen, Yang Du, Changhui Diao, Xiuheng Liu, Lei Wang.

  Enhancer of zeste homolog 2 (EZH2), a well-known methyltransferase, mediates histone H3 lysine 27 trimethylation (H3K27me3) and plays a crucial role in several kidney disease models. However, its role in renal ischemia/reperfusion (I/R) injury still remains unclear. In this study, we found that EZH2 was positively related to renal I/R injury and inhibition of EZH2 with DZNeP alleviated I/R injury and blocked the activation of oxidative stress and pyroptosis in vivo. Similarly, inhibition of EZH2 with either DZNeP or si-RNA also exerted an inhibitory effect on hypoxia/reoxygenation (H/R)-induced oxidative stress and pyroptosis in vitro. Moreover, further study revealed that ablation of reactive oxygen species (ROS) with N-acetyl-cysteine (NAC) suppressed pyroptosis in human renal proximal tubular epithelial cell line cells exposed to H/R stimulation. Furthermore, Nox4, which was positively related to the generation of ROS, was upregulated during H/R process, while it could be reversed by EZH2 inhibition. Consistently, Nox4-mediated ROS generation was attenuated upon inhibition of EZH2 with DZNeP or si-RNA. Additionally, the transcriptional activity of Nox4 was enhanced by the activation of ALK5/Smad2/3 signaling pathway, which was abolished by ALK5 knockdown in vitro. Finally, EZH2 inhibition blocked H/R and I/R-activated ALK5/Smad2/3 pathway and also resulted in an obvious decrease in the transcriptional activity and protein expression levels of Nox4. In conclusion, our results proved that EZH2 inhibition alleviated renal pyroptosis by blocking Nox4-dependent ROS generation through ALK5/Smad2/3 signaling pathway, indicating that EZH2 could be a potential therapeutic target for renal I/R injury.

Keywords:  Nox4; enhancer of zeste homolog 2; ischemia‐reperfusion injury; oxidative stress; pyroptosis; reactive oxygen species

DOI:  https://doi.org/10.1096/fj.201901816R
J Clin Invest. 2020 Jan 07. pii: 128190. [Epub ahead of print]

Prevention of Connexin43 remodeling protects against duchenne muscular dystrophy cardiomyopathy.

Eric Himelman, Mauricio A Lillo, Julie Nouet, J Patrick Gonzalez, Qingshi Zhao, Lai-Hua Xie, Hong Li, Tong Liu, Xander Ht Wehrens, Paul D Lampe, Glenn I Fishman, Natalia Shirokova, Jorge E Contreras, Diego Fraidenraich.

  Aberrant expression of the cardiac gap junction protein connexin-43 (Cx43) has been suggested to play a role in the development of cardiac disease in the mdx mouse model of Duchenne muscular dystrophy (DMD), however a mechanistic understanding of this association is lacking. Here, we identified a reduction of phosphorylation of Cx43 serines S325/S328/S330 in human and mouse DMD hearts. We hypothesized that hypo-phosphorylation of Cx43 serine-triplet triggers pathological Cx43 redistribution to the lateral sides of cardiomyocytes (remodeling). Therefore, we generated knock-in mdx mice in which the Cx43 serine-triplet was replaced with either phospho-mimicking glutamic acids (mdxS3E) or non-phosphorylatable alanines (mdxS3A). The mdxS3E but not mdxS3A mice were resistant to Cx43 remodeling with a corresponding reduction of Cx43 hemichannel activity. MdxS3E cardiomyocytes displayed improved intracellular Ca2+ signaling and a reduction of NOX2/reactive oxygen species (ROS) production. Furthermore, mdxS3E mice were protected against inducible arrhythmias, related lethality and the development of cardiomyopathy. Inhibition of microtubule polymerization by colchicine reduced both NOX2/ROS and oxidized CaMKII, increased S325/S328/S330 phosphorylation and prevented Cx43 remodeling in mdx hearts. Together, these results demonstrate a mechanism of dystrophic Cx43-remodeling and suggest that targeting Cx43 may be a therapeutic strategy to prevent heart dysfunction and arrhythmias in DMD patients.

Keywords:  Cardiology; Cardiovascular disease; Cell Biology; Neuromuscular disease

DOI:  https://doi.org/10.1172/JCI128190