Expression profiling analysis of NOX4 mutant mouse model
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE20400 | GEO | 2011/01/01
SECONDARY ACCESSION(S): PRJNA125533
REPOSITORIES: GEO
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Project description:NADPH oxidases are the only know enzyme family that has reactive oxygen species (ROS, e.g. superoxide and hydrogen peroxide) as their main metabolic product. They are therefore a prima candidate gene family to define a molecular source for ROS in physiology and pathophysiology. The type 4 NADPH oxidase (NOX4) is the most abundant isoform with the highest basal expression levels.The Nox4 knockout mouse was designed using a cre/lox recombination technique which allows to create either constitutive (type III recombination) and conditional (type I recombination) knockout alleles. cDNA microarray experiments of brain, kidney, muscle and pancreas of four male mutant mice versus a RNA pool of four male wild type mice. For each animal two technical replicates were performed including a dye swap experiment. All animals with the same age of 17 weeks on a C57BL/6 background.
2011-01-01 | E-GEOD-20400 | biostudies-arrayexpress
Project description:Hypoxic stress is a feature of rapidly growing thyroid tumors. Cancer progression is thought to be driven by reactive oxygen species (ROS) induced hypoxia adaptation. NADPH oxidases (NOXs), which produce ROS as their primary and sole function, has become of particular interest in thyroid malignancy. NOX4 was demonstrated to be upregulated in papillary thyroid cancers, functioning as a mitochondrial energetic sensor to modulate ATP levels, mediating overproduction of ROS induced by IL-δ, inversely correlating to thyroid differentiation. In this study, we analyzed hypoxia-treated BCPAP cells transfected with siRNA against NOX4. Results provides insight into the role of NOX4 in hypoxia adaptation.
2020-10-31 | GSE160491 | GEO
Project description:Within the family of NADPH oxidases, Nox4 is unique as it is predominantly localized in the endoplasmic reticulum, has constitutive activity and generates H2O2. We hypothesize that these features are consequences of a so far unidentified Nox4-interacting protein. Interacting proteins were screened by quantitative SILAC-Co-immunoprecipitation in HEK293 cells stably overexpressing Nox4. By this technique, several interacting proteins were identified with calnexin showing the most robust interaction.
2016-02-17 | PXD003514 | Pride
Project description:Tightly regulated and cell-specific NADPH-oxidases (Nox) represent one of the major sources of reactive oxygen species (ROS) signaling molecules that are involved in tissue development and stem cell self-renewal. We have characterized the role of Nox4 in osteo-progenitors during postnatal bone development. Nox4 expression in bone and ROS generation were increased during early osteoblast differentiation and bone development. Stromal osteoblastic cell self-renewal, proliferation and ROS production were significantly lower in samples from whole body Nox4 knockout mice (Nox4-/-) and conditional knockout (CKO) mice with depletion of Nox4 in the limb bud mesenchyme compared with those from control mice (Nox4fl/fl), but they were reversed after 9 passages. In both sexes, bone volume, trabecular number and bone mineral density were significantly lower in 3-week old CKO and Nox4-/- mice compared with Nox4fl/fl controls. This was reflected in serum levels of bone formation markers alkaline phosphatase (ALP) and procollagen 1 intact N-terminal propeptide (P1NP). However, under-developed bone formation in 3-week old CKO and Nox4-/- mice quickly caught up to levels of control mice by 6-week of age, remained no different at 13-week of age, and was reversed in 32-week old male mice. Osteoclastogenesis showed no differences among groups, however, CTX1 reflecting osteoclast activity was significantly higher in 3-week old male CKO and Nox4-/- mice compared with control mice, and significantly lower in 32-week old Nox4-/- mice compared with control mice. These data suggest that Nox4 expression and ROS signaling in bone and osteoblastic cells coordinately play a role on osteoblast differentiation, proliferation and maturation.
2022-06-22 | GSE195454 | GEO
Project description:Within the family of NADPH oxidases, Nox4 is unique as it is predominantly localized in the endoplasmic reticulum, has constitutive activity and generates H2O2. We hypothesize that these features are consequences of a so far unidentified Nox4-interacting protein. Interacting proteins were screened by quantitative SILAC-Co-immunoprecipitation in HEK293 cells stably overexpressing Nox4. By this technique, several interacting proteins were identified with calnexin showing the most robust interaction.
2017-03-29 | MSV000080772 | MassIVE
Project description:Within the family of NADPH oxidases, Nox4 is unique as it is predominantly localized in the endoplasmic reticulum, has constitutive activity and generates H2O2. We hypothesize that these features are consequences of a so far unidentified Nox4-interacting protein. Blue native gel electrophoresis of solubilized membranes from HEK293 cells (overexpressing Nox4) separated macromolecular complexes containing Nox4. The combination of native gel electrophoresis and quantitative mass spectrometry was applied to identify proteins that co-migrate with Nox4. In addition, the data set contains information about macromolecular protein complexes in human cellular membranes that are stable to the solubilization with digitonin.
2017-04-07 | PXD003509 | Pride
Project description:Differentiation of fibroblasts to myofibroblasts is governed by the transforming growth factor beta (TGF-β) through a mechanism involving redox signaling and generation of reactive oxygen species (ROS). Myofibroblasts synthesize proteins of the extracellular matrix and display a contractile phenotype. Myofibroblasts are predominant contributors of wound healing and several pathological states, including fibrotic diseases and cancer. Inhibition of the ROS-generating enzyme NADPH oxidase 4 (NOX4) has been proposed to mitigate fibroblast to myofibroblast differentiation and to offer a therapeutic option for the treatment of fibrotic diseases. In this study, we addressed the role of NOX4 in physiological wound healing and in TGF-β-induced myofibroblast differentiation. We explored the phenotypic changes induced by TGF-β in primary skin fibroblasts isolated from Nox4-deficient mice by immunofluorescence, Western blotting and RNA sequencing. Mice deficient for Cyba, the gene coding for p22phox, a key subunit of NOX4 were used for confirmatory experiments as well as human primary skin fibroblasts. In vivo, the wound healing was similar in wild-type and Nox4-deficient mice. In vitro, despite a strong upregulation following TGF-β treatment, Nox4 did not influence skin myofibroblast differentiation. Nevertheless, up-regulation of the mitochondrial protein Ucp2 and the stress-response protein Hddc3, as well as down-regulation of Islr were observed in Nox4-deficient fibroblasts. Altogether, we provide extensive evidence challenging a profibrotic role of NOX4 in skin fibroblasts and show that Nox4 regulates Ucp2 and Hddc3 expression, suggesting the presence of a so far undescribed redox crosstalk between NOX4 and redox homeostasis in fibroblasts.
2023-03-15 | GSE197562 | GEO
Project description:The regulation of protein function by reversible oxidation is increasingly recognized as a key mechanism for the control of cellular signalling, modulating crucial biological processes such cell differentiation. In this scenario, the NADPH oxidases must occupy a prominent position. Our results show that haematopoietic stem and progenitor cells (HSPCs) express three p22phox-dependent NADPH oxidases members (Nox1, Nox2 and Nox4). By deleting the p22phox coding gene (Cyba), here we have analysed the importance of this family of enzymes during in vivo haematopoiesis. Cyba-/- mice show a myeloid bias in detriment of B-cells differentiation, and an enrichment of HSPCs populations. By means of haematopoietic transplant experiments we have also tried to dissect the specific role of the NADPH oxidases. While the absence of Nox1 or Nox2 provides an advantage of reconstitution, the lack of Nox4 rendered the opposite result, what suggests a functional specificity among the different NADPH oxidases. Regarding signalling pathways, the activation of AKT and STAT5 is hampered in Cyba-/- cells, the later due to the downregulation of STAT5 protein. This is in line with the diminished response to IL-7 shown by our results, what could explain defective B-cells development observed in Cyba-/- mice, supporting the requirement of NADPH oxidase activity for terminal B-cell differentiation.
2020-01-29 | GSE131725 | GEO
Project description:PDAC cells acquire metabolic changes that augment NADPH production and cytosolic redox homeostasis. Here we show that high NADPH levels drive activity of NADPH oxidase 4 (NOX4) expressed in the endoplasmic reticulum (ER) membrane. NOX4 produces H2O2 metabolized by Peroxiredoxin 4 (PRDX4) in the ER lumen. Using functional genomics and subsequent in vitro and in vivo validations, we find that PDAC cell lines with high NADPH levels are dependent on PRDX4 for their growth and survival. PRDX4 addiction is associated with increased reactive oxygen species, a DNA-PKcs-governed DNA damage response and radiosensitivity, which can be rescued by depletion of NOX4 or NADPH. As such, this study has identified NOX4 as a protein which paradoxically converts the reducing power of the cytosol to an ER-specific oxidative stress vulnerability in PDAC that may be therapeutically exploited by targeting PRDX4.
2021-05-25 | GSE166165 | GEO
Project description:Reactive oxygen species (ROS) play important roles as second messengers in a wide array of cellular processes including differentiation of stem cells. We identified Nox4 as the major ROS generating enzyme whose expression is induced during differentiation of embryoid body (EB) into cells of all three germ layers. The role of Nox4 was examined using iPS cells generated from Nox4 -/- mouse. Differentiation markers showed significantly reduced expression levels consistent with the importance of Nox4-generated ROS during this process. From transcriptomic analyses, we found insulin-like growth factor 2 (IGF2), a member of a gene family extensively involved in embryonic development, as one of the most down-regulated genes. Indeed, addition of IGF2 to culture partly restored the differentiation efficiency of Nox4-/- iPS cells. Our results reveal an important signaling axis mediated by ROS in control of crucial events during differentiation of pluripotent stem cells.
2021-07-28 | GSE180912 | GEO