P47phox Research Articles

NETs biomarkers in saliva and serum OSCC patients: One hypothesis, two conclusions

PurposeThe actual role of neutrophils and neutrophil extracellular traps (NETs) in the course of cancer has not been clearly defined. The aim of this study was to determine the clinical usefulness of NETs biomarkers in saliva in confrontation with the blood serum and tumor tissue as a potential prognostic and therapeutic target in patients with oral squamous cell carcinoma (OSCC). Material and methodsExpression of myeloperoxidase (MPO), and histones H2A, H2B, H3 in the tumor tissue, was investigated using immunohistochemistry. The expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits: p47-phox, p67-phox (neutrophil cytosolic factor 2, NCF2) and panRac, as well as citrullinated histone H3 (CitH3) in peripheral blood neutrophil lysates, was assessed via Western blot. ELISA tests were employed to measure the concentrations of circulating free DNA (cfDNA) and MPO in saliva only, and NOX1, NCF2, DNASE1 in saliva and serum. ResultsExtracellular expression of MPO and histones was localized within tumor tissue. Significantly lower expression of p67-phox, panRac, and CitH3 was determined in OSCC patients. Considerably lower concentrations of NOX1, NCF2, and DNASE1 in the saliva samples of cancer patients were observed. However, the levels of NOX1, NCF2, and DNASE1 in the serum of patients with cancer were substantially higher. ConclusionsThe results obtained from the saliva of cancer patients suggest an impairment of the immunological homeostasis within the oral cavity related to NET formation, the causes of which should be sought in deficient activation of NADPH oxidase.

Ncf1 Governs Immune Niches in the Lung to Mediate Pulmonary Inflammation in Mice.

Neutrophil cytosolic factor 1 (Ncf1) is a major genetic factor associated with autoimmune diseases and has been identified as a key player in autoimmune mediated inflammation. We addressed the role of Ncf1 in an antigen-induced pulmonary inflammation model, and found that the Ncf1m1j mutation, causing a deficient reactive oxygen species response, alleviated disease. The Ncf1m1j mutation was associated with a reduced inflammatory cell infiltration in airways, but had limited effect on mucus secretion, antibody production and lung fibrosis. The disease remission in the Ncf1 mutated mice was reversed when functional Ncf1 was transgenically expressed in alveolar macrophages, suggesting that the cellular inflammation was depended on functional Ncf1 in alveolar macrophages. By determining cytokine and chemokine profiles in lung and serum, we found that Ncf1 deficiency allowed an increased expression of Th1 cytokines, including TNF-α, IFN-γ and IL-12. Since also epithelial cytokines were found to be regulated by Ncf1, we tested the effect of Ncf1 in IL-33 and IL-25 induced lung inflammation models. Mice with the Ncf1m1j mutation showed less sensitivity to IL-33, but not IL-25, induced lung inflammation, in a macrophage independent manner. The mice with deficient Ncf1 showed a reduced eosinophil infiltration and group 2 innate lymphoid cell (ILC2) activation. The production of IFN-γ in CD4+ T cells was increased, whereas IL-5 and IL-13 in ILC2 were decreased. Importantly, anti-IFN-γ antibody treatment of Ncf1 deficient mice increased eosinophil infiltration and rescued ILC2 activation in the lung. We conclude that Ncf1 deficiency enhances Th1 response, deactivates ILC2, and protects against pulmonitis.

NOX Subunit Gene Polymorphisms and the Risk of Chronic Artery Disease in Type 2 Diabetes Patients

BackgroundOxidative stress is a key feature of type 2 diabetes (T2D) and coronary artery disease (CAD). NADPH oxidase (NOX) is the main producer of reactive oxygen species and therefore genes encoding NOX subunits may contribute to the disease susceptibility. The present study investigated whether polymorphisms of genes encoding cytochrome a (CYBA), Rac family small GTPase 1 (RAC1) and neutrophil cytosolic factor 4 (NCF4) jointly contribute to the risk for CAD in T2D patients of Central Russia. MethodsA total of 1579 T2D patients (591 males and 988 females) including 448 patients with CAD (133 males and 315 females) were recruited for the study. Genotyping was performed by MALDI-TOF mass spectrometry with the use of MassArray Analyzer 4 platform (Agena Bioscience). ResultsA gender-stratified analysis showed that the genotypes rs10238136-A/T-T/T RAC1 (OR 1.96, 95 CI 1.08-3.58, P=0.029) and rs4821544-C/C NCF4 (OR 1.71, 95 CI 1.12-2.59, P=0.013) were associated with the increased risk of CAD exclusively in females. SNP rs rs4821544 was also associated with increased levels of triglycerides (P=0.03), whereas carriers of minor allele rs4821544-C of NCF4 gene had higher glycosylated hemoglobin level (P=0.032). In males, association of genotypes rs4673-G/A-A/A CYBA with susceptibility to CAD (OR 1.72, 95CI 1.09-2.70, P=0.018) and was established. SNP rs4673 CYBA was also associated with increased plasma hydrogen peroxide level (P=0.02). Conclusions: The study demonstrates that genes encoding NOX subunits (CYBA, RAC1, and NCF4) jointly contribute to the risk of CAD in T2D patients, and their effects on disease susceptibility are gender-specific.

NCF1/2/4 Are Prognostic Biomarkers Related to the Immune Infiltration of Kidney Renal Clear Cell Carcinoma

Neutrophil cytoplasmic factor 1/2/4 (NCF1/2/4) belongs to the NADPH oxidase complex, which is a cytoplasmic component, and its polymorphism is the main factor related to autoimmune diseases, which is probably caused by the regulation of peroxide. They also play a role in tumor growth and metastasis. This research is aimed at evaluating the biological function and prognostic role of NCF1, NCF2, and NCF4 genes in kidney renal clear cell carcinoma (KIRC) by using multiple online bioinformatics website, including Oncomine, GEPIA, UALCAN, Kaplan–Meier Plotter, TIMER, TISIDB, cBioPortal, LinkedOmics, GeneMANIA, and DAVID databases. The mRNA levels of NCFs were higher in KIRC tissues than in normal tissues. The overexpression of NCFs was significantly correlated with advanced pathological grades and individual cancer stages in KIRC. Meanwhile, the expressions of NCFs played an important role in the tumorigenesis and progression of KIRC. Prognostic value analysis suggested that high transcription levels of NCF1/4 were associated with poor overall survival in KIRC patients. In addition, results from the LinkedOmics database showed that the KEGG pathway related to NCFs mainly focused on immune activation and immune regulation function. NCF genetic alterations, including copy number amplification, missense mutation, and deep deletion, could be found through the cBioPortal database. Further, NCF expression was significantly correlated with infiltration levels of various immune cells as well as immune signatures. Protein-protein interaction network and enrichment analysis of NCF1/2/4 in KIRC showed that NCF coexpressed genes mainly associated with diverse immune marker sets showed significance. Overall, these results indicated that NCFs could be prognostic biomarkers as well as effective targets for diagnosis in KIRC.

Utility of Immunohistochemistry and Immunofluorescence in Determining the Pathogenic Variants of Chronic Granulomatous Disease.

Chronic granulomatous disease (CGD) is a primary immunodeficiency disorder of phagocytes due to defects in any of the five subunits of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. An initial diagnosis of CGD is made by flow cytometry-based dihydrorhodamine assay or nitro blue tetrazolium test, which is further confirmed by molecular assays. Expression of five subunits of NADPH oxidase components by either flow cytometric or western blot analysis provides clues toward the potential gene targets which are subsequently confirmed by various genetic assays. Immunohistochemistry (IHC) and immunofluorescence (IF) have never been earlier used to determine the expression of different subunits of NADPH oxidase system. We evaluated the utility of IHC and IF in determining the underlying pathogenic variants of CGD. Twelve genetically confirmed cases of CGD, comprising of biopsy specimens (n = 6), tissue blocks from autopsy cases (n = 3), and cellblocks of cell pellet prepared from peripheral blood (n = 4) were included. IHC for p67phox and p47phox subunits and IF for cytochrome b558 were performed. All 4 cases with pathogenic variation of NCF2 gene showed loss of expression for p67phox subunit. Two cases with pathogenic variation of NCF1 gene showed loss of expression for p47phox subunit. Five cases, except a single case with CYBB gene pathogenic variation, showed loss of expression for cytochrome b558 on IF. Thus, loss of expression consistently matched with the underlying genetic defects assessed by sequencing. Our results confirm our hypothesis that IHC and IF are two rapid, economical, pathologist-friendly techniques providing pertinent information regarding the underlying pathogenic variants and such immuno-analysis can be easily performed on the tissue.

The downregulation of NADPH oxidase Nox4 during hypoxia in hemangioendothelioma cells: a possible role of p22 phox on Nox4 protein stability

NADPH oxidase (Nox) 4 produces H2O2 by forming a heterodimer with p22 phox and is involved in hemangioendothelioma development through monocyte chemoattractant protein-1 (MCP-1) upregulation. Here, we show that Nox4 protein levels were maintained by p22 phox in hemangioendothelioma cells and Nox4 protein stability was dependent on p22 phox coexpression. Conversely, the degradation of Nox4 monomer was enhanced by p22 phox knockdown. Under hypoxic conditions in hemangioendothelioma cells, p22 phox was downregulated at the mRNA and protein levels. Downregulation of p22 phox protein resulted in the enhanced degradation of Nox4 protein in hypoxia-treated hemangioendothelioma cells. In contrast, Nox2, a Nox isoform, was not altered at the protein level under hypoxic conditions. Nox2 exhibited a higher affinity for p22 phox compared with Nox4, suggesting that when coexpressed with Nox4 in the same cells, Nox2 acts as a competitor. Nox2 knockdown restored Nox4 protein levels partially reduced by hypoxic treatment. Thus, Nox4 protein levels were attenuated in hypoxia-treated cells resulting from p22 phox depletion. MCP-1 secretion was decreased concurrently with hypoxia-induced Nox4 downregulation compared with that under normoxia.

Human SLE variant NCF1-R90H promotes kidney damage and murine lupus through enhanced Tfh2 responses induced by defective efferocytosis of macrophages

ObjectivesWe previously identified a hypomorphic variant, p.Arg90His (p.R90H) of neutrophil cytosolic factor 1 (NCF1, a regulatory subunit of phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 complex), as an putative...

Astragaloside IV prevents endothelial dysfunction by improving oxidative stress in streptozotocin‑induced diabetic mouse aortas

Oxidative stress serves a role in endothelial dysfunction exhibited by patients with diabetes mellitus. Astragaloside IV (AS-IV) is a major active ingredient of Radix Astragali, which is considered to exhibit vasoprotective effects through unknown mechanisms. Thus, the current study was performed to investigate the protective effects of AS-IV in streptozotocin (STZ)-induced endothelial dysfunction and to explore whether antioxidant mechanisms were involved. The protective effects of AS-IV on the endothelium-dependent relaxation and contraction of aortic rings were determined by isometric tension recordings. NADPH subunits and endothelial nitric oxide synthase (eNOS) expression was identified via western blotting. Superoxide dismutase and malondialdehyde levels were assayed using ELISA. Furthermore, the generation of reactive oxygen species (ROS) and nitric oxide (NO) was detected via dihydroethidium and 4,5-diaminofluorescein diacetate staining, respectively. The results revealed that STZ-injected mice exhibited increased aortic endothelium-dependent vasoconstriction and decreased vasorelaxation to acetylcholine. However, AS-IV treatment reversed these effects. NG-nitro-L-arginine was subsequently used to completely inhibit impaired relaxation. Accordingly, impaired NO generation was restored following AS-IV treatment by increasing eNOS phosphorylation levels. Furthermore, ROS formation was also depressed following AS-IV treatment compared with that in STZ-injected mice. AS-IV also decreased the expression of various NADPH subunits, including human neutrophil cytochrome b light chain, neutrophil cytosolic factor 1, NADPH oxidase (NOX)2, NOX4 and Rac-1. The results of the current study may provide novel evidence that diabetes-induced vascular injury arises from either the inhibition of eNOS or the activation of NOX-derived ROS generation. In addition, the results warrant further investigation into the application of AS-IV treatment, leading to the improvement of oxidative stress, in patients with diabetes exhibiting endothelial dysfunction.

Granuloma Formation in a Cyba-Deficient Model of Chronic Granulomatous Disease Is Associated with Myeloid Hyperplasia and the Exhaustion of B-Cell Lineage.

Haematopoiesis is a paradigm of cell differentiation because of the wide variety and overwhelming number of mature blood cells produced daily. Under stress conditions, the organism must adapt to a boosted demand for blood cells. Chronic granulomatous disease (CGD) is a genetic disease caused by inactivating mutations that affect the phagocyte oxidase. Besides a defective innate immune system, CGD patients suffer from recurrent hyper-inflammation episodes, circumstances upon which they must face emergency haematopoiesis. The targeting of Cybb and Ncf1 genes have produced CGD animal models that are a useful surrogate when studying the pathophysiology and treatment of this disease. Here, we show that Cyba−/− mice spontaneously develop granuloma and, therefore, constitute a CGD animal model to complement the existing Cybb−/− and Ncf1−/− models. More importantly, we have analysed haematopoiesis in granuloma-bearing Cyba−/− mice. These animals showed a significant loss of weight, developed remarkable splenomegaly, bone marrow myeloid hyperplasia, and signs of anaemia. Haematological analyses showed a sharped decrease of B-cells and a striking development of myeloid cells in all compartments. Collectively, our results show that granuloma inflammatory lesions dramatically change haematopoiesis homeostasis. Consequently, we suggest that besides their defective innate immunity, the alteration of haematopoiesis homeostasis upon granuloma may contribute to the dismal outcome of CGD.

Neutrophil-derived reactive oxygen species promote tumor colonization

A single-nucleotide polymorphism of neutrophil cytosolic factor 1 (Ncf1), leading to an impaired generation of reactive oxygen species (ROS), is a causative genetic factor for autoimmune disease. To study a possible tumor protection effect by the Ncf1 mutation in a manner dependent on cell types, we used experimental mouse models of lung colonization assay by B16F10 melanoma cells. We observed fewer tumor foci in Ncf1 mutant mice, irrespective of αβT, γδT, B-cell deficiencies, or of a functional Ncf1 expression in CD68-positive monocytes/macrophages. The susceptibility to tumor colonization was restored by the human S100A8 (MRP8) promoter directing a functional Ncf1 expression to granulocytes. This effect was associated with an increase of both ROS and interleukin 1 beta (IL-1β) production from lung neutrophils. Moreover, neutrophil depletion by anti-Ly6G antibodies increased tumor colonization in wild type but failed in the Ncf1 mutant mice. In conclusion, tumor colonization is counteracted by ROS-activated and IL-1β-secreting tissue neutrophils.

Targeting MicroRNA-192-5p, a Downstream Effector of NOXs (NADPH Oxidases), Reverses Endothelial DHFR (Dihydrofolate Reductase) Deficiency to Attenuate Abdominal Aortic Aneurysm Formation.

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Inhibition of NOX1 Mitigates Blood Pressure Increases in Elastin Insufficiency.

Elastin (ELN) insufficiency leads to the cardiovascular hallmarks of the contiguous gene deletion disorder, Williams-Beuren syndrome, including hypertension and vascular stiffness. Previous studies showed that Williams-Beuren syndrome deletions, which extended to include the NCF1 gene, were associated with lower blood pressure (BP) and reduced vascular stiffness. NCF1 encodes for p47phox, the regulatory component of the NOX1 NADPH oxidase complex that generates reactive oxygen species (ROS) in the vascular wall. Dihydroethidium and 8-hydroxyguanosine staining of mouse aortas confirmed that Eln heterozygotes (Eln+/- ) had greater ROS levels than the wild-types (Eln+/+ ), a finding that was negated in vessels cultured without hemodynamic stressors. To analyze the Nox effect on ELN insufficiency, we used both genetic and chemical manipulations. Both Ncf1 haploinsufficiency (Ncf1+/- ) and Nox1 insufficiency (Nox1-/y ) decreased oxidative stress and systolic BP in Eln+/- without modifying vascular structure. Chronic treatment with apocynin, a p47phox inhibitor, lowered systolic BP in Eln+/- , but had no impact on Eln+/+ controls. In vivo dosing with phenylephrine (PE) produced an augmented BP response in Eln+/- relative to Eln+/+ , and genetic modifications or drug-based interventions that lower Nox1 expression reduced the hypercontractile response to PE in Eln+/- mice to Eln+/+ levels. These results indicate that the mechanical and structural differences caused by ELN insufficiency leading to oscillatory flow can perpetuate oxidative stress conditions, which are linked to hypertension, and that by lowering the Nox1-mediated capacity for vascular ROS production, BP differences can be normalized.

Weighted Gene Co-expression Network Analysis Identifies Crucial Genes Mediating Progression of Carotid Plaque.

BackgroundSurface rupture of carotid plaque can cause severe cerebrovascular disease, including transient ischemic attack and stroke. The aim of this study was to elucidate the molecular mechanism governing carotid plaque progression and to provide candidate treatment targets for carotid atherosclerosis.MethodsThe microarray dataset GSE28829 and the RNA-seq dataset GSE104140, which contain advanced plaque and early plaque samples, were utilized in our analysis. Differentially expressed genes (DEGs) were screened using the “limma” R package. Gene modules for both early and advanced plaques were identified based on co-expression networks constructed by weighted gene co-expression network analysis (WGCNA). Gene Ontology (GO) and Kyoto Encyclopedia of Genes Genomes (KEGG) analyses were employed in each module. In addition, hub genes for each module were identified. Crucial genes were identified by molecular complex detection (MCODE) based on the DEG co-expression network and were validated by the GSE43292 dataset. Gene set enrichment analysis (GSEA) for crucial genes was performed. Sensitivity analysis was performed to evaluate the robustness of the networks that we constructed.ResultsA total of 436 DEGs were screened, of which 335 were up-regulated and 81 were down-regulated. The pathways related to inflammation and immune response were determined to be concentrated in the black module of the advanced plaques. The hub gene of the black module was ARHGAP18 (Rho GTPase activating protein 18). NCF2 (neutrophil cytosolic factor 2), IQGAP2 (IQ motif containing GTPase activating protein 2) and CD86 (CD86 molecule) had the highest connectivity among the crucial genes. All crucial genes were validated successfully, and sensitivity analysis demonstrated that our results were reliable.ConclusionTo the best of our knowledge, this study is the first to combine DEGs and WGCNA to establish a DEG co-expression network in carotid plaques, and it proposes potential therapeutic targets for carotid atherosclerosis.

Natural Loss-of-Function Mutations in Qa2 and NCF1 Cause the Spread of Mannan-Induced Psoriasis

A basis for the genetic predisposition to psoriasis is a single locus, PSORS1, within the major histocompatibility complex I region. This murine major histocompatibility complex locus encodes nonclassical molecules such as Qa2. We hypothesized that a natural loss-of-function variant of Qa2 gene clusters promotes psoriasis. In this study, we have developed a mannan-induced psoriasis model with the double deficiency of Qa2 and ROS owing to natural mutations of Qa2 gene clusters and the Ncf1 gene in the C57BL/6 background, respectively. We report three key findings in mice with mannan-induced psoriasis. A complete deficiency of Qa2 resulted in the expansion of IL-17‒producing γδ T cells and group 3 innate lymphoid cells in draining lymph nodes, leading to ear psoriasis. A single copy of Qa2-encoding genes was enough to protect against mannan-induced psoriasis, and such a protection was erased by a mutated Ncf1. Double defects with Qa2 and Ncf1 elicited a spread of exaggerated ear psoriasis to the nails, and the deficiency of γδ T cells reduced the severity of nail psoriasis. Collectively, these findings in mice provide evidence for the importance of Ncf1 mutations and Qa2 gene clusters, possibly corresponding to the PSORS1 locus in the spread of psoriasis.

Genotype Analysis of ABCC1, NCF4 and CBR3 Polymorphism and the Association With Childhood Acute Lymphoblastic Leukemia in Mexican Childhood Population.

Background: The identification of genetic risk factors for Acute Lymphoblastic Leukemia (ALL), are increasingly urgent and necessary. Objective: The purpose of this study is to determine the association of the genetic polymorphisms ABCC1 rs3743527, NCF4 rs1883112 and CBR3 rs1056892 with ALL. Methods: DNA samples were obtained in 71 children with ALL (from 2 to 18 years) and in 71 controls without ALL, to determine the polymorphisms by real-time polymerase chain reaction (qPCR), using specific TaqMan probes in a StepOne® thermal cycler (Applied Biosystems, United States). Results: The results of the Odds Ratio analysis show that in the rs1883112 polymorphism of the NCF4 gene, the heterozygous allele has a risk effect for ALL (OR = 3.1870, CI = 1.8880–7.9383 and p = 0.0002), in turn the mutated genotype (AA) is associated with a protective effect (OR = 0.26, 0.1248 to 0.5434 and p = 0.0003). On the other hand, the CBR3 rs1056892 polymorphism shows a significant association of risk to ALL, in the presence of the HT genotype (OR = 2.77, IC = 1.3837 to 5.5651 and p = 0.004) and the mutated genotype of this polymorphism has a significant association with protection to ALL in the HM genotype (OR = 0.52, IC = 0.2639 to 1.0304 and p = 0.05). While the inheritance models of the polymorphisms let us see that of the rs1883112 polymorphism of the NCF4 polymorphism; the HT genotype of the codominant model shows a protective effect against ALL (OR = 0.4117, IC = 0.1718 to 0.9866 and p = 0.04), the recessive model shows us and confirms what we already saw in table number 3, being that there is an association with protective effect in the HM genotype (OR = 0.2604, IC = 0.1248 to 0.5434 and p = 0.0003). In the polymorphism rs1056892 of the CBR3 gene, a protection association was found in the heterozygous allele of the codominant model (OR = 0.3448, IC = 0.1375 to 0.8896 and p = 0.0274). In addition, the recessive inheritance model for the HM genotype shows a protective effect to ALL, (OR = 0.52, CI = 0.9919 to 3.8638 and p = 0.05). Conclusion: There is an evident impact of the NCF4 rs1883112 and CBR3 rs1056892 polymorphisms with an increased risk of susceptibility to ALL; Likewise, through the codominant inheritance model, the effect of the variation of the CBR3 rs1056892 gene as a protective factor against ALL was evaluated.

Copy number variation of the NCF1 gene in patients of acne vulgaris

ObjectiveThe objective of this study was to evaluate the influence of neutrophil cytosolic factor 1 gene copy number variation (CNV) on acne vulgaris in a sample of Egyptian population to explore whether this CNV affects disease occurrence or increases its risk.BackgroundAcne vulgaris is a common skin condition that affects the majority of the population during adolescence. Acne is characterized by a variable combination of comedones, pustules, inflammation, and scarring. Oxidative stress has been considered to play a role in acne. Neutrophil cytosolic factor 1 (NCF1) is a component of NADPH oxidase enzyme, which is responsible for the production of reactive oxygen species from phagocytes. It is reported that increased or decreased copy numbers of NCF1 gene affects many diseases such as rheumatoid arthritis and systemic lupus erythematosus.Patients and methodsThis study included 50 patients divided into two groups: 25 acne patients and 25 apparently healthy controls. Both groups were investigated for NCF1 CNV by quantitative real-time PCR.ResultsWe found a statistically significant increased CNV of the NCF1 gene in the acne group compared with the control group (P = 0.028).ConclusionOur study suggests that an increased number of copies of the NCF1 gene may be a predisposing factor for acne vulgaris development. Moreover, decreased NCF1 copy numbers may be protective for acne.

Identification of Potential Biomarkers Associated with Acute Myocardial Infarction by Weighted Gene Coexpression Network Analysis.

Background In the general population, acute myocardial infarction (AMI) represents a significant cause of mortality. This study is aimed at identifying novel diagnostic biomarkers to aid in treating and diagnosing AMI. Methods The Gene Expression Omnibus (GEO) database was explored to extract two microarray datasets, GSE66360 and GSE48060, which were subsequently merged into a single cohort. Both AMI and control samples were analyzed for differentially expressed genes (DEGs), which were subsequently subjected to weighed gene coexpression network analysis (WGCNA) to identify the most significant module. Gene Ontology (GO) and pathway analyses subsequently carried out the most significant gene modules along with construction of a protein-protein interaction network (PPI). Cytoscape plugin cytoHubba allowed for the prediction of the top 4 key genes according to the network maximal clique centrality (MCC) algorithm. The expression levels and diagnostic value of the four key genes were additionally verified in the GSE62646 dataset. Results A WCGNA analysis revealed 878 DEGs which were clustered into 6 modules. The module with the most significance in AMI was colored blue. Subsequent GO and KEGG pathway enrichment analysis on blue module genes revealed that they were primarily enriched in the inflammation-related pathways. These findings, in combination with PPI and coexpression networks, resulted in the identification of the top four genes by cytoHubba, which included leukocyte immunoglobulin-like receptor B2 (LILRB2), toll-like receptor 2 (TLR2), neutrophil cytosolic factor 2 (NCF2), and S100A9. Among them, LILRB2, NCF2, and S100A9 were validated in the GSE62646 dataset. Conclusions The results suggested that LILRB2, NCF2, and S100A9 could be potential gene biomarkers for AMI.

Investigation of causative genetic defects in patients with primary immunodeficiency by next generation sequencing

Aim: Inborn errors of immunity are rare diseases presented with a broad range of clinical symptoms. There are more than 450 causative genetic defects and the genetic diagnosis is very important for the patients. Use of next generation sequencing facilitated the molecular and genetic identification of these diseases for last 10 years. We aimed to search disease-causing defects in patients with primary immunodeficiencies (PIDs) by next generation sequencing. Materials and Methods: The study included 12 PID patients without genetic diagnosis. We performed whole exome sequencing for the investigation of genetic defects and Sanger sequencing for variant validation.Results: We found seven different disease-causing mutations in 6 patients with a diagnosis rate of 50%. There were three known pathogenic variants in CYBA, SBDS and RAG2 genes. We identified two additional new causative variants in NCF2 and SBDS genes and two novel mutations in PGM3 and SAMD9L genes.Conclusion: The result revealed that NGS-based methods especially whole exome sequencing can be used efficiently for genetic diagnosis of primary immunodeficiency diseases.

Arterial hypertension in children with Williams-Beuren syndrome

Williams-Beuren Syndrome (WBS) is a multisystemic disorder with prevalence of 1/7500 [1], affects both genders equally and it is caused by a microdeletion of 26-28 genes, including elastin and NCF1 genes, of the chromosome region 7q11.23 [1,2]. The disease is transmitted in an autosomal dominant fashion, but almost all cases are the result of de novo mutations [1].

P47phox deficiency improves cognitive impairment and attenuates tau hyperphosphorylation in mouse models of AD

BackgroundAlzheimer’s disease (AD) is characterized by progressive memory loss and cognitive impairment. The aggregation of amyloid β (Aβ) and hyperphosphorylated tau protein are two major pathological features of AD. Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase, NOX) has been indicated in Aβ pathology; however, whether and how it affects tau pathology are not yet clear.MethodsThe role of NOX2 in cognitive function, amyloid plaque formation, and tau hyperphosphorylation were examined in APP/PS1 transgenic mice mated with p47phox-deficient mice (with deletion of the gene of neutrophil cytosolic factor 1, Ncf1) and/or in p47phox-deficient mice receiving intracerebroventricular (ICV) injection of streptozotocin (STZ). The cognitive and non-cognitive functions in these mice were assessed by Morris water maze, Rotarod test, open field, and elevated plus maze. Aβ levels, amyloid plaques, p47phox expression, and astrocyte activation were evaluated using immunofluorescence staining, ELISA, and/or Western blotting. Cultured primary neuronal cells were treated with okadaic acid or conditioned media (CM) from high glucose-stimulated primary astrocytes. The alteration in tau pathology was determined using Western blotting and immunofluorescence staining.ResultsDeletion of the gene coding for p47phox, the organizer subunit of NOX2, significantly attenuated cognitive impairment and tau pathology in these mice. p47phox deficiency decreased the activation of astrocytes but had no effect on Aβ levels and amyloid plaque formation in the brains of aged APP/PS1 mice, which displayed markedly increased expression of p47phox in neurons and astrocytes. Cell culture studies found that neuronal p47phox deletion attenuated okadaic acid-induced tau hyperphosphorylation at specific sites in primary cultures of neurons. CM from high glucose-treated WT astrocytes increased tau hyperphosphorylation in primary neurons, whereas this effect was absent from p47phox-deficient astrocytes.ConclusionsThese results suggest that p47phox is associated with cognitive function and tau pathology in AD. p47phox expressed in neurons contributes to tau hyperphosphorylation directly, while p47phox in astrocytes affect tau hyperphosphorylation by activating astrocytes indirectly. Our results provide new insights into the role of NOX2 in AD and indicate that targeted inhibition of p47phox may be a new strategy for the treatment of AD.