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1.
Int J Mol Sci ; 22(6)2021 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-33799876

RESUMO

The basic helix-loop-helix/Per-ARNT-SIM (bHLH-PAS) proteins are a family of transcription factors regulating expression of a wide range of genes involved in different functions, ranging from differentiation and development control by oxygen and toxins sensing to circadian clock setting. In addition to the well-preserved DNA-binding bHLH and PAS domains, bHLH-PAS proteins contain long intrinsically disordered C-terminal regions, responsible for regulation of their activity. Our aim was to analyze the potential connection between disordered regions of the bHLH-PAS transcription factors, post-transcriptional modifications and liquid-liquid phase separation, in the context of disease-associated missense mutations. Highly flexible disordered regions, enriched in short motives which are more ordered, are responsible for a wide spectrum of interactions with transcriptional co-regulators. Based on our in silico analysis and taking into account the fact that the functions of transcription factors can be modulated by posttranslational modifications and spontaneous phase separation, we assume that the locations of missense mutations inducing disease states are clearly related to sequences directly undergoing these processes or to sequences responsible for their regulation.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Predisposição Genética para Doença/genética , Proteínas Intrinsicamente Desordenadas/genética , Receptores de Hidrocarboneto Arílico/genética , Proteínas Repressoras/genética , Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Humanos , Proteínas Intrinsicamente Desordenadas/metabolismo , Mutação de Sentido Incorreto , Processamento Pós-Transcricional do RNA , Receptores de Hidrocarboneto Arílico/metabolismo , Proteínas Repressoras/metabolismo , Transcrição Genética
2.
Gene ; 778: 145485, 2021 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-33581269

RESUMO

Recent genome-wide association studies (GWAS) highlighted the importance of genetic variations on SLC22A3 and MIA3 genes in developing coronary heart disease (CHD) among different ethnicities. However, the influence of these variations is not recognized within the Iranian population. Hence, in the present study, we aim to investigate two key single nucleotide polymorphisms (SNPs) on CHD incidence in this population. For this purpose, from Tehran Cardiometabolic Genetic Study (TCGS), 453 individuals with CHD were selected as a case and 453 individuals as a control that matched their age and gender. After quality control of two selected SNPs, rs2048327 (SLC22A3) and rs17465637 (MIA3), we used genotyps resulted from chip-typing technology and conducted the logistic regression analysis adjusted for non-genetic risk factors to detect the possible association of these SNPs with the CHD development. Our findings demonstrated the rs2048327-G and rs17465637-C can significantly increase the risk of CHD development about two times in only males and females, respectively. Interestingly, in the male carriers of the risk allele (G) of rs2048327, the low high-density lipoprotein (HDL) level can significantly predispose them to develop coronary heart disease in the future. According to our results, paying more attention to gender and genetic markers can help more efficient coronary heart disease screening and diagnosis.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , HDL-Colesterol/sangue , Doença das Coronárias/genética , Proteínas de Transporte de Cátions Orgânicos/genética , Polimorfismo de Nucleotídeo Único , Idoso , Estudos de Casos e Controles , Doença das Coronárias/sangue , Feminino , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Humanos , Irã (Geográfico) , Modelos Logísticos , Masculino , Pessoa de Meia-Idade
3.
Ecotoxicol Environ Saf ; 211: 111947, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33503546

RESUMO

The chicken (Gallus gallus), which has three aryl hydrocarbon receptor (AHR) isoforms (ckAHR1, ckAHR2, and ckAHR1ß) and two AHR nuclear translocator (ARNT) isoforms (ckARNT1 and ckARNT2), is highly sensitive to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and can serve as an avian model to gain an understanding of the mechanism underlying dioxin toxicity. To elucidate the mechanism of TCDD-induced immunotoxicity in avian species, we treated chicken embryos in ovo with graded concentrations of TCDD (1.5, 2.5, 3.0, 3.3, 3.5, and 4.0 µM). Initially, we measured mRNA expression levels of ckAHR and ckARNT isoforms and analyzed the T cell populations and transcriptome in the thymuses of TCDD-treated chicken embryos. Quantitative polymerase chain reaction analysis revealed that mRNA expressions of ckAHR1 and ckARNT2 were dominant in the thymus. Severe weight loss and thymus atrophy were observed in the TCDD-treated embryos. Immunophenotyping analyses demonstrated significant increases in CD4+CD8-CD25+ and CD4+CD8+CD25+ regulatory T cells (Tregs) populations following TCDD exposure, suggesting that TCDD suppresses T cell-mediated immune responses in chicken embryos. In addition, thymic transcriptome analyses intimated that alteration of the signaling pathways related to erb-b2 receptor tyrosine kinase 4 (ERBB4) and wnt family member 5A (WNT5A), and bone morphogenetic protein (BMP) may be associated with the TCDD-induced thymus atrophy. We also observed significantly altered expression levels of genes including interleukine 13 receptor subunit alpha 2 (IL13RA2), transforming growth factor beta 1 (TGFß1), collagen type III alpha 1 chain (COL3A1), and collagen type IX alpha 3 chain (COL9A3), implying immunosuppression, fibrosis development, and collagen deposition. Collectively, these findings suggest that TCDD exposure activates the ckAHR1-ckARNT2 signaling pathway and suppresses immune responses through the prompted differentiation to CD4+CD8-CD25+ and CD4+CD8+CD25+ Tregs and altered expressions of immune-related genes in the thymus of chicken embryos.


Assuntos
Poluentes Ambientais/toxicidade , Dibenzodioxinas Policloradas/toxicidade , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos , Linfócitos T CD8-Positivos/metabolismo , Diferenciação Celular , Embrião de Galinha , Galinhas/metabolismo , Sistema Imunitário/efeitos dos fármacos , Isoformas de Proteínas/genética , Receptores de Hidrocarboneto Arílico/metabolismo , Transdução de Sinais , Linfócitos T , Transcriptoma
4.
Gene ; 771: 145361, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33338508

RESUMO

Hypoxia is a critical, but frequently overlooked problem, which commonly exists in Chinese mitten crab rearing. However, little information is available on the molecular mechanisms of the detrimental effects of hypoxia in this species. In the present study, crabs were subjected to acute hypoxia stress (DO 1.0 mg/L), followed by reoxygenation (DO 6.8 mg/L). Hepatopancreas from five groups of crabs (three to four crabs per group), including normoxia, hypoxia for one and six hours, and reoxygenation for one and 12 h, were used for transcriptome sequencing. The pooled total RNA of all samples were utilized to reconstruct a reference transcriptome with PacBio RS II sequencing, obtaining 49.19 G clean data, with a mean length of 2,180 bp. Seventeen cDNA libraries were constructed and sequenced to identify differentially expressed genes (DEGs) among the different samples (FDR < 0.05 and |log2 fold change| ≥1). A total of 103 and 251 DEGs were identified when exposed to hypoxia for one and six hours, respectively. Totally 462 and 673 DEGs were identified during reoxygenation at 1 and 12 h, respectively. Among these DEGs, two transcripts with complete ORFs were identified to encode hypoxia-inducible factor 1 (Es-Hif-1α/ß), which is a transcriptional activator of various genes correlated to the cellular adaptive responses to hypoxia. Es-Hif-1a/ß expressions were significantly upregulated when exposed to hypoxia treatment, and no significant difference was observed for Es-Hif-1α between hypoxic treatment for 6 h and reoxygenation. The significant KEGG enrichment revealed that the DEGs under hypoxia were mainly enriched in "PPAR signaling pathway", "Gap junction" and "Phototransduction-fly". Compared with crabs in normoxia, even with 12 h of reoxygenation, the hepatopancreas transcriptome under hypoxia remained severely affected, implying its adverse effect on growth and development, or increased susceptibility to diseases.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Braquiúros/genética , Perfilação da Expressão Gênica/veterinária , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Animais , Proteínas de Artrópodes/genética , Hipóxia Celular , Regulação da Expressão Gênica , Hepatopâncreas/química , Análise de Sequência de RNA/veterinária , Estresse Fisiológico
5.
Ecotoxicol Environ Saf ; 201: 110835, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32563159

RESUMO

The activation of the aryl hydrocarbon receptor (AHR) occurs through the binding of dioxin-like compounds (DLCs) or natural ligands. In this pathway, the AHR-ARNT (AHR nuclear translocator) heterodimer serves to regulate critical physiological functions, such as immune responses and the metabolism of xenobiotics. Birds have three AHR isoforms (AHR1, AHR1ß, and AHR2) and two ARNT isoforms (ARNT1 and ARNT2). However, how AHR and ARNT dimerization pair in birds regulates the AHR signaling pathway in an isoform-specific manner remains unknown. In this study, we initially sought to clarify the major chicken AHR-ARNT (ckAHR-ckARNT) pairs by estimating the mRNA tissue distributions of various ckAHR and ckARNT isoforms. Our results indicated that the ckAHR1-ckARNT1 represented the major dimerization pair in most tissues except the brain. We then measured the transactivation potencies of various ckAHR-ckARNT pairs by natural ligands and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), in in vitro reporter gene assays using COS-7 and LMH cell lines. Our results from the in vitro assays demonstrated that the ckAHR1-ckARNT1 pair was strongly activated by the five natural ligands, namely, 6-formylindolo [3,2-b]carbazole, L-kynurenin, kynurenic acid, indoxyl-3-sulfate, and 1,3,7-tribromodibenzo-p-dioxin, but not by TCDD. In in silico ligand docking simulations with ckAHR1 homology models, all the natural ligands showed a interaction pattern that was distinct from that observed with anthropogenic DLCs, including TCDD. In conclusion, our findings indicate that the ckAHR1-ckARNT1 may be the most important dimerization pair in most tissues for regulating the physiological functions driven by natural ligands, although it was less reactive to TCDD.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Galinhas/metabolismo , Dibenzodioxinas Policloradas/metabolismo , Multimerização Proteica , Receptores de Hidrocarboneto Arílico/metabolismo , Xenobióticos/metabolismo , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Células COS , Linhagem Celular Tumoral , Chlorocebus aethiops , Simulação por Computador , Ligantes , Simulação de Acoplamento Molecular , Ligação Proteica , Isoformas de Proteínas , Receptores de Hidrocarboneto Arílico/genética , Transdução de Sinais , Especificidade da Espécie , Transfecção
6.
PLoS Genet ; 16(5): e1008757, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32379754

RESUMO

In the last decades in vitro studies highlighted the potential for crosstalk between Hypoxia-Inducible Factor-(HIF) and glucocorticoid-(GC) signalling pathways. However, how this interplay precisely occurs in vivo is still debated. Here, we use zebrafish larvae (Danio rerio) to elucidate how and to what degree hypoxic signalling affects the endogenous glucocorticoid pathway and vice versa, in vivo. Firstly, our results demonstrate that in the presence of upregulated HIF signalling, both glucocorticoid receptor (Gr) responsiveness and endogenous cortisol levels are repressed in 5 days post fertilisation larvae. In addition, despite HIF activity being low at normoxia, our data show that it already impedes both glucocorticoid activity and levels. Secondly, we further analysed the in vivo contribution of glucocorticoids to HIF activity. Interestingly, our results show that both glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) play a key role in enhancing it. Finally, we found indications that glucocorticoids promote HIF signalling via multiple routes. Cumulatively, our findings allowed us to suggest a model for how this crosstalk occurs in vivo.


Assuntos
Glucocorticoides/farmacologia , Fator 1 Induzível por Hipóxia/fisiologia , Receptor Cross-Talk/fisiologia , Peixe-Zebra , Animais , Animais Geneticamente Modificados , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Embrião não Mamífero , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/genética , Glucocorticoides/metabolismo , Fator 1 Induzível por Hipóxia/metabolismo , Larva/genética , Larva/metabolismo , Receptor Cross-Talk/efeitos dos fármacos , Receptores de Glucocorticoides/metabolismo , Receptores de Glucocorticoides/fisiologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Proteínas Supressoras de Tumor/genética , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética
7.
Mol Carcinog ; 59(8): 897-907, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32319143

RESUMO

Lactate dehydrogenase isozyme (LDH) is a tetramer constituted of two isoforms, LDHA and LDHB, the expression of which is associated with cell metabolism and cancer progression. Our previous study reveals that CC-chemokine ligand-18 (CCL18) is involved in progression of prostate cancer (PCa).This study aims to investigate how CCL18 regulates LDH isoform expression, and therefore, contributes to PCa progression. The data revealed that the expression of LDHA was upregulated and LDHB was downregulated in PCa cells by CCL18 at both messenger RNA and protein levels. The depletion of CCR8 reduced the ability of CCL18 to promote the proliferation, migration, and lactate production of PCa cells. Depletion of a CCR8 regulated transcription factor, ARNT, significantly reduced the expression of LDHA. In addition, The Cancer Genome Atlas dataset analyses revealed a positive correlation between CCR8 and ARNT expression. Two dimension difference gel electrophoresis revealed that the LDHA/LDHB ratio was increased in the prostatic fluid of patients with PCa and PCa tissues. Furthermore, increased LDHA/LDHB ratio was associated with poor clinical outcomes of patients with PCa. Together, our results indicate that the CCR8 pathway programs LDH isoform expression in an ARNT dependent manner and that the ratio of LDHA/LDHB has the potential to serve as biomarkers for PCa diagnosis and prognosis.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Biomarcadores Tumorais/metabolismo , Quimiocinas CC/metabolismo , Regulação Neoplásica da Expressão Gênica , L-Lactato Desidrogenase/metabolismo , Neoplasias da Próstata/patologia , Receptores CCR8/metabolismo , Apoptose , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Biomarcadores Tumorais/genética , Proliferação de Células , Quimiocinas CC/genética , Humanos , Isoenzimas , L-Lactato Desidrogenase/genética , Masculino , Prognóstico , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Receptores CCR8/genética , Taxa de Sobrevida , Células Tumorais Cultivadas
8.
Infect Immun ; 88(4)2020 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-31964743

RESUMO

Invasive aspergillosis (IA) is a life-threatening infection that affects an increasing number of patients undergoing chemotherapy or allo-transplantation, and recent studies have shown that genetic factors contribute to disease susceptibility. In this two-stage, population-based, case-control study, we evaluated whether 7 potentially functional single nucleotide polymorphisms (SNPs) within the ARNT2 and CX3CR1 genes influence the risk of IA in high-risk hematological patients. We genotyped selected SNPs in a cohort of 500 hematological patients (103 of those had been diagnosed with proven or probable IA), and we evaluated their association with the risk of developing IA. The association of the most interesting markers of IA risk was then validated in a replication population, including 474 subjects (94 IA and 380 non-IA patients). Functional experiments were also performed to confirm the biological relevance of the most interesting markers. The meta-analysis of both populations showed that carriers of the ARNT2 rs1374213G, CX3CR1 rs7631529A, and CX3CR1 rs9823718G alleles (where the RefSeq identifier appears as a subscript) had a significantly increased risk of developing IA according to a log-additive model (P value from the meta-analysis [P Meta] = 9.8 · 10-5, P Meta = 1.5 · 10-4, and P Meta =7.9 · 10-5, respectively). Haplotype analysis also confirmed the association of the CX3CR1 haplotype with AG CGG with an increased risk of IA (P = 4.0 · 10-4). Mechanistically, we observed that monocyte-derived macrophages (MDM) from subjects carrying the ARNTR2 rs1374213G allele or the GG genotype showed a significantly impaired fungicidal activity but that MDM from carriers of the ARNT2 rs1374213G and CX3CR1 rs9823718G or CX3CR1 rs7631529A alleles had deregulated immune responses to Aspergillus conidia. These results, together with those from expression quantitative trait locus (eQTL) data browsers showing a strong correlation of the CX3CR1 rs9823718G allele with lower levels of CX3CR1 mRNA in whole peripheral blood (P = 2.46 · 10-7) and primary monocytes (P = 4.31 · 10-7), highlight the role of the ARNT2 and CX3CR1 loci in modulating and predicting IA risk and provide new insights into the host immune mechanisms involved in IA development.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Aspergillus/imunologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Receptor 1 de Quimiocina CX3C/genética , Predisposição Genética para Doença , Aspergilose Pulmonar Invasiva/genética , Polimorfismo de Nucleotídeo Único , Estudos de Casos e Controles , Genótipo , Doenças Hematológicas/complicações , Humanos , Medição de Risco
9.
Exp Cell Res ; 388(2): 111845, 2020 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-31945318

RESUMO

BACKGROUND: Hypoxia-inducible factor (HIF)-2α associates with poor outcome in neuroblastoma and glioblastoma, and gain-of-function mutations in the EPAS1 gene (encoding HIF-2α) have been reported in paragangliomas and pheochromocytomas. Specific targeting of a druggable hydrophobic pocket in the HIF-2α PAS-B domain with PT2385 have demonstrated promising clinical results for clear cell renal cell carcinoma (ccRCC). Here, we investigated the effect of PT2385-mediated inhibition of ARNT dependent HIF-2 activity. METHODS: Neuroblastoma patient-derived xenograft (PDX) cells were treated with PT2385 and analyzed for HIF-2-dependent gene expression, HIF activity, HIF-2α protein localization, response to chemotherapy and orthotopic tumor growth in vivo. Two-sided student t-test was used. RESULTS: We detected high levels of HIF-2α protein in perivascular niches in neuroblastoma PDXs in vivo and at oxygenated conditions in PDX-derived cell cultures in vitro, particularly in the cytoplasmic fraction. Nuclear HIF-2α expression was reduced following PT2385 treatment, but surprisingly, virtually no effects on tumor growth in vivo or expression of canonical HIF downstream target genes in vitro were observed. In coherence, RNA sequencing of PT2385-treated PDX cells revealed a virtually unaffected transcriptome. Treatment with PT2385 did not affect cellular response to chemotherapy. In contrast, HIF-2α protein knockdown resulted in profound downregulation of target genes. CONCLUSIONS: The lack of effect from PT2385 treatment in combination with high cytoplasmic HIF-2α expression at normoxia suggest that HIF-2α have additional roles than acting as an ARNT dependent transcription factor. It is important to further unravel the conditions at which HIF-2α has transcriptional and non-transcriptional roles in neuroblastoma.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/antagonistas & inibidores , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Indanos/farmacologia , Neuroblastoma/patologia , Sulfonas/farmacologia , Transcriptoma/efeitos dos fármacos , Animais , Apoptose , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Biomarcadores Tumorais/genética , Proliferação de Células , Feminino , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Neuroblastoma/genética , Neuroblastoma/metabolismo , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
10.
Environ Sci Pollut Res Int ; 27(9): 9227-9235, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31916168

RESUMO

A yeast reporter gene assay system with improved performance for dioxin detection was established. Since yeast reporter gene assays are relatively simple, easy to handle, and inexpensive, they have been used for various assessments of environmental contaminants. We previously constructed a yeast assay strain expressing the aryl hydrocarbon receptor (AhR) and AhR nuclear translocator (Arnt) carrying the lacZ reporter gene, for detection of dioxins. In the present study, genes encoding cell wall mannoproteins and ATP-binding cassette transporters in the yeast assay strains were deleted in order to increase the substance influx and prevent its efflux. We also established an assay procedure for protoplasts of these yeasts. These modifications improved the detection limit 40-fold and reduced the duration of the assay by 40%. By combining the yeast protoplast and a rapid sample preparation technique using disposal multilayer solid-phase extraction columns to remove unintended aryl hydrocarbons, this yeast reporter gene assay system detected the ligand activities of dioxins and related compounds in 1 g of forest soil containing dioxins at a concentration 10 times lower than the Japanese environmental standard for dioxins in soil.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Dioxinas , Dibenzodioxinas Policloradas/química , Receptores de Hidrocarboneto Arílico/genética , Saccharomyces cerevisiae/química , Translocador Nuclear Receptor Aril Hidrocarboneto/química , Dioxinas/química , Genes Reporter , Protoplastos , Receptores de Hidrocarboneto Arílico/química
11.
PLoS One ; 14(12): e0225332, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31800592

RESUMO

BACKGROUND AND AIM: Non-alcoholic steatohepatitis (NASH) is predicted to become the most common cause of cirrhosis and liver failure. Risk factors include obesity, insulin resistance and diabetes. Macrophages and other myeloid cells play crucial roles in initiating and driving inflammation. Aryl hydrocarbon Receptor Nuclear Translocator (ARNT) is a transcription factor which binds to a range of partners to mediate responses to environmental signals, including the diet. In people with diabetes it is decreased in liver. We hypothesised that myeloid cell ARNT activity may contribute to the development of liver pathology. METHODS: Floxed-ARNT mice were bred with LysM-Cre mice to generate mice with reduced ARNT in myeloid cells. Animals were fed a high fat diet (HFD) and liver pathology was assessed. Histology, mRNA, fat accumulation and metabolism were studied. RESULTS: Animals with reduced myeloid ARNT developed steatohepatitis on a HFD, with additional alterations of metabolism and fat deposition. Steatohepatitis was accompanied by hepatic macrophage infiltration and expression of both M1 and M2 markers. Expression of mRNAs for Cxcl1, Mcp-1, Tnf-α and Tgf-ß1 were increased. Human livers from controls and people with NASH were tested; ARNT mRNA was decreased by 80% (p = 0.0004). CONCLUSIONS: Decreased myeloid ARNT may play a role in the conversion from non-alcoholic fatty liver to steatohepatitis. Increasing ARNT may be a therapeutic strategy to reduce NASH.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Células Mieloides/metabolismo , Hepatopatia Gordurosa não Alcoólica/genética , Adulto , Animais , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Quimiocina CXCL1/genética , Quimiocina CXCL1/metabolismo , Dieta Hiperlipídica/efeitos adversos , Feminino , Deleção de Genes , Humanos , Metabolismo dos Lipídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
12.
Sci Rep ; 9(1): 18405, 2019 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-31804579

RESUMO

The interaction of genetic susceptibility and dietary habits in cardiovascular disease (CVD) remains undetermined. The purpose of this study was to investigate whether a Mediterranean dietary style modified the genetic risk of developing CVD in a Chinese cohort. A total of 2098 subjects with dietary information from a Chinese community cohort (CVDFACTS) were enrolled. Candidate genes, including SNP markers rs1333049 (CDKN2B, 9p21.3), rs17465637 (MIA3, 1q41) and rs501120 (CXCL12, 10q11.21), were genotyped to analyze the association with future CVD. The impact of dietary pattern was also analyzed according to adherence to the diet using the Mediterranean Diet Score (MDS). After an average follow-up of 7.8 years, only the C risk allele of rs1333049 at chromosome 9p21.3 was associated with a higher risk of MI with either an additive [HR = 1.78, 95% CI:1.23-2.5] or a recessive model [HR = 2.40, 95% CI: 1.42-4.04], and the CC genotype had a higher risk of developing MI (p = 0.009, log-rank test). There was no significant difference in the association of the lipid profile with future CV outcomes among the MDS tertiles. However, the high MI risk of the CC genotype in individuals consuming a less healthy diet (MDS1) (HR: 6.39, 95% CI: 1.74-23.43) significantly decreased to 2.38 (95% CI: 0.57-10.04) in individuals consuming a healthier diet (MDS3), indicating that a healthier dietary pattern (higher MDS) modified the risk of developing MI in carriers of variants in CDKN2B. In conclusion, genetic variants of CDKN2B at 9p21 were significantly associated with future MI risk in a Chinese cohort, and the genetic risk of MI could be modified by a healthier diet.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Quimiocina CXCL12/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Dieta Mediterrânea , Infarto do Miocárdio/genética , Adulto , Idoso , Alelos , Translocador Nuclear Receptor Aril Hidrocarboneto/sangue , Grupo com Ancestrais do Continente Asiático , Quimiocina CXCL12/sangue , Cromossomos Humanos Par 9/química , Estudos de Coortes , Inibidor de Quinase Dependente de Ciclina p15/sangue , Feminino , Expressão Gênica , Frequência do Gene , Predisposição Genética para Doença , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/etnologia , Infarto do Miocárdio/patologia , Polimorfismo de Nucleotídeo Único , Fatores de Risco
13.
Endocr Regul ; 53(4): 237-249, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31734653

RESUMO

OBJECTIVE: The aim of the present study was to examine the effect of glucose deprivation on the expression of genes encoded glucocorticoid receptor (NR3C1) and some related proteins (NR3C2, AHR, NRIP1, NNT, ARHGAP35, SGK1, and SGK3) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1/inositol requiring enzyme 1) for evaluation of their possible significance in the control of glioma growth through endoplasmic reticulum stress signaling mediated by IRE1 and glucose deprivation. METHODS: The expression of NR3C1, NR3C2, AHR, NRIP1, NNT, ARHGAP35, SGK1, and SGK3 genes in U87 glioma cells transfected by empty vector pcDNA3.1 (control cells) and cells without ERN1 signaling enzyme function (transfected by dnERN1) under glucose deprivation was studied by real time quantitative polymerase chain reaction. RESULTS: It was shown that the expression level of NR3C2, AHR, SGK1, SGK3, and NNT genes was up-regulated in control U87 glioma cells under glucose deprivation condition in comparison with the control cells growing with glucose. At the same time, the expression of NRIP1 gene is down-regulated in these glioma cells under glucose deprivation, but NR3C1 and ARHGAP35 genes was resistant to this experimental condition. We also showed that inhibition of ERN1 signaling enzyme function significantly modified the response of most studied gene expressions to glucose deprivation condition. Thus, effect of glucose deprivation on the expression level of NR3C2, AHR, and SGK1 genes was significantly stronger in ERN1 knockdown U87 glioma cells since the expression of NNT gene was resistant to glucose deprivation condition. Moreover, the inhibition of ERN1 enzymatic activities in U87 glioma cells led to up-regulation of ARHGAP35 gene expression and significant down-regulation of the expression of SGK3 gene in response to glucose deprivation condition. CONCLUSIONS: Results of this study demonstrated that glucose deprivation did not change the expression level of NR3C1 gene but it significantly affected the expression of NR3C2, AHR, NRIP, SGK1, SGK3, and NNT genes in vector-transfected U87 glioma cells in gene specific manner and possibly contributed to the control of glioma growth since the expression of most studied genes in glucose deprivation condition was significantly dependent on the functional activity of IRE1 signaling enzyme.


Assuntos
Neoplasias Encefálicas/genética , Endorribonucleases/genética , Glioma/genética , Glucose/deficiência , Proteínas Serina-Treonina Quinases/genética , Receptores de Glucocorticoides/genética , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Endorribonucleases/deficiência , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Glioma/patologia , Glucose/farmacologia , Fatores de Troca do Nucleotídeo Guanina/genética , Humanos , Proteínas Imediatamente Precoces/genética , Proteínas Mitocondriais/genética , NADP Trans-Hidrogenase Específica para A ou B/genética , Proteína 1 de Interação com Receptor Nuclear/genética , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Repressoras/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
14.
Endocrinology ; 160(12): 2825-2836, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31580427

RESUMO

The transcription factor aryl hydrocarbon receptor nuclear translocator (ARNT)/hypoxia-inducible factor (HIF)-1ß (ARNT/HIF1ß) plays a key role in maintaining ß-cell function and has been shown to be one of the most downregulated transcription factors in islets from patients with type 2 diabetes. We have shown a role for ARNT/HIF1ß in glucose sensing and insulin secretion in vitro and no defects in in vivo glucose homeostasis. To gain a better understanding of the role of ARNT/HIF1ß in the development of diabetes, we placed control (+/+/Cre) and ß-cell-specific ARNT/HIF1ß knockout (fl/fl/Cre) mice on a high-fat diet (HFD). Unlike the control (+/+/Cre) mice, HFD-fed fl/fl/Cre mice had no impairment in in vivo glucose tolerance. The lack of impairment in HFD-fed fl/fl/Cre mice was partly due to an improved islet glucose-stimulated NADPH/NADP+ ratio and glucose-stimulated insulin secretion. The effects of the HFD-rescued insulin secretion in fl/fl/Cre islets could be reproduced by treating low-fat diet (LFD)-fed fl/fl/Cre islets with the lipid signaling molecule 1-monoacylglcyerol. This suggests that the defects seen in LFD-fed fl/fl/Cre islet insulin secretion involve lipid signaling molecules. Overall, mice lacking ARNT/HIF1ß in ß-cells have altered lipid signaling in vivo and are resistant to an HFD's ability to induce diabetes.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Diabetes Mellitus Experimental/metabolismo , Células Secretoras de Insulina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Diabetes Mellitus Experimental/etiologia , Dieta Hiperlipídica , Diglicerídeos , Glucose/metabolismo , Homeostase , Secreção de Insulina , Masculino , Camundongos Knockout , NADP/metabolismo
15.
Infect Immun ; 87(11)2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31451620

RESUMO

Cutaneous leishmaniasis is characterized by vascular remodeling. Following infection with Leishmania parasites, the vascular endothelial growth factor A (VEGF-A)/VEGF receptor 2 (VEGFR-2) signaling pathway mediates lymphangiogenesis, which is critical for lesion resolution. Therefore, we investigated the cellular and molecular mediators involved in VEGF-A/VEGFR-2 signaling using a murine model of infection. We found that macrophages are the predominant cell type expressing VEGF-A during Leishmania major infection. Given that Leishmania parasites activate hypoxia-inducible factor 1α (HIF-1α) and this transcription factor can drive VEGF-A expression, we analyzed the expression of HIF-1α during infection. We showed that macrophages were also the major cell type expressing HIF-1α during infection and that infection-induced VEGF-A production is mediated by ARNT/HIF activation. Furthermore, mice deficient in myeloid ARNT/HIF signaling exhibited larger lesions without differences in parasite numbers. These data show that L. major infection induces macrophage VEGF-A production in an ARNT/HIF-dependent manner and suggest that ARNT/HIF signaling may limit inflammation by promoting VEGF-A production and, thus, lymphangiogenesis during infection.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Leishmaniose Cutânea/metabolismo , Macrófagos/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Antígenos CD/genética , Antígenos CD/metabolismo , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Células Cultivadas , Regulação da Expressão Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Leishmania major , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/patologia , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais , Pele/metabolismo , Pele/parasitologia , Fator A de Crescimento do Endotélio Vascular/genética
16.
J Inherit Metab Dis ; 42(5): 898-908, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31276219

RESUMO

Exome sequencing has recently identified mutations in the gene TANGO2 (transport and Golgi organization 2) as a cause of developmental delay associated with recurrent crises involving rhabdomyolysis, cardiac arrhythmias, and metabolic derangements. The disease is not well understood, in part as the cellular function and subcellular localization of the TANGO2 protein remain unknown. Furthermore, the clinical syndrome with its heterogeneity of symptoms, signs, and laboratory findings is still being defined. Here, we describe 11 new cases of TANGO2-related disease, confirming and further expanding the previously described clinical phenotype. Patients were homozygous or compound heterozygous for previously described exonic deletions or new frameshift, splice site, and missense mutations. All patients showed developmental delay with ataxia, dysarthria, intellectual disability, or signs of spastic diplegia. Of importance, we identify two subjects (aged 12 and 17 years) who have never experienced any overt episode of the catabolism-induced metabolic crises typical for the disease. Mitochondrial complex II activity was mildly reduced in patients investigated in association with crises but normal in other patients. In one deceased patient, post-mortem autopsy revealed heterotopic neurons in the cerebral white matter, indicating a possible role for TANGO2 in neuronal migration. Furthermore, we have addressed the subcellular localization of several alternative isoforms of TANGO2, none of which were mitochondrial but instead appeared to have a primarily cytoplasmic localization. Previously described aberrations in Golgi morphology were not observed in cultured skin fibroblasts.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/deficiência , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Deficiências do Desenvolvimento/genética , Metabolismo Energético/genética , Deficiência Intelectual/genética , Mitocôndrias/genética , Adolescente , Translocador Nuclear Receptor Aril Hidrocarboneto/fisiologia , Ataxia/genética , Paralisia Cerebral/genética , Criança , Pré-Escolar , Disartria/genética , Exoma , Éxons , Feminino , Humanos , Masculino , Mutação , Linhagem , Fenótipo , Sequenciamento Completo do Exoma
17.
Clin Sci (Lond) ; 133(12): 1353-1365, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31189688

RESUMO

Pregnancies complicated by severe fetal growth restriction with abnormal umbilical artery Doppler velocimetry (FGRadv) are at substantial risk for adverse perinatal and long-term outcomes. Impaired angiogenesis of the placental vasculature in these pregnancies results in a sparse, poorly branched vascular tree, which structurally contributes to the abnormally elevated fetoplacental vascular resistance that is clinically manifested by absent or reversed umbilical artery Doppler indices. Previous studies have shown that aryl hydrocarbon receptor nuclear translocator (ARNT) is a key mediator of proper placental angiogenesis, and within placental endothelial cells (ECs) from human FGRadv pregnancies, low expression of ARNT leads to decreased vascular endothelial growth factor A (VEGFA) expression and deficient tube formation. Thus, the aim of the present study was to determine the effect of VEGFA administration or ARNT overexpression on angiogenic potential of FGRadv ECs. ECs were isolated and cultured from FGRadv or gestational age-matched control placentas and subjected to either vehicle vs VEGFA treatment or transduction with adenoviral-CMV (ad-CMV) vs adenoviral-ARNT (ad-ARNT) constructs. They were then assessed via wound scratch and tube formation assays. We found that VEGFA administration nominally improved FGRadv EC migration (P<0.01) and tube formation (P<0.05). ARNT overexpression led to significantly enhanced ARNT expression in FGRadv ECs (P<0.01), to a level similar to control ECs. Despite this, FGRadv EC migration (P<0.05) and tube formation (P<0.05) were still only partially rescued. This suggests that although ARNT does play a role in fetoplacental EC migration, other factors in addition to ARNT are likely also important in placental angiogenesis.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/metabolismo , Células Endoteliais/metabolismo , Retardo do Crescimento Fetal/metabolismo , Neovascularização Patológica , Placenta/irrigação sanguínea , Adulto , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Estudos de Casos e Controles , Movimento Celular , Células Cultivadas , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Feminino , Retardo do Crescimento Fetal/genética , Retardo do Crescimento Fetal/patologia , Retardo do Crescimento Fetal/fisiopatologia , Humanos , Gravidez , Transdução de Sinais , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologia
18.
Curr Eye Res ; 44(10): 1157-1169, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31090463

RESUMO

Aim: The aim of this study was to investigate the possible mechanisms of ocular damage induced by pinealectomy (PNX) and preeclampsia (PE), and to determine the cellular and molecular effects of melatonin treatment on oxidative stress, DNA damage, molecular chaperone responses, induction of apoptosis and angiogenesis in the fetal eye of both PNX and PNX+PE animals. Material and Methods: We analysed therapeutic potential of melatonin on fetal eye damage in PNX and PNX+PE animals using Malondialdehyde (MDA), Random Amplified Polymorphic DNA (RAPD), qRT-PCR and Western blot assays. Results: Our study presents three preliminary findings: (a) in fetal eye tissues, PNX and PNX+PE significantly induce oxidative damage to both DNA and protein contents, leading to a dramatic increase in caspase-dependent apoptotic signalling in both mitochondrial and death receptor pathways; (b) the same conditions trigger hypoxia biomarkers in addition to significant overexpression of HIF1-α, HIF1-ß, MMP9 and VEGF genes in the fetal eye; (c) finally, melatonin regulates not only the expression of genes encoding antioxidant enzymes and increase in DNA damage as well as lipid peroxidation but also limits programmed cell death processes in the fetal eye of PNX and PNX+PE animals . Furthermore, melatonin can relatively modulate genes in the HIF1 family, TNF-α and VEGF, thus acting as a direct anti-angiogenic molecule. In conclusion, both PNX and PNX+PE induce ocular damage at both cellular and molecular levels in fetal eye tissue of rats. Conclusion: Our results clearly indicate the potential of melatonin as a preventative therapeutic intervention for fetal ocular damage triggered by both PNX and PNX+PE.


Assuntos
Apoptose , Dano ao DNA , Olho/irrigação sanguínea , Melatonina/deficiência , Neovascularização Patológica/patologia , Estresse Oxidativo/fisiologia , Pré-Eclâmpsia/fisiopatologia , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Western Blotting , Olho/embriologia , Feminino , Regulação da Expressão Gênica/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Peroxidação de Lipídeos , Malondialdeído/metabolismo , Metaloproteinase 9 da Matriz/genética , Melatonina/fisiologia , Neovascularização Patológica/metabolismo , Pinealectomia , Gravidez , Técnica de Amplificação ao Acaso de DNA Polimórfico , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Fator A de Crescimento do Endotélio Vascular/genética
19.
Zygote ; 27(1): 49-53, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30714556

RESUMO

SummaryIn eutherian mammals, the placenta plays a critical role in embryo development by supplying nutrients and hormones and mediating interaction with the mother. To establish the fine connection between mother and embryo, the placenta needs to be formed normally, but the mechanism of placental differentiation is not fully understood. We previously revealed that mouse prolyl oligopeptidase (POP) plays a role in trophoblast stem cell (TSC) differentiation into two placental cell types, spongiotrophoblasts (SpT) and trophoblast giant cells. Here, we focused on SpT differentiation and attempted to elucidate a molecular mechanism. For Ascl2, Arnt, and Egfr genes that are indispensable for SpT formation, we found that a POP-specific inhibitor, SUAM-14746, significantly decreased Ascl2 expression, which was consistent with a significant decrease in expression of Flt1, a gene downstream of Ascl2. Although this downregulation was unlikely to be mediated by the PI3K-Akt pathway, our results indicated that POP controls TSC differentiation into SpT by regulating the Ascl2 gene.


Assuntos
Placenta/citologia , Serina Endopeptidases/genética , Trofoblastos/citologia , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Diferenciação Celular/genética , Receptores ErbB/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Camundongos , Gravidez , Prolina/análogos & derivados , Prolina/farmacologia , Serina Endopeptidases/metabolismo , Inibidores de Serino Proteinase/farmacologia , Tiazolidinas/farmacologia , Trofoblastos/fisiologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genética
20.
Neuropediatrics ; 50(2): 122-125, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30650451

RESUMO

CASE: We report a 15-year-old Indian girl born to a consanguineous couple, who presented with epilepsy, developmental delay, neuroregression, and episodes of alternating hemiparesis. In addition, she had one episode of rhabdomyolysis at the age of 7 years. Extensive genetic and metabolic work up through the years was unrevealing. Eventually a trio whole exome sequencing (WES) revealed homozygous single nucleotide variants in TANGO2 gene. DISCUSSION: TANGO2 related recurrent metabolic crises with encephalomyopathy and cardiac arrhythmias were described very recently and only 15 cases were reported in literature at the time of writing. Alternating hemiplegia of childhood which was seen in our patient, has not been described in previous patients with TANGO2 mutation, and thereby expands the emerging phenotypic spectrum of this novel entity. This report also reiterates the utility of WES in diagnosing newly recognized neurogenetic conditions.


Assuntos
Translocador Nuclear Receptor Aril Hidrocarboneto/genética , Variação Genética/genética , Hemiplegia/diagnóstico por imagem , Hemiplegia/genética , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Adolescente , Feminino , Homozigoto , Humanos
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