RESUMO
In this study, we aimed to identify novel genes involved in experimental and human asthma, importance of which has not yet been recognized. In an ovalbumin-induced murine model of asthma, we applied microarray gene expression analysis at different time points after allergen challenges. Advanced statistical methods were used to relate gene expression changes to cellular processes and to integrate our results into multiple levels of information available in public databases. At 4 h after the first allergen challenge, gene expression pattern reflected mainly an acute, but non-atopic, inflammatory response and strong chemotactic activity. At 24 h after the third allergen challenge, gene set enrichment analysis revealed significant over-representation of gene sets corresponding to T(h)2-type inflammation models. Among the top down-regulated transcripts, an anti-oxidant enzyme, paraoxonase-1 (PON1), was identified. In human asthmatic patients, we found that serum PON1 activity was reduced at exacerbation, but increased parallel with improving asthma symptoms. PON1 gene polymorphisms did not influence the susceptibility to the disease. Our observations suggest that an altered PON1 activity might be involved in the pathogenesis of asthma, and serum PON1 level might be used for following up the effect of therapy.
Assuntos
Arildialquilfosfatase/fisiologia , Asma/genética , Perfilação da Expressão Gênica , Animais , Arildialquilfosfatase/genética , Modelos Animais de Doenças , Regulação para Baixo , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Helminthic infections, which are particularly common in the developing world, are associated with the accumulation of mucosal mast cells (MMCs) in the epithelial layer of the gut. Although intestinal parasite infection models argue that IL-18 plays a role in MMC differentiation and function, the direct effect of IL-18 on MMCs is still not well understood. To clarify the role of IL-18 in mast cell biology, we analyzed gene expression changes in MMCs in vitro. DNA microarray technology uncovered a group of chemokines regulated by IL-18, among which Ccl1 (I-309, TCA-3) showed the highest up-regulation. Ccl1 induction was only transient in mast cells and was characteristic for both immature and mature MMCs, but not for connective tissue-type mast cells. IL-18 exerts its Ccl1-inducing effect in MMCs primarily via the activation of NFkappaB. Moreover, IL-18 was effective both in the absence and the presence of IgE-antigen complex. The Ccl1 receptor (CCR8) is known to be expressed by T(h)2 cells and is involved in their recruitment. Our present findings suggest that IL-18 may contribute to mast cell-influenced Th2 responses by inducing Ccl1 production.
Assuntos
Quimiocina CCL1/metabolismo , Interleucina-18/metabolismo , Mastócitos/metabolismo , Células Th1/metabolismo , Células Th2/metabolismo , Animais , Quimiocina CCL1/antagonistas & inibidores , Quimiocina CCL1/genética , Dinitrofenóis/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Imunidade nas Mucosas , Interleucina-18/genética , Interleucina-18/imunologia , Interleucina-18/farmacologia , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Análise em Microsséries , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Albumina Sérica/metabolismo , Transdução de SinaisRESUMO
One main area of pharmacogenomics is the discovery of new drugs and drug targets with molecular genetic or genomic methods; the other is the study of how genomic differences influence the variability in patients responses to drugs. In this review the authors summarise the most important results of this latter issue. Despite the availability of three major classes of therapeutic agents for asthma, it has been estimated that as many as half of asthmatic patients do not respond to treatment with beta2-agonists, leukotriene modifiers or inhaled corticosteroids. Moreover, in some individuals asthma therapy has been associated with serious adverse drug reactions. An estimated 60 to 80% of variability in individual responses to therapy may have genetic bases. All of the currently available data on asthma pharmacogenomics originated from genetic variations in genes of the drug treatment target or target pathways. Results of genetic association studies that investigate responses to beta2-agonist, leukotriene modifier and corticosteroid therapy will be summarised and recent findings in the literature highlighted. Although, at present pharmacogenomics can explain only a fraction of the adverse drug responses, hopefully these results mark the clinical use of genotyping at an individual level as adjunct to pharmacotherapy for asthma and many other diseases.
Assuntos
Antiasmáticos/farmacologia , Antiasmáticos/uso terapêutico , Asma/tratamento farmacológico , Administração por Inalação , Agonistas de Receptores Adrenérgicos beta 2 , Agonistas Adrenérgicos beta/farmacologia , Antiasmáticos/efeitos adversos , Asma/metabolismo , Desenho de Fármacos , Variação Genética , Glucocorticoides/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Humanos , Antagonistas de Leucotrienos/farmacologia , Farmacogenética , Polimorfismo de Nucleotídeo ÚnicoRESUMO
MicroRNAs (miRs) are involved in the pathogenesis of several neoplasms; however, there are no data on their expression patterns and possible roles in adrenocortical tumors. Our objective was to study adrenocortical tumors by an integrative bioinformatics analysis involving miR and transcriptomics profiling, pathway analysis, and a novel, tissue-specific miR target prediction approach. Thirty-six tissue samples including normal adrenocortical tissues, benign adenomas, and adrenocortical carcinomas (ACC) were studied by simultaneous miR and mRNA profiling. A novel data-processing software was used to identify all predicted miR-mRNA interactions retrieved from PicTar, TargetScan, and miRBase. Tissue-specific target prediction was achieved by filtering out mRNAs with undetectable expression and searching for mRNA targets with inverse expression alterations as their regulatory miRs. Target sets and significant microarray data were subjected to Ingenuity Pathway Analysis. Six miRs with significantly different expression were found. miR-184 and miR-503 showed significantly higher, whereas miR-511 and miR-214 showed significantly lower expression in ACCs than in other groups. Expression of miR-210 was significantly lower in cortisol-secreting adenomas than in ACCs. By calculating the difference between dCT(miR-511) and dCT(miR-503) (delta cycle threshold), ACCs could be distinguished from benign adenomas with high sensitivity and specificity. Pathway analysis revealed the possible involvement of G2/M checkpoint damage in ACC pathogenesis. To our knowledge, this is the first report describing miR expression patterns and pathway analysis in sporadic adrenocortical tumors. miR biomarkers may be helpful for the diagnosis of adrenocortical malignancy. This tissue-specific target prediction approach may be used in other tumors too.
Assuntos
Adenoma/genética , Neoplasias do Córtex Suprarrenal/genética , Carcinoma Adrenocortical/genética , Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , MicroRNAs/genética , Transdução de Sinais/genética , Adenoma/diagnóstico , Neoplasias do Córtex Suprarrenal/diagnóstico , Carcinoma Adrenocortical/diagnóstico , Adulto , Algoritmos , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/genética , Sistemas de Liberação de Medicamentos/métodos , Feminino , Previsões/métodos , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Pessoa de Meia-Idade , Modelos Biológicos , Especificidade de Órgãos/genéticaRESUMO
This study was undertaken to investigate the association of Chlamydophila pneumoniae infection (CPI) with asthma and allergy. One hundred forty-one patients with asthma aged 3-21 years, 125 healthy controls aged 3-21 years, and 62 allergic but nonasthmatic patients aged 4-20 years participated in this study. C. pneumoniae-specific antibodies were measured by ELISA. There were no significant differences in the percentage of patients positive for C. pneumoniae-specific antibodies between the three groups. Significantly more allergic asthmatic patients were positive for C. pneumoniae-specific IgA and IgA + IgG than nonallergic asthmatic patients, and this difference remained significant after adjustment for age and gender: adjusted odds ratio (OR) = 5.9 (1.7-26.2) and p = 0.01 for IgA, and OR = 5.2 (1.6-25.8) and p = 0.02 for IgA + IgG. The prevalence of the C. pneumoniae-specific IgA and the IgA + IgG positivity also was significantly lower in the nonallergic asthmatic group than in the allergic and control groups (p < 0.005). No food/drug-allergic patient was positive for C. pneumoniae-specific IgA, whereas 41.6% of the inhalative-allergic patients were positive for this antibody (p = 0.002). In our population CPI does not associate directly with asthma and allergy, but chronic or recurrent infection is associated with allergic asthma and inhalative allergy as opposed to nonallergic asthma and noninhalative allergy.
Assuntos
Anticorpos Antibacterianos/sangue , Asma/complicações , Infecções por Chlamydophila/complicações , Chlamydophila pneumoniae/imunologia , Hipersensibilidade/complicações , Pneumonia Bacteriana/complicações , Adolescente , Adulto , Antiasmáticos/uso terapêutico , Asma/tratamento farmacológico , Asma/epidemiologia , Asma/imunologia , Asma/microbiologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Infecções por Chlamydophila/epidemiologia , Infecções por Chlamydophila/imunologia , Infecções por Chlamydophila/microbiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Hungria/epidemiologia , Hipersensibilidade/epidemiologia , Hipersensibilidade/imunologia , Hipersensibilidade/microbiologia , Masculino , Razão de Chances , Pneumonia Bacteriana/epidemiologia , Pneumonia Bacteriana/imunologia , Pneumonia Bacteriana/microbiologia , Prevalência , Recidiva , Estudos Retrospectivos , Índice de Gravidade de DoençaRESUMO
Many studies detect elevated numbers of mast cells in tumors, but it is still controversial whether they are beneficial or detrimental for tumor cells. Furthermore, many tumors, such as melanomas, produce large quantities of transforming growth factor (TGF)-beta and during tumorigenesis the apoptotic and growth-inhibitory effects of TGF-betas are lost. Based on these data we investigated the gene expression changes in TGF-betaI-treated human mast cells with DNA microarray and detected 45 differentially regulated genes, among them T-cell immunoglobulin and mucin domain-containing protein 3 (TIM-3). As the major sources of TIM-3 ligand galectin-9 are not tumor cells, but rather mast cells, this raises the possibility of an autocrine mechanism resulting in local immunosuppression through the elevated TIM-3 expression by TGF-betaI. Interestingly, not only melanoma tissue sections contained TIM-3-positive mast cells, but we detected this protein also in melanoma cells. Furthermore, TIM-3 was expressed in both WM35 and HT168-M1 melanoma cell lines at a higher level than in isolated epidermal melanocytes, which can contribute to the lower adhering capacity of tumor cells. In conclusion, the immunoregulatory molecule TIM-3 in TGF-beta-stimulated mast cells and melanoma cells may support the survival of this tumor type.
Assuntos
Mastócitos/metabolismo , Melanoma/metabolismo , Receptores Virais/metabolismo , Neoplasias Cutâneas/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Regulação para Cima , Células Cultivadas , Epiderme/metabolismo , Galectinas/metabolismo , Receptor Celular 2 do Vírus da Hepatite A , Humanos , Ligantes , Mastócitos/efeitos dos fármacos , Mastócitos/patologia , Melanócitos/metabolismo , Melanoma/patologia , Proteínas de Membrana , Análise de Sequência com Séries de Oligonucleotídeos , Reprodutibilidade dos Testes , Neoplasias Cutâneas/patologiaRESUMO
Several data indicate a connection between Chlamydophila pneumoniae infection and asthma. Although C. pneumoniae is a common cause of infection, not all infected patients develop asthma. This suggests that certain individuals may be genetically predisposed to the chronic effects of C. pneumoniae infection on airway functions. We investigated the possible modifying effect of different polymorphisms on C. pneumoniae infection and on the susceptibility to asthma in 318 children, among those 144 had asthma and 174 had no asthmatic symptoms. C. pneumoniae-specific antibodies were measured by ELISA. Tumor necrosis factor-alpha (TNFalpha), monocyte chemoattractant protein-1 (MCP-1), and RANTES (regulated on activation normal T cell expressed and secreted) genotypes were determined by PCR-restriction fragment length polymorphism (RFLP). There were no significant differences in the percentage of children positive for C. pneumoniae-specific antibodies between cases and controls. None of the genotypes was associated with altered susceptibility to C. pneumoniae infection. Among asthmatic children carrying the TNFalpha -308A allele, there were significantly more patients positive for C. pneumoniae-specific IgG, than among control children carrying the same allele (20.1% versus 9.2% of asthmatic versus control children, respectively; p = 0.002; odds ratio = 3.52 (1.52-7.53); p = 0.005). This study indicates the possible roles of polymorphisms in the immune system in the susceptibility to asthma in children infected with C. pneumoniae.
Assuntos
Asma , Infecções por Chlamydophila , Chlamydophila pneumoniae/imunologia , Polimorfismo Genético , Regiões Promotoras Genéticas , Fator de Necrose Tumoral alfa/genética , Adolescente , Anticorpos/sangue , Asma/etiologia , Asma/genética , Asma/imunologia , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Criança , Pré-Escolar , Infecções por Chlamydophila/complicações , Infecções por Chlamydophila/imunologia , Suscetibilidade a Doenças , Feminino , Genótipo , Humanos , Masculino , Análise de Regressão , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Serum levels of MCP-1 were measured in children with and without asthma in order to determine a possible correlation between the MCP-1-2518A/G polymorphism, serum levels of MCP-1 and asthma. Two groups of subjects -160 children with asthma and 158 healthy children were screened with a PCR-based genotyping assay. Serum MCP-1 level was measured by ELISA. The -2518G allele occurred at a significantly higher frequency in asthmatic children than in controls. The mean serum MCP-1 level was significantly lower in the asthmatic than in the control children. There was no significant association between the MCP-1 genotypes and the serum MCP-1 levels.
Assuntos
Asma/sangue , Quimiocina CCL2/sangue , Adolescente , Asma/complicações , Quimiocina CCL2/genética , Criança , Pré-Escolar , Infecções por Chlamydophila/complicações , Ensaio de Imunoadsorção Enzimática , Feminino , Glucocorticoides/uso terapêutico , Humanos , Masculino , Polimorfismo Genético , Índice de Gravidade de DoençaRESUMO
Very little is known about the effect of vitrification on gene functions after warming. The goals of our study were to compare the gene expression patterns, and identify those most affected. For this, 8-cell stage embryos were collected from ICR mice and vitrified with solid surface vitrification technique, while maintaining equal numbers of embryos as control. Total RNAs were extracted and two rounds of amplification were employed. Finally three micrograms of contrasting RNA samples were hybridized on the Agilent Mouse 22 K oligonucleotide slides and the results were analyzed with subsequent verification by independent real-time PCR analyses. The two rounds of amplification with 5 ng tRNA input have yielded 15-16 microg of cRNA. The analyses of repeated hybridizations showed 20,183 genes/ESTs as common signatures, and unsupervised analysis identified 628 differentially expressed (P < 0.01) genes. However, with at least 1.5-fold change considerations, 183 genes were differentially expressed (P < 0.01) out of which 107 were upregulated. The independent analysis with real-time PCR and unamplified samples fully confirmed the results of microarray, indicating the linearity of amplification. Furthermore, this novel gene expression study for vitrified embryos identified many new candidate genes with overrepresentation in some important biological processes. Thus, it is possible to conclude that the expression pattern reflected a broad spectrum of consequences of vitrification on embryos, with most effects on metabolism, regulatory role and stress response genes and allowed the identification of new candidate marker genes for cryosurvival.