RESUMO
BACKGROUND: There is a spectacular rise in the global prevalence of type 2 diabetes mellitus (T2DM) due to the worldwide obesity epidemic. However, a significant proportion of T2DM patients are non-obese and they also have an increased risk of cardiovascular diseases. As the Goto-Kakizaki (GK) rat is a well-known model of non-obese T2DM, the goal of this study was to investigate the effect of non-obese T2DM on cardiac alterations of the transcriptome in GK rats. METHODS: Fasting blood glucose, serum insulin and cholesterol levels were measured at 7, 11, and 15 weeks of age in male GK and control rats. Oral glucose tolerance test and pancreatic insulin level measurements were performed at 11 weeks of age. At week 15, total RNA was isolated from the myocardium and assayed by rat oligonucleotide microarray for 41,012 genes, and then expression of selected genes was confirmed by qRT-PCR. Gene ontology and protein-protein network analyses were performed to demonstrate potentially characteristic gene alterations and key genes in non-obese T2DM. RESULTS: Fasting blood glucose, serum insulin and cholesterol levels were significantly increased, glucose tolerance and insulin sensitivity were significantly impaired in GK rats as compared to controls. In hearts of GK rats, 204 genes showed significant up-regulation and 303 genes showed down-regulation as compared to controls according to microarray analysis. Genes with significantly altered expression in the heart due to non-obese T2DM includes functional clusters of metabolism (e.g. Cyp2e1, Akr1b10), signal transduction (e.g. Dpp4, Stat3), receptors and ion channels (e.g. Sln, Chrng), membrane and structural proteins (e.g. Tnni1, Mylk2, Col8a1, Adam33), cell growth and differentiation (e.g. Gpc3, Jund), immune response (e.g. C3, C4a), and others (e.g. Lrp8, Msln, Klkc1, Epn3). Gene ontology analysis revealed several significantly enriched functional inter-relationships between genes influenced by non-obese T2DM. Protein-protein interaction analysis demonstrated that Stat is a potential key gene influenced by non-obese T2DM. CONCLUSIONS: Non-obese T2DM alters cardiac gene expression profile. The altered genes may be involved in the development of cardiac pathologies and could be potential therapeutic targets in non-obese T2DM.
Assuntos
Glicemia/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Regulação da Expressão Gênica/fisiologia , Expressão Gênica/fisiologia , Miocárdio/metabolismo , Transcrição Gênica/fisiologia , Transcriptoma , Animais , Coração/fisiopatologia , Masculino , Mesotelina , Síndrome Metabólica/metabolismo , RatosRESUMO
BACKGROUND: Diabetic patients have an increased risk of developing cardiovascular diseases, which are the leading cause of death in developed countries. Although multivitamin products are widely used as dietary supplements, the effects of these products have not been investigated in the diabetic heart yet. Therefore, here we investigated if a preparation of different minerals, vitamins, and trace elements (MVT) affects the cardiac gene expression pattern in experimental diabetes. METHODS: Two-day old male Wistar rats were injected with streptozotocin (i.p. 100 mg/kg) or citrate buffer to induce diabetes. From weeks 4 to 12, rats were fed with a vehicle or a MVT preparation. Fasting blood glucose measurement and oral glucose tolerance test were performed at week 12, and then total RNA was isolated from the myocardium and assayed by rat oligonucleotide microarray for 41012 oligonucleotides. RESULTS: Significantly elevated fasting blood glucose concentration and impaired glucose tolerance were markedly improved by MVT-treatment in diabetic rats at week 12. Genes with significantly altered expression due to diabetes include functional clusters related to cardiac hypertrophy (e.g. caspase recruitment domain family, member 9; cytochrome P450, family 26, subfamily B, polypeptide; FXYD domain containing ion transport regulator 3), stress response (e.g. metallothionein 1a; metallothionein 2a; interleukin-6 receptor; heme oxygenase (decycling) 1; and glutathione S-transferase, theta 3), and hormones associated with insulin resistance (e.g. resistin; FK506 binding protein 5; galanin/GMAP prepropeptide). Moreover the expression of some other genes with no definite cardiac function was also changed such as e.g. similar to apolipoprotein L2; brain expressed X-linked 1; prostaglandin b2 synthase (brain). MVT-treatment in diabetic rats showed opposite gene expression changes in the cases of 19 genes associated with diabetic cardiomyopathy. In healthy hearts, MVT-treatment resulted in cardiac gene expression changes mostly related to immune response (e.g. complement factor B; complement component 4a; interferon regulatory factor 7; hepcidin). CONCLUSIONS: MVT-treatment improved diagnostic markers of diabetes. This is the first demonstration that MVT-treatment significantly alters cardiac gene expression profile in both control and diabetic rats. Our results and further studies exploring the mechanistic role of individual genes may contribute to the prevention or diagnosis of cardiac complications in diabetes.
Assuntos
Diabetes Mellitus Experimental/genética , Cardiomiopatias Diabéticas/genética , Coração/efeitos dos fármacos , Minerais/farmacologia , Miocárdio/metabolismo , RNA Mensageiro/metabolismo , Oligoelementos/farmacologia , Transcriptoma/efeitos dos fármacos , Vitaminas/farmacologia , Animais , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Under physiological and pathological conditions, extracellular vesicles (EVs) are present in the extracellular compartment simultaneously with soluble mediators. We hypothesized that cytokine effects may be modulated by EVs, the recently recognized conveyors of intercellular messages. In order to test this hypothesis, human monocyte cells were incubated with CCRF acute lymphoblastic leukemia cell line-derived EVs with or without the addition of recombinant human TNF, and global gene expression changes were analyzed. EVs alone regulated the expression of numerous genes related to inflammation and signaling. In combination, the effects of EVs and TNF were additive, antagonistic, or independent. The differential effects of EVs and TNF or their simultaneous presence were also validated by Taqman assays and ELISA, and by testing different populations of purified EVs. In the case of the paramount chemokine IL-8, we were able to demonstrate a synergistic upregulation by purified EVs and TNF. Our data suggest that neglecting the modulating role of EVs on the effects of soluble mediators may skew experimental results. On the other hand, considering the combined effects of cytokines and EVs may prove therapeutically useful by targeting both compartments at the same time.
Assuntos
Citocinas/metabolismo , Exossomos/metabolismo , Linhagem Celular Tumoral , Quimiocina CCL2/metabolismo , Análise por Conglomerados , Citocinas/genética , Humanos , Interleucina-8/genética , Interleucina-8/metabolismo , RNA Mensageiro/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Regulação para Cima/efeitos dos fármacosRESUMO
The currently available medical treatment options of adrenocortical cancer (ACC) are limited. In our previous meta-analysis of adrenocortical tumor genomics data, ACC was associated with reduced retinoic acid production and retinoid X receptor-mediated signaling. Our objective has been to study the potential antitumoral effects of 9-cis retinoic acid (9-cisRA) on the ACC cell line NCI-H295R and in a xenograft model. Cell proliferation, hormone secretion, and gene expression have been studied in the NCI-H295R cell line. A complex bioinformatics approach involving pathway and network analysis has been performed. Selected genes have been validated by real-time qRT-PCR. Athymic nude mice xenografted with NCI-H295R have been used in a pilot in vivo xenograft model. 9-cisRA significantly decreased cell viability and steroid hormone secretion in a concentration- and time-dependent manner in the NCI-H295R cell line. Four major molecular pathways have been identified by the analysis of gene expression data. Ten genes have been successfully validated involved in: (1) steroid hormone secretion (HSD3B1, HSD3B2), (2) retinoic acid signaling (ABCA1, ABCG1, HMGCR), (3) cell-cycle damage (GADD45A, CCNE2, UHRF1), and the (4) immune response (MAP2K6, IL1R2). 9-cisRA appears to directly regulate the cell cycle by network analysis. 9-cisRA also reduced tumor growth in the in vivo xenograft model. In conclusion, 9-cisRA might represent a promising new candidate in the treatment of hormone-secreting adrenal tumors and adrenocortical cancer.
Assuntos
Neoplasias do Córtex Suprarrenal/tratamento farmacológico , Antineoplásicos/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Tretinoína/farmacologia , Alitretinoína , Animais , Antineoplásicos/uso terapêutico , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Biologia Computacional/métodos , Regulação Neoplásica da Expressão Gênica/imunologia , Regulação Neoplásica da Expressão Gênica/fisiologia , Hormônios Esteroides Gonadais/metabolismo , Humanos , Camundongos , Camundongos Nus , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/fisiologia , Tretinoína/uso terapêutico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Candida parapsilosis is a human fungal pathogen with increasing global significance. Understanding how macrophages respond to C. parapsilosis at the molecular level will facilitate the development of novel therapeutic paradigms. The complex response of murine macrophages to infection with C. parapsilosis was investigated at the level of gene expression using an Agilent mouse microarray. We identified 155 and 511 differentially regulated genes at 3 and 8h post-infection, respectively. Most of the upregulated genes encoded molecules involved in immune response and inflammation, transcription, signaling, apoptosis, cell cycle, electron transport and cell adhesion. Typical of the classically activated macrophages, there was significant upregulation of genes coordinating the production of inflammatory cytokines such as TNF, IL-1 and IL-15. Further, we used both primary murine macrophages and macrophages differentiated from human peripheral mononuclear cells to confirm the upregulation of the TNF-receptor family member TNFRSF9 that is associated with Th1 T-helper cell responses. Additionally, the microarray data indicate significant differences between the response to C. parapsilosis infection and that of C. albicans.
Assuntos
Candida/fisiologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Transcriptoma , Animais , Células Cultivadas , Humanos , Macrófagos/patologia , Camundongos , Fagocitose , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/genética , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/metabolismoRESUMO
UNLABELLED: Whole-body polarized light therapy has been primarily investigated in various clinical observations and in a few in vitro model systems. AIMS: In the present study, clinical and molecular effects of whole-body polarized light treatment on children suffering from recurrent respiratory infection were studied. METHODS: Incidence and duration of respiratory symptoms as well as the length of appropriate antibiotic therapy have been measured. Simultaneously, genome-wide gene expression pattern was examined by whole genome cDNA microarray in peripheral lymphocytes of children. RESULTS: Twenty of twenty five children showed a marked clinical improvement, while in five of twenty five had poor or no changes. Gene expression pattern of the peripheral lymphocytes of the patients was compared in favorable and poor responders. Lymphocytes of the children with a documented improved clinical response to polarized light therapy showed a decrease in the expression of chemokine genes, such as CXCL1, CXCL2, IL-8 and in that of the tumor necrosis alpha (TNFα) gene. On the contrary, a rapid elevation was found in the expression of gene encoding for CYP4F2, a leukotriene-B(4)-metabolizing enzyme. In children with poor clinical response to polarized light therapy, no similar changes were detected in the gene expression pattern of the lymphocytes. CONCLUSIONS: Improved clinical symptoms and modified gene expression profile of lymphocytes reveals anti-inflammatory effect upon whole body polarized light irradiation.
Assuntos
Epigênese Genética , Perfilação da Expressão Gênica , Fototerapia , Infecções Respiratórias/metabolismo , Infecções Respiratórias/terapia , Estações do Ano , Quimiocinas/metabolismo , Criança , Pré-Escolar , Doença Crônica , Citocinas/metabolismo , Feminino , Estudo de Associação Genômica Ampla , Humanos , Lactente , Inflamação/terapia , Masculino , Fototerapia/métodos , Recidiva , Infecções Respiratórias/genética , Infecções Respiratórias/imunologia , Infecções Respiratórias/prevenção & controle , Infecções Respiratórias/transmissãoRESUMO
MicroRNAs are involved in the pathogenesis of several tumors, however, there have been no data on microRNA expression in pheochromocytomas to date. The objective of our study was to perform microRNA expression profiling in sporadic and hereditary benign, and recurring adrenomedullary tumors. Furthermore, the applicability of formalin-fixed paraffin-embedded tissue samples for the analysis of microRNA expression in pheochromocytomas was examined. MicroRNA expression data of three matched frozen and formalin-fixed paraffin-embedded samples were correlated. A total of 21 formalin-fixed paraffin-embedded samples (sporadic benign, multiple endocrine neoplasia 2, von Hippel-Lindau disease, sporadic recurring) were subjected to microRNA expression profiling using microarrays. MicroRNAs with significant differences in expression were validated and sample sizes were extended including tumors from neurofibromatosis type 1 patients by real-time quantitative reverse-transcription PCR (n=33). MicroRNA target prediction was carried out by TargetScan and MicroCosm Targets. Pathway analysis of targets was performed by Ingenuity Pathway Analysis and DIANA mirPath. Furthermore, microRNA expression profiles of a malignant pheochromocytoma and a pair of primary and recurrent tumors were studied by TaqMan Human MicroRNA Cards. MicroRNA expression correlated well between frozen and formalin-fixed paraffin-embedded samples (70-92%). Microarray analysis revealed 16 significantly differentially expressed microRNAs. Five of these were validated by real-time RT-PCR. miR-139-3p, miR-541 and miR-765 were significantly differentially expressed between sporadic benign and von Hippel-Lindau-related pheochromocytomas. Significantly higher expression of miR-885-5p and miR-1225-3p was found in multiple endocrine neoplasia type 2 and sporadic recurring pheochromocytomas, respectively. Pathway analysis revealed the possible involvement of Notch- and G-protein-coupled receptor signaling in tumor recurrence. MicroRNA expression profiles in the primary recurrent and recurring malignant comparisons have been similar. In conclusion, we have proved that formalin-fixed paraffin-embedded samples can be used for the analysis of microRNA expression in pheochromocytomas. MicroRNA expression patterns differ between various sporadic, hereditary and recurring tumors and miR-1225-3p may be useful for identifying recurring pheochromocytomas.
Assuntos
Neoplasias das Glândulas Suprarrenais/genética , Perfilação da Expressão Gênica , MicroRNAs/genética , Feocromocitoma/genética , Adulto , Análise por Conglomerados , Feminino , Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Pessoa de Meia-Idade , Neoplasia Endócrina Múltipla Tipo 2a/complicações , Neoplasia Endócrina Múltipla Tipo 2a/genética , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Doença de von Hippel-Lindau/complicações , Doença de von Hippel-Lindau/genéticaRESUMO
Lipopolysaccharide (LPS) is the main component of Gram-negative bacteria that - upon infection - activates the host immune system and is crucial in fighting pathogens as well as in the induction of sepsis. In the present study we addressed the question whether the key structural components of LPS equally take part in the activation of different macrophage immune responses. By genomic modifications of Escherichia coli MG1655, we constructed a series of strains harboring complete and truncated forms of LPS in their cell wall. These strains were exposed to RAW 264.7 macrophages, after which phagocytosis, fast release of pre-synthesized TNF and activation of NF-kappaB signal transduction pathway were quantified. According to our results the core and lipid A moieties are involved in immune recognition. The most ancient part, lipid A is crucial in evoking immediate TNF release and activation of NF-kappaB. The O-antigen inhibits phagocytosis, leading to immune evasion.
Assuntos
Escherichia coli/imunologia , Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Animais , Linhagem Celular , Escherichia coli/genética , Genoma Bacteriano , Lipídeo A/genética , Lipídeo A/imunologia , Lipopolissacarídeos/genética , Macrófagos/microbiologia , Camundongos , NF-kappa B/biossíntese , Antígenos O/genética , Antígenos O/imunologia , Fagocitose/imunologia , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Inflammatory activation of monocytes is an essential part of both innate immune responses and the pathogenesis of conditions such as atherosclerosis. However, the mechanisms which modulate the response of monocytes to inflammatory stimuli are still poorly understood. Here, we report that tribbles-2 (trb-2) is a novel regulator of inflammatory activation of monocytes. Down-regulation of trb-2 levels potentiates LPS-induced IL-8 production via enhanced activation of the extracellular signal-regulated kinase and jun kinase mitogen-activated protein kinase (MAPK) pathways. In keeping with this, the endogenous level of trb-2 expression in human primary monocytes is inversely correlated to the cell's ability to produce IL-8. We show that trb-2 is a binding partner and a negative regulator of selected MAPKs. The potential in vivo relevance of these findings is highlighted by the observation that modified low-density lipoprotein profoundly down-regulates trb-2 expression, which may, in turn, significantly contribute to the inflammatory processes in the development of vascular disease. Taken together, our results define trb-2 as a potent novel regulator of monocyte biology, controlling the activation of these cells.
Assuntos
Interleucina-8/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lipoproteínas LDL/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Monócitos/metabolismo , Aterosclerose/etiologia , Proteínas Quinases Dependentes de Cálcio-Calmodulina , Linhagem Celular , Células Cultivadas , Regulação da Expressão Gênica , Humanos , Imunidade Inata , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Lipoproteínas LDL/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Monócitos/citologia , Ligação Proteica , RNA Interferente Pequeno/genética , Transdução de Sinais/imunologiaRESUMO
Adipose tissue is one of the main sources of inflammatory mediators, with interleukin-6 (IL-6) among them. Although high systemic levels of inflammatory mediators are cachectogenic and/or anorexic, today it is a widely propagated thesis that in the background of obesity, a low level of chronic inflammation can be found, with IL-6 being one of the many suggested mediators. This paper reviews the studies describing elevated IL-6 levels in obese patients and the role of adipocytes and adipose-tissue macrophages in the production of IL-6. The secretion of IL-6 is regulated by several physiologic or pathologic factors: hormones, cytokines, diet, physical activity, stress, hypoxia, and others. Adipose tissue-derived IL-6 may have an effect on metabolism through several mechanisms, including adipose tissue-specific gene expression, triglyceride release, lipoprotein lipase downregulation, insulin sensitivity, and so on. Having a better understanding of these mechanisms may contribute to the prevention and treatment of obesity.
Assuntos
Tecido Adiposo/metabolismo , Interleucina-6 , Obesidade/imunologia , Adipócitos/metabolismo , Aterosclerose/metabolismo , Dieta , Exercício Físico , Humanos , Hipóxia/metabolismo , Inflamação/imunologia , Resistência à Insulina , Interleucina-6/sangue , Interleucina-6/imunologiaRESUMO
Inflammatory activation of monocytes is a hallmark event in cardiovascular disease. Activated monocytes migrate into atherosclerotic lesions, differentiate into macrophages and ingest lipids to become foam cells. These, in turn, through interaction with other inflammatory cell types contribute to plaque instability and are thought to play a key role in the development of acute coronary syndromes. In the current manuscript we investigated whether inflammatory activation of monocyte THP-1 cells influences their ability to take-up chemically modified LDL. We have also studied whether tribbles proteins, which have been shown to regulate the activation of inflammatory signal processing networks, have a modulatory role in the uptake of modified LDL by monocyte. Here, we show that activation of THP-1 cells by LPS potentiates LDL uptake. The greatest effect of LPS was seen after 16 h, compared to acute stimulation. Specific MAPK pathways are involved in this potentiation. Inhibition of both the p38 and ERK pathways led to reduced LPS uptake, specifically in LPS stimulated cells. Expression of tribbles, regulators of MAPK signalling, was dynamically modulated by LPS activation. However, neither suppression of tribbles expression by transient transfection of specific siRNAs nor transient overexpression of these proteins led to changes in the capacity of THP-1 cells to take up modified LDL. Therefore, we conclude that LPS potentiation of LDL uptake of THP-1 cells is MAPK dependent but is not mediated by tribbles.
Assuntos
LDL-Colesterol/metabolismo , Regulação da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Monócitos/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina , Proteínas de Ciclo Celular/biossíntese , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Regulação para Baixo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/imunologia , Inflamação/patologia , Peptídeos e Proteínas de Sinalização Intracelular/deficiência , Lipopolissacarídeos/farmacologia , Monócitos/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Proteínas Repressoras/biossíntese , Proteínas Repressoras/genéticaRESUMO
Cholestasis predisposes to hypersensitivity to LPS, leading to potential septic complications. We set out to characterize the involvement of Kupffer cell (KC) activation in the hepatic microcirculatory and structural consequences of obstructive jaundice in the presence and absence of acute endotoxemia. The hepatic microcirculatory consequences of 3-day extrahepatic bile duct ligation (BDL) were assessed in rats. The contributions of changes in hepatic perfusion, leukocyte influx, and proinflammatory cytokine release to the development of hepatic structural damage were also determined. Furthermore, the corresponding consequences of BDL in combination with acute (2-h) endotoxemia (1 mg kg(-1) LPS, i.v.) were compared with those observed after LPS alone. In a second series, the same protocols were applied in identical groups of rats where the KC function was inhibited with 24-h gadolinium chloride pretreatment (10 mg kg(-1), i.v.). Bile duct ligation induced minor inflammatory reactions but caused a marked reduction in hepatic sinusoidal perfusion and severe histological damage. LPS treatment, however, elicited an approximately 5-fold increase in leukocyte adherence in the central venules and pronounced IL-6 and TNF-alpha release, but without significant structural damage. The combination of BDL with LPS enhanced the perfusion failure, leukocyte sticking/deposition, and proinflammatory cytokine release; most of these changes can be effectively ameliorated by gadolinium chloride. In conclusion, when obstructive jaundice is followed by a second hit of LPS, perfusion failure, liver inflammation, and structural damage are enhanced, the KCs playing a decisive role in this scenario. Therapeutic strategies aimed at KC blockade can potentially reduce the risk of inflammatory complications in cholestasis.
Assuntos
Endotoxemia/fisiopatologia , Icterícia Obstrutiva/complicações , Células de Kupffer/efeitos dos fármacos , Microcirculação/efeitos dos fármacos , Animais , Ductos Biliares , Gadolínio/farmacologia , Interleucina-6/sangue , Icterícia Obstrutiva/tratamento farmacológico , Ligadura , Fígado/patologia , Masculino , Microcirculação/fisiopatologia , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/sangueRESUMO
Preeclampsia is a disease of the mother, fetus, and placenta, and the gaps in our understanding of the complex interactions among their respective disease pathways preclude successful treatment and prevention. The placenta has a key role in the pathogenesis of the terminal pathway characterized by exaggerated maternal systemic inflammation, generalized endothelial damage, hypertension, and proteinuria. This sine qua non of preeclampsia may be triggered by distinct underlying mechanisms that occur at early stages of pregnancy and induce different phenotypes. To gain insights into these molecular pathways, we employed a systems biology approach and integrated different "omics," clinical, placental, and functional data from patients with distinct phenotypes of preeclampsia. First trimester maternal blood proteomics uncovered an altered abundance of proteins of the renin-angiotensin and immune systems, complement, and coagulation cascades in patients with term or preterm preeclampsia. Moreover, first trimester maternal blood from preterm preeclamptic patients in vitro dysregulated trophoblastic gene expression. Placental transcriptomics of women with preterm preeclampsia identified distinct gene modules associated with maternal or fetal disease. Placental "virtual" liquid biopsy showed that the dysregulation of these disease gene modules originates during the first trimester. In vitro experiments on hub transcription factors of these gene modules demonstrated that DNA hypermethylation in the regulatory region of ZNF554 leads to gene down-regulation and impaired trophoblast invasion, while BCL6 and ARNT2 up-regulation sensitizes the trophoblast to ischemia, hallmarks of preterm preeclampsia. In summary, our data suggest that there are distinct maternal and placental disease pathways, and their interaction influences the clinical presentation of preeclampsia. The activation of maternal disease pathways can be detected in all phenotypes of preeclampsia earlier and upstream of placental dysfunction, not only downstream as described before, and distinct placental disease pathways are superimposed on these maternal pathways. This is a paradigm shift, which, in agreement with epidemiological studies, warrants for the central pathologic role of preexisting maternal diseases or perturbed maternal-fetal-placental immune interactions in preeclampsia. The description of these novel pathways in the "molecular phase" of preeclampsia and the identification of their hub molecules may enable timely molecular characterization of patients with distinct preeclampsia phenotypes.
Assuntos
Doenças Placentárias , Pré-Eclâmpsia , Adulto , Biomarcadores/sangue , Feminino , Humanos , Doenças Placentárias/sangue , Doenças Placentárias/genética , Doenças Placentárias/fisiopatologia , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/genética , Pré-Eclâmpsia/fisiopatologia , Gravidez , Proteômica , Biologia de Sistemas , Trofoblastos/metabolismo , Trofoblastos/patologiaRESUMO
Glioblastoma is the most common intracranial malignancy that constitutes about 50 % of all gliomas. Despite aggressive, multimodal therapy consisting of surgery, radiation, and chemotherapy, the outcome of patients with glioblastoma remains poor with 5-year survival rates of <10 %. Resistance to conventional therapies is most likely caused by several factors. Alterations in the functions of local immune mediators may represent a critical contributor to this resistance. The tumor microenvironment contains innate and adaptive immune cells in addition to the cancer cells and their surrounding stroma. These various cells communicate with each other by means of direct cell-cell contact or by soluble factors including cytokines and chemokines, and act in autocrine and paracrine manners to modulate tumor growth. There are dynamic interactions among the local immune elements and the tumor cells, where primarily the protective immune cells attempt to overcome the malignant cells. However, by developing somatic mutations and epigenetic modifications, the glioblastoma tumor cells acquire the capability of counteracting the local immune responses, and even exploit the immune cells and products for their own growth benefits. In this review, we survey those immune mechanisms that likely contribute to glioblastoma pathogenesis and may serve as a basis for novel treatment strategies.
Assuntos
Neoplasias Encefálicas/patologia , Glioblastoma/patologia , Linfócitos do Interstício Tumoral/patologia , Macrófagos/patologia , Microglia/patologia , Evasão Tumoral/imunologia , Microambiente Tumoral , Barreira Hematoencefálica , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/terapia , Vacinas Anticâncer/uso terapêutico , Comunicação Celular , Citocinas/metabolismo , Citotoxicidade Imunológica , Epigênese Genética , Vesículas Extracelulares , Glioblastoma/genética , Glioblastoma/imunologia , Glioblastoma/metabolismo , Glioblastoma/terapia , Humanos , Imunoterapia/métodos , Inflamação , Mutação , Células Mieloides/patologia , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Células-Tronco Neoplásicas/patologia , Fator de Crescimento Transformador beta/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Glioblastoma is the most aggressive form of gliomas and is associated with short survival. Recent advancements in molecular genetics resulted in the identification of glioma genomic, epigenomic and transcriptomic hallmarks, and multidimensional data allowed clustering of glioblastomas into molecular subtypes. Parallel with these developments, much scientific attention has been attracted by the exploration of two functional processes linked to mitochondrial regulation. One of these processes involves genomic and mitochondrial gene mutations, mitochondrial protein expression modifications and altered metabolic regulation that define glioblastoma. The second mitochondrially-centered process involves complex molecular interactions and pathways that influence the extrinsic or the intrinsic mechanisms of apoptosis regulation and may underlie the uncontrolled spreading, recurrence and drug resistance of glioblastoma. While the available data are not yet comprehensive, these two complex processes represent important aspects of tumor cell biology, which may provide complementary opportunities for therapeutic manipulations of this highly resistant tumor type.
Assuntos
Apoptose , Neoplasias Encefálicas , Metabolismo Energético/fisiologia , Glioblastoma , Mitocôndrias/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/fisiopatologia , Glioblastoma/genética , Glioblastoma/patologia , Glioblastoma/fisiopatologia , Humanos , Mitocôndrias/genética , MutaçãoRESUMO
Glioblastoma is the most common intracranial malignancy and constitutes about 50 % of all gliomas. Both inter-tumor and intra-tumor histological heterogeneity had been recognized by the early 1980-ies. Recent works using novel molecular platforms provided molecular definitions of these tumors. Based on comprehensive genomic sequence analyses, The Cancer Genome Atlas Research Network (TCGA) cataloged somatic mutations and recurrent copy number alterations in glioblastoma. Robust transcriptome and epigenome studies also revealed inter-tumor heterogeneity. Integration and cluster analyses of multi-dimensional genomic data lead to a new classification of glioblastoma tumors into subtypes with distinct biological features and clinical correlates. However, multiple observations also revealed tumor area-specific patterns of genomic imbalance. In addition, genetic alterations have been identified that were common to all areas analyzed and other alterations that were area specific. Analyses of intra-tumor transcriptome variations revealed that in more than half of the examined cases, fragments from the same tumor mass could be classified into at least two different glioblastoma molecular subgroups. Intra-tumor heterogeneity of molecular genetic profiles in glioblastoma may explain the difficulties encountered in the validation of oncologic biomarkers, and contribute to a biased selection of patients for single target therapies, treatment failure or drug resistance. In this paper, we summarize the currently available literature concerning inter- and intra-tumor molecular heterogeneity of glioblastomas, and call attention to the importance of this topic in relation to the growing efforts in routine molecular diagnostics and personalized therapy.
Assuntos
Variação Genética/genética , Glioblastoma/genética , Glioblastoma/patologia , Glioblastoma/classificação , Humanos , PrognósticoRESUMO
AIM: The adrenolytic agent mitotane is widely used in the treatment of adrenocortical cancer; however, its mechanism of action is poorly elucidated. We have studied mitotane-induced mRNA expression changes in the NCI-H295R adrenocortical cancer cell line. MATERIALS & METHODS: Cell viability and hormone assays were used to select the optimal mitotane concentration effectively inhibiting hormone secretion without affecting cell viability. RNA isolated from cultures treated for 48 and 72 h was subjected to Agilent 4×44K microarray platforms. Microarray results were validated by quantitative reverse-transcription PCR. RESULTS: Altogether, 117 significantly differentially expressed genes were detected at 48 h and 72 h (p < 0.05) in mitotane-treated samples relative to controls. Three significantly underexpressed genes involved in steroid hormone biosynthesis (HSD3B1, HSD3B2 and CYP21A2) and four significantly overexpressed genes (GDF15, ALDH1L2, TRIB3 and SERPINE2) have been validated. CONCLUSION: Gene-expression changes might be involved in the adrenal action of mitotane and in the inhibition of hormone secretion.
Assuntos
Neoplasias do Córtex Suprarrenal/genética , Antineoplásicos Hormonais/farmacologia , Expressão Gênica/efeitos dos fármacos , Hormônios/genética , Mitotano/farmacologia , Córtex Suprarrenal/efeitos dos fármacos , Neoplasias do Córtex Suprarrenal/tratamento farmacológico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Expressão Gênica/genética , Humanos , Análise em Microsséries/métodos , RNA Mensageiro/genéticaRESUMO
OBJECTIVES: To investigate the consequences of treatment with an exogenous glucocorticoid agonist (methylprednisolone) and antagonist (RU-38486) on the local and systemic responses in L-arginine-induced acute pancreatitis in rats. METHODS: The methylprednisolone and RU-38486 were administered just before pancreatitis induction. Plasma amylase activity, interleukin 6 activity, pancreatic weight/body weight ratio, plasma macrophage migration inhibitory factor (MIF) concentration, and pancreatic nuclear transcription factor (NF) kappaB activity were determined. The extents of pancreas, liver, and lung injuries were assessed by histology. RESULTS: Acute pancreatitis resulted in NF-kappaB activation and proinflammatory cytokine release in rats. In the glucocorticoid agonist group, plasma amylase and interleukin 6 levels were significantly decreased as compared with those of RU-38486 and nontreated groups. Antagonist treatment led to significantly higher MIF production at 8 and 12 hours after L-arginine injection as compared with the agonist-treated and nontreated groups. Glucocorticoid agonist treatment significantly decreased the level of NF-kappaB 24 hours after pancreatitis induction. Histological investigations showed protective effect of agonist treatment on acute pancreatitis-induced tissue damage in the pancreas and lung. CONCLUSIONS: These results corroborated the importance of MIF in acute pancreatitis. The glucocorticoid-dependent mechanisms seem to play a crucial role in the control of the inflammatory response and tissue damage in L-arginine-induced experimental acute pancreatitis.
Assuntos
Glucocorticoides/antagonistas & inibidores , Glucocorticoides/uso terapêutico , Metilprednisolona/uso terapêutico , Mifepristona/uso terapêutico , Pancreatite/induzido quimicamente , Pancreatite/tratamento farmacológico , Doença Aguda , Amilases/sangue , Animais , Arginina/toxicidade , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Inflamação/fisiopatologia , Interleucina-6/sangue , Células L , Fatores Inibidores da Migração de Macrófagos/sangue , Masculino , Camundongos , Ratos , Ratos WistarRESUMO
Migration and proliferation of smooth muscle cells are key to a number of physiological and pathological processes, including wound healing and the narrowing of the vessel wall. Previous work has shown links between inflammatory stimuli and vascular smooth muscle cell proliferation and migration through mitogen-activated protein kinase (MAPK) activation, although the molecular mechanisms of this process are poorly understood. Here we report that tribbles-1, a recently described modulator of MAPK activation, controls vascular smooth muscle cell proliferation and chemotaxis via the Jun kinase pathway. Our findings demonstrate that this regulation takes place via direct interactions between tribbles-1 and MKK4/SEK1, a Jun activator kinase. The activity of this kinase is dependent on tribbles-1 levels, whereas the activation and the expression of MKK4/SEK1 are not. In addition, tribbles-1 expression is elevated in human atherosclerotic arteries when compared with non-atherosclerotic controls, suggesting that this protein may play a role in disease in vivo. In summary, the data presented here suggest an important regulatory role for trb-1 in vascular smooth muscle cell biology.