RESUMEN
High throughput omics approaches provide an unprecedented opportunity for dissecting molecular mechanisms in cancer biology. Here we present deep profiling of whole proteome, phosphoproteome and transcriptome in two high-grade glioma (HGG) mouse models driven by mutated RTK oncogenes, PDGFRA and NTRK1, analyzing 13,860 proteins and 30,431 phosphosites by mass spectrometry. Systems biology approaches identify numerous master regulators, including 41 kinases and 23 transcription factors. Pathway activity computation and mouse survival indicate the NTRK1 mutation induces a higher activation of AKT downstream targets including MYC and JUN, drives a positive feedback loop to up-regulate multiple other RTKs, and confers higher oncogenic potency than the PDGFRA mutation. A mini-gRNA library CRISPR-Cas9 validation screening shows 56% of tested master regulators are important for the viability of NTRK-driven HGG cells, including TFs (Myc and Jun) and metabolic kinases (AMPKa1 and AMPKa2), confirming the validity of the multiomics integrative approaches, and providing novel tumor vulnerabilities.
Asunto(s)
Neoplasias Encefálicas/genética , Perfilación de la Expresión Génica , Glioma/genética , Proteómica , Proteínas Quinasas Activadas por AMP/metabolismo , Animales , Neoplasias Encefálicas/metabolismo , Modelos Animales de Enfermedad , Retroalimentación Fisiológica , Glioma/metabolismo , Ratones , Mutación , Proteína Oncogénica p65(gag-jun)/metabolismo , Fosfopéptidos/metabolismo , Fosfoproteínas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/genética , Receptor trkA/genética , Transducción de Señal , Biología de Sistemas , Regulación hacia ArribaRESUMEN
Breast cancer (BC) is the most common female malignancy in the world. Triple-negative breast cancer (TNBC) is a subtype of BC characterized by the lack of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor-2 (HER-2), resulting in the limited therapeutic options. Due to the aggressive behaviors at early stage, TNBC exhibits poorer outcomes compared to other BC subtypes. Hematogenous metastasis, which spreads cancerous cells to lungs and/or bones, plays a pivotal role in the progression of TNBC. Therefore, it is of great importance to study the anti-angiogenesis regulation mechanism for finding new treatment options for TNBC. Arsenic trioxide (ATO) exhibits anti-cancer effect on solid tumors, including TNBC. However, the roles and the molecular mechanism of ATO in the anti-angiogenesis of TNBC remain less well documented. Our data showed that ATO restrained the expression and secretion of vascular endothelial growth factor (VEGF) and impaired the angiogenic ability in TNBC cells. In addition, ATO suppressed the angiogenic ability in TNBC by inhibiting the interaction of the enhancer of zeste homolog 2 (EZH2) with p65, downregulating the nuclear factor-κB (NF-κB) activity, hence contributing to the regulation of IL-6/Stat3 signaling pathway. All of our findings would help to better understand the mechanism of ATO anti-angiogenesis in TNBC, thus highlighting the therapeutic potential of ATO in TNBC by targeting angiogenesis.
Asunto(s)
Trióxido de Arsénico/farmacología , Neovascularización Patológica/metabolismo , Neoplasias de la Mama Triple Negativas/metabolismo , Trióxido de Arsénico/metabolismo , Línea Celular Tumoral , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/fisiología , Femenino , Células Endoteliales de la Vena Umbilical Humana , Humanos , FN-kappa B/metabolismo , Proteína Oncogénica p65(gag-jun)/metabolismo , Receptor ErbB-2/metabolismo , Receptores de Estrógenos/efectos de los fármacos , Receptores de Estrógenos/metabolismo , Factor de Transcripción STAT3/metabolismo , Transducción de Señal/efectos de los fármacos , Neoplasias de la Mama Triple Negativas/patología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Schwann cells (SCs), the glia of the peripheral nervous system, play an essential role in nerve regeneration. Upon nerve injury, SCs are reprogrammed into unique "repair SCs," and these cells remove degenerating axons/myelin debris, promote axonal regrowth, and ultimately remyelinate regenerating axons. The AP-1 transcription factor JUN is promptly induced in SCs upon nerve injury and potently mediates this injury-induced SC plasticity; however, the regulation of these JUN-dependent SC injury responses is unclear. Previously, we produced mice with a SC-specific deletion of O-GlcNAc transferase (OGT). This enzyme catalyzes O-GlcNAcylation, a posttranslational modification that is influenced by the cellular metabolic state. Mice lacking OGT in SCs develop a progressive demyelinating peripheral neuropathy. Here, we investigated the nerve repair process in OGT-SCKO mutant mice and found that the remyelination of regenerating axons is severely impaired. Gene expression profiling of OGT-SCKO SCs revealed that the JUN-dependent SC injury program was elevated in the absence of injury and failed to shut down at the appropriate time after injury. This aberrant JUN activity results in abnormalities in repair SC function and redifferentiation and prevents the timely remyelination. This aberrant nerve injury response is normalized in OGT-SCKO mice with reduced Jun gene dosage in SCs. Mechanistically, OGT O-GlcNAcylates JUN at multiple sites, which then leads to an attenuation of AP-1 transcriptional activity. Together, these results highlight the metabolic oversight of the nerve injury response via the regulation of JUN activity by O-GlcNAcylation, a pathway that could be important in the neuropathy associated with diabetes and aging.
Asunto(s)
Enfermedades Desmielinizantes/metabolismo , Regeneración Nerviosa , Proteína Oncogénica p65(gag-jun)/metabolismo , Células de Schwann/metabolismo , Nervio Ciático/lesiones , Nervio Ciático/metabolismo , Factor de Transcripción AP-1/metabolismo , Acilación/genética , Envejecimiento/genética , Envejecimiento/metabolismo , Envejecimiento/patología , Animales , Axones/metabolismo , Enfermedades Desmielinizantes/genética , Enfermedades Desmielinizantes/patología , Neuropatías Diabéticas/genética , Neuropatías Diabéticas/metabolismo , Neuropatías Diabéticas/patología , Eliminación de Gen , Ratones , Ratones Noqueados , N-Acetilglucosaminiltransferasas/genética , N-Acetilglucosaminiltransferasas/metabolismo , Proteína Oncogénica p65(gag-jun)/genética , Células de Schwann/patología , Nervio Ciático/patología , Factor de Transcripción AP-1/genéticaRESUMEN
Recently, the NLRP3 inflammasome activation in the eyes has been known to be associated with the pathogenesis of age-related macular degeneration. The aim of this study was to investigate the protective effects of cyanidin-3-glucoside (C3G), an important anthocyanin with great potential for preventing eye diseases, against 4-hydroxyhexenal- (HHE-) induced inflammatory damages in human retinal pigment epithelial cells, ARPE-19. We noticed that C3G pretreatment to the ARPE-19 cells rescued HHE-induced antiproliferative effects. Cell apoptosis ratio induced by HHE was also decreased by C3G, measured by flow cytometry. The activation of NLRP3 inflammasome induced by HHE was found with increases of caspase-1 activity, proinflammatory cytokine releases (IL-1ß and IL-18), and NLRP3 inflammasome-related gene expressions (NLRP3, IL-1ß, IL-18, and caspase-1). The C3G showed potent inhibitive effects on these NLRP3 inflammasome activation hallmarks induced by HHE. Moreover, we noticed that the C3G's pretreatment leads to a delayed and a decreased JNK activation in HHE-challenged ARPE-19 cells. Finally, using a luciferase reporter gene assay system, we demonstrated that HHE-induced activation protein- (AP-) 1 transcription activity was abolished by C3G pretreatment in a dose-dependent manner. Taken together, these data showed that HHE leads to inflammatory damages to ARPE-19 cells while C3G has great protective effects, highlighting future potential applications of C3G against AMD-associated inflammation.
Asunto(s)
Antocianinas/farmacología , Antiinflamatorios/farmacología , Oftalmopatías/tratamiento farmacológico , Glucósidos/farmacología , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Epitelio Pigmentado de la Retina/inmunología , Aldehídos/metabolismo , Caspasa 1/metabolismo , Línea Celular , Citocinas/metabolismo , Humanos , Mediadores de Inflamación/metabolismo , MAP Quinasa Quinasa 4/metabolismo , Proteína Oncogénica p65(gag-jun)/metabolismo , Epitelio Pigmentado de la Retina/efectos de los fármacos , Transducción de Señal , Factor de Transcripción AP-1/metabolismoRESUMEN
BACKGROUND: The diagnosis of early phase lung adenocarcinoma (LADC) is associated with therapeutic strategy, effect, and survival time. However, the sensitive biomarkers of early phase LADC are still unclear. This study aimed to identify protein-coding genes that can be used as biomarkers of early stage LADC. METHODS: Gene microarray analysis was performed to identify key hub genes that show different expression in lung adenocarcinoma compared to normal tissues. The microarray data of lung adenocarcinoma in stages IA, IB, IIA, IIB, and normal tissues (GSE10072) were downloaded from a free online database, Gene Expression Omnibus (GEO). RESULTS: A total of 572 differentially expressed genes (DEGs) were identified between early phase lung adenocarcinoma and normal tissues using R software. Database for Annotation, Visualization and Integrated Discovery online tools were used to obtain Gene Ontology analysis and the Kyoto Encyclopedia of Genes and Genomes was used to analyze DEGs. Cytoscape software was used to express the protein-protein interaction network. We found that some cancer-related Gene Ontology terms and pathways (e.g. cell adhesion, cell surface receptor signaling pathway, PI3K-Akt signaling pathway) were significantly enriched in DEGs. CONCLUSION: Protein-coding genes JUN, FYN, CAV1, and SFN may play vital roles in the progress of early-stage lung adenocarcinoma. Consequently, through bioinformatics analysis, the key genes could be established to provide more potential references for the therapeutic targets of lung adenocarcinoma.
Asunto(s)
Adenocarcinoma/genética , Biología Computacional , Redes Reguladoras de Genes/genética , Neoplasias Pulmonares/genética , Proteínas de Neoplasias/genética , Proteínas 14-3-3/genética , Adenocarcinoma/patología , Adenocarcinoma del Pulmón , Biomarcadores de Tumor/genética , Caveolina 1/genética , Bases de Datos Genéticas , Exorribonucleasas/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Neoplasias Pulmonares/patología , Análisis por Micromatrices , Estadificación de Neoplasias , Proteína Oncogénica p65(gag-jun)/genética , Mapas de Interacción de Proteínas/genética , Proteínas Proto-Oncogénicas c-fyn/genética , Transducción de Señal/genética , Programas InformáticosRESUMEN
Cumulative evidence demonstrates that most RNAs exhibit specific subcellular distribution. However, the mechanisms regulating this phenomenon and its functional consequences are still under investigation. Here, we reveal that cadherin complexes at the apical zonula adherens (ZA) of epithelial adherens junctions recruit the core components of the RNA-induced silencing complex (RISC) Ago2, GW182, and PABPC1, as well as a set of 522 messenger RNAs (mRNAs) and 28 mature microRNAs (miRNAs or miRs), via PLEKHA7. Top canonical pathways represented by these mRNAs include Wnt/ß-catenin, TGF-ß, and stem cell signaling. We specifically demonstrate the presence and silencing of MYC, JUN, and SOX2 mRNAs by miR-24 and miR-200c at the ZA. PLEKHA7 knockdown dissociates RISC from the ZA, decreases loading of the ZA-associated mRNAs and miRNAs to Ago2, and results in a corresponding increase of MYC, JUN, and SOX2 protein expression. The present work reveals a mechanism that directly links junction integrity to the silencing of a set of mRNAs that critically affect epithelial homeostasis.
Asunto(s)
Uniones Adherentes/metabolismo , Cadherinas/metabolismo , Células Epiteliales/metabolismo , ARN Mensajero/metabolismo , Complejo Silenciador Inducido por ARN/metabolismo , Uniones Adherentes/genética , Animales , Células CACO-2 , Cadherinas/genética , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Perros , Humanos , Células de Riñón Canino Madin Darby , MicroARNs/genética , MicroARNs/metabolismo , Proteína Oncogénica p65(gag-jun)/genética , Proteína Oncogénica p65(gag-jun)/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Mensajero/genética , Complejo Silenciador Inducido por ARN/genética , Factores de Transcripción SOXB1/genética , Factores de Transcripción SOXB1/metabolismoRESUMEN
Therapies that target signalling molecules that are mutated in cancers can often have substantial short-term effects, but the emergence of resistant cancer cells is a major barrier to full cures. Resistance can result from secondary mutations, but in other cases there is no clear genetic cause, raising the possibility of non-genetic rare cell variability. Here we show that human melanoma cells can display profound transcriptional variability at the single-cell level that predicts which cells will ultimately resist drug treatment. This variability involves infrequent, semi-coordinated transcription of a number of resistance markers at high levels in a very small percentage of cells. The addition of drug then induces epigenetic reprogramming in these cells, converting the transient transcriptional state to a stably resistant state. This reprogramming begins with a loss of SOX10-mediated differentiation followed by activation of new signalling pathways, partially mediated by the activity of the transcription factors JUN and/or AP-1 and TEAD. Our work reveals the multistage nature of the acquisition of drug resistance and provides a framework for understanding resistance dynamics in single cells. We find that other cell types also exhibit sporadic expression of many of these same marker genes, suggesting the existence of a general program in which expression is displayed in rare subpopulations of cells.
Asunto(s)
Reprogramación Celular/efectos de los fármacos , Reprogramación Celular/genética , Resistencia a Antineoplásicos/efectos de los fármacos , Resistencia a Antineoplásicos/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Melanoma/genética , Melanoma/patología , Animales , Línea Celular Tumoral , Proteínas de Unión al ADN/metabolismo , Epigénesis Genética/efectos de los fármacos , Receptores ErbB/metabolismo , Femenino , Marcadores Genéticos/efectos de los fármacos , Marcadores Genéticos/genética , Humanos , Hibridación Fluorescente in Situ , Indoles/farmacología , Masculino , Proteínas Nucleares/metabolismo , Proteína Oncogénica p65(gag-jun)/metabolismo , Factores de Transcripción SOXE/deficiencia , Factores de Transcripción SOXE/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Análisis de la Célula Individual , Sulfonamidas/farmacología , Factores de Transcripción de Dominio TEA , Factor de Transcripción AP-1/metabolismo , Factores de Transcripción/metabolismo , Transcripción Genética/efectos de los fármacos , Vemurafenib , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
AIM: To investigate the role of Δ133p53 isoform in nuclear factor-κB (NF-κB) inhibitor pyrrolidine dithiocarbamate (PDTC)-mediated growth inhibition of MKN45 gastric cancer cells. METHODS: The growth rate of MKN45 cells after treatment with different concentrations of only PDTC or PTDC in combination with cisplatin was detected by the CCK-8 assay. mRNA expression levels of Δ133p53, p53ß, and the NF-κB p65 subunit and p65 protein levels were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence, respectively. Growth of MKN45 cells was significantly inhibited by PDTC alone in a dose-dependent manner (P < 0.01). Moreover, the inhibitory effect of cisplatin was remarkably enhanced in a dose-dependent manner by co-treatment with PDTC (P < 0.01). RESULTS: RT-PCR analysis revealed that mRNA expression of p65 was curbed significantly in a dose-dependent manner by treatment with only PDTC (P < 0.01), and this suppressive effect was further enhanced when co-treated with cisplatin (P < 0.01). With respect to the other p53 isoforms, mRNA level of Δ133p53 was significantly reduced in a dose-dependent manner by treatment with only PDTC or PTDC in combination with cisplatin (P < 0.01), whereas p53ß mRNA expression was not altered by PDTC treatment (P > 0.05). A similar tendency of change in p65 protein expression, as observed for the corresponding mRNA, was detected by immunofluorescence analysis (P < 0.01). Pearson correlation analysis demonstrated that Δ133p53 and p65 mRNA expression levels were positively related, while no significant relationship was observed between those of p65 and p53ß (r = 0.076, P > 0.01). CONCLUSION: Δ133p53 isoform (not p53ß) is required in PDTC-induced inhibition of MKN45 gastric cancer cells, indicating that disturbance in the cross-talk between p53 and NF-κB pathways is a promising target in pharmaceutical research for the development of treatment strategies for gastric cancer.
Asunto(s)
FN-kappa B/antagonistas & inhibidores , Proteína Oncogénica p65(gag-jun)/metabolismo , Pirrolidinas/farmacología , Neoplasias Gástricas/metabolismo , Tiocarbamatos/farmacología , Proteína p53 Supresora de Tumor/metabolismo , Línea Celular Tumoral , Humanos , FN-kappa B/metabolismo , Isoformas de Proteínas/metabolismo , Pirrolidinas/uso terapéutico , Neoplasias Gástricas/tratamiento farmacológico , Tiocarbamatos/uso terapéuticoRESUMEN
Oral submucous fibrosis (OSF) is potentially premalignant with progressive and irreversible extracellular matrix deposition accompanied by epithelial atrophy and like other fibrotic disorders, is primarily a TGF-ß driven disease. OSF is caused by prolonged chewing of areca nut. Our previous studies reported a pivotal role for TGF-ß activation and its effects contributing to OSF. However, the mechanism for activation of TGF-ß signaling in OSF is still unknown. In this study we demonstrate activation of TGF-ß signaling with sub-cytotoxic dose of areca nut in epithelial cells and discovered a key role for pJNK in this process. In good correlation; pJNK was detected in OSF tissues but not in normal tissues. Moreover, activation of JNK was found to be dependent on muscarinic acid receptor induced Ca2+/CAMKII as well as ROS. JNK dependent phosphorylation of ATF2/c-Jun transcription factors resulted in TGF-ß transcription and its signaling. pATF2/p-c-Jun were enriched on TGF-ß promoter and co-localized in nuclei of epithelial cells upon areca nut treatment. In corroboration, OSF tissue sections also had nuclear pATF2 and p-c-Jun. Our results provide comprehensive mechanistic details of TGF-ß signaling induced by etiological agent areca nut in the manifestation of fibrosis which can lead to new therapeutic modalities for OSF.
Asunto(s)
Factor de Transcripción Activador 2/metabolismo , Areca/química , MAP Quinasa Quinasa 4/metabolismo , Mucosa Bucal , Neoplasias de la Boca , Nueces/química , Proteína Oncogénica p65(gag-jun)/metabolismo , Extractos Vegetales/farmacología , Lesiones Precancerosas , Factor de Crecimiento Transformador beta/metabolismo , Línea Celular Transformada , Femenino , Fibrosis , Humanos , Masculino , Mucosa Bucal/metabolismo , Mucosa Bucal/patología , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Extractos Vegetales/química , Lesiones Precancerosas/tratamiento farmacológico , Lesiones Precancerosas/metabolismo , Lesiones Precancerosas/patologíaRESUMEN
T cell antigen receptor (TCR) signaling drives distinct responses depending on the differentiation state and context of CD8(+) T cells. We hypothesized that access of signal-dependent transcription factors (TFs) to enhancers is dynamically regulated to shape transcriptional responses to TCR signaling. We found that the TF BACH2 restrains terminal differentiation to enable generation of long-lived memory cells and protective immunity after viral infection. BACH2 was recruited to enhancers, where it limited expression of TCR-driven genes by attenuating the availability of activator protein-1 (AP-1) sites to Jun family signal-dependent TFs. In naive cells, this prevented TCR-driven induction of genes associated with terminal differentiation. Upon effector differentiation, reduced expression of BACH2 and its phosphorylation enabled unrestrained induction of TCR-driven effector programs.
Asunto(s)
Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Linfocitos T CD8-positivos/fisiología , Factor de Transcripción AP-1/metabolismo , Virus Vaccinia/inmunología , Vaccinia/inmunología , Inmunidad Adaptativa , Animales , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Linfocitos T CD8-positivos/virología , Diferenciación Celular/genética , Células Cultivadas , Elementos de Facilitación Genéticos/genética , Regulación de la Expresión Génica , Memoria Inmunológica/genética , Activación de Linfocitos/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína Oncogénica p65(gag-jun) , Transducción de Señal/genética , Factor de Transcripción AP-1/genéticaRESUMEN
HMGA2, CDK4, and JUN genes have been described as frequently coamplified with MDM2 in atypical lipomatous tumor, well-differentiated liposarcoma, and dedifferentiated liposarcoma. We studied the frequency of amplification of these genes in a series of 48 dedifferentiated liposarcomas and 68 atypical lipomatous tumors/well-differentiated liposarcomas. We correlated their amplification status with clinicopathological features and outcomes. Histologically, both CDK4 (P=0.007) and JUN (P=0.005) amplifications were associated with dedifferentiated liposarcoma, whereas amplification of the proximal parts of HMGA2 (5'-untranslated region (UTR) and exons 1-3) was associated with atypical lipomatous tumor/well-differentiated liposarcoma (P=0.01). CDK4 amplification was associated with axial tumors. Amplification of 5'-UTR and exons 1-3 of HMGA2 was associated with primary status and grade 1. Shorter overall survival was correlated with: age >64 years (P=0.03), chemotherapy used in first intent (P<0.001), no surgery (P=0.003), grade 3 (P<0.001), distant metastasis (P<0.001), node involvement (P=0.006), and CDK4 amplification (P=0.07). In multivariate analysis, distant metastasis (HR=8.8) and grade 3 (HR=18.2) were associated with shorter overall survival. A shorter recurrence-free survival was associated with dedifferentiated liposarcoma (P<0.001), grade 3 (P<0.001), node involvement (P<0.001), distant metastasis (P=0.02), recurrent status (P=0.009), axial location (P=0.001), and with molecular features such as CDK4 (P=0.05) and JUN amplification (P=0.07). Amplification of 5'-UTR and exons 1-3 (P=0.08) and 3'-UTR (P=0.01) of HMGA2 were associated with longer recurrence-free survival. Distant metastasis was associated with shorter recurrence-free survival (HR=5.8) in multivariate analysis. Dedifferentiated liposarcoma type was associated with axial location, grade 3 and recurrent status. In conclusion, we showed that the amplification of HMGA2 was associated with the atypical lipomatous tumor/well-differentiated liposarcoma histological type and a good prognosis, whereas CDK4 and JUN amplifications were associated with dedifferentiated liposarcoma histology and a bad prognosis. In addition, we also provided the first description of the molecular evolution of a well-differentiated liposarcoma into four successive dedifferentiated liposarcoma relapses, which was consistent with our general observations.
Asunto(s)
Quinasa 4 Dependiente de la Ciclina/genética , Amplificación de Genes , Genes jun/genética , Proteína HMGA2/genética , Liposarcoma/genética , Liposarcoma/patología , Neoplasias de los Tejidos Blandos/genética , Adulto , Anciano , Biomarcadores de Tumor/genética , Hibridación Genómica Comparativa , Supervivencia sin Enfermedad , Femenino , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Estimación de Kaplan-Meier , Liposarcoma/mortalidad , Masculino , Persona de Mediana Edad , Proteína Oncogénica p65(gag-jun)/genética , Pronóstico , Neoplasias de los Tejidos Blandos/mortalidad , Neoplasias de los Tejidos Blandos/patologíaRESUMEN
Hypoxia-inducible factor 1α (HIF1α) is a transcription factor that regulates the adaptation of cells to hypoxic microenvironments, for example inside solid tumours. Stabilisation of HIF1α can also occur in normoxic conditions in inflamed tissue or as a result of inactivating mutations in negative regulators of HIF1α. Aberrant overexpression of HIF1α in many different cancers has led to intensive efforts to develop HIF1α-targeted therapies. However, the role of HIF1α is still poorly understood in chronic inflammation that predisposes the colon to carcinogenesis. We have previously reported that the transcription of HIF1α is upregulated and that the protein is stabilised in inflammatory lesions that are caused by the non-steroidal anti-inflammatory drug (NSAID) sulindac in the mouse proximal colon. Here, we exploited this side effect of long-term sulindac administration to analyse the role of HIF1α in colon inflammation using mice with a Villin-Cre-induced deletion of Hif1α exon 2 in the intestinal epithelium (Hif1α(ΔIEC)). We also analysed the effect of sulindac sulfide on the aryl hydrocarbon receptor (AHR) pathway in vitro in colon cancer cells. Most sulindac-treated mice developed visible lesions, resembling the appearance of flat adenomas in the human colon, surrounded by macroscopically normal mucosa. Hif1α(ΔIEC) mice still developed lesions but they were smaller than in the Hif1α-floxed siblings (Hif1α(F/F)). Microscopically, Hif1α(ΔIEC) mice had significantly less severe colon inflammation than Hif1α(F/F) mice. Molecular analysis showed reduced MIF expression and increased E-cadherin mRNA expression in the colon of sulindac-treated Hif1α(ΔIEC) mice. However, immunohistochemistry analysis revealed a defect of E-cadherin protein expression in sulindac-treated Hif1α(ΔIEC) mice. Sulindac sulfide treatment in vitro upregulated Hif1α, c-JUN and IL8 expression through the AHR pathway. Taken together, HIF1α expression augments inflammation in the proximal colon of sulindac-treated mice, and AHR activation by sulindac might lead to the reduction of E-cadherin protein levels through the mitogen-activated protein kinase (MAPK) pathway.
Asunto(s)
Neoplasias del Colon/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/fisiología , Inflamación , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Cadherinas/metabolismo , Línea Celular Tumoral , Neoplasias del Colon/patología , Modelos Animales de Enfermedad , Exones , Femenino , Eliminación de Gen , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/deficiencia , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Inmunohistoquímica , Interleucina-8/metabolismo , Mucosa Intestinal/patología , Sistema de Señalización de MAP Quinasas , Masculino , Ratones , Proteína Oncogénica p65(gag-jun)/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Sulindac/uso terapéutico , Regulación hacia ArribaRESUMEN
The down-modulation of the ß-catenin antagonist Chibby 1 (CBY1) associated with the BCR-ABL1 fusion gene of chronic myeloid leukemia (CML) contributes to the aberrant activation of ß-catenin, particularly in leukemic stem cells (LSC) resistant to tyrosine kinase (TK) inhibitors. It is, at least partly, driven by transcriptional events and gene promoter hyper-methylation. Here we demonstrate that it also arises from reduced protein stability upon binding to 14-3-3σ adapter protein. CBY1/14-3-3σ interaction in BCR-ABL1+ cells is mediated by the fusion protein TK and AKT phosphorylation of CBY1 at critical serine 20, and encompasses the 14-3-3σ binding modes I and II involved in the binding with client proteins. Moreover, it is impaired by c-Jun N-terminal kinase (JNK) phosphorylation of 14-3-3σ at serine 186, which promotes dissociation of client proteins. The ubiquitin proteasome system UPS participates in reducing stability of CBY1 bound with 14-3-3σ through enhanced SUMOylation. Our results open new routes towards the research on molecular pathways promoting the proliferative advantage of leukemic hematopoiesis over the normal counterpart.
Asunto(s)
Proteínas 14-3-3/metabolismo , Biomarcadores de Tumor/metabolismo , Proteínas Portadoras/biosíntesis , Exorribonucleasas/metabolismo , Regulación Leucémica de la Expresión Génica/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/metabolismo , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/biosíntesis , Procesamiento Proteico-Postraduccional , Secuencias de Aminoácidos , Benzamidas/farmacología , Proteínas Portadoras/genética , Regulación hacia Abajo , Proteínas de Fusión bcr-abl/metabolismo , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Proteína Oncogénica p65(gag-jun) , Fosforilación , Complejo de la Endopetidasa Proteasomal/metabolismo , Unión Proteica , Mapeo de Interacción de Proteínas , Estabilidad Proteica , Proteínas Proto-Oncogénicas c-akt/metabolismo , Pirazoles/farmacología , Fracciones Subcelulares/metabolismo , Sumoilación , beta Catenina/antagonistas & inhibidoresRESUMEN
A number of genes regulate regeneration of peripheral axons, but their ability to drive axon growth and regeneration in the central nervous system (CNS) remains largely untested. To address this question we overexpressed eight transcription factors and one small GTPase alone and in pairwise combinations to test whether combinatorial overexpression would have a synergistic impact on CNS neuron neurite growth. The Jun oncogene/signal transducer and activator of transcription 6 (JUN/STAT6) combination increased neurite growth in dissociated cortical neurons and in injured cortical slices. In injured cortical slices, JUN overexpression increased axon growth to a similar extent as JUN and STAT6 together. Interestingly, JUN overexpression was not associated with increased growth associated protein 43 (GAP43) or integrin alpha 7 (ITGA7) expression, though these are predicted transcriptional targets. This study demonstrates that JUN overexpression in cortical neurons stimulates axon growth, but does so independently of changes in expression of genes thought to be critical for JUNs effects on axon growth. We conclude that JUN activity underlies this CNS axonal growth response, and that it is mechanistically distinct from peripheral regeneration responses, in which increases in JUN expression coincide with increases in GAP43 expression.
Asunto(s)
Axones/metabolismo , Corteza Cerebral/crecimiento & desarrollo , Proteína Oncogénica p65(gag-jun)/metabolismo , Animales , Antígenos CD/genética , Antígenos CD/metabolismo , Axones/fisiología , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/metabolismo , Proteína GAP-43/genética , Proteína GAP-43/metabolismo , Regulación del Desarrollo de la Expresión Génica , Células HEK293 , Humanos , Cadenas alfa de Integrinas/genética , Cadenas alfa de Integrinas/metabolismo , Regeneración Nerviosa , Neurogénesis , Proteína Oncogénica p65(gag-jun)/genética , Ratas , Ratas Sprague-Dawley , Factor de Transcripción STAT6/genética , Factor de Transcripción STAT6/metabolismoRESUMEN
The capacity of tumour cells to maintain continual overgrowth potential has been linked to the commandeering of normal self-renewal pathways. Using an epithelial cancer model in Drosophila melanogaster, we carried out an overexpression screen for oncogenes capable of cooperating with the loss of the epithelial apico-basal cell polarity regulator, scribbled (scrib), and identified the cell fate regulator, Abrupt, a BTB-zinc finger protein. Abrupt overexpression alone is insufficient to transform cells, but in cooperation with scrib loss of function, Abrupt promotes the formation of massive tumours in the eye/antennal disc. The steroid hormone receptor coactivator, Taiman (a homologue of SRC3/AIB1), is known to associate with Abrupt, and Taiman overexpression also drives tumour formation in cooperation with the loss of Scrib. Expression arrays and ChIP-Seq indicates that Abrupt overexpression represses a large number of genes, including steroid hormone-response genes and multiple cell fate regulators, thereby maintaining cells within an epithelial progenitor-like state. The progenitor-like state is characterised by the failure to express the conserved Eyes absent/Dachshund regulatory complex in the eye disc, and in the antennal disc by the failure to express cell fate regulators that define the temporal elaboration of the appendage along the proximo-distal axis downstream of Distalless. Loss of scrib promotes cooperation with Abrupt through impaired Hippo signalling, which is required and sufficient for cooperative overgrowth with Abrupt, and JNK (Jun kinase) signalling, which is required for tumour cell migration/invasion but not overgrowth. These results thus identify a novel cooperating oncogene, identify mammalian family members of which are also known oncogenes, and demonstrate that epithelial tumours in Drosophila can be characterised by the maintenance of a progenitor-like state.
Asunto(s)
Carcinogénesis , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Sistema de Señalización de MAP Quinasas/genética , Neoplasias Glandulares y Epiteliales/genética , Proteínas Nucleares/genética , Animales , Proliferación Celular , Modelos Animales de Enfermedad , Proteínas de Drosophila/metabolismo , Neoplasias del Ojo/genética , Neoplasias del Ojo/patología , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana , Neoplasias Experimentales/genética , Neoplasias Experimentales/patología , Neoplasias Glandulares y Epiteliales/patología , Proteínas Nucleares/metabolismo , Proteína Oncogénica p65(gag-jun)/genética , Proteína Oncogénica p65(gag-jun)/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismoRESUMEN
BACKGROUND: Influenza viruses have been associated with various neurological and muscular symptoms. The aim of this study was to evaluate the pediatric neurological and muscular manifestations of influenza B during a 5-month epidemic at a single center. METHODS: We retrospectively reviewed the medical records of 355 pediatric patients with laboratory-confirmed influenza B infection. RESULTS: Neurological and muscular symptoms were exhibited by 28 patients (7.9%). The mean age was 48.7 ± 25.2 months. The mean time between respiratory symptoms and neurological symptoms was 2.2 ± 1.5 days. The most common symptom was seizure (19/28, 67.9%), followed by myositis (5/28, 17.9%), increased intracerebral pressure (1/28, 3.6%), delirium (1/28, 3.6%), and severe headache (1/28, 3.6%). There was one severe case of meningitis with myocarditis (1/28, 3.6%). All seizures were febrile: 15 simple febrile seizures (78.9%), three complex febrile seizures (15.8%), and one febrile status epilepticus (5.3%). The mean age of nine patients with their first seizures was 37.9 ± 22.2 months, which was older than the typical age of onset for febrile seizure. All the patients, except one, were treated with oseltamivir. There were no deaths or chronic debilitating sequelae. CONCLUSIONS: The neurological and muscular complications of influenza B infection in children are relatively mild, and febrile seizure is the most common. However, clinicians should be alert to the possibility of rare severe complications during influenza B outbreaks.
Asunto(s)
Virus de la Influenza B/patogenicidad , Gripe Humana/complicaciones , Enfermedades Musculares/etiología , Enfermedades del Sistema Nervioso/etiología , Niño , Preescolar , Femenino , Humanos , Lactante , Virus de la Influenza B/genética , Masculino , Enfermedades Musculares/diagnóstico , Enfermedades Musculares/virología , Enfermedades del Sistema Nervioso/clasificación , Enfermedades del Sistema Nervioso/diagnóstico , Enfermedades del Sistema Nervioso/virología , Proteína Oncogénica p65(gag-jun)/genética , Proteína Oncogénica p65(gag-jun)/metabolismo , Estudios RetrospectivosRESUMEN
Anaplastic large cell lymphoma (ALCL) is an aggressive non-Hodgkin's lymphoma found in children and young adults. ALCLs frequently carry a chromosomal translocation that results in expression of the oncoprotein nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). The key molecular downstream events required for NPM-ALK-triggered lymphoma growth have been only partly unveiled. Here we show that the activator protein 1 family members JUN and JUNB promote lymphoma development and tumor dissemination through transcriptional regulation of platelet-derived growth factor receptor-ß (PDGFRB) in a mouse model of NPM-ALK-triggered lymphomagenesis. Therapeutic inhibition of PDGFRB markedly prolonged survival of NPM-ALK transgenic mice and increased the efficacy of an ALK-specific inhibitor in transplanted NPM-ALK tumors. Notably, inhibition of PDGFRA and PDGFRB in a patient with refractory late-stage NPM-ALK(+) ALCL resulted in rapid, complete and sustained remission. Together, our data identify PDGFRB as a previously unknown JUN and JUNB target that could be a highly effective therapy for ALCL.
Asunto(s)
Linfoma Anaplásico de Células Grandes , Proteínas Nucleares , Proteínas Tirosina Quinasas , Proteínas Tirosina Quinasas Receptoras , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas , Receptor beta de Factor de Crecimiento Derivado de Plaquetas , Adulto , Quinasa de Linfoma Anaplásico , Animales , Benzamidas , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Mesilato de Imatinib , Linfoma Anaplásico de Células Grandes/tratamiento farmacológico , Linfoma Anaplásico de Células Grandes/metabolismo , Linfoma Anaplásico de Células Grandes/patología , Ratones , Ratones Transgénicos , Terapia Molecular Dirigida , Estadificación de Neoplasias , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Nucleofosmina , Proteína Oncogénica p65(gag-jun)/genética , Proteína Oncogénica p65(gag-jun)/metabolismo , Piperazinas/administración & dosificación , Proteínas Tirosina Quinasas/genética , Proteínas Tirosina Quinasas/metabolismo , Pirimidinas/administración & dosificación , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Proteínas Tirosina Quinasas Receptoras/genética , Proteínas Tirosina Quinasas Receptoras/metabolismo , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/antagonistas & inhibidores , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/genética , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/antagonistas & inhibidores , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Inducción de Remisión , Trasplante de Células Madre , Factor de Transcripción AP-1/genética , Factor de Transcripción AP-1/metabolismo , Translocación GenéticaRESUMEN
The AP1 transcription factor Batf3 is required for homeostatic development of CD8α(+) classical dendritic cells that prime CD8 T-cell responses against intracellular pathogens. Here we identify an alternative, Batf3-independent pathway in mice for CD8α(+) dendritic cell development operating during infection with intracellular pathogens and mediated by the cytokines interleukin (IL)-12 and interferon-γ. This alternative pathway results from molecular compensation for Batf3 provided by the related AP1 factors Batf, which also functions in T and B cells, and Batf2 induced by cytokines in response to infection. Reciprocally, physiological compensation between Batf and Batf3 also occurs in T cells for expression of IL-10 and CTLA4. Compensation among BATF factors is based on the shared capacity of their leucine zipper domains to interact with non-AP1 factors such as IRF4 and IRF8 to mediate cooperative gene activation. Conceivably, manipulating this alternative pathway of dendritic cell development could be of value in augmenting immune responses to vaccines.
Asunto(s)
Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Células Dendríticas/citología , Células Dendríticas/metabolismo , Factores Reguladores del Interferón/metabolismo , Animales , Presentación de Antígeno , Antígenos CD/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/química , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/deficiencia , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Antígenos CD8/inmunología , Antígenos CD8/metabolismo , Antígeno CTLA-4/metabolismo , Diferenciación Celular , Línea Celular Tumoral , Linaje de la Célula , Células Dendríticas/inmunología , Femenino , Fibrosarcoma/inmunología , Fibrosarcoma/metabolismo , Fibrosarcoma/patología , Regulación de la Expresión Génica , Cadenas alfa de Integrinas/metabolismo , Factores Reguladores del Interferón/deficiencia , Factores Reguladores del Interferón/genética , Interleucina-10/metabolismo , Interleucina-12/inmunología , Interleucina-12/metabolismo , Leucina Zippers , Masculino , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Proteína Oncogénica p65(gag-jun)/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , Proteínas Represoras/deficiencia , Proteínas Represoras/genética , Linfocitos T Colaboradores-Inductores/citología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/metabolismo , Toxoplasma/inmunologíaRESUMEN
Neuroinflammation is thought to play a pathogenic role in many neurodegenerative disorders including amyotrophic lateral sclerosis (ALS). In this study we demonstrate that the expression of nitric oxide (NO) synthase-2 (NOS2), and cyclooxygenase (COX)-2 induced by lipopolysaccharide (LPS) with interferon-γ is higher in microglial-enriched cultures from G93A-SOD1 mice, an ALS animal model, than from wild type mice. The levels of CCAAT/enhancer binding protein ß (C/EBPß), a transcription factor that regulates proinflammatory gene expression, are also upregulated in activated G93A-SOD1 microglial cells. In vivo, systemic lipopolysaccharide also induces an exacerbated neuroinflammatory response in G93A-SOD1 mice versus wild type mice, with increased expression of glial fibrillary acidic protein (GFAP), CD11b, nitric oxide synthase-2, cyclooxygenase-2, proinflammatory cytokines, and C/EBPß. Finally, we report that C/EBPß is expressed by microglia in the spinal cord of ALS patients. This is the first demonstration to our knowledge of microglial C/EBPß expression in human disease. Altogether these findings indicate that G93A-SOD1 expression results in an exacerbated pattern of neuroinflammation and suggest that C/EBPß is a candidate to regulate the expression of potentially neurotoxic genes in microglial cells in ALS.
Asunto(s)
Esclerosis Amiotrófica Lateral/metabolismo , Esclerosis Amiotrófica Lateral/patología , Proteínas Potenciadoras de Unión a CCAAT/metabolismo , Regulación de la Expresión Génica/genética , Microglía/patología , Anciano , Anciano de 80 o más Años , Esclerosis Amiotrófica Lateral/genética , Análisis de Varianza , Animales , Animales Recién Nacidos , Proteínas Potenciadoras de Unión a CCAAT/genética , Antígeno CD11b/metabolismo , Células Cultivadas , Ciclooxigenasa 2/metabolismo , Modelos Animales de Enfermedad , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Proteína Ácida Fibrilar de la Glía/metabolismo , Humanos , Interferón-alfa/farmacología , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microglía/efectos de los fármacos , Persona de Mediana Edad , Óxido Nítrico Sintasa de Tipo II/metabolismo , Proteína Oncogénica p65(gag-jun)/metabolismo , ARN Mensajero/metabolismo , Médula Espinal/metabolismo , Médula Espinal/patología , Superóxido Dismutasa/genéticaRESUMEN
In this study, we have assessed the ability of two TAT-fused peptides PYC36D-TAT and JNKI-1D-TAT (JNKI-1 or XG-102), which respectively inhibit jun proto-oncogene (c-Jun) and c-Jun N-terminal kinase (JNK) activation, to reduce infarct volume and improve functional outcome (adhesive tape removal) after transient focal cerebral ischemia in Spontaneously Hypertensive (SH) rats. PYC36D-TAT and JNKI-1D-TAT peptide batches used for experiments were tested in vitro and protected cortical neurons against glutamate excitotoxicity. Rats were treated intravenously with three different doses of PYC36D-TAT (7.7, 76, or 255 nmol/kg), JNKI-1D-TAT (255 nmol/kg), D-TAT peptide (255 nmol/kg), or saline (vehicle control), 10 minutes after reperfusion after 90 minutes of middle cerebral artery occlusion (MCAO). Contrary to other stroke models, no treatment significantly reduced infarct volume or improved functional score measurements compared with vehicle-treated animals when assessed 48 hours after MCAO. Additionally, assessment of the JNKI-1D-TAT peptide, when administered 1 or 2 hours after reperfusion after 90 minutes of MCAO, also did not improve histological or functional outcomes at 48 hours after occlusion. This study is the first to evaluate the efficacy of PYC36D-TAT and JNKI-1D-TAT using the SH rat, which has recently been shown to be more sensitive to AMPA receptor activation rather than to NMDA receptor activation after cerebral ischemia, and which may have contributed to the negative findings.