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Laterality is a crucial physiological process intricately linked to the cilium-centrosome complex during embryo development. Defects in the process can result in severe organ mispositioning. Coiled-coil domain containing 141 (CCDC141) has been previously known as a centrosome-related gene, but its role in left-right (LR) asymmetry has not been characterized. In this study, we utilize the zebrafish model and human exome analysis to elucidate the function of ccdc141 in laterality defects. The knockdown of ccdc141 in zebrafish disrupts early LR signaling pathways, cilia function, and Kupffer's vesicle (KV) formation. Unlike ccdc141-knockdown embryos exhibiting aberrant LR patterns, ccdc141-null mutants show no apparent abnormality, suggesting a genetic compensation response effect. In parallel, we observe a marked reduction in α-tubulin acetylation levels in the ccdc141 crispants. The treatment with histone deacetylase (HDAC) inhibitors, particularly the HDAC6 inhibitor, rescues the ccdc141 crispant phenotypes. Furthermore, exome analysis of 70 patients with laterality defects reveals an increased burden of CCDC141 mutations, with in-vivo studies verifying the pathogenicity of the patient mutation CCDC141-R123G. Our findings highlight the critical role of CCDC141 in ciliogenesis and demonstrate that CCDC141 mutations lead to abnormal LR patterns. Thus, we identify CCDC141 as a causative gene for laterality defects.
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Conotruncal heart defects (CTD), subtypes of congenital heart disease, result from abnormal cardiac outflow tract development (OFT). FOXC1 and FOXC2 are closely related members of the forkhead transcription factor family and play essential roles in the development of OFT. We confirmed their expression pattern in mouse and human embryos, identifying four variants in FOXC1 and three in FOXC2 by screening these two genes in 605 patients with sporadic CTD. Western blot demonstrated expression levels, while Dual-luciferase reporter assay revealed affected transcriptional abilities for TBX1 enhancer in two FOXC1 variants and three FOXC2 variants. This might result from the altered DNA-binding abilities of mutant proteins. These results indicate that functionally impaired FOXC1 and FOXC2 variants may contribute to the occurrence of CTD.
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Fatores de Transcrição Forkhead , Cardiopatias Congênitas , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Humanos , Cardiopatias Congênitas/genética , Cardiopatias Congênitas/metabolismo , Animais , Camundongos , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismoRESUMO
Deep vein thrombosis (DVT) of the lower extremities is one of the most common peripheral vascular diseases, with significant complications and sequelae. Metabolomics aims to identify small molecules in biological samples. It can serve as a promising method for screening compounds that can be used for early disease detection, diagnosis, treatment response prediction, and prognosis. In addition, high-throughput metabolomics screening can yield significant insights into the pathophysiological pathways of DVT. Currently, the metabolomic profiles of DVT have yielded inconsistent expression patterns. This article examines the recent advancements in metabolomic studies of DVT and analyzes the factors that may influence the results.
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Trombose Venosa , Humanos , Trombose Venosa/etiologia , Extremidade Inferior/irrigação sanguínea , Prognóstico , Progressão da Doença , Fatores de RiscoRESUMO
Defects in laterality pattern can result in abnormal positioning of the internal organs during the early stages of embryogenesis, as manifested in heterotaxy syndrome and situs inversus, while laterality defects account for 3~7% of all congenital heart defects (CHDs). However, the pathogenic mechanism underlying most laterality defects remains unknown. In this study, we recruited 70 laterality defect patients with CHDs to identify candidate disease genes by exome sequencing. We then evaluated rare, loss-of-function (LOF) variants, identifying candidates by referring to previous literature. We chose TRIP11, DNHD1, CFAP74, and EGR4 as candidates from 776 LOF variants that met the initial screening criteria. After the variants-to-gene mapping, we performed function research on these candidate genes. The expression patterns and functions of these four candidate genes were studied by whole-mount in situ hybridization, gene knockdown, and gene rescue methods in zebrafish models. Among the four genes, trip11, dnhd1, and cfap74 morphant zebrafish displayed abnormalities in both cardiac looping and expression patterns of early signaling molecules, suggesting that these genes play important roles in the establishment of laterality patterns. Furthermore, we performed immunostaining and high-speed cilia video microscopy to investigate Kupffer's vesicle organogenesis and ciliogenesis of morphant zebrafish. Impairments of Kupffer's vesicle organogenesis or ciliogenesis were found in trip11, dnhd1, and cfap74 morphant zebrafish, which revealed the possible pathogenic mechanism of their LOF variants in laterality defects. These results highlight the importance of rare, LOF variants in identifying disease-related genes and identifying new roles for TRIP11, DNHD1, and CFAP74 in left-right patterning. Additionally, these findings are consistent with the complex genetics of laterality defects.
Assuntos
Cardiopatias Congênitas , Síndrome de Heterotaxia , Animais , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo , Padronização Corporal/genética , Cardiopatias Congênitas/metabolismo , Síndrome de Heterotaxia/genética , Síndrome de Heterotaxia/metabolismo , Cílios/genética , Cílios/metabolismoRESUMO
BACKGROUND: TBX1 (T-box transcription factor 1) is a major candidate gene that likely contributes to the etiology of velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS). Although the haploinsufficiency of TBX1 in both mice and humans results in congenital cardiac malformations, little has been elucidated about its upstream regulation. We aimed to explore the transcriptional regulation and dysregulation of TBX1. METHODS: Different TBX1 promoter reporters were constructed. Luciferase assays and electrophoretic mobility shift assays (EMSAs) were used to identify a cis-regulatory element within the TBX1 promoter region and its trans-acting factor. The expression of proteins was identified by immunohistochemistry and immunofluorescence. Variants in the cis-regulatory element were screened in conotruncal defect (CTD) patients. In vitro functional assays were performed to show the effects of the variants found in CTD patients on the transactivation of TBX1. RESULTS: We identified a cis-regulatory element within intron 1 of TBX1 that was found to be responsive to GATA6 (GATA binding protein 6), a transcription factor crucial for cardiogenesis. The expression patterns of GATA6 and TBX1 overlapped in the pharyngeal arches of human embryos. Transfection experiments and EMSA indicated that GATA6 could activate the transcription of TBX1 by directly binding with its GATA cis-regulatory element in vitro. Furthermore, sequencing analyses of 195 sporadic CTD patients without the 22q11.2 deletion or duplication identified 3 variants (NC_000022.11:g.19756832C > G, NC_000022.11:g.19756845C > T, and NC_000022.11:g. 19756902G > T) in the non-coding cis-regulatory element of TBX1. Luciferase assays showed that all 3 variants led to reduced transcription of TBX1 when incubated with GATA6. CONCLUSIONS: Our findings showed that TBX1 might be a direct transcriptional target of GATA6, and variants in the non-coding cis-regulatory element of TBX1 disrupted GATA6-mediated transactivation.
Assuntos
Síndrome de DiGeorge , Cardiopatias Congênitas , Animais , Síndrome de DiGeorge/genética , Fator de Transcrição GATA6 , Humanos , Camundongos , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo , Fatores de Transcrição/genética , Ativação Transcricional/genéticaRESUMO
The endothelial-to-mesenchymal transition (EndMT) is a critical process that occurs during the development of the outflow tract (OFT). Malformations of the OFT can lead to the occurrence of conotruncal defect (CTD). SOX7 duplication has been reported in patients with congenital CTD, but its specific role in OFT development remains poorly understood. To decipher this, histological analysis showed that SRY-related HMG-box 7 (SOX7) was regionally expressed in the endocardial endothelial cells and in the mesenchymal cells of the OFT, where EndMT occurs. Experiments, using in vitro collagen gel culture system, revealed that SOX7 was a negative regulator of EndMT that inhibited endocardial cell (EC) migration and resulted in decreased number of mesenchymal cells. Forced expression of SOX7 in endothelial cells blocked further migration and improved the expression of the adhesion protein vascular endothelial (VE)-cadherin (VE-cadherin). Moreover, a VE-cadherin knockdown could partly reverse the SOX7-mediated repression of cell migration. Luciferase and electrophoretic mobility shift assay (EMSA) demonstrated that SOX7 up-regulated VE-cadherin by directly binding to the gene's promoter in endothelial cells. The coding exons and splicing regions of the SOX7 gene were also scanned in the 536 sporadic CTD patients and in 300 unaffected controls, which revealed four heterozygous SOX7 mutations. Luciferase assays revealed that two SOX7 variants weakened the transactivation of the VE-cadherin promoter. In conclusion, SOX7 inhibited EndMT during OFT development by directly up-regulating the endothelial-specific adhesion molecule VE-cadherin. SOX7 mutations can lead to impaired EndMT by regulating VE-cadherin, which may give rise to the molecular mechanisms associated with SOX7 in CTD pathogenesis.
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Antígenos CD/metabolismo , Caderinas/metabolismo , Endocárdio/embriologia , Cardiopatias Congênitas/embriologia , Fatores de Transcrição SOXF/metabolismo , Animais , Antígenos CD/genética , Caderinas/genética , Movimento Celular , Embrião de Mamíferos , Endocárdio/citologia , Endotélio/crescimento & desenvolvimento , Transição Epitelial-Mesenquimal/fisiologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas , Ratos , Fatores de Transcrição SOXF/genéticaRESUMO
B cells have recently emerged as playing regulatory role in autoimmune diseases. We have previously demonstrated that human peripheral blood CD19+CD24hiCD27+ B cells have regulatory function both in healthy donors and in patients with autoimmune disease. However, the mechanism of this regulation is still not fully understood. In this study, microarrays were utilized to compare gene expression of CD19+CD24hiCD27+ B cells (regulatory B cells, Bregs) with CD19+CD24loCD27- B cells (non-Bregs) in human peripheral blood. We found that heat shock protein 70 (HSP70) expression was significantly upregulated in Bregs. In vitro studies explored that HSP70 inhibition impaired the regulatory function of peripheral blood Bregs. In mouse models of autoimmune disease, using HSP70-deficient mice or HSP70 inhibitors, Bregs suppressed effector cells and rescued disease-associated phenotypes that were dependent on HSP70. Mechanistically, Bregs secreted HSP70, directly suppressing effector cells, such as T effect cells. These findings reveal that HSP70 is a novel factor that modulates Breg function and suggest that enhancing Breg-mediated production of HSP70 could be a viable therapy for autoimmune disease.
Assuntos
Doenças Autoimunes/etiologia , Doenças Autoimunes/metabolismo , Linfócitos B/imunologia , Linfócitos B/metabolismo , Suscetibilidade a Doenças , Proteínas de Choque Térmico HSP70/metabolismo , Imunomodulação , Transferência Adotiva , Adulto , Idoso , Animais , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/terapia , Linfócitos B Reguladores/imunologia , Linfócitos B Reguladores/metabolismo , Colite/diagnóstico , Colite/etiologia , Colite/metabolismo , Colite/terapia , Citocinas/antagonistas & inibidores , Citocinas/metabolismo , Sulfato de Dextrana/efeitos adversos , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica , Proteínas de Choque Térmico HSP70/genética , Humanos , Imuno-Histoquímica , Imunofenotipagem , Masculino , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Pessoa de Meia-IdadeRESUMO
PURPOSE: This retrospective study aimed to determine the prognostic factors associated with overall survival after intracranial local and distant recurrence in patients undergoing stereotactic radiosurgery (SRS) for brain metastases. PATIENTS AND METHODS: Clinical characteristics and therapeutic parameters of 251 patients, who were treated with initial stereotactic radiosurgery for brain metastases and later experienced intracranial recurrence, were analyzed to identify prognostic factors of post-recurrence overall survival (PROS). A Cox proportional hazard model was applied for univariate and multivariate analyses. RESULTS: Among the 251 patients, the median post-recurrence overall survival was 8 months, and the six-month PROS rate was 60.2%. The interval from initial radiosurgery treatment to intracranial recurrence (hazard ratio [HR]:0.970), the number of brain recurrent tumors (HR:1.245), the number of extracranial metastatic organs (HR:1.183), recursive partition analysis (RPA) (HR:1.778), and Eastern Cooperative Oncology Group Performance Status (ECOG PS) (HR:2.442) were identified as independent prognostic factors. The patients who received local treatment for solitary brain recurrence achieved better survival (the median survival time after recurrence was 22 months). In patients without extracranial metastasis, the median post-recurrence overall survival of the local treatment group was longer than that in the whole brain radiation therapy (WBRT) group (P<0.001) and the systemic therapy group (P<0.001). CONCLUSION: A shorter interval from initial stereotactic radiosurgery to recurrence, an increasing number of brain recurrences and extracranial metastatic organs, and poor RPA and ECOG PS values are associated with poor post-recurrence prognosis. When the number of brain recurrent tumors and extracranial metastatic organs was limited, local treatment including stereotactic radiosurgery, surgery or intensity-modulated radiation therapy (IMRT) improved the post-recurrence overall survival.
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B lymphocytes are the source of autoantibodies against the thyroid-stimulating hormone receptor (TSHR) in Graves' disease (GD). Characterization of autoimmune B-cell expression profiles might enable a better understanding of GD pathogenesis. To reveal this, the expression levels of long noncoding RNAs (lncRNAs) and mRNAs (genes) in purified B cells from patients with newly diagnosed GD and healthy individuals were compared using microarrays, which elucidated 604 differentially expressed lncRNAs (DE-lncRNAs) and 410 differentially expressed genes (DEGs). GO and pathway analyses revealed that the DEGs are mainly involved in immune response. A protein-protein interaction network presented experimentally validated interactions among the DEGs. Two independent algorithms were used to identify the DE-lncRNAs that regulate the DEGs. Functional annotation of the deregulated lncRNA-mRNA pairs identified 14 pairs with mRNAs involved in cell proliferation. The lncRNAs TCONS_00022357-XLOC_010919 and n335641 were predicted to regulate TCL1 family AKT coactivator A (TCL1A), and the lncRNA n337845 was predicted to regulate SH2 domain containing 1A (SH2D1A). TCL1A and SH2D1A are highly involved in B-cell proliferation. The differential expression of both genes was validated by qRT-PCR. In conclusion, lncRNA and mRNA expression profiles of B cells from patients with GD indicated that the lncRNA-mRNA pairs n335641-TCL1A, TCONS_00022357-XLOC_010919-TCL1A, and n337845-SH2D1A may participate in GD pathogenesis by modulating B-cell proliferation and survival. Therefore, the identified lncRNA and mRNA may represent novel biomarkers and therapeutic targets for GD.
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Conotruncal heart defects (CTD) are an important subtype of congenital heart disease that occur due to abnormality in the development of the cardiac outflow tract (OFT). FOXH1 is a transcription factor that participates in the morphogenesis of the right ventricle and OFT. In this study, we confirmed the expression of FOXH1 in mouse and human embryos during OFT development. We also scanned the coding exons and splicing regions of the FOXH1 gene in 605 patients with sporadic CTD and 300 unaffected controls, from which we identified seven heterozygous FOXH1 gene mutations. According to bioinformatics analysis results, they were predicted potentially deleterious at conserved amino acid sites. Western blot was used to show that all the variants decreased the expression of FOXH1 protein, while dual-luciferase reporter assay showed that six of them, with an exception of p.P35R, had enhanced abilities to modulate the expression of MEF2C, which interacts with NKX2.5 and is involved in cardiac growth. The electrophoretic mobility shift assays result showed that two mutations altered DNA-binding abilities of mutant FOXH1 proteins. Phenotype heterogeneity was found in patients with the same mutation. These results indicate that FOXH1 mutations lead to disease-causing functional changes that contribute to the occurrence of CTD.
Assuntos
Fatores de Transcrição Forkhead/genética , Cardiopatias Congênitas/genética , Proteína Homeobox Nkx-2.5/genética , Fatores de Transcrição MEF2/genética , Animais , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Predisposição Genética para Doença , Cardiopatias Congênitas/patologia , Humanos , Lactente , Recém-Nascido , Masculino , Camundongos , Mutação de Sentido Incorreto/genéticaRESUMO
PURPOSE: Traditionally, radiosurgery was considered less effective for patients with cystic brain metastases. However, comparisons of prognosis between cystic and solid brain metastases in cancer patients have been seldom reported. We aimed to compare the survival between cystic and solid brain metastases and assess risk factors for overall survival after brain metastases (BMOS) in patients who underwent radiosurgery treatment. PATIENTS AND METHODS: The Kaplan-Meier method and multivariate Cox regression analysis were used to compare survival time and evaluate risk factors for BMOS. RESULTS: A total of 356 patients (including 498 brain metastases) were analyzed in our study, including 67 patients (67/356, 18.8%) with 75 cystic brain metastases. There is no statistical significance in BMOS between patients with cystic (17 months, range: 3-64 months) and solid (17.5 months, range: 1-65 months) brain metastases (P=0.148). However, the volume of cystic brain metastases decreased more slowly than solid brain metastases (P<0.05). The results indicated that high recursive partitioning analysis classification (P=0.006), large volume of brain metastases (P=0.006), and different primary lesion (especially gastrointestinal tract tumor) (P=0.001) were associated with poor prognosis in patients with brain metastases. CONCLUSION: There is no difference in prognosis and local control between patients with cystic and solid brain metastases who underwent radiosurgery. However, the rate and speed of tumor shrinkage were lower in cystic brain metastases after radiotherapy. Patients with larger brain metastases had shorter survival time, regardless of cystic or solid brain metastases.
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OBJECTIVES: The purpose of this research was to study the outcome of brain metastases in a cohort of patients undergoing a second course of stereotactic image-guided robotic radiosurgery and to identify predictors corelated with survival. METHODS: A total of 63 patients with primary malignancies underwent a second course of CyberKnife radiosurgery for intracranial progression, including recurrence and new metastases after initial stereotactic radiosurgery (SRS). Overall survival (OS) and control rate were calculated by the Kaplan-Meier method. A Cox proportional hazards model was used to analyze predictive factors for survival. RESULTS: With a median follow-up duration of 12 months after second SRS, the median OS of the second course of radiosurgery was 18 months. On multivariate analysis, the sum of total planned target volume (hazard ratio, 2.112; 95% confidence interval, 1.069-4.173) and minimum dose (hazard ratio, 1.990, 95% confidence interval, 1.017-3.892) were significantly associated with OS. Median intracranial progression-free survival was 23 months. The 6-month and 12-month local control rates of the targets were 97.0% and 94.4%, respectively. Univariate analysis showed that only tumor number significantly influenced intracranial progression-free survival (P = 0.012). Nine patients (14.2%) developed brain necrosis. Median time to brain necrosis in regions in which brain necrosis occurred after a single course of SRS was not reached, compared with 16 months for those treated with repeat SRS (P = 0.041). CONCLUSIONS: A second course of CyberKnife radiosurgery seems to be an effective salvage option for brain progression after initial SRS. The total planned target volume shows prediction for OS. Tumor volume of initial SRS may influence selection of the potential population that may benefit from salvage radiosurgery.
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Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/secundário , Radiocirurgia , Radioterapia Guiada por Imagem , Reirradiação , Procedimentos Cirúrgicos Robóticos , Adulto , Idoso , Neoplasias Encefálicas/mortalidade , Progressão da Doença , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Necrose/etiologia , Lesões por Radiação , Radiocirurgia/métodos , Dosagem Radioterapêutica , Radioterapia Guiada por Imagem/métodos , Estudos Retrospectivos , Procedimentos Cirúrgicos Robóticos/métodos , Terapia de Salvação , Análise de Sobrevida , Resultado do TratamentoRESUMO
BACKGROUND & AIMS: B cells infiltrate tumors, but little is known about how they affect tumor growth and progression. microRNA15A (MIR15A or miRNA15A) and microRNA16-1 (MIR16-1 or miRNA16-1) regulate cell proliferation, apoptosis, and drug resistance. We investigated their involvement in B-cell-mediated immune suppression by colorectal tumors. METHODS: Mice with disruptions of the gene cluster that encodes MIR15A and MIR16-1 (knockout mice), and control (C57BL/B6) mice were given azoxymethane with dextran sodium sulfate (AD) to induce formation of colorectal tumors. Mice were given anti-CD20 to delete B cells, or injections of agomir to increase MIR15A and MIR16-1. Proliferation of CD8+T cells was measured by carboxyfluorescein-succinimidyl-ester analysis. Colon tissues were collected from mice and analyzed by flow cytometry, microRNA (miRNA) sequencing, and for cytokine production. Intestinal epithelial cells (IECs) were isolated and transfected with miRNA mimics, to identify their targets. We analyzed miRNA expression patterns and quantified B cells in colorectal cancer tissue microarrays derived from 90 patients who underwent surgical resection, from July 2006 through April 2008, in Shanghai, China; expression data were compared with clinical outcomes. RESULTS: Tumors that developed in knockout mice following administration of AD were larger and contained greater numbers of B cells than tumors that grew in control mice. Most of the B cells in the tumors were positive for immunoglobulin A (IgA+). IgA+ B cells expressed high levels of immune regulatory molecules (programmed death ligand 1, interleukin 10, and transforming growth factor beta), and repressed the proliferation and activation of CD8+ T cells. Levels of MIR15A and MIR16-1 were reduced in colon tumors from mice, compared with nontumor colon tissue. Incubation of IECs with IL17A reduced expression of MIR15A and MIR16-1. Transgenic expression of MIR15A and MIR16-1 in IECs decreased activation of NF-κB and STAT1 by reducing expression of I-kappaB kinases; this resulted in reduced production of chemokine (C-X-C motif) ligands 9 and 10 and decreased chemotaxis of IgA+ B cells. Tumors in mice injected with AD and agomir grew more slowly than tumors in mice not given in agomir and contained fewer IgA+ B cells. We found a negative correlation between levels of MIR15A and MIR16-1 and numbers of IgA+B cells in human colorectal tumor tissues; high levels of MIR15A and MIR16-1 and low numbers of IgA+B cells were associated with longer survival times of patients. CONCLUSIONS: We found increased levels of MIR15A and MIR16-1 to reduce numbers of IgA+ B cells in colorectal tumor tissues and correlate with increased survival time of patients. In mice that lack MIR15A and MIR16-1, colon tumors grow more rapidly and contain increased numbers of IgA+ B cells. MIR15A and MIR16-1 appear to activate signaling pathways required for B-cell-mediated immune suppression.
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Linfócitos B Reguladores/metabolismo , Quimiotaxia de Leucócito , Neoplasias Colorretais/metabolismo , Linfócitos do Interstício Tumoral/metabolismo , MicroRNAs/metabolismo , Transdução de Sinais , Evasão Tumoral , Animais , Azoximetano , Linfócitos B Reguladores/imunologia , Proliferação de Células , Quimiocina CXCL10/imunologia , Quimiocina CXCL10/metabolismo , Quimiocina CXCL9/imunologia , Quimiocina CXCL9/metabolismo , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/genética , Neoplasias Colorretais/imunologia , Sulfato de Dextrana , Regulação Neoplásica da Expressão Gênica , Genótipo , Humanos , Quinase I-kappa B/metabolismo , Linfócitos do Interstício Tumoral/imunologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , MicroRNAs/genética , NF-kappa B/metabolismo , Fenótipo , Fator de Transcrição STAT1/metabolismo , Fatores de Tempo , Carga TumoralRESUMO
Signal transducer and activator of transcription 6 (STAT6) is involved in epithelial cell growth. However, little is known regarding the STAT6 phosphorylation status in Graves' disease (GD) and its role in thyroid epithelial cells (TECs). In this study, we found that STAT6 phosphorylation (p-STAT6) was significantly increased in TECs from both GD patients and experimental autoimmune Graves' disease mice and that STAT6 deficiency ameliorated GD symptoms. Autocrine IL-4 signalling in TECs activated the phosphorylation of STAT6 via IL-4 R engagement, and the downstream targets of STAT6 were Bcl-xL and cyclin D1. Thus, the IL-4-STAT6-Bcl-xL/cyclin D1 pathway is crucial for TEC hyperplasia, which aggravates GD. More importantly, in vitro and in vivo experiments demonstrated that STAT6 phosphorylation inhibited by AS1517499 decreased TEC hyperplasia, thereby reducing serum T3 and T4 and ameliorating GD. Thus, our study reveals that in addition to the traditional pathogenesis of GD, in which autoantibody TRAb stimulates thyroid-stimulating hormone receptors and consequently produces T3, T4, TRAb could also trigger TECs producing IL-4, and IL-4 then acts in an autocrine manner to activate p-STAT6 signalling and stimulate unrestricted cell growth, thus aggravating GD. These findings suggest that STAT6 inhibitors could be potent therapeutics for treating GD.
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Doença de Graves/metabolismo , Doença de Graves/patologia , Fator de Transcrição STAT6/deficiência , Índice de Gravidade de Doença , Células Epiteliais da Tireoide/metabolismo , Células Epiteliais da Tireoide/patologia , Animais , Ciclina D1/metabolismo , Humanos , Hiperplasia , Interleucina-4/metabolismo , Camundongos Endogâmicos BALB C , Modelos Biológicos , Fosforilação/efeitos dos fármacos , Pirimidinas/farmacologia , Receptores de Interleucina/metabolismo , Fator de Transcrição STAT6/metabolismo , Células Epiteliais da Tireoide/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Proteína bcl-X/metabolismoRESUMO
Biological response modifiers (BRMs) emerge as a lay of new compounds or approaches used in improving cancer immunotherapy. Evidences highlight that cytokines, Toll-like receptor (TLR) signaling, and noncoding RNAs are of crucial roles in modulating antitumor immune response and cancer-related chronic inflammation, and BRMs based on them have been explored. In particular, besides some cytokines like IFN-α and IL-2, several Toll-like receptor (TLR) agonists like BCG, MPL, and imiquimod are also licensed to be used in patients with several malignancies nowadays, and the first artificial small noncoding RNA (microRNA) mimic, MXR34, has entered phase I clinical study against liver cancer, implying their potential application in cancer therapy. According to amounts of original data, this chapter will review the regulatory roles of TLR signaling, some noncoding RNAs, and several key cytokines in cancer and cancer-related immune response, as well as the clinical cases in cancer therapy based on them.
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Aminoquinolinas/uso terapêutico , Antineoplásicos/uso terapêutico , Regulação Neoplásica da Expressão Gênica/imunologia , Fatores Imunológicos/uso terapêutico , Imunoterapia/métodos , Neoplasias/terapia , Ensaios Clínicos como Assunto , Humanos , Imiquimode , Interferon-alfa/genética , Interferon-alfa/imunologia , Interleucina-2/genética , Interleucina-2/imunologia , Mycobacterium bovis/genética , Mycobacterium bovis/imunologia , Neoplasias/genética , Neoplasias/imunologia , Neoplasias/patologia , RNA não Traduzido/genética , RNA não Traduzido/imunologia , RNA não Traduzido/uso terapêutico , Receptores de Trombopoetina/genética , Receptores de Trombopoetina/imunologia , Transdução de Sinais , Receptores Toll-Like/agonistas , Receptores Toll-Like/genética , Receptores Toll-Like/imunologiaRESUMO
UNLABELLED: Increasing evidence in recent years has suggested that B cells act as a crucial regulator in autoimmune diseases. However, little is known about their role in autoimmune hepatitis (AIH) and the underlying regulatory mechanisms. In this study, we show that B cells ameliorated experimental AIH (EAH) by suppressing CD4+ T-cell responses and that CD11b expression on B cells was required for the regulatory function of B cells. In vitro studies reveal that the suppressive function of CD11b was mediated by the impairment of T-cell antigen receptor (TCR) signaling transduction and the promotion of TCR down-regulation. Moreover, we show that the increased CD11b expression on B cells was interleukin (IL)-10 dependent and that additional IL-10 stimulation promoted CD11b expression on B cells, thereby enhancing B-cell regulatory effects. CONCLUSION: These findings reveal a previously unrecognized role for CD11b in B-cell regulatory function and its protective effect on EAH.
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Linfócitos B/imunologia , Antígeno CD11b/fisiologia , Linfócitos T CD4-Positivos/imunologia , Hepatite Autoimune/imunologia , Animais , Antígenos CD20/imunologia , Antígeno CD11b/análise , Hepatite Autoimune/prevenção & controle , Interferon gama/biossíntese , Interleucina-10/fisiologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BLRESUMO
PURPOSE: MicroRNAs (miRNA) are involved in and are controlled by epigenetic regulation, and thereby form a reciprocal regulatory circuit. Using next-generation sequencing (NGS)-based miRNA profiling, this study aimed to discover esophageal squamous cell carcinoma (ESCC)-specific miRNAs and miRNA-related epigenetic modulations. EXPERIMENTAL DESIGN: NGS-based miRNA profiles were generated for four pairs of ESCC tissues and adjacent normal tissues. In situ hybridization was used to assess miRNA expression and its correlation with prognosis. miRNA-related DNA methylations were identified using bisulfite genomic sequencing, and the role of DNA methyltransferase 1 (DNMT1) was investigated using RNA interference. miRNA targets were screened by mRNA sequencing, and functional validation was performed in vitro and in vivo. RESULTS: NGS-based miRNA profiling identified 78 differentially expressed miRNAs in ESCC. Among them, microRNA126-3p (miR-126) was significantly downregulated, and its downregulation correlated with poor ESCC prognosis. Downregulation of miR-126 was due to promoter hypermethylation of its host gene, Egfl7. DNMT1 was aberrantly upregulated in ESCC and responsible for the hypermethylation of Egfl7. Intriguingly, DNMT1 was suppressed by overexpression of miR-126, indicating the existence of a regulatory feedback circuit. ADAM9 was identified as a key target of miR-126. Ectopic expression of miR-126 or silencing of ADAM9 reduced ESCC cell proliferation and migration by inhibiting epidermal growth factor receptor-AKT signaling. CONCLUSIONS: Our results indicate that miR-126 is a potential prognostic indicator for ESCC and suggest that a novel "DNMT1-miR-126 epigenetic circuit" is involved in ESCC progression. Consequently, miR-126-based epigenetic modulations may provide a basic rationale for new approaches to antitumor therapeutics.
Assuntos
Carcinoma de Células Escamosas/genética , DNA (Citosina-5-)-Metiltransferases/metabolismo , Epigênese Genética , Neoplasias Esofágicas/genética , Regulação Neoplásica da Expressão Gênica/genética , MicroRNAs/genética , Transdução de Sinais , Proteínas ADAM/metabolismo , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células , DNA (Citosina-5-)-Metiltransferase 1 , Metilação de DNA , Receptores ErbB/metabolismo , Neoplasias Esofágicas/mortalidade , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago , Genes Supressores de Tumor , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Hibridização In Situ , Estimativa de Kaplan-Meier , Proteínas de Membrana/metabolismo , MicroRNAs/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Análise Serial de TecidosRESUMO
Dextran sodium sulphate (DSS) induced colitis is commonly used to simulate human ulcerative colitis (UC). However, the mucosal immune responses related to the severity of disease have not been comprehensively documented. We used different concentration of DSS, induced various severities of colitis, and simultaneously examined the frequency of immune cells, antibodies and cytokine production. We found that T regulatory cells (Tregs), B cells, and IgA secretion increased on the recovery phase of mild colitis, accompanied by CD11b(+) cells, interleukin (IL)-6 and tumor necrosis factor (TNF)-α accumulated mildly. While during severe and irreversible colitis, the CD11b(+) cells, IL-6, and TNF-α infiltrated severely with Tregs, B cells, and IgA increased inconspicuously. These results demonstrate that Tregs, B cells, and IgA may play a significant role in maintaining the homeostasis of gut, by suppressing CD11b(+) cells and the pro-inflammatory cytokines.
Assuntos
Colite/induzido quimicamente , Sulfato de Dextrana/farmacologia , Animais , Linfócitos B/imunologia , Antígeno CD11b/metabolismo , Colite/imunologia , Sulfato de Dextrana/administração & dosagem , Homeostase , Imunoglobulina A/metabolismo , Interleucina-6/metabolismo , Camundongos Endogâmicos C57BL , Linfócitos T Reguladores/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Paclitaxel (PTX) is the front-line chemotherapeutic agent against human non-small cell lung cancer (NSCLC). However, its therapeutic efficacy is restricted by the increasing frequency of chemotherapeutic resistance in NSCLC. Accumulating evidence has shown the potential role of microRNAs (miRNAs) in the chemotherapeutic sensitivity of cancer cells. Previously it was reported that microRNA-7 (miR-7) acts as an important tumor suppressor in NSCLC. Therefore, the present study was conducted to determine the regulatory role of miR-7 in PTX chemotherapy for NSCLC. Four NSCLC cell lines were used to analyze the correlation of the PTX-sensitivity and endogenoaus miR-7 expression. miR-7 expression was up- and downregulated using miR-7 mimics and inhibitors respectively, and the role of miR-7 in sensitizing NSCLC cells to PTX was assessed by cell viability and apoptosis assays. The molecular mechanism of PTX sensitivity was determined by quantitative polymerase chain reaction and western blotting. It was found that the sensitivity of NSCLC cells to PTX was dependent on endogenous miR-7. Upregulation of miR-7 enhanced the PTX-sensitivity of NSCLC cells by suppressing cell proliferation and promoting cell apoptosis, while the inhibition of miR-7 abrogated the antiproliferative proapoptotic effects of PTX. Pretreatment of miR-7 mimics enhanced the PTX-mediated downregulation of epidermal growth factor receptor (EGFR) in NSCLC cells. These results have identified miR-7 as a potential EGFR-targeting sensitizer in PTX therapy. These data may facilitate the development of novel chemotherapeutic approaches for NSCLC.
RESUMO
Systemic lupus erythematosus (SLE) is characterized by prominent autoinflammatory tissue damage associated with impaired removal of dying cells and DNA. Self DNA-containing immune complexes are able to activate both innate and adaptive immune responses and play an important role in the maintenance and exacerbation of autoimmunity in SLE. In this study, we used DNA from lymphocytes that have undergone activation-induced cell death (ALD-DNA) and analyzed its role on the activation and differentiation of B cells from normal BALB/c mice as well as lupus-prone MRL+/+ and MRL/lpr mice. We found that ALD-DNA directly increased the expression of costimulatory molecules and the survival of naïve B cells in vitro. Although ALD-DNA alone had little effect on the proliferation of naïve B cells, it enhanced LPS-activated B cell proliferation in vitro and in vivo. In addition, ALD-DNA increased plasma cell numbers and IgG production in LPS-stimulated cultures of naïve B cells, in part via enhancing IL-6 production. Importantly, B cells from lupus mice were hyperresponsive to ALD-DNA and/or LPS relative to normal control B cells in terminal plasma cell differentiation, as evidenced by increases in CD138+ cell numbers, IgM production, and mRNA levels of B lymphocyte-induced maturation protein-1 (Blimp-1) and the X-box binding protein 1 (XBP1). Furthermore, ALD-DNA enhanced CD40-activated naïve B cell proliferation. Collectively, these data indicate that self DNA can serve as a DAMP (damage-associated molecular pattern) that cooperates with signals from both innate and adaptive immunity to promote polyclonal B cell activation, a common characteristic of autoimmune diseases.