RESUMO
Human papillomavirus 18 (HPV18) is a highly malignant HPV genotype among high-risk HPVs, characterized by the difficulty of detecting it in precancerous lesions and its high prevalence in adenocarcinomas. The cellular targets and molecular mechanisms underlying its infection remain unclear. In this study, we aimed to identify the cells targeted by HPV18 and elucidate the molecular mechanisms underlying HPV18 replication. Initially, we established a lentiviral vector (HPV18LCR-GFP vector) containing the HPV18 long control region promoter located upstream of EGFP. Subsequently, HPV18LCR-GFP vectors were transduced into patient-derived squamocolumnar junction organoids, and the presence of GFP-positive cells was evaluated. Single-cell RNA sequencing of GFP-positive and GFP-negative cells was conducted. Differentially expressed gene analysis revealed that 169 and 484 genes were significantly upregulated in GFP-positive and GFP-negative cells, respectively. Pathway analysis showed that pathways associated with cell cycle and viral carcinogenesis were upregulated in GFP-positive cells, whereas keratinization and mitophagy/autophagy-related pathways were upregulated in GFP-negative cells. siRNA-mediated luciferase reporter assay and HPV18 genome replication assay validated that, among the upregulated genes, ADNP, FHL2, and NPM3 were significantly associated with the activation of the HPV18 early promoter and maintenance of the HPV18 genome. Among them, NPM3 showed substantially higher expression in HPV-related cervical adenocarcinomas than in squamous cell carcinomas, and NPM3 knockdown of HPV18-infected cells downregulated stem cell-related genes. Our new experimental model allows us to identify novel genes involved in HPV18 early promoter activities. These molecules might serve as therapeutic targets in HPV18-infected cervical lesions.
Assuntos
Adenocarcinoma , Infecções por Papillomavirus , Neoplasias do Colo do Útero , Feminino , Humanos , Papillomavirus Humano 18/genética , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Adenocarcinoma/genética , Organoides/patologiaRESUMO
Human papillomavirus (HPV) is a major cause of cervical cancer. As the natural history of HPV-associated cervical lesions is HPV genotype-dependent, it is important to understand the characteristics of these genotypes and to manage them accordingly. Among high-risk HPVs, HPV16 and 18 are particularly aggressive, together accounting for 70% of HPV genotypes detected in cervical cancer. Other than HPV16 and 18, HPV31, 33, 35, 45, 52, and 58 are also at a high risk of progression to cervical intraepithelial neoplasia (CIN)3 or higher. Recent studies have shown that the natural history of HPV16, 18, 52, and 58, which are frequently detected in Japan, depends on the HPV genotype. For example, HPV16 tends to progress in a stepwise fashion from CIN1 to CIN3, while HPV52 and 58 are more likely to persist in the CIN1 to CIN2 state. Among the high-risk HPVs, HPV18 has some peculiar characteristics different from those of other high-risk HPV types; the detection rate in precancerous lesions is much lower than those of other high-risk HPVs, and it is frequently detected in highly malignant adenocarcinoma and small cell carcinoma. Recent findings demonstrate that HPV18 may be characterized by latent infection and carcinogenesis in stem cell-like cells. In this context, this review outlines the natural history of HPV-infected cervical lesions and the characteristics of each HPV genotype.
Assuntos
Infecções por Papillomavirus , Displasia do Colo do Útero , Neoplasias do Colo do Útero , Humanos , Feminino , Neoplasias do Colo do Útero/virologia , Infecções por Papillomavirus/virologia , Displasia do Colo do Útero/virologia , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Genótipo , Papillomavirus HumanoRESUMO
Patients with cervical cancer benefiting from immune checkpoint inhibitors (ICIs) are limited. Recently, PD-L1 amplification has been attracted attention as a reliable marker of ICIs. A 47-year-old woman with stage IIB cervical cancer experienced disease progression during postoperative adjuvant chemotherapy. Cancer genomic profiling revealed that the tumor was microsatellite stable with PD-L1 amplification, therefore, nivolumab was administered by enrolling in the BELIEVE trial. Despite nivolumab treatment, remarkable disease progression was observed. At the beginning of nivolumab treatment, the patient already had multiple liver metastases with severe systemic inflammation as indicated by a high neutrophil-to-lymphocyte ratio (NLR), both of which are negative predictive markers for ICI. Despite the presence of PD-L1 amplification, nivolumab was ineffective in cancer progression, which may be attributable to the presence of liver metastasis and high NLR. ICI is recommended to be administered at an early stage of cancer treatment to enhance its effectiveness.
Assuntos
Antineoplásicos Imunológicos , Antígeno B7-H1 , Nivolumabe , Neoplasias do Colo do Útero , Antineoplásicos Imunológicos/administração & dosagem , Antineoplásicos Imunológicos/uso terapêutico , Antígeno B7-H1/genética , Biomarcadores Tumorais/genética , Quimioterapia Adjuvante , Progressão da Doença , Feminino , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/secundário , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/tratamento farmacológico , Recidiva Local de Neoplasia/genética , Nivolumabe/administração & dosagem , Nivolumabe/uso terapêutico , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/cirurgiaRESUMO
Loss of heterozygosity (LOH) has been reported to occur in HLA regions in cervical intraepithelial neoplasia (CIN) and cervical cancer. However, the details of how this is related to the progression of CIN have been unclear. In this study, we examined the human papillomavirus (HPV) antigen-presenting capacity of people with CIN and the significance of LOH of HLA class I in the progression of CIN. It was shown that differences in antigen-presenting capacity among each case depended on HLA types, not HPV genotypes. Focusing on the HLA type, there was a positive correlation between antigen-presenting capacity against HPV and the frequency of allelic loss. Furthermore, the lost HLA-B alleles had a higher HPV antigen-presenting capacity than intact alleles. In addition, frequency of LOH of HLA class I was significantly higher in advanced CIN (CIN2-3) than in cervicitis or early-stage CIN (CIN1): around half of CIN2-3 had LOH of any HLA class I. Moreover, the antigen-presenting capacity against E5, which is the HPV proteins that facilitate viral escape from this immune surveillance by suppressing HLA class I expression, had the most significant impact on the LOH in HLA-B. This study suggests that HPV evades immune surveillance mechanisms when host cells lose the capacity for antigen presentation by HLA class I molecules, resulting in long-term infection and progression to advanced lesions.
Assuntos
Antígenos de Histocompatibilidade Classe I , Perda de Heterozigosidade , Infecções por Papillomavirus , Displasia do Colo do Útero , Neoplasias do Colo do Útero , Humanos , Displasia do Colo do Útero/imunologia , Displasia do Colo do Útero/genética , Displasia do Colo do Útero/virologia , Displasia do Colo do Útero/patologia , Feminino , Neoplasias do Colo do Útero/imunologia , Neoplasias do Colo do Útero/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe I/genética , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/genética , Apresentação de Antígeno/imunologia , Adulto , Alelos , Papillomaviridae/imunologia , Vigilância Imunológica , Pessoa de Meia-Idade , GenótipoRESUMO
OBJECTIVES: This study sought to systematically evaluate the ability of a high-resolution mapping system (Rhythmia, Boston Scientific, Marlborough, Massachusetts) to rapidly and accurately localize residual endocardial and epicardial conduction after mitral isthmus (MI) ablation, facilitating MI block. BACKGROUND: Achieving conduction block across the mitral isthmus (MI) is challenging. METHODS: Fifty consecutive patients undergoing MI ablation after pulmonary vein isolation were enrolled. After initial endocardial radiofrequency (RF) ablation across the lateral MI, high-resolution activation mapping of the MI with simultaneous coronary sinus (CS) mapping was performed to verify block or localize residual conduction across the MI during left atrial (LA) appendage and CS pacing. Propagation maps were used to identify residual conduction across the MI as endocardial, via the CS or Marshall tract. RESULTS: In all 50 patients, after the initial endocardial ablation across the MI, repeat high-resolution mapping of the LA and CS was obtained (median: 3,329 mapped points; 4.0 min of mapping time). The initial endocardial MI ablation resulted in block in 9 of 50 patients (18%). In the remaining 41 patients, the propagation map identified residual conduction in 4 patterns: 1) only endocardial gap in 12 patients (29%); 2) only CS connection in 10 patients (24%); 3) both endocardial and CS connections in 14 patients (34%); and 4) Marshall tract connection in 5 patients (12%). In 8 patients, the propagation map revealed residual conduction, despite differential atrial pacing suggesting bidirectional block. Focal ablation at the identified residual conduction site (median: 0.7 min of RF) resulted in block in 49 of 50 (98%) patients. CONCLUSIONS: High-resolution propagation maps of the LA/CS rapidly and accurately localize residual endocardial and epicardial conduction across the MI. Focal ablation with short RF time at the identified gap(s) achieved complete block across MI in 98% of cases.
Assuntos
Fibrilação Atrial , Ablação por Cateter , Veias Pulmonares , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/cirurgia , Endocárdio , Frequência Cardíaca , Humanos , Veias Pulmonares/cirurgiaRESUMO
[Figure: see text].
Assuntos
Consumo de Bebidas Alcoólicas/prevenção & controle , Fibrilação Atrial/cirurgia , Ablação por Cateter/métodos , Veias Pulmonares/cirurgia , Idoso , Consumo de Bebidas Alcoólicas/epidemiologia , Fibrilação Atrial/epidemiologia , Fibrilação Atrial/fisiopatologia , Comorbidade , Feminino , Seguimentos , Humanos , Incidência , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Fatores de Tempo , Resultado do TratamentoRESUMO
BACKGROUND: Recent advances in fibrosis biology have identified transforming growth factor (TGF)-beta type I receptor-mediated activation of Smads as playing a central part in the development of fibrosis. However, to date, there have been few studies that examined the localisation and distribution of receptor-activated Smads protein (R-Smads: Smad2 and 3) during the fibrosis progression. AIMS: To histopathologically assess the time-course change of the localisation and distribution of the Smads protein in pulmonary fibrosis. METHODS: Pulmonary fibrosis was induced by intranasal injection of bleomycin (0.3 U/mouse). Lungs were isolated 2, 5, 7, 9 and 14 days after bleomycin treatment. Histological changes in the lungs were evaluated by haematoxylin-eosin stain or Masson's trichrome stain, and scored. TGF-beta1, Smad3 and phosphorylated Smad2 localisations in lung tissues were determined by immunohistochemistry. RESULTS: The bleomycin treatment led to considerable pulmonary fibrotic changes accompanied by marked increase in TGF-beta1 expression in infiltrating macrophages. With the progression in fibrosis (day 7-14), marked increases in Smad3-positive and pSmad2-positive cells were observed. There were intense Smad3-positive and pSmad2-positive signals localised to the nuclei of the infiltrating macrophages and to type II epithelial cells, and less intense signals in fibroblasts and hyperplastic alveolar/bronchiolar epithelial cells. CONCLUSIONS: The time-course data of TGF-beta1 and R-Smads indicate that progressive enhancement of TGF-beta1 signalling via R-Smad is activated in the process of fibrosis progression.
Assuntos
Fibrose Pulmonar/metabolismo , Proteínas Smad Reguladas por Receptor/metabolismo , Animais , Antibióticos Antineoplásicos , Bleomicina , Modelos Animais de Doenças , Progressão da Doença , Técnicas Imunoenzimáticas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Transdução de Sinais , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Fator de Crescimento Transformador beta/metabolismoRESUMO
OBJECTIVES: We examined the prevalence of chronic obstructive pulmonary disease (COPD) diagnosed and at-risk status, and public awareness of COPD among adults in Japan, as well as respondent characteristics and health outcomes compared with controls. METHODS: Regression models used 2012 National Health and Wellness Survey in Japan data to compare COPD-diagnosed, at-risk, and healthy adults (aged ≥18) on demographics, health behaviors, health-related quality of life (HRQoL), productivity and healthcare resource use. RESULTS: Among n = 29,978 respondents, diagnosed COPD prevalence was 0.9%; 26.9% were at-risk. Relative to controls, those at-risk and diagnosed with COPD had significantly greater healthcare resource use, with lower productivity and HRQoL. Fewer than 20% of respondents were aware of COPD. CONCLUSIONS: Over 25% of adult Japanese respondents were at-risk for COPD and had health outcomes impairments relative to controls. Efforts to increase awareness among the general public are needed.
Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Avaliação de Resultados em Cuidados de Saúde , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Qualidade de Vida , Adulto , Idoso , Eficiência , Feminino , Comportamentos Relacionados com a Saúde , Recursos em Saúde/estatística & dados numéricos , Inquéritos Epidemiológicos , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Análise de RegressãoRESUMO
BACKGROUND: A large number of chronic obstructive pulmonary disease (COPD) patients in Japan remain undiagnosed, primarily due to the underuse of spirometry. Two studies were conducted to see whether the COPD Assessment Test (CAT) in primary care has the potential to identify those patients who need spirometry for a diagnosis of COPD and to determine whether patients with cardiovascular disease had airflow limitation, which could be detected by CAT. MATERIALS AND METHODS: Two multicenter, noninterventional, prospective studies (studies 1 and 2) were conducted across Japan. Patients in both studies were ≥40 years old with a smoking history. Those in study 1 were seen in primary care and had experienced repeated respiratory tract infections, but had no diagnosis of COPD. Patients in study 2 were identified in cardiovascular disease clinics when routinely visiting for their cardiovascular disease. All patients completed the CAT prior to lung-function testing by hand-held spirometry. The presence of airflow limitation was defined as a forced expiratory volume in 1 second (FEV1)/FEV6 ratio<0.73. RESULTS: A total of 3,062 subjects completed the CAT (2,067 in study 1, 995 in study 2); 88.8% were male, and the mean age (±standard deviation) was 61.5±11.6 years. Airflow limitation was found in 400 (19.4%) patients in study 1, and 269 (27.0%) in study 2. The CAT score in patients with airflow limitation was significantly higher than in patients without airflow limitation in both studies: 8.6 (95% confidence interval [CI] 7.9-9.2) versus 7.4 (95% CI 7.1-7.6) in study 1, and 8.3 (95% CI 7.5-9.2) versus 6.4 (95% CI 6.0-6.8) in study 2 (both P<0.001). CONCLUSION: These findings suggest that the CAT has the potential to identify patients with cardiovascular disease or a history of frequent chest infections who need spirometry to diagnose COPD.
Assuntos
Pulmão/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Fumar/efeitos adversos , Inquéritos e Questionários , Doença Aguda , Adulto , Idoso , Bronquite/diagnóstico , Bronquite/epidemiologia , Bronquite/fisiopatologia , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/epidemiologia , Doenças Cardiovasculares/fisiopatologia , Diagnóstico Precoce , Feminino , Volume Expiratório Forçado , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Atenção Primária à Saúde , Estudos Prospectivos , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Fatores de Risco , Fumar/epidemiologia , EspirometriaRESUMO
BACKGROUND AND OBJECTIVES: Cardiovascular disease (CVD) and chronic obstructive pulmonary disease (COPD) commonly coexist and share common risk factors. The prevalence of COPD in outpatients with a smoking history and CVD in Japan is unknown. The aim of this study was to determine the proportion of Japanese patients with a smoking history being treated for CVD who have concurrent airflow limitation compatible with COPD. A secondary objective was to test whether the usage of lung function tests performed in the clinic influenced the diagnosis rate of COPD in the patients identified with airflow limitation. METHODS: In a multicenter observational prospective study conducted at 17 centers across Japan, the prevalence of airflow limitation compatible with COPD (defined as forced expiratory volume (FEV)1/FEV6 <0.73, by handheld spirometry) was investigated in cardiac outpatients ≥40 years old with a smoking history who routinely visited the clinic for their CVD. Each patient completed the COPD Assessment Test prior to spirometry testing. RESULTS: Data were available for 995 patients with a mean age of 66.6±10.0 years, of whom 95.5% were male. The prevalence of airflow limitation compatible with COPD was 27.0% (n=269), and 87.7% of those patients (n=236) did not have a prior diagnosis of COPD. The prevalence of previously diagnosed airflow limitation was higher in sites with higher usage of lung function testing (14.0%, 15.2% respectively) compared against sites where it is performed seldom (11.1%), but was still low. CONCLUSION: The prevalence of airflow limitation in this study indicates that a quarter of outpatients with CVD have COPD, almost all of whom are undiagnosed. This suggests that it is important to look routinely for COPD in CVD outpatients.
Assuntos
Doenças Cardiovasculares/epidemiologia , Pulmão/fisiopatologia , Pacientes Ambulatoriais/estatística & dados numéricos , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Adulto , Idoso , Doenças Cardiovasculares/diagnóstico , Comorbidade , Feminino , Volume Expiratório Forçado , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prevalência , Estudos Prospectivos , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Fatores de Risco , Fumar/efeitos adversos , Fumar/epidemiologia , EspirometriaRESUMO
Bronchial asthma is an inflammatory disease of the airways. The Sec14l3 gene, encoding a 45-kDa secretory protein, is specifically expressed in airway epithelium. Here, we report on the kinetics of Sec14l3 expression following allergic inflammation of the lung. Brown Norway rats were sensitized by intraperitoneal injection of ovalbumin, followed by challenge with aerosolized ovalbumin after a 3-week interval. This animal model showed many features similar to human allergic asthma: an increase in inflammatory cells such as eosinophils, lymphocytes and neutrophils in bronchoalveolar lavage (BAL) fluid and histopathological alteration of lung tissue, exhibiting infiltration of these inflammatory cells and degeneration and necrosis of alveolar epithelium. These parameters reached their maximal level 24h after allergen challenge. In contrast, quantitative polymerase chain reaction analyses demonstrated a rapid and significant reduction of Sec14l3 mRNA in lung tissue and maximum reduction (to 1.4% of the control) was observed at 24h. Pretreatment with dexamethasone significantly suppressed both the Sec14I3 mRNA reduction and all of the inflammatory changes. The 45-kDa secretory protein was identified in the supernatant of BAL fluids. Two-dimensional gel images of the supernatant proteome also revealed down-regulation of the protein following inflammation (to approximately 30% of the control at 24h). Thus, Sec14l3 expression is highly and inversely associated with the progression of airway inflammation. Sec14l3 mRNA and protein may function in the homeostasis of airway epithelial cells under normal conditions.
Assuntos
Proteínas de Transporte/genética , Regulação da Expressão Gênica/imunologia , Hipersensibilidade/genética , Alérgenos/imunologia , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Dexametasona/imunologia , Dexametasona/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hipersensibilidade/imunologia , Hipersensibilidade/patologia , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/patologia , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RatosRESUMO
Activin receptor-like kinase 5 (ALK5) is a type I receptor of transforming growth factor (TGF)-beta. ALK5 inhibition has been reported to attenuate the tissue fibrosis including pulmonary fibrosis, renal fibrosis and liver fibrosis. To elucidate the inhibitory mechanism of ALK5 inhibitor on pulmonary fibrosis in vivo, we performed the histopathological assessment, gene expression analysis of extracellular matrix (ECM) genes and immunohistochemistry including receptor-activated Smads (R-Smads; Smad2/3), CTGF, myofibroblast marker (alpha-smooth muscle actin; aSMA) and type I collagen deposition in the lung using Bleomycin (BLM)-induced pulmonary fibrosis model. ALK5 inhibitor, SB-525334 (10 mg/kg or 30 mg/kg) was orally administered at twice a day. Lungs were isolated 5, 7, 9 and 14 days after BLM treatment. BLM treatment led to significant pulmonary fibrotic changes accompanied by significant upregulation of ECM mRNA expressions, Smad2/3 nuclear translocation, CTGF expression, myofibroblast proliferation and type I collagen deposition. SB-525334 treatment attenuated the histopathological alterations in the lung, and significantly decreased the type I and III procollagen and fibronectin mRNA expression. Immunohistochemistry revealed that SB-525334 treatment showed significant attenuation in Smad2/3 nuclear translocation, decrease in CTGF-expressing cells, myofibroblast proliferation and type I collagen deposition. These results suggest that ALK5 inhibition attenuates R-Smads activation thereby attenuates pulmonary fibrosis.
Assuntos
Receptores de Ativinas Tipo I/antagonistas & inibidores , Receptores de Ativinas Tipo I/metabolismo , Bleomicina/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Fibrose Pulmonar/enzimologia , Fibrose Pulmonar/patologia , Receptores de Fatores de Crescimento Transformadores beta/antagonistas & inibidores , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Transporte Ativo do Núcleo Celular , Animais , Proliferação de Células , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo , Proteínas da Matriz Extracelular/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas Imediatamente Precoces/metabolismo , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Serina-Treonina Quinases , Fibrose Pulmonar/induzido quimicamente , RNA Mensageiro/genética , Receptor do Fator de Crescimento Transformador beta Tipo I , Proteína Smad2/metabolismo , Proteína Smad3/metabolismoRESUMO
The in vivo conversion ratio of N1-methylnicotinamide (NMN) to N1-methyl-2-pyridone-5-carboxamide (2-PY) and N1-methyl-4-pyridone-3-carboxamide (4-PY) as a parameter for the estimation of aldehyde oxidase level in rats was examined. NMN and its pyridones (2-PY and 4-PY) are usually detected in the urine of rats. When we measured the ratio of the amount of pyridones to the total amount of NMN and pyridones (RP value) in the urine of rats, marked intraspecies variations were observed. The variation in RP value among strains was closely related to the differences of liver aldehyde oxidase activity measured with NMN as a substrate. RP values after administration of NMN to different strains of rats confirmed the existence of strain differences of aldehyde oxidase activity in vivo. We demonstrated that measurements of NMN and its pyridones usually excreted in the urine can be used to predict the in vivo level of aldehyde oxidase.