RESUMO
Fibroblastic foci, known to be the leading edge of fibrosis development in idiopathic pulmonary fibrosis (IPF), are composed of fibrogenic myofibroblasts. Autophagy has been implicated in the regulation of myofibroblast differentiation. Insufficient mitophagy, the mitochondria-selective autophagy, results in increased reactive oxygen species, which may modulate cell signaling pathways for myofibroblast differentiation. Therefore, we sought to investigate the regulatory role of mitophagy in myofibroblast differentiation as a part of IPF pathogenesis. Lung fibroblasts were used in in vitro experiments. Immunohistochemical evaluation in IPF lung tissues was performed. PARK2 was examined as a target molecule for mitophagy regulation, and a PARK2 knockout mouse was employed in a bleomycin-induced lung fibrosis model. We demonstrated that PARK2 knockdown-mediated mitophagy inhibition was involved in the mechanism for activation of the platelet-derived growth factor receptor (PDGFR)/PI3K/AKT signaling pathway accompanied by enhanced myofibroblast differentiation and proliferation, which were clearly inhibited by treatment with both antioxidants and AG1296, a PDGFR inhibitor. Mitophagy inhibition-mediated activation of PDGFR signaling was responsible for further autophagy suppression, suggesting the existence of a self-amplifying loop of mitophagy inhibition and PDGFR activation. IPF lung demonstrated reduced PARK2 with concomitantly increased PDGFR phosphorylation. Furthermore, bleomycin-induced lung fibrosis was enhanced in PARK2 knockout mice and subsequently inhibited by AG1296. These findings suggest that insufficient mitophagy-mediated PDGFR/PI3K/AKT activation, which is mainly attributed to reduced PARK2 expression, is a potent underlying mechanism for myofibroblast differentiation and proliferation in fibroblastic foci formation during IPF pathogenesis.
Assuntos
Fibrose Pulmonar Idiopática/patologia , Mitofagia/fisiologia , Miofibroblastos/patologia , Ubiquitina-Proteína Ligases/metabolismo , Animais , Western Blotting , Diferenciação Celular/fisiologia , Imunofluorescência , Humanos , Fibrose Pulmonar Idiopática/metabolismo , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Knockout , Microscopia Eletrônica de Transmissão , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores do Fator de Crescimento Derivado de Plaquetas/metabolismo , Transdução de Sinais/fisiologiaRESUMO
A 48-year-old man developed general fatigue, dyspnea, and fever at an altitude of 1562 m from the morning of the first day of a 3-day hike. Despite pharyngeal discomfort and mild general fatigue, he felt that the symptoms were not sufficient to abandon his plan. He usually required 1.5 hours to reach Tokusawa (6.4 km from the starting point at an altitude of 1500 m), but this time he required 2.5 hours and slept briefly upon arrival at Tokusawa due to extreme fatigue and respiratory discomfort. His symptoms became aggravated, so he presented at a mountain clinic with oxygen saturation at 80% and body temperature of 37.6ºC. He was diagnosed with hypoxemia due to pneumonia and/or other disease(s) and was evacuated to a hospital where a chest computed tomography scan revealed ground glass opacity and infiltrative shadows. He was treated for pneumonia, but another doctor discovered during follow-up that the patient had sprayed 300 mL of a waterproofing aerosol on mountain equipment in a nonventilated, enclosed area of his home on the night before starting out on the hike. Therefore, waterproofing spray was considered to have caused pulmonary damage. Self-reporting or appropriate questionnaires are the only means of identifying this type of injury. The differential diagnosis of pulmonary problems in an outdoor setting should include toxic aerosol exposure from waterproofing spray.
Assuntos
Lesão Pulmonar/diagnóstico , Recreação , Aerossóis , Diagnóstico Diferencial , Humanos , Japão , Lesão Pulmonar/diagnóstico por imagem , Lesão Pulmonar/etiologia , Masculino , Pessoa de Meia-IdadeRESUMO
Cigarette smoke (CS)-induced cellular senescence has been implicated in the pathogenesis of chronic obstructive pulmonary disease, and SIRT6, a histone deacetylase, antagonizes this senescence, presumably through the attenuation of insulin-like growth factor (IGF)-Akt signaling. Autophagy controls cellular senescence by eliminating damaged cellular components and is negatively regulated by IGF-Akt signaling through the mammalian target of rapamycin (mTOR). SIRT1, a representative sirtuin family, has been demonstrated to activate autophagy, but a role for SIRT6 in autophagy activation has not been shown. Therefore, we sought to investigate the regulatory role for SIRT6 in autophagy activation during CS-induced cellular senescence. SIRT6 expression levels were modulated by cDNA and small interfering RNA transfection in human bronchial epithelial cells (HBECs). Senescence-associated ß-galactosidase staining and Western blotting of p21 were performed to evaluate senescence. We demonstrated that SIRT6 expression levels were decreased in lung homogenates from chronic obstructive pulmonary disease patients, and SIRT6 expression levels correlated significantly with the percentage of forced expiratory volume in 1 s/forced vital capacity. CS extract (CSE) suppressed SIRT6 expression in HBECs. CSE-induced HBEC senescence was inhibited by SIRT6 overexpression, whereas SIRT6 knockdown and mutant SIRT6 (H133Y) without histone deacetylase activity enhanced HBEC senescence. SIRT6 overexpression induced autophagy via attenuation of IGF-Akt-mTOR signaling. Conversely, SIRT6 knockdown and overexpression of a mutant SIRT6 (H133Y) inhibited autophagy. Autophagy inhibition by knockdown of ATG5 and LC3B attenuated the antisenescent effect of SIRT6 overexpression. These results suggest that SIRT6 is involved in CSE-induced HBEC senescence via autophagy regulation, which can be attributed to attenuation of IGF-Akt-mTOR signaling.
Assuntos
Autofagia/fisiologia , Brônquios/patologia , Senescência Celular/fisiologia , Células Epiteliais/patologia , Fator de Crescimento Insulin-Like I/fisiologia , Doença Pulmonar Obstrutiva Crônica/patologia , Sirtuínas/fisiologia , Fumaça/efeitos adversos , Acetilação , Proteína 5 Relacionada à Autofagia , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Volume Expiratório Forçado , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Associadas aos Microtúbulos/fisiologia , Mutação , Processamento de Proteína Pós-Traducional , Proteínas Proto-Oncogênicas c-akt/fisiologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Interferência de RNA , RNA Interferente Pequeno/farmacologia , Transdução de Sinais/fisiologia , Sirtuínas/antagonistas & inibidores , Sirtuínas/genética , Serina-Treonina Quinases TOR/fisiologia , Nicotiana , Capacidade VitalRESUMO
Programmed cell death ligand-1 (PD-L1) has recently gained considerable attention for its role in tumor immune escape. Here, we identify a miR-197/CKS1B/STAT3-mediated PD-L1 network in chemoresistant non-small-cell lung cancer (NSCLC), independent of immunoinhibitory signals. miR-197 is downregulated in platinum-resistant NSCLC specimens, resulting in the promotion of chemoresistance, tumorigenicity, and pulmonary metastasis in vitro and in vivo. Mechanistic investigations reveal that a miR-197-mediated CKS1B/STAT3 axis exerts tumor progression regulated by various oncogenic genes (Bcl-2, c-Myc, and cyclin D1), and PD-L1 is a putative biomarker of this axis. Furthermore, we demonstrate that a miR-197 mimic sensitizes PD-L1(high) drug-resistant cells to chemotherapy. These results indicate that the biological interaction between PD-L1 and chemoresistance occurs through the microRNA regulatory cascade. More importantly, expression levels of miR-197 are inversely correlated with PD-L1 expression (n = 177; P = 0.026) and are associated with worse overall survival (P = 0.015). Our discoveries suggest that the miR-197/CKS1B/STAT3-mediated network can drive tumor PD-L1 expression as a biomarker of this cascade, and miR-197 replacement therapy may be a potential treatment strategy for chemoresistant NSCLC.
Assuntos
Antineoplásicos/uso terapêutico , Antígeno B7-H1/metabolismo , Quinases relacionadas a CDC2 e CDC28/metabolismo , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos , Neoplasias Pulmonares/tratamento farmacológico , MicroRNAs/metabolismo , Fator de Transcrição STAT3/metabolismo , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Linhagem Celular Tumoral , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , MicroRNAs/genética , Análise de SobrevidaRESUMO
Mitochondria are dynamic organelles that continuously change their shape through fission and fusion. Disruption of mitochondrial dynamics is involved in disease pathology through excessive reactive oxygen species (ROS) production. Accelerated cellular senescence resulting from cigarette smoke exposure with excessive ROS production has been implicated in the pathogenesis of chronic obstructive pulmonary disease (COPD). Hence, we investigated the involvement of mitochondrial dynamics and ROS production in terms of cigarette smoke extract (CSE)-induced cellular senescence in human bronchial epithelial cells (HBEC). Mitochondrial morphology was examined by electron microscopy and fluorescence microscopy. Senescence-associated ß-galactosidase staining and p21 Western blotting of primary HBEC were performed to evaluate cellular senescence. Mitochondrial-specific superoxide production was measured by MitoSOX staining. Mitochondrial fragmentation was induced by knockdown of mitochondrial fusion proteins (OPA1 or Mitofusins) by small-interfering RNA transfection. N-acetylcysteine and Mito-TEMPO were used as antioxidants. Mitochondria in bronchial epithelial cells were prone to be more fragmented in COPD lung tissues. CSE induced mitochondrial fragmentation and mitochondrial ROS production, which were responsible for acceleration of cellular senescence in HBEC. Mitochondrial fragmentation induced by knockdown of fusion proteins also increased mitochondrial ROS production and percentages of senescent cells. HBEC senescence and mitochondria fragmentation in response to CSE treatment were inhibited in the presence of antioxidants. CSE-induced mitochondrial fragmentation is involved in cellular senescence through the mechanism of mitochondrial ROS production. Hence, disruption of mitochondrial dynamics may be a part of the pathogenic sequence of COPD development.
Assuntos
Brônquios/patologia , Senescência Celular/efeitos dos fármacos , Células Epiteliais/patologia , Mitocôndrias/patologia , Nicotiana/efeitos adversos , Espécies Reativas de Oxigênio/metabolismo , Western Blotting , Brônquios/efeitos dos fármacos , Brônquios/metabolismo , Células Cultivadas , Dinaminas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , GTP Fosfo-Hidrolases/antagonistas & inibidores , GTP Fosfo-Hidrolases/genética , GTP Fosfo-Hidrolases/metabolismo , Humanos , Técnicas Imunoenzimáticas , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Microscopia Eletrônica , Proteínas Associadas aos Microtúbulos/antagonistas & inibidores , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas Mitocondriais/antagonistas & inibidores , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , RNA Interferente Pequeno/genéticaRESUMO
Autophagy, a process that helps maintain homeostatic balance between the synthesis, degradation, and recycling of organelles and proteins to meet metabolic demands, plays an important regulatory role in cellular senescence and differentiation. Here we examine the regulatory role of autophagy in idiopathic pulmonary fibrosis (IPF) pathogenesis. We test the hypothesis that epithelial cell senescence and myofibroblast differentiation are consequences of insufficient autophagy. Using biochemical evaluation of in vitro models, we find that autophagy inhibition is sufficient to induce acceleration of epithelial cell senescence and myofibroblast differentiation in lung fibroblasts. Immunohistochemical evaluation of human IPF biospecimens reveals that epithelial cells show increased cellular senescence, and both overlaying epithelial cells and fibroblasts in fibroblastic foci (FF) express both ubiquitinated proteins and p62. These findings suggest that insufficient autophagy is an underlying mechanism of both accelerated cellular senescence and myofibroblast differentiation in a cell-type-specific manner and is a promising clue for understanding the pathogenesis of IPF.
Assuntos
Autofagia , Fibrose Pulmonar Idiopática/fisiopatologia , Proteínas Adaptadoras de Transdução de Sinal/biossíntese , Diferenciação Celular/fisiologia , Senescência Celular/fisiologia , Estresse do Retículo Endoplasmático/fisiologia , Células Epiteliais/patologia , Células Epiteliais/fisiologia , Humanos , Miofibroblastos/citologia , Proteína Sequestossoma-1 , Tunicamicina/farmacologia , Ubiquitina/biossínteseRESUMO
BACKGROUND: Marked accumulation of alveolar macrophages (AM) conferred by apoptosis resistance has been implicated in pathogenesis of chronic obstructive pulmonary disease (COPD). Apoptosis inhibitor of macrophage (AIM), has been shown to be produced by mature tissue macrophages and AIM demonstrates anti-apoptotic property against multiple apoptosis-inducing stimuli. Accordingly, we attempt to determine if AIM is expressed in AM and whether AIM is involved in the regulation of apoptosis in the setting of cigarette smoke extract (CSE) exposure. METHODS: Immunohistochemical evaluations of AIM were performed. Immunostaining was assessed by counting total and positively staining AM numbers in each case (n = 5 in control, n = 5 in non-COPD smoker, n = 5 in COPD). AM were isolated from bronchoalveolar lavage fluid (BALF). The changes of AIM expression levels in response to CSE exposure in AM were evaluated. Knock-down of anti-apoptotic Bcl-xL was mediated by siRNA transfection. U937 monocyte-macrophage cell line was used to explore the anti-apoptotic properties of AIM. RESULTS: The numbers of AM and AIM-positive AM were significantly increased in COPD lungs. AIM expression was demonstrated at both mRNA and protein levels in isolated AM, which was enhanced in response to CSE exposure. AIM significantly increased Bcl-xL expression levels in AM and Bcl-xL was involved in a part of anti-apoptotic mechanisms of AIM in U937 cells in the setting of CSE exposure. CONCLUSIONS: These results suggest that AIM expression in association with cigarette smoking may be involved in accumulation of AM in COPD.
Assuntos
Proteínas Reguladoras de Apoptose/biossíntese , Regulação da Expressão Gênica , Macrófagos Alveolares/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Idoso , Antígenos CD/biossíntese , Antígenos CD/genética , Antígenos de Diferenciação de Linfócitos T/biossíntese , Antígenos de Diferenciação de Linfócitos T/genética , Proteínas Reguladoras de Apoptose/genética , Líquido da Lavagem Broncoalveolar , Células Cultivadas , Feminino , Células HEK293 , Humanos , Lectinas Tipo C/biossíntese , Lectinas Tipo C/genética , Macrófagos Alveolares/patologia , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/patologia , Células U937RESUMO
TLR3, one of the TLRs involved in the recognition of infectious pathogens for innate and adaptive immunity, primarily recognizes viral-associated dsRNA. Recognition of dsRNA byproducts released from apoptotic and necrotic cells is a recently proposed mechanism for the amplification of toxicity, suggesting a pivotal participation of TLR3 in viral infection, as well as in lung diseases where apoptosis plays a critical role, such as asthma and chronic obstructive pulmonary disease. In addition to metabolic control, insulin signaling was postulated to be protective by inhibiting apoptosis. Therefore, we explored the role of insulin signaling in protecting against TLR3-mediated apoptosis of human bronchial epithelial cells. Significant TLR3-mediated apoptosis was induced by polyinosinic-polycytidylic acid, a dsRNA analog, via caspase-8-dependent mechanisms. However, insulin efficiently inhibited TLR3/polyinosinic-polycytidylic acid-induced human bronchial epithelial cell apoptosis via PI3K/Akt and ERK pathways, at least in part, via upregulation of cellular FLIPs and through protein synthesis-independent mechanisms. These results indicate the significance of TLR3-mediated dsRNA-induced apoptosis in the pathogenesis of apoptosis-driven lung disease and provide evidence for a novel protective role of insulin.
Assuntos
Apoptose/imunologia , MAP Quinases Reguladas por Sinal Extracelular/fisiologia , Insulina/fisiologia , Sistema de Sinalização das MAP Quinases/imunologia , Fosfatidilinositol 3-Quinase/fisiologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Mucosa Respiratória/imunologia , Receptor 3 Toll-Like/antagonistas & inibidores , Brônquios/enzimologia , Brônquios/imunologia , Brônquios/patologia , Células Cultivadas , Humanos , Mucosa Respiratória/enzimologia , Mucosa Respiratória/patologia , Receptor 3 Toll-Like/fisiologiaRESUMO
Cigarette smoke induces damage to proteins and organelles by oxidative stress, resulting in accelerated epithelial cell senescence in the lung, which is implicated in chronic obstructive pulmonary disease (COPD) pathogenesis. Although the detailed molecular mechanisms are not fully understood, cellular energy status is one of the most crucial determinants for cell senescence. Creatine kinase (CK) is a constitutive enzyme, playing regulatory roles in energy homeostasis of cells. Among two isozymes, brain-type CK (CKB) is the predominant CK in lung tissue. In this study, we investigated the role of CKB in cigarette smoke extract (CSE)-induced cellular senescence in human bronchial epithelial cells (HBECs). Primary HBECs and Beas2B cells were used. Protein carbonylation was evaluated as a marker of oxidative protein damage. Cellular senescence was evaluated by senescence-associated ß-galactosidase staining. CKB inhibition was examined by small interfering RNA and cyclocreatine. Secretion of IL-8, a hallmark of senescence-associated secretary phenotype, was measured by ELISA. CKB expression levels were reduced in HBECs from patients with COPD compared with that of HBECs from nonsmokers. CSE induced carbonylation of CKB and subsequently decreased CKB protein levels, which was reversed by a proteasome inhibitor. CKB inhibition alone induced cell senescence, and further enhanced CSE-induced cell senescence and IL-8 secretion. CSE-induced oxidation of CKB is a trigger for proteasomal degradation. Concomitant loss of enzymatic activity regulating energy homeostasis may lead to the acceleration of bronchial epithelial cell senescence, which is implicated in the pathogenesis of COPD.
Assuntos
Brônquios/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Creatina Quinase Forma BB/metabolismo , Células Epiteliais/efeitos dos fármacos , Fumaça/efeitos adversos , Fumar/efeitos adversos , Brônquios/enzimologia , Brônquios/imunologia , Brônquios/patologia , Células Cultivadas , Creatina Quinase Forma BB/antagonistas & inibidores , Creatina Quinase Forma BB/genética , Ciclina B1/metabolismo , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/enzimologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Humanos , Imuno-Histoquímica , Interleucina-8/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma , Carbonilação Proteica/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Ubiquitinação , beta-Galactosidase/metabolismoRESUMO
Reepithelialization of remodeled air spaces with bronchial epithelial cells is a prominent pathological finding in idiopathic pulmonary fibrosis (IPF) and is implicated in IPF pathogenesis. Recent studies suggest that epithelial senescence is a risk factor for development of IPF, indicating such reepithelialization may be influenced by the acceleration of cellular senescence. Among the sirtuin (SIRT) family, SIRT6, a class III histone deacetylase, has been demonstrated to antagonize senescence. We evaluated the senescence of bronchiolization in association with SIRT6 expression in IPF lung. Senescence-associated ß-galactosidase staining and immunohistochemical detection of p21 were performed to evaluate cellular senescence. As a model for transforming growth factor (TGF)-ß-induced senescence of abnormal reepithelialization, we used primary human bronchial epithelial cells (HBEC). The changes of SIRT6, p21, and interleukin (IL)-1ß expression levels in HBEC, as well as type I collagen expression levels in fibroblasts, were evaluated. In IPF lung samples, an increase in markers of senescence and SIRT6 expression was found in the bronchial epithelial cells lining cystically remodeled air spaces. We found that TGF-ß induced senescence in primary HBEC by increasing p21 expression, and, whereas TGF-ß also induced SIRT6, it was not sufficient to inhibit cellular senescence. However, overexpression of SIRT6 efficiently inhibited TGF-ß-induced senescence via proteasomal degradation of p21. TGF-ß-induced senescent HBEC secreted increased amounts of IL-1ß, which was sufficient to induce myofibroblast differentiation in fibroblasts. These findings suggest that accelerated epithelial senescence plays a role in IPF pathogenesis through perpetuating abnormal epithelial-mesenchymal interactions, which can be antagonized by SIRT6.
Assuntos
Brônquios/patologia , Senescência Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Fibrose Pulmonar Idiopática/patologia , Sirtuínas/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Diferenciação Celular/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Células Epiteliais/efeitos dos fármacos , Humanos , Fibrose Pulmonar Idiopática/metabolismo , Interleucina-1beta/metabolismo , Miofibroblastos/citologia , Miofibroblastos/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/metabolismo , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , beta-Galactosidase/metabolismoRESUMO
A chemotherapeutic regimen for advanced thymic carcinoma has not yet been established. We describe a patient with advanced thymic mucoepidermoid carcinoma who achieved a complete response to combination chemotherapy with cisplatin (Randa) and irinotecan hydrochloride (Campto). A 74-year-old man was admitted to our hospital because of chest pain, general fatigue, appetite loss and weight loss. Chest computed tomography examinations revealed an anterior mediastinal tumour (5.5cmx3.5cmx9.5cm) that had invaded the subcutis through the sternum. The patient was treated with three courses of cisplatin and irinotecan hydrochloride followed by radiotherapy; he has since exhibited a complete response for 3 months.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma Mucoepidermoide/tratamento farmacológico , Neoplasias do Timo/tratamento farmacológico , Idoso , Camptotecina/administração & dosagem , Camptotecina/análogos & derivados , Carcinoma Mucoepidermoide/radioterapia , Cisplatino/administração & dosagem , Terapia Combinada , Humanos , Irinotecano , Masculino , Neoplasias do Timo/radioterapiaRESUMO
A 64-year-old woman presented skin lesions on her face, upper extremities and finger erythema (heliotropism and Gottron's sign). She had weakness in her lower extremities. She was given a diagnosis of dermatomyositis (DM), because the serum examination showed that a myositis-specific antibody was positive whereas Jo-1 antibody was negative. The findings of chest X-ray and computed tomography showed that she had limited small cell lung cancer, but no interstitial pneumonia. She was treated with standard chemotherapy consisting of cisplatin and etoposide with accelerated hyperfractionation radiotherapy. She showed partial response to the treatment, whereas the skin lesions and muscle weakness deteriorated accompanied with bone marrow suppression due to chemotherapy. Skin and muscle biopsy were performed and pathological findings showed typical perivasculitis infiltrated with lymphocytes in muscle and skin. With the recovery of bone marrow suppression and partial response due to chemotherapy, the skin lesions improved and creatine kinase became normalized. She was given a diagnosis of paraneoplastic DM. Since severe leukocytopenia paralleled the deterioration of DM, the decrease of peripheral white blood cell counts, especially regulatory T cell counts, may be associated with DM activity.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Dermatomiosite/complicações , Leucopenia/induzido quimicamente , Neoplasias Pulmonares/tratamento farmacológico , Síndromes Paraneoplásicas/complicações , Carcinoma de Pequenas Células do Pulmão/tratamento farmacológico , Antineoplásicos Fitogênicos/administração & dosagem , Cisplatino/administração & dosagem , Etoposídeo/administração & dosagem , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Cases of non-small-cell lung cancer (NSCLC) carrying the somatic mutation of epidermal growth factor receptor (EGFR) have been shown to be hyperresponsive to the EGFR tyrosine kinase inhibitor gefitinib (IRESSA). If EGFR mutations can be observed in serum DNA, this could serve as a noninvasive source of information on the genotype of the original tumor cells that could influence treatment and the ability to predict patient response to gefitinib. Serum genomic DNA was obtained from Japanese patients with NSCLC before first-line gefitinib monotherapy. Scorpion Amplified Refractory Mutation System technology was used to detect EGFR mutations. Wild-type EGFR was detected in all of the 27 serum samples. EGFR mutations were detected in 13 of 27 (48.1%) patients and two major EGFR mutations were identified (E746_A750del and L858R). The EGFR mutations were seen significantly more frequently in patients with a partial response than in patients with stable disease or progressive disease (P = 0.046, Fisher's exact test). The median progression-free survival was significantly longer in patients with EGFR mutations than in patients without EGFR mutations (200 versus 46 days; P = 0.005, log-rank test). The median survival was 611 days in patients with EGFR mutations and 232 days in patients without EGFR mutations (P > 0.05). In pairs of tumor and serum samples obtained from 11 patients, the EGFR mutation status in the tumors was consistent with those in the serum of 8 of 11 (72.7%) of the paired samples. Thus, EGFR mutations were detectable using Scorpion Amplified Refractory Mutation System technology in serum DNA from patients with NSCLC. These results suggest that patients with EGFR mutations seem to have better outcomes with gefitinib treatment, in terms of progression-free survival, overall survival, and response, than those patients without EGFR mutations.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , DNA de Neoplasias/genética , Receptores ErbB/genética , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Quinazolinas/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Pulmonar de Células não Pequenas/sangue , Análise Mutacional de DNA/métodos , DNA de Neoplasias/sangue , Progressão da Doença , Intervalo Livre de Doença , Feminino , Seguimentos , Gefitinibe , Genótipo , Humanos , Neoplasias Pulmonares/sangue , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Mutação Puntual , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade , Deleção de Sequência , Taxa de Sobrevida , Resultado do TratamentoRESUMO
Molecular targeted drugs have been developed and have come to play a part in the standard treatment of cancers. However, issues such as the optimum dose, selection of patients, and verification of the molecular targets remain to be discussed, because unexpected clinical problems related to the clinical efficacy, adverse events, and development of resistance have appeared. Therefore, proof of principle (POP) studies and pharmacodynamic or pharmacogenomic research to explore new bio-markers for the drugs are essential for clinical progress. On the other hand, the higher cost of development and care must also be discussed as new problems.
Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/farmacocinética , Resistencia a Medicamentos Antineoplásicos , Neoplasias/tratamento farmacológico , Farmacogenética , Antineoplásicos/efeitos adversos , Relação Dose-Resposta a Droga , Determinação de Ponto Final , Humanos , Neoplasias/patologiaRESUMO
BACKGROUND: Community-acquired pneumonia (CAP) has high morbidity and mortality. Unfortunately, the pathogen detection rate using conventional culture methods is relatively low. We compared comprehensive real-time polymerase chain reaction (real-time PCR) analysis of nasopharyngeal swab specimens (NPS) and sputum samples against conventional methods for ability to detect causative pathogens of CAP. METHODS: We prospectively enrolled adult CAP patients, including those with prior antibiotic use, from December 2012 to May 2014. For each patient, causative pathogens were investigated conventionally and by real-time PCR that can identify 6 bacterial and 11 viral pathogens. RESULTS: Patients numbered 92 (mean age, 63 years; 59 male), including 30 (33%) with prior antibiotic use. Considering all patients, identification of causative pathogens by real-time PCR was significantly more frequent than by conventional methods in all patients (72% vs. 57%, p = 0.018). In patients with prior antibiotic use, identification rates also differed significantly (PCR, 77%; conventional, 50%; p = 0.027). Mixed infections were more frequent according to real-time PCR than conventional methods (26% vs. 4%, p < 0.001). By the real-time PCR, Streptococcus pneumoniae was most frequently identified (38%) as a causative pathogen, followed by Haemophilus influenzae (37%) and Mycoplasma pneumoniae (5%). PCR also identified viral pathogens (21%), with sensitivity enhanced by simultaneous examination of both NPS and sputum samples rather than only NPS samples. CONCLUSIONS: Real-time PCR of NPS and sputum samples could better identify bacterial and viral pathogens in CAP than conventional methods, both overall and in patients with prior antibiotic treatment.
Assuntos
Infecções Comunitárias Adquiridas/diagnóstico , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Pneumonia/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/classificação , Bactérias/isolamento & purificação , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Nasofaringe/microbiologia , Nasofaringe/virologia , Estudos Prospectivos , Sensibilidade e Especificidade , Escarro/microbiologia , Escarro/virologia , Vírus/classificação , Vírus/isolamento & purificaçãoRESUMO
Cigarette smoke (CS)-induced mitochondrial damage with increased reactive oxygen species (ROS) production has been implicated in COPD pathogenesis by accelerating senescence. Mitophagy may play a pivotal role for removal of CS-induced damaged mitochondria, and the PINK1 (PTEN-induced putative kinase 1)-PARK2 pathway has been proposed as a crucial mechanism for mitophagic degradation. Therefore, we sought to investigate to determine if PINK1-PARK2-mediated mitophagy is involved in the regulation of CS extract (CSE)-induced cell senescence and in COPD pathogenesis. Mitochondrial damage, ROS production, and cell senescence were evaluated in primary human bronchial epithelial cells (HBEC). Mitophagy was assessed in BEAS-2B cells stably expressing EGFP-LC3B, using confocal microscopy to measure colocalization between TOMM20-stained mitochondria and EGFP-LC3B dots as a representation of autophagosome formation. To elucidate the involvement of PINK1 and PARK2 in mitophagy, knockdown and overexpression experiments were performed. PINK1 and PARK2 protein levels in lungs from patients were evaluated by means of lung homogenate and immunohistochemistry. We demonstrated that CSE-induced mitochondrial damage was accompanied by increased ROS production and HBEC senescence. CSE-induced mitophagy was inhibited by PINK1 and PARK2 knockdown, resulting in enhanced mitochondrial ROS production and cellular senescence in HBEC. Evaluation of protein levels demonstrated decreased PARK2 in COPD lungs compared with non-COPD lungs. These results suggest that PINK1-PARK2 pathway-mediated mitophagy plays a key regulatory role in CSE-induced mitochondrial ROS production and cellular senescence in HBEC. Reduced PARK2 expression levels in COPD lung suggest that insufficient mitophagy is a part of the pathogenic sequence of COPD.
Assuntos
Senescência Celular , Células Epiteliais/patologia , Mitofagia , Proteínas Quinases/fisiologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Ubiquitina-Proteína Ligases/fisiologia , Adulto , Idoso , Autofagia , Brônquios/citologia , Feminino , Humanos , Imuno-Histoquímica , Pulmão/fisiopatologia , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Mitocôndrias/patologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Espécies Reativas de Oxigênio/metabolismo , Fumar/efeitos adversos , Produtos do TabacoRESUMO
Respiratory infection is a major cause of exacerbation in chronic obstructive pulmonary disease (COPD). Infectious contributions to exacerbations remain incompletely described. We therefore analyzed respiratory tract samples by comprehensive real-time polymerase chain reaction (PCR) in combination with conventional methods. We evaluated multiple risk factors for prolonged hospitalization to manage COPD exacerbations, including infectious agents. Over 19 months, we prospectively studied 46 patients with 50 COPD exacerbations, collecting nasopharyngeal swab and sputum samples from each. We carried out real-time PCR designed to detect six bacterial species and eleven viruses, together with conventional procedures, including sputum culture. Infectious etiologies of COPD exacerbations were identified in 44 of 50 exacerbations (88%). Infections were viral in 17 of 50 exacerbations (34%). COPD exacerbations caused by Gram-negative bacilli, including enteric and nonfermenting organisms, were significantly associated with prolonged hospitalization for COPD exacerbations. Our results support the use of a combination of real-time PCR and conventional methods for determining both infectious etiologies and risk of extended hospitalization.
Assuntos
Progressão da Doença , Doença Pulmonar Obstrutiva Crônica/complicações , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções Respiratórias/etiologia , Infecções Respiratórias/microbiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Escarro/microbiologiaRESUMO
A 63-year-old woman was diagnosed with advanced lung adenocarcinoma complicated by Trousseau's syndrome characterized by non-bacterial thrombotic endocarditis, asymptomatic brain infarction, deep venous thrombosis, and low-grade disseminated intravascular coagulation (DIC). The patient's DIC rapidly became widespread, and multiple micropulmonary embolisms led to severe respiratory failure. She received a blood transfusion and anticoagulant treatment with heparin and recombinant human soluble thrombomodulin, which modestly ameliorated her symptoms, and additional chemotherapy led to tumor shrinkage with concomitant resolution of Trousseau's syndrome. Although there are no established medical approaches for managing Trousseau's syndrome, intensive anticoagulant treatment may be effective for improving the patients' general condition in order for them to be able to undergo subsequent combination chemotherapy.
Assuntos
Adenocarcinoma/complicações , Adenocarcinoma/tratamento farmacológico , Anticoagulantes/uso terapêutico , Coagulação Intravascular Disseminada/complicações , Coagulação Intravascular Disseminada/tratamento farmacológico , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/tratamento farmacológico , Síndromes Paraneoplásicas/complicações , Síndromes Paraneoplásicas/tratamento farmacológico , Adenocarcinoma/diagnóstico , Adenocarcinoma de Pulmão , Coagulação Intravascular Disseminada/diagnóstico , Feminino , Heparina/uso terapêutico , Humanos , Neoplasias Pulmonares/diagnóstico , Pessoa de Meia-Idade , Síndromes Paraneoplásicas/diagnóstico , Resultado do Tratamento , Trombose Venosa/complicações , Trombose Venosa/diagnóstico , Trombose Venosa/tratamento farmacológicoRESUMO
Thymomas are associated with a wide spectrum of autoimmune paraneoplastic diseases. Here we report the case of 31-year-old male with invasive thymoma, myasthenia gravis, polymyositis, and acute fulminant myocarditis that presented with cardiogenic shock requiring intra-aortic balloon pumping and percutaneous cardiopulmonary support. Corticosteroid therapy was effective. To our knowledge, this is the first case of thymoma with acute fulminant cardiomyositis that was successfully treated by assisted circulation and corticosteroids, despite a poor prognosis.
RESUMO
We present a case of organizing pneumonia complicated by pneumothorax in association with cyst formation that developed during corticosteroid treatment. Although it has been reported that the check-valve mechanism is a plausible cause of cyst and pneumothorax formation in patients with organizing pneumonia, the details of the corresponding pathological changes that occur in air-trapping have not been elucidated. A pathological examination of lung specimens obtained with video-assisted thoracoscopic surgery suggested that granulation tissues plugging the bronchiole lumens might be a potential cause of the check-valve mechanism in this case. In this report, we also reviewed eight other cases of organizing pneumonia with pneumothorax or cyst formation.