Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010.

In our republic, occupational exposure to asbestos has been restricted to jobs concerned with disposal of products, materials and buildings containing asbestos, and research work studying asbestos fibres. The reason consists in health risks as all kinds of asbestos belong to high-risk carcinogens. The asbestos fibre related diseases should be reported. All data connected are kept in the National Institute of Public Health (NIPH) in Prague. The purpose of this study was to find out how much people are aware of the risks related to asbestos and find people who suffered from asbestos dust-induced disease, i.e. mesothelioma of pleura. The quantitative method was used to summarise information on asbestos-induced disease occurence in 1970-2010 available in the National Institute of Public Health in Prague. Along with that, the work maps the population awareness of asbestos risks. The thesis itself can be divided into two parts. The first one contains information based on data collection while the other part deals with data of NIPH in Prague. Two hypotheses were predicted: Hypothesis H1: People are aware of the material containing asbestos. Hypothesis H2: People are aware of negative impact of asbestos on human health. Both hypotheses were proved. Three hypotheses were suggested in the other part: Hypothesis H3: Latency, the time between first exposure to manifestation of disease, is never less than 20 years. Hypothesis H4: Incidence of mesothelioma of pleura and peritoneum is higher in people aged 60-69. Hypothesis H5: Smoking affects the course of the disease negatively. Hypotheses H3 and H4 were proved. Hypothesis H5 cannot be neither proved or disproved. Although the number of smokers among mesothelioma patients was higher, there is no evidence of negative effect of smoking on the disease. To prove this hypothesis, the data colllection should be larger. Findings of the study proved the time of exposure to asbestos fibres does not affect the occurence of this disease. The findings can be used in further research studies.

Abstract Reactive oxygen species (ROS) can cause severe damage to cells and organs but they are also important mediators of inflammatory responses and cellular signalling. Due to the significant role of ROS, the cells have evolved a broad antioxidative system to regulate the concentration of these species. Peroxiredoxins (Prxs) are enzymes that participate in the regulation of the cellular redox-homeostasis by detoxifying hydrogen peroxide. Prxs are not classified as conventional antioxidant enzymes and their physiological role, whether protective or regulatory, is still unclear. The aim of this project was to study the localization and regulation of Prxs in normal human lung and also their role in selected lung disorders (pulmonary sarcoidosis, pleural mesothelioma, lung carcinomas and chronic obstructive disorder, COPD). Additionally the expression of thioredoxin (Trx) and thioredoxin reductase (TrxR) was analysed in the lung of smokers and COPD patients. These enzymes are important reductants in cell and Prxs are one of their targets. Lung is an important organ in the field of ROS and antioxidant research since it is especially vulnerable to exogenous oxidative stress caused by pollutants, cigarette smoke and also by high oxygen pressure. The results showed that all six human Prxs were expressed in healthy human lung but in a cell-specific manner. The most prominent expression was detected in the epithelium and in macrophages, the cells most prone to oxidative stress. There were also differences in subcellular locations of Prxs. The expression of Prxs in non-malignant lung diseases (pulmonary sarcoidosis and COPD) and in smoker's lung was very similar with that in normal lung. Higher expression of Prx V and VI was detected in a subpopulation of macrophages sampled from COPD patients' lung. In contrast, Trx expression was induced in the bronchial epithelium of smoker's lung. Differences in the expression compared to normal lung were seen in lung malignancies (pleural mesothelioma and lung carcinomas). Interestingly, different Prxs were highly expressed in different types of carcinomas. In pleural mesothelioma, all Prxs except Prx IV were highly expressed when compared to normal pleura, in adenocarcinoma Prxs I, II, VI and especially IV, and in squamous cell carcinoma Prxs I, II and IV were upregulated. Tests performed on cultured cells in vitro revealed only a minor increase in the Prx expression after severe oxidant stress in malignant lung cell line originating from alveolar type II pneumocytes (A549) or non-malignant cell line derived from bronchial epithelium. None of the tested growth factors or cytokines affected Prx expression or oxidation state, but severe oxidant stress influenced remarkably the oxidation state of the Prxs.

Abstract Reactive nitrogen species (RNS) and reactive oxygen species (ROS) have been linked with the pathogenesis of lung malignancies and chronic obstructive pulmonary disease (COPD). In vitro studies indicated that mesothelioma and lung carcinoma cell lines synthesize nitric oxide synthases (NOS) mRNA. The Comet-assay indicated that asbestos fibers caused DNA single -strand breaks in mesothelial cells, and this effect was enhanced by glutathione depletion. The use of FPG in the Comet assay indicated that the asbestos induced DNA strand breaks were oxidant mediated. In vivo non-neoplastic pleura was mostly negative for inducible NOS (iNOS), while inflamed pleura was positive. The immunohistochemical expression of iNOS was detected in 74% and 96% of malignant mesotheliomas and metastatic pleural adenocarcinomas, respectively. Epithelial and mixed mesotheliomas expressed more often intense iNOS immunoreactivity compared to the sarcomatoid subtype. Normal mesothelial cells showed occasional positivity for endothelial NOS (eNOS), but reactive mesothelial cells were strongly stained. eNOS was found in 89% of mesotheliomas. Vascular endothelial growth factor (VEGF) was identified in 47%, a VEGF receptor FLK1 in 69% and the VEGF receptor, FLT1, in 71% of mesotheliomas. FLK1 or FLT1 immunoreactivities were more often seen in epithelioid and biphasic mesotheliomas than in sarcomatoid mesotheliomas. In lung samples of non-smokers, smokers and COPD patients, the levels of nitrotyrosine were higher in alveolar macrophages of smokers and COPD patients than in the non-smokers and in the alveolar epithelium of smokers and COPD patients than in the non-smokers. The iNOS expression was weak in the bronchial and alveolar epithelium in all groups but eNOS was most prominently expressed in alveolar macrophages while neuronal NOS (nNOS) was negative in all of the major cell types of the lung. Bronchial metaplasia-dysplasia-sequence was clearly positive for iNOS, nNOS and nitrotyrosine. Thus, smoking can cause protein nitration also in normal lung. Prominent iNOS and nNOS immunoreactivity in metaplasia-dysplasia-lesions suggests a divergent role of NOSs in carcinogenesis and destruction of alveolar epithelium in emphysematous lung. In lung cancer samples, iNOS was detected in 40% cases, while 89% and 81% cases were positive for eNOS and nNOS, respectively. Intense eNOS staining was seen more often in adenocarcinomas than in squamous cells carcinomas, and iNOS immunoreactivity was seen more often in grade I-II tumors than in grade III tumors. The patients with tumors showing high expression of iNOS, eNOS and nNOS, exhibited better survival, but this was not an independent prognostic factor.

Le mésothéliome pleural malin (MPM) est un cancer rare et de mauvais pronostic. Le diagnostic de MPM restant difficile à ce jour, nous avons tout d'abord évalué l'intérêt de l'analyse du transcriptome, à partir des cellules présentes dans le liquide pleural, à des fins diagnostiques. Si cette approche parait faisable, ses contraintes techniques la rendent incompatibles avec une utilisation en routine. Notre deuxième axe de travail s'est inscrit dans le domaine du traitement local du MPM, avec l'évaluation de l'administration intrapleurale (IP) de deux molécules de chimiothérapie : le pemetrexed et le lipoplatine. Les résultats sur modèle animal montrent que les profils pharmacocinétiques de ces deux molécules sont significativement différents entre une administration intraveineuse et une administration IP. Dans l'optique d'identifier de nouvelles cibles thérapeutiques, nous avons évalué la voie de l'adrénomédulline (AM) dans le MPM. L'AM et ses récepteurs sont conjointement exprimés dans les biopsies pleurales de MPM. In vitro, l'AM augmente les capacités de prolifération, de migration et d'invasion des cellules de MPM, par l'intermédiaire de la voie des MAPK. L'inhibition de l'AM ou de ses récepteurs apparait comme une stratégie thérapeutique prometteuse dans le MPM, de part ses effets directs sur les cellules néoplasiques, mais aussi ses effets indirects, via une inhibition de l'angiogénèse et de la lymphangiogénèse tumorale (in vivo). Enfin, nous avons évalué de nouveaux critères de jugement pour les essais cliniques de phase II. Nous avons montré que le taux de survie sans progression à 9 semaines est le critère le plus performant pour prédire la survie globale. / Malignant pleural mesothelioma (MPM) is a rare cancer with poor prognosis. As MPM diagnosis remains difficult today, we first assessed the potential value of transcriptome analysis, from cells in pleural fluid, with diagnostic purpose. If this approach looks feasible, its technical constraints make it incompatible with routine practice. The second axis of our work concerned MPM local treatment, with the assessment of intrapleural (IP) administration of two chemotherapy drugs: pemetrexed and lipoplatin. Results obtained in an animal model show that the pharmacokinetic profiles of these two drugs are significantly different between intravenous and IP administration. With the goal of identifying new therapeutic targets, we explored the adrenomedullin (AM) pathway in MPM. AM and its receptors are jointly expressed in pleural biopsies from MPM. Moreover we demonstrated in vitro that AM increases the proliferation, migration and invasion abilities of MPM cells, through the MAPK signaling pathway. Inhibition of AM or its receptors appears as a promising therapeutic strategy, because of its direct effects on malignant cells, but also its indirect effects, via tumor angiogenesis and lymphangiogenesis inhibition (in vivo). Finally, we assessed new endpoints for phase II clinical studies. We showed that the progression-free survival rate at 9 weeks is the most performant criterion to predict overall survival.

Le mésothéliome est une tumeur maligne rare des séreuses, au pronostic sombre, dont le facteur de risque principal connu est l’exposition à l’amiante. Il se déclare avec un temps de latence d’environ 30 ans après le début de l’exposition, atteint principalement la plèvre et touche majoritairement les hommes. L’analyse de cette pathologie, reconnue comme une maladie professionnelle et pour laquelle une Déclaration Obligatoire de maladie a été instituée en janvier 2012, s’est focalisée jusqu’alors essentiellement sur l’homme, tant sur le plan épidémiologique que sur le plan biologique. Il apparaît pourtant dans la littérature internationale des différences notables par sexe.Ces constations posent la question d’investiguer d’autres facteurs de risque, en particulier l’hypothèse d’une prédisposition génétique, les mécanismes biologiques intervenant dans les voies de la carcinogenèse du mésothéliome étant partiellement connus. Ceci incite à étudier les facteurs épidémiologiques, cliniques, biologiques et immunohistochimiques, prédisposant chez la femme au développement du mésothéliome pleural et péritonéal, et d’évaluer leurs impacts pronostiques, à partir des cas diagnostiqués entre 1998 et 2013, issus de la plus importante base de données française. Elle incite également à établir un état des lieux épidémiologique actualisé par sexe en France pour le mésothéliome pleural et péritonéal ; ces données étant inexistantes pour la France pour le péritoine. / Mesothelioma is a rare malignant serous tumor with a poor prognosis of which asbestos exposure is the major known risk factor. It occurs with a latent period about 30 years after exposure, reaches mainly the pleura and affects mainly the men. The study of this pathology, recognized as an occupationnal disease and for which a Mandatory Declaration of Disease was introduced in January 2012, focused until then mainly on men, both epidemiologically and biologically. However, there are significant gender differences in the international literature.These results raise the question of investigating other risk factors, in particular the hypothesis of genetic predisposition, the biological mechanisms involved in mesothelioma carcinogenesis pathways being partially known. This encourages the study of epidemiological, clinical, biological and immunohistochemical factors predisposing women to the development of pleural and peritoneal mesothelioma, and to evaluate their prognostic impacts, based on cases diagnosed between 1980 and 2015, from the largest French database. It also encourages the establishment of an updated epidemiological inventory by gender in France for pleural and peritoneal mesothelioma; these data are non-existent for peritoneum.

京都大学 / 0048 / 新制・論文博士 / 博士(社会健康医学) / 乙第13292号 / 論社医博第13号 / 新制||社医||10(附属図書館) / (主査)教授 川上 浩司, 教授 平井 豊博, 教授 伊達 洋至 / 学位規則第4条第2項該当 / Doctor of Public Health / Kyoto University / DFAM

Le mésothéliome pleural malin (MPM) est une tumeur primitive de la plèvre, rare, très agressive, avec un pronostic sombre. Nous avons souhaité au cours de ce travail de thèse, identifier de nouveaux biomarqueurs du MPM en testant l’influence de l’inactivation des membres de la famille RASSF/Hippo sur la survie des 448 patients inclus dans l’essai clinique MAPS (IFCT-GFPC-0701). Nous souhaitions également comprendre quelles fonctions et signalisations essentielles à l’homéostasie cellulaire, auxquelles participe la voie de signalisation RASSF/Hippo, sont perturbées lors du processus de transformation des cellules mésothéliales. L’inactivation des membres de la voie a été étudiée par PCR spécifique de méthylation (MS-PCR) et leur influence sur la survie des 448 patients inclus dans l’essai clinique MAPS testée en analyse uni- et multivariée avant d’être validée par boostrap. D’autre part, nous avons mimé in cell, l’inactivation par ARN interférence de plusieurs membres de la voie Hippo dans des cellules de lignées mésothéliales humaines (MSTO-211H, H2452, H28 et H2052). Nous avons pu identifier plusieurs biomarqueurs du MPM : i) la kinase MST1 dont l’inactivation est un facteur de mauvais pronostic, ii) l’amphiréguline dont l’expression cytoplasmique est au contraire un facteur de bon pronostic et enfin iii) le CD44 dont l’expression élevée constitue un outil diagnostique du MPM. Les approches in cell, nous ont permis de démontrer que les altérations de la voie RASSF/Hippo induisent une activité inappropriée de l’effecteur terminal YAP : le moins bon pronostic des patients présentant une inactivation de MST1 s’explique ainsi par le fait qu’en régulant l’activité de YAP, MST1 contrôle la balance apoptose/prolifération et prévient l’invasion et la croissance sans adhésion. En son absence, ces processus cellulaires sont dérégulés. Ce travail démontre l’importance de l’axe CD44/RASSF1A/MST1 dans le contrôle d’une activité appropriée de YAP et de l’homéostasie des cellules mésothéliales. La compréhension des désordres cellulaires induits par la dérégulation de le voie RASSF/Hippo, désigne YAP comme cible thérapeutique potentielle chez les patients atteints de MPM et présentant des altérations de cette voie de signalisation. / Malignant pleural mesothelioma (MPM) is a rare, very aggressive, primary tumor with a poor prognosis. During this thesis, we wanted to identify new biomarkers of MPM by testing the influence of the RASSF/Hippo pathway inactivation on the survival of the 448 patients included in the clinical trial MAPS (IFCT- GFPC-0701). We also wanted to understand which functions and signals essential to cellular homeostasis, linked to RASSF/Hippo signaling pathway, are disturbed during the mesothelial cell transformation process. Inactivation of RASSF/Hippo members was studied by methylation-specific PCR (MS-PCR) and their influence on the survival of the 448 patients included in the MAPS clinical trial tested in uni- and multivariate analysis before being validated by bootstrap. We also mimed in cell, by RNA interference, several members of the Hippo pathway inactivation in human mesothelial cells lines (MSTO-211H, H2452, H28 and H2052). We have identified several biomarkers of MPM: i) MST1 kinase whose inactivation is a factor of poor prognosis, ii) amphiregulin whose cytoplasmic expression is on the contrary a factor of good prognosis and finally iii) CD44 whose high expression is a diagnostic tool for MPM. In cell we demonstrate that RASSF/Hippo pathway alterations induce an inappropriate activity of YAP, one Hippo end effector: the poorer prognosis of patients with inactivation of MST1 is thus explained by the fact that, by regulating YAP activity, MST1 controls the apoptosis/proliferation balance and prevents invasion and growth without adhesion from mesothelial cells. In its absence, these cellular processes are deregulated. This work finally demonstrates the importance of the CD44/RASSF1A/MST1 axis in controlling appropriate YAP activity and mesothelial cell homeostasis. The understanding of the cellular disorders induced by the of the RASSF/Hippo pathway deregulation designates YAP as a potential therapeutic target in patients with MPM and presenting alterations of this signaling pathway.

Introdução Trata-se de um estudo descritivo das sentenças de segunda instância do Poder Judiciário que envolva o problema do trabalhador exposto ao risco do amianto como uma questão de saúde pública. Objetivo Avaliar as decisões judiciais a respeito da exposição dos trabalhadores ao risco do amianto como causa de doença do trabalho a partir da análise de acórdãos judiciais de 1999 até 2009. Métodos Os objetivos foram alcançados mediante pesquisa em acórdãos judiciais publicados no TRT (Tribunal Regional do Trabalho) da 2ª Região, TRT (Tribunal Regional do Trabalho) da 15ª Região e TST (Tribunal Superior do Trabalho), que são tribunais pertencentes à Justiça do Trabalho, bem como no TJSP (Tribunal de Justiça de São Paulo), STJ (Superior Tribunal de Justiça) e STF (Supremo Tribunal Federal), pertencentes à Justiça Comum, a partir do momento histórico em que houve a promulgação da Constituição Federal de 1988, considerando-se o ano da publicação ou do registro dos acórdãos de 1999 a 2009. Os acórdãos foram selecionados por meio da internet nos sites dos Tribunais em referência, que disponibilizam a busca on line. Resultados Os resultados indicam que de um total de 119 decisões judiciais analisadas, considerando-se todos os critérios de inclusão e exclusão, apenas em 42 casos houve a comprovação do nexo de causalidade entre a exposição do trabalhador ao risco do amianto e a doença adquirida no trabalho, o que evidencia que o trabalhador desincumbiuse de seu ônus probatório em pequena parte dos acórdãos. Conclusões Pela avaliação dos acórdãos judiciais de 1999 até 2009 a respeito da exposição dos trabalhadores ao risco do amianto como causa de acidente do trabalho, doença profissional ou morte, somente em 35,3 por cento das decisões foram declarados procedentes os pedidos dos trabalhadores / Introduction - This is a descriptive study of Judiciary second appeal decisions involving the problem of the risk of workers when exposed to asbestos as a public health issue. Objective - To evaluate judicial decisions regarding the exposure of workers to the risk of asbestos as a cause of occupational disease from the analysis of Judicial Decisions from 1999 to 2009. Methods - The objectives have been achieved by researching judicial decisions published by the TRT (Regional Court of Labor) of the 2nd Region, by the TRT (Regional Court of Labor) of the 15th Region and by the TST (Superior Labor Court), which are courts of the Labor Justice Department, as well as the ones by the TJSP (Court of Justice of São Paulo), by the STJ (Higher Justice Court) and by the STF (Supreme Federal Court), of the Common Justice, from the historical moment in which the 1988 Constitution was promulgated, considering the year of publication or record of the Judiciary Decisions from 1999 to 2009. The judicial decisions were selected with the use of the internet in the sites of the Courts previously mentioned, which made possible to make this search on line. Results - The results indicate that from a total of 119 judicial decisions, which were analyzed, and also taking in consideration all the inclusion and exclusion criteria, only in 42 cases there was proof of a causal link between the worker\'s risky exposure to asbestos and the disease acquired at work, which shows that the employee is discharged of the evidential burden on a small portion of the decisions. Conclusions - For the evaluation of the judicial decisions from 1999 to 2009 regarding the exposure of workers to the risk of asbestos as a cause of work accidents, and occupational disease or death, only 35.3 per cent of decisions were in favor of the workers claim