Analysis of outcome and cancer stem cells status in patients with resectable pancreatic adenocarcinoma

RESUMO: Actualmente, a única possibilidade de cura para doentes com adenocarcinoma do pâncreas (PDAC) é a ressecção cirúrgica, no início deste estudo, perguntamo-nos se os predictores clínico-patológicos clássicos de prognostico poderiam ser validados em uma grande cohort de doentes com cancro do pâncreas ressecável e se outros predictores clínicos poderiam ter um papel na decisão de que doentes beneficiariam de ressecção cirúrgica. No capítulo 2, observamos que até 30% dos doentes morrem no primeiro ano após a ressecção cirúrgica, pelo que o nosso objectivo foi determinar factores pré-operatórios que se correlacionam com mortalidade precoce após ressecação cirúrgica com recurso a um instrumento estatisticamente validado, o Charlson-Age Comorbidity Index (CACI), determinamos que um CACI score superior a 4 foi preditivo de internamentos prolongados (p <0,001), complicações pós-operatórias (p = 0,042), e mortalidade em 1 ano pós- ressecção cirúrgica (p <0,001). Um CACI superior a 6 triplicou a mortalidade no primeiro ano pós-cirurgia e estes doentes têm menos de 50% de probabilidade de estarem vivos um ano após a cirurgia. No capítulo 3, o nosso objectivo foi identificar uma proteína de superfície que se correlacionasse estatisticamente com o prognostico de doentes com adenocarcinoma do pâncreas e permitisse a distinção de subgrupos de doentes de acordo com as suas diferenças moleculares, perguntamo-nos ainda se essa proteína poderia ser um marcador de células-estaminais. No nosso trabalho anterior observamos que as células tumorais na circulação sanguínea apresentavam genes com características bifenotípica epitelial e mesenquimal, enriquecimento para genes de células estaminais (ALDH1A1 / ALDH1A2 e KLF4), e uma super-expressão de genes da matriz extracelular (colagénios, SPARC, e DCN) normalmente identificados no estroma de PDAC. Após a avaliação dos tumores primários com RNA-ISH, muitos dos genes identificados, foram encontrados co-localizando em uma sub-população de células na região basal dos ductos pancreáticos malignos. Além disso, observamos que estas células expressam o marcador SV2A neuroendócrino, e o marcador de células estaminais ALDH1A1/2. Em comparação com tumores negativos para SV2, os doentes com tumores SV2 positivos apresentaram níveis mais baixos de CA 19-9 (69% vs. 52%, p = 0,012), tumores maiores (> 4 cm, 23% vs. 10%, p = 0,0430), menor invasão de gânglios linfáticos (69% vs. 86%, p = 0,005) e tumores mais diferenciados (69% vs. 57%, p = 0,047). A presença de SV2A foi associada com uma sobrevida livre de doença mais longa (HR: 0,49 p = 0,009) bem como melhor sobrevida global (HR: 0,54 p = 0,018). Em conjunto, esta informação aponta para dois subtipos diferentes de adenocarcinoma do pâncreas, e estes subtipos co-relacionam estatisticamente com o prognostico de doentes, sendo este subgrupo definido pela presença do clone celular SV2A / ALDH1A1/2 positivo com características neuroendócrinas. No Capítulo 4, a expressão de SV2A no cancro do pâncreas foi validado em linhas celulares primárias. Demonstramos a heterogeneidade do adenocarcinoma do pâncreas de acordo com características clonais neuroendócrinas. Ao comparar as linhas celulares expressando SV2 com linhas celulares negativas, verificamos que as linhas celulares SV2+ eram mais diferenciadas, diferindo de linhas celulares SV2 negativas no que respeita a mutação KRAS, proliferação e a resposta à quimioterapia. No capítulo 5, perguntamo-nos se o clone celular SV2 positivo poderia explicar a resistência a quimioterapia observada em doentes. Observamos um aumento absoluto de clones celulares expressando SV2A, em múltiplas linhas de evidência - doentes, linhas de células primárias e xenotransplantes. Embora, tenhamos sido capazes de demonstrar que o adenocarcinoma do pâncreas é uma doença heterogénea, consideramos que a caracterização genética destes clones celulares expressando SV2A é de elevada importância. Pretendemos colmatar esta limitação com as seguintes estratégias: Após o tratamento com quimioterapia neoadjuvante na nossa coorte, realizamos microdissecação a laser das amostras primarias em parafina, de forma a analisar mutações genéticas observadas no adenocarcinoma pancreático; em segundo lugar, pretendemos determinar consequências de knockdown da expressão de SV2A em nossas linhas celulares seguindo-se o tratamento com gemicitabina para determinação do papel funcional de SV2A; finalmente, uma vez que os nossos esforços anteriores com um promotor - repórter e SmartFlare ™ falharam, o próximo passo será realizar RNA-ISH PrimeFlow™ seguido de FACS e RNA-seq para caracterização deste clone celular. Em conjunto, conseguimos provar com várias linhas de evidência, que o adenocarcinoma pancreático é uma doença heterogénea, definido por um clone de células que expressam SV2A, com características neuroendócrinas. A presença deste clone no tecido de doentes correlaciona-se estatisticamente com o prognostico da doença, incluindo sobrevida livre de doença e sobrevida global. Juntamente com padrões de proliferação e co-expressão de ALDH1A1/2, este clone parece apresentar um comportamento de células estaminais e está associado a resistência a quimioterapia, uma vez que a sua expressão aumenta após agressão química, quer em doentes, quer em linhas de células primárias.----------------------------- ABSTRACT: Currently, the only chance of cure for patients with pancreatic adenocarcinoma is surgical resection, at the beginning of my thesis studies, we asked if the classical clinicopathologic predictors of outcome could be validated in a large cohort of patients with early stage pancreatic cancer and if other clinical predictors could have a role on deciding which patients would benefit from surgery. In chapter 2, we found that up to 30% of patients die within the first year after curative intent surgery for pancreatic adenocarcinoma. We aimed at determining pre-operative factors that would correlate with early mortality following resection for pancreatic cancer using a statistically validated tool, the Charlson-Age Comorbidity Index (CACI). We found that a CACI score greater than 4 was predictive of increased length of stay (p<0.001), post-operative complications (p=0.042), and mortality within 1-year of pancreatic resection (p<0.001). A CACI score of 6 or greater increased 3-fold the odds of death within the first year. Patients with a high CACI score have less than 50% likelihood of being alive 1 year after surgery. In chapter 3 we aimed at identifying a surface protein that correlates with patient’s outcome and distinguishes sub-groups of patients according to their molecular differences and if this protein could be a cancer stem cell marker. The most abundant class of circulating tumor cells identified in our previous work was found to have biphenotypic features of epithelial to mesenchymal transition, enrichment for stem-cell associated genes (ALDH1A1/ALDH1A2 and KLF4), and an overexpression of extracellular matrix genes (Collagens, SPARC, and DCN) normally found in the stromal microenvironment of PDAC primary tumors. Upon evaluation of matched primary tumors with RNA-ISH, many of the genes identified were found to co-localize in a sub-population of cells at the basal region of malignant pancreatic ducts. In addition, these cells expressed the neuroendocrine marker SV2A, and the stem cell marker ALDH1A1/2. Compared to SV2 negative tumors, patients with SV2 positive tumors were more likely to present with lower CA 19-9 (69% vs. 52%, p = 0.012), bigger tumors (size > 4 cm, 23% vs. 10%, p= 0.0430), less nodal involvement (69% vs. 86%, p = 0.005) and lower histologic grade (69% vs. 57%, p = 0.047). The presence of SV2A expressing cells was associated with an improved disease free survival (HR: 0.49 p=0.009) and overall survival (HR: 0.54 p=0.018) and correlated linearly with ALDH1A2. Together, this information points to two different sub-types of pancreatic adenocarcinoma, and these sub-types correlated with patients’ outcome and were defined by the presence of a SV2A/ ALDH1A1/2 expressing clone with neuroendocrine features. In Chapter 4, SV2A expression in cancer was validated in primary cell lines. We were able to demonstrate pancreatic adenocarcinoma heterogeneity according to neuroendocrine clonal features. When comparing SV2 expressing cell lines with SV2 negative cell lines, we found that SV2+ cell lines were more differentiated and differ from SV2 negative cell lines regarding KRAS mutation, proliferation and response to chemotherapy. In Chapter 5 we aimed at determining if this SV2 positive clone could explain chemoresistance observed in patients. We found an absolute increase in SV2A expressing cells, with multiple lines of evidence, in patients, primary cell lines and xenografts. Although, we have been able to show evidence that pancreatic adenocarcinoma is a heterogeneous disease, our findings warrant further investigation. To further characterize SV2A expressing clones after treatment with neoadjuvant chemotherapy in our cohort, we have performed laser capture microdissection of the paraffin embedded tissue in this study and will analyze the tissue for known genetic mutations in pancreatic adenocarcinoma; secondly, we want to know what will happen after knocking down SV2A expression in our cell lines followed by treatment with gemcitabine to determine if SV2A is functionally important; finally, since our previous efforts with a promoter – reporter and SmartFlare™ have failed, we will utilize a novel PrimeFlow™ RNA-ISH assay followed by FACS and RNA sequencing to further characterize this cellular clone. Overall our data proves, with multiple lines of evidence, that pancreatic adenocarcinoma is a heterogeneous disease, defined by a clone of SV2A expressing cells, with neuroendocrine features. The presence of this clone in patients’ tissue correlates with patient’s disease free survival and overall survival. Together with patterns of proliferation and ALDH1A1/2 co-expression, this clone seems to present a stem-cell-like behavior and is associated with chemoresistance, since it increases after chemotherapy, both in patients and primary cell lines.

Teamwork and Gendered Work Cultures: The Case of Finland

In this article I focus on women workers’ experiences of transformation from line work to teamworking in Finnish clothing companies in the 1990s and also show what happened after this transformation in the clothing branch. The undertone of it is rather melancholic. Following an initial period of intensive and successful development, clothing work was moved from Finland to countries of cheap labour, such as Estonia, Latvia, Lithuania and Russia, and even China. In this type of network manufacturing, the development of modern information and communication technologies played a central role. My aim is to present the standpoint of women clothing workers in this process. The main body of the empirical data of my study consists of dialogues with clothing workers, union representatives, supervisors and managers. I also make use of my fieldwork notes, memos and research diaries from three companies over a period of five years. Furthermore, in the background lie the action research material from Scandinavian type work conferences and the survey material of an extensive mail inquiry that covered the whole branch in Finland. My own research started in 1991 as a mail inquiry and then continued as a case study in companies from 1992 to 2000, by employing action research and ethnographic methodologies.

The First WORKS Conference on Transformation of Work in a Global Knowledge Economy

The relationship between the changes of the global economy and individual working conditions formed the background of the first WORKS conference “The transformation of work in a global knowledge economy: towards a conceptual framework”, held in Chania, Greece from 21st – 22nd September, 2006 and attended by around 50 European researchers. Experts from academia and trade unions from all over the world were invited to give insights into their field of research, contributing to one of the main topics of the conference: (i) globalisation and organisational restructuring, (ii) workers’ organisation, the quality of working life and the gender dimension and (iii) global experiences and recommendations.

The recognition of work

The following article argues that recognition structures in work relations differ significantly in the sphere of paid work in contrast to unpaid work in private spheres. According to the systematic approach on recognition of Axel Honneth three different levels of recognition are identified: the interpersonal recognition, organisational recognition and societal recognition. Based on this framework it can be stated that recognition structures in the sphere of paid work and in private spheres differ very much. Whereas recognition in private spheres depends very much on personal relations, thus on the interpersonal level, recognition in employment relationships can be moreover built on organisational structures. Comparing recognition structures in both fields it becomes apparent, that recognition in field of employment can be characterised as much more concrete, comparable and measurable. Therefore, it can be concluded that the structural differences of recognition contribute to the high societal and individual importance of employment in contrast to unpaid work in private spheres.

Into a new phase of the research on restructuring of work in the knowledge society: the Third WORKS General Assembly in Sofia (Bulgaria)

The WORKS Project started two years ago (2005), involving the efforts of research institutes of 13 European countries with the main purpose of improving the understanding of the major changes in work in the knowledge-based society, taking account both of global forces and the regional diversity within Europe. This research meeting in Sofia (Bulgaria) aimed to present synthetically the massive amount of data collected in the case studies (occupational and organisational) and with the quantitative research during last year.

The globalisation in the clothing sector and its implications for work organisation: a view from the Portuguese case

The clothing sector in Portugal is still seen, in many aspects as a traditional sector with some average characteristics, such as: low level of qualifications, less flexible labour legislation and stronger unionisation, very low salaries and low capability of investment in innovation and new technology. Is, nevertheless, a very important sector in terms of labour market, with increased weight in the exporting structure. Globalisation and delocalisation are having a strong impact in the organisation of work and in occupational careers in the sector. With the pressure of global competitiveness in what concerns time and prices, very few companies are able to keep a position in the market without changes in organisation of work and workers. And those that can perform good responses to such challenges are achieving a better economical stability. The companies have found different ways to face this reality according to size, capital and position. We could find two main paths: one where companies outsource a part or the entire production to another territory (for example, several manufacturing tasks), close and/or dismissal the workers. Other path, where companies up skilled their capacities investing, for example, in design, workers training, conception and introduction of new or original products. This paper will present some results from the European project WORKS – Work organisation and restructuring in the knowledge society (6th Framework Programme), focusing the Portuguese case studies in several clothing companies in what concern implications of global context for the companies in general and for the workers in particular, in a comparative analysis with some other European countries.

International Conference on “Foresight Studies on Work in the Knowledge Society“ in Monte de Caparica (FCT-UNL) on 19-20 October 2006

On 19 and 20 October 2006, the Research Centre on Enterprise and Work Organisation (IET) organised the first international conference on “Foresight Studies on Work in the Knowledge Society”. It took place at the auditorium of the new Library of FCT-UNL and had the support of the research project “CodeWork@VO” (financed by FCT-MCTES and co-ordinated by INESC, Porto). The conference related to the European research project “Work Organisation and Restructuring in the Knowledge Society” (WORKS), which is financed by the European Commission. The main objective of the conference was to analyse and discuss research findings on the trends of work structures in the knowledge society, and to debate on new work organisation models and new forms of work supported by ICT.

Management Structure and Work Team Processes; Responsibilities and Responsiveness

This paper explores the management structure of the team-based organization. First it provides a theoretical model of structures and processes of work teams. The structure determines the team’s responsibilities in terms of authority and expertise about specific regulation tasks. The responsiveness of teams to these responsibilities are the processes of teamwork, in terms of three dimensions, indicating to what extent teams indeed use the space provided to them. The research question that this paper addresses is to what extent the position of responsibilities in the team-based organization affect team responsiveness. This is done by two hypotheses. First, the effect of the so-called proximity of regulation tasks is tested. It is expected that the responsibility for tasks positioned higher in the organization (i.e. further from the team) generally has a negative effect on team responsiveness, whereas tasks positioned lower in the organization (i.e. closer to the team) will have a positive effect on the way in which teams respond. Second, the relationship between the number of tasks for which the team is responsible with team responsiveness is tested. Theory suggests that teams being responsible for a larger number of tasks perform better, i.e. show higher responsiveness. These hypotheses are tested by a study of 109 production teams in the automotive industry. The results show that, as the theory predicts, increasing numbers of responsibilities have positive effects on team responsiveness. However, the delegation of expertise to teams seems to be the most important predictor of responsiveness. Also, not all regulation tasks show to have effects on team responsiveness. Most tasks do not show to have any significant effect at all. A number of tasks affects team responsiveness positively, when their responsibility is positioned lower in the organization, but also a number of tasks affects team responsiveness positively when located higher in the organization, i.e. further from the teams in the production. The results indicate that more attention can be paid to the distribution of responsibilities, in particular expertise, to teams. Indeed delegating more expertise improve team responsiveness, however some tasks might be located better at higher organizational levels, indicating that there are limitations to what responsibilities teams can handle.

Working in IT projects – Options and Limits of Work Design

Flexible forms of work like project work are gaining importance in industry and services. Looking at the research on project work, the vast majority of present literature is on project management, but increasingly, problems concerning the quality of work and the efficiency of project teams become visible. The question now is how project work can be structured in order to simultaneously provide efficient and flexible work and healthy working conditions ensuring the development of human resources for a long time. Selected results of publicly funded research into project work will be presented based on case studies in 7 software development /IT consulting project teams (N=34). A set of different methods was applied: interviews with management/project managers, group interviews on work constraints, a monthly diary about well-being and critical incidences in the course of the project, and a final evaluation questionnaire on project outcomes focusing on economic and health aspects. Findings reveal that different types of projects exist with varying degree of team members’ autonomy and influence on work structuring. An effect of self-regulation on mental strain could not be found. The results emphasize, that contradicting requirements and insufficient organizational resources with respect to the work requirements lead to an increased work intensity or work obstruction. These contradicting requirements are identified as main drivers for generating stress. Finally, employees with high values on stress for more than 2 months have significantly higher exhaustion rates than those with only one month peaks. Structuring project work and taking into account the dynamics of project work, there is a need for an active role of the project team in contract negotiation or the detailed definition of work – this is not only a question of individual autonomy but of negotiation the range of option for work structuring. Therefore, along with the sequential definition of the (software) product, the working conditions need to be re-defined.

The transformation of work? A quantitative evaluation of changes in work in Portugal

This report is made for the Work Package 15 of WORKS project and tries to develop more information on the Portuguese situation in the work structures changes in the recent years. It starts with an analysis of socio- economical indicators (Macro economical indicators, Employment indicators, Consumption, Technology at the workplace, Productivity), and then approaches the situation in terms of work flexibility in its dimensions of time use and New forms of work organisation. It traces employment in business functions with a sectoral and occupational approach, and analyses the occupational change in South Europe with particular relevance to Portugal (skill utilisation and job satisfaction, occupational and industrial mobility, quantitative evaluation of the shape of employment in Europe. Finaly are analysed the globalisation indicators.

Noise management at work

Every single day millions of European workers are exposed to noise at work. One in five of European’s workers have to raise his voice to be heard for at least half of the time that there are at work and 7% suffer from work-related hearing impairment. In Europe noise-induced hearing loss is the most common reported occupational disease. This paper deals with issues related with noise management at work, especially regarding the compliance with the new noise European Directive, (2003/10/EC).

Work organisation and restructuring in the knowledge society

It is generally agreed that major changes in work are taking place in the organisation of work as corporate structures are transformed in the context of economic globalisation and rapid technological change. But how can these changes be understood? And what are the impacts on social institutions and on workers and their families? The WORKS project brought together 17 research institutes in 13 European countries to investigate these important issues through a comprehensive four year research programme.

Futures of work and skills: some foresight exercises in Europe and USA

The Knowledge-based society brought a new way of living and working. The increasing decline of work in primary sector and traditional industries, related with the significant increase of employment in the service sector and in the knowledge work, changed the way companies and individuals establish their relations, the way work and life is organised. These changes are usual and fast and so the feeling of insecurity and unpredictability become more and more sharp. In this context, foresight exercises are necessary tools helping in the identification of the key variables and main trends of evolution. This report will present some foresight studies about work and skills in Europe and USA, in order to contribute to think about possible evolutions and trends.

Assessing scenarios on the future of work

In this paper will be discussed different types of scenarios and the aims for using scenarios. Normaly they are being used by organisations due to the need to anticipate processes, to support policy-making and to understand the complexities of relations. Such organisations can be private companies, R&D organisations and networks of organisations, or even by some public administration institutions. Some cases will be discussed as the methods for ongoing scenario-building process (Shell Internacional). Scenarios should anticipate possible relations among social actors as in the Triple Helix Model, and is possible to develop strategic intelligence in the innovation process that would enable the construction of scenarios. Such processes can be assessed. The focus will be made in relation to the steps chosen for the WORKS scenarios. In this case is there a model of work changes that can be used for foresight? Differences according to sectors were found, as well on other dimensions. Problems of assessment are analysed with specific application to the scenario construction methods.

Fragmentation? The future of work in Europe in a global economy: the WORKS final International Conference debate

WORKS final conference report