Malignant pleural mesothelioma (MPM) is an aggressive tumour that grows in the pleural cavity. MPM spheroids released in the pleural fluid can form new tumour foci. Cell–cell, cell–extracellular matrix (ECM) interactions in 2D and 3D impact malignant cell behaviour during cell adhesion, migration, proliferation and epithelial–mesenchymal transition (EMT). In this study, epithelioid, biphasic and sarcomatoid MPM cell types as well as benign mesothelial cells were tested with regards to the above phenotypes. Fibronectin (FN) and homologous cell‐derived extracellular matrix (hcd‐ECM) treated substratum differentially affected the above phenotypes. 3D MPM spheroid invasion was higher in FN‐collagen matrices in the epithelioid and biphasic cells, while 3D cell cultures of epithelioid and sarcomatoid MPM cells in FN‐collagen showed a higher contractility compared to hcd‐ECM‐collagen. Cell aggregates demonstrated invasive behaviour in hcd‐ECM matrices alone. Our results suggest that ECM and the dimensionality affect malignant cell behaviour during cell culture studies.

Background/Aim: Malignant pleural mesothelioma (MPM) is a therapy-resistant neoplasm of the pleura. Standard chemotherapy consists of a combination of cisplatin (CPDD) and pemetrexed (PEM). The aim of this study was to assess whether inhibition of aerobic glycolysis by 2-deoxyglucose (2DG) would enhance the effects of standard chemotherapy. Materials and Methods: MeT-5A, M14K, MSTO and ZL34 cell lines were used. Cell viability with 2DG and cell proliferation and spheroid formation with CPDD+PEM alone and with 2-DG were tested. Results: Viability with 2-DG was dose-dependent. Cell proliferation with CPDD+PEM on 2D surface was reduced in all cell types, 2-DG inclusion demonstrated a synergistic effect in MSTO and ZL34 cells. Spheroid growth in 3D with CPDD+PEM or CPDD+PEM+2-DG lowered spheroid growth in all cell types. Conclusion: 2-DG synergizes with CPDD+PEM in lowering MPM cell proliferation in 2D to <20%. In 3D MPM spheroid growth 2-DG synergism with CPDD+PEM treatment is not maintained.Malignant pleural mesothelioma (MPM) is a therapyresistant neoplasm of the pleura arising from asbestos exposure with a median survival after diagnosis of around one year (1). In a cohort of 19,000 MPM cases it was shown that with no treatment, the overall survival was 4.8 months, while with chemotherapy the overall survival was reported to be 11.3 months (2). The most frequent histotype of MPM is epithelioid followed by biphasic and sarcomatoid. The sarcomatoid histotype is considered to be the most aggressive followed by the biphasic and epithelioid type (3). Usually due to late diagnosis the tumor is unresectable, therefore the main treatment is chemotherapy by the combination of cisplatin (CPDD) and pemetrexed (PEM) (1).A characteristic feature of MPM compared to other solid tumors, is that it develops as a sheet in the pleural membrane (in a 2-dimentional like fashion) wrapping and congesting the lung parenchyma (4). On the other hand, the spontaneous formation of spheroid structures in explant culture indicates an inherent property of the primary cells to recapitulate a morphological 3-dimentional character, that may be associated with local dissemination of MPM in the pleural cavity. Previously, pleural fluid cytology demonstrated the presence of 3-dimentional spheroid like cell aggregates with coiled cords of cells, small papillae with a central core and at times tissue fragments with pseudoacinar formation (5). Therefore, when assessing the efficacy of chemotherapeutic agents in vitro, it is reasonable to do so both in 2-D and 3-D cultures.CPDD and PEM treatment have been shown to directly induce the expression of multidrug resistance ABC transporter proteins like ABCG2 in biphasic and sarcomatoid mesothelioma cell models. The induction of such transporters results in the expulsion of chemotherapeutic agents that renders MPM chemo-resistant (6). Furthermore, cell-extracellular matrix (ECM) interactions apart from determining tumor cell implantation to novel sites, also induce the expression of multidrug resistance ABC t...

Malignant pleural mesothelioma (MPM) is an aggressive cancer with poor prognosis. The main treatment for MPM is doublet chemotherapy with Cisplatin and Pemetrexed, while ongoing trials test the efficacy of pemetrexed monotherapy. However, there is lack of evidence regarding the effects of Cisplatin and Pemetrexed on MPM cell phenotypes, especially in three-dimensional (3D) cell cultures. In this study, we evaluated the effects Cisplatin and Pemetrexed on cell viability using homologous cell derived extracellular matrix (hECM) as substratum and subsequently in the following 3D cell culture phenotypes: tumor spheroid formation, tumor spheroid invasion, and collagen gel contraction. We used benign mesothelial MeT-5A cells as controls and the MPM cell lines M14K (epithelioid), MSTO (biphasic), and ZL34 (sarcomatoid). Cell viability of all cell lines was significantly decreased with all treatments. Mean tumor spheroid perimeter was reduced after treatment with Pemetrexed or the doublet therapy in all cell lines, while Cisplatin reduced the mean spheroid perimeter of MeT-5A and MSTO cells. Doublet treatment reduced the invasive capacity of spheroids of cell lines into collagenous matrices, while Cisplatin lowered the invasion of the MSTO and ZL34 cell lines, and Pemetrexed lowered the invasion of MeT-5A and ZL34 cell lines. Treatment with Pemetrexed or the combination significantly reduced the collagen gel contraction of all cell lines, while Cisplatin treatment affected only the MeT-5A and M14K cells. The results of the current study can be used as an in vitro 3D platform for testing novel drugs against MPM for ameliorating the effects of first line chemotherapeutics.

an allogeneic mixed-lymphocyte reaction (MLR). Lymphocyte proliferation (flow-cytometric measurement of CFSE) and cytokine production (multiplex bead array) were assessed at day 5. The proportion of lymphocytes primed to produce IFNg was measured at day 7 (intracellular staining). Results UPM-stimulation increased DC expression of the maturation marker CD83 (p = 0.0038) and chemokine receptor CCR7 (p = 0.0018). It had no effect on CD40 or MHC Class I expression. UPM-stimulation of DCs also significantly increased CD8 lymphocyte proliferation (p = 0.020), and the production of IFNg, TNFa and IL-13 by CD8 lymphocytes in MLR at day 5 (all p < 0.05; Table 1). The proportion of CD8 lymphocytes primed to produce IFNg was also increased by UPM-stimulation of DCs (p = 0.034). Conclusion No evidence of an impaired Tc1 response was seen with UPM-stimulated DCs, in contrast to our previous findings with CD4 T lymphocytes. This may be because CD8 lymphocytes are more primed to respond and produce cytokines at baseline. However, UPM-treatment of DCs did significantly increase DC expression of CCR7, which directs DCs to lymph nodes, and increased the priming of Tc1 and Tc2 responses in the absence of any other stimulation. Inhalation of UPM may give rise to pathological CD8 responses to otherwise innocuous novel antigens. Introduction Increasing evidence suggests accelerated ageing as a pathogenic mechanism in COPD. Methods and results Telomere length in circulating WBC, a marker of biological ageing, was assessed in 200 ex-smoker COPD patients (108 male, age 61.5 ± 6.4 years, FEV 1 45.6 ± 17.1% predicted), 50 ex-smokers with normal lung function (27 male, age 59.9 ± 7.3 years, FEV 1 109.1 ± 13.4%predicted) and 50 non-smoker healthy subjects (27 male, age 59.3 ± 8.3 years, FEV 1 113.2 ± 13.1% predicted). TL was assessed by qPCR and expressed as relative T/S ratio. S49 TELOMERE ATTRITION IN CIRCULATING WHITE BLOOD CELLS IN COPD RELATES TO LUNG FUNCTION AND OUTCOMESTL was shorter in COPD (0.77 ± 0.2 relative T/S ratio) than in both ex-smokers (0.83 ± 0.2 relative T/S ratio) and nonsmokers (0.84 ± 0.2 relative T/S ratio) (p < 0.05). Furthermore TL correlated negatively with age (r -0.17, p 0.007), emphysema score (r -0.217, p 0.001), number of exacerbations in the previous year to inclusion in the study (r -0.129, p 0.04), number of hospitalisations over 3 years follow-up (r -0.167, p 0.004) and positively with FEV 1 (r 0.135, p = 0.03) and arterial oxygen saturation (r 0.161, p 0.01). Conclusion COPD patients have evidence of premature ageing (shortened TL) compared to normal subjects irrespective of their smoking history. TL relates to FEV 1 , SatO 2 , exacerbation rate and hospitalisations.

In this study, we investigated the self-reported (questionnaire-based) prevalence of Obstructive Sleep Apnoea Syndrome (OSAS) and the prevalence of Chronic Obstructive Pulmonary Diseases (COPD) in the context of demographics and adherence to the Mediterranean diet in the general population of Alonissos, a non-profit line island in Greece (i.e., with scarce boat transportation to the mainland). In this cross-sectional study, 236 inhabitants of Alonissos participated (circa 10% of the island’s population), and 115 males and 121 females were evaluated with appropriate questionnaires for OSAS, COPD, and adherence to the Mediterranean diet and subsequently underwent spirometry testing to establish COPD diagnosis. The self-reported prevalence of OSAS and COPD was 9.44% and 18.8%, respectively. However, only 8.99% of the participants were diagnosed with COPD based on their spirometry testing. Adherence to the Mediterranean Diet was moderate. The high prevalence of COPD and OSAS in this underprivileged island in terms of healthcare access highlights the need for improvements in health promotion and primary healthcare provision in non-profit line Greek islands.