Effective combination of liposome-targeted chemotherapy and PD-L1 blockade of murine colon cancer

Therapeutic cancer drug efficacy can be limited by insufficient tumor penetration, rapid clearance, systemic toxicity and (acquired) drug resistance. The poor therapeutic index due to inefficient drug penetration and rapid drug clearance and toxicity can be improved by using a liposomal platform. Drug resistance for instance against pemetrexed, can be reduced by combination with docetaxel. Here, we developed a specific liposomal formulation to simultaneously deliver docetaxel and pemetrexed to enhance efficacy and safety. Hydrophobic docetaxel and hydrophilic pemetrexed were co-encapsulated into pH-sensitive liposomes using a thin-film hydration method with high efficiency. The physicochemical properties, toxicity, and immunological effects of liposomes were examined in vitro. Biodistribution, anti-tumor efficacy, and systemic immune response were evaluated in vivo in combination with PD-L1 immune checkpoint therapy using two murine colon cancer models. In cellular experiments, the liposomes exhibited strong cytotoxicity and induced immunogenic cell death. In vivo, the treatment with the liposome-based drug combination inhibited tumor development and stimulated immune responses. Liposomal encapsulation significantly reduced systemic toxicity compared to the delivery of the free drug. Tumor control was strongly enhanced when combined with anti-PDL1 immunotherapy in immunocompetent mice carrying syngeneic MC38 or CT26 colon tumors. We showed that treatment with liposome-mediated chemotherapy of docetaxel and pemetrexed combined with anti-PD-L1 immunotherapy is a promising strategy for the treatment of colon cancers.Horizon 2020 (H2020

Preclinical evaluation of EpCAM-binding designed ankyrin repeat proteins (DARPins) as targeting moieties for bimodal near-infrared fluorescence and photoacoustic imaging of cancer

PURPOSE Fluorescence-guided surgery (FGS) can play a key role in improving radical resection rates by assisting surgeons to gain adequate visualization of malignant tissue intraoperatively. Designed ankyrin repeat proteins (DARPins) possess optimal pharmacokinetic and other properties for in vivo imaging. This study aims to evaluate the preclinical potential of epithelial cell adhesion molecule (EpCAM)-binding DARPins as targeting moieties for near-infrared fluorescence (NIRF) and photoacoustic (PA) imaging of cancer. METHODS EpCAM-binding DARPins Ac2, Ec4.1, and non-binding control DARPin Off7 were conjugated to IRDye 800CW and their binding efficacy was evaluated on EpCAM-positive HT-29 and EpCAM-negative COLO-320 human colon cancer cell lines. Thereafter, NIRF and PA imaging of all three conjugates were performed in HT-29_luc2 tumor-bearing mice. At 24 h post-injection, tumors and organs were resected and tracer biodistributions were analyzed. RESULTS Ac2-800CW and Ec4.1-800CW specifically bound to HT-29 cells, but not to COLO-320 cells. Next, 6 nmol and 24 h were established as the optimal in vivo dose and imaging time point for both DARPin tracers. At 24 h post-injection, mean tumor-to-background ratios of 2.60 ± 0.3 and 3.1 ± 0.3 were observed for Ac2-800CW and Ec4.1-800CW, respectively, allowing clear tumor delineation using the clinical Artemis NIRF imager. Biodistribution analyses in non-neoplastic tissue solely showed high fluorescence signal in the liver and kidney, which reflects the clearance of the DARPin tracers. CONCLUSION Our encouraging results show that EpCAM-binding DARPins are a promising class of targeting moieties for pan-carcinoma targeting, providing clear tumor delineation at 24 h post-injection. The work described provides the preclinical foundation for DARPin-based bimodal NIRF/PA imaging of cancer

The Cost of Flexibility at the Margin. Comparing the Wage Penalty for Fixed-term Contracts in Germany and Spain using Quantile Regression

Germany and Spain are typically regarded as 'rigid' economies, yet both have had different experiences of fixed-term jobs. Using quantile regression we find that in West Germany the earnings of permanent and fixed-term workers are most similar among high earners and most dissimilar among low earners. In Spain, the wage penalty shows little variation across the distribution of wages. This pattern was also found for different occupational groups, although there are clear differences in the absolute wage penalty across occupations. In conclusion we caution against generalizing findings from Spain to other 'rigid' European labour markets. Copyright 2007 The Authors. Journal compilation CEIS, Fondazione Giacomo Brodolini and Blackwell Publishing Ltd. 2007.

Labour Markets, Gender and Institutions: Essays in honour of Gunther Schmid

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BASS 4: a software system for ergonomic design and evaluation of working hours

OBJECTIVE: To extend an existing computer programme for the evaluation and design of shift schedules (BASS 3) by integrating workload as well as economic aspects. METHODS: The redesigned prototype BASS 4 includes a new module with a suitable and easily applicable screening method (EBA) for the assessment of the intensity of physical, emotional and cognitive workload components and their temporal patterns. Specified criterion functions based on these ratings allow for an adjustment of shift and rest duration according to the intensity of physical and mental workload. Furthermore, with regard to interactive effects both workload and temporal conditions, e.g. time of day, are taken into account. In a second new module, important economic aspects and criteria have been implemented. Different ergonomic solutions for scheduling problems can now also be evaluated with regard to their economic costs. RESULTS: The new version of the computer programme (BASS 4) can now simultaneously take into account numerous ergonomic, legal, agreed and economic criteria for the design and evaluation of working hours. CONCLUSIONS: BASS 4 can now be used as an instrument for the design and the evaluation of working hours with regard to legal, ergonomic and economic aspects at the shop floor as well as in administrative (e.g. health and safety inspection) and research problems.<br>OBJETIVOS: Expandir um programa computacional existente para planejamento e avaliação dos horários de turnos (BASS 3) por meio da incorporação da carga de trabalho e características econômicas. MÉTODOS: O protótipo BASS 4 contém um novo módulo com um método de triagem (EBA) conveniente e de fácil aplicação para a avaliação da intensidade dos componentes físico, emocional e cognitivo da carga de trabalho e seus padrões temporais. O uso de critérios específicos com base nestas avaliações possibilita ajustar a duração do turno e do descanso de acordo com a intensidade da carga de trabalho física e mental. Além disso, quanto aos efeitos interativos, tanto a carga de trabalho como os aspectos temporais, p. e., hora do dia, são considerados. Foram introduzidos em um outro módulo características e critérios econômicos de relevância. O novo programa permite também que sejam avaliadas diferentes soluções ergonômicas para problemas de planejamento segundo os custos financeiros. RESULTADOS: A nova versão do programa (BASS 4) tem a capacidade agora de processar simultaneamente vários critérios econômicos, ergonômicos, legais e acordados para o planejamento e avaliação do horário de trabalho. CONCLUSÕES: O BASS 4 pode ser usado agora como um instrumento para planejamento e avaliação dos horários de trabalho, incluindo-se características econômicas, ergonômicas e legais, no setor de produção e em questões administrativas (p.e. fiscalização da saúde e segurança) e relacionados à pesquisa

A multimodal molecular imaging approach targeting urokinase plasminogen activator receptor for the diagnosis, resection and surveillance of urothelial cell carcinoma

With a 5-year recurrence rate of 30–78%, urothelial cell carcinoma (UCC) rates amongst the highest of all solid malignancies. Consequently, after transurethral resection, patients are subjugated to life-long endoscopic surveillance. A multimodal near-infrared (NIR) fluorescence-based imaging strategy can improve diagnosis, resection and surveillance, hence increasing quality of life. Methods: Expression of urokinase plasminogen activator receptor (uPAR) and epithelial cell adhesion molecule (EpCAM) are determined on paraffin-embedded human UCC using immunohistochemistry and on UCC cell lines by flow cytometry. MNPR-101, a humanised monoclonal antibody targeting uPAR is conjugated to IRDye800CW and binding is validated in vitro using surface plasmon resonance and cell-based binding assays. In vivo NIR fluorescence and photoacoustic three-dimensional (3D) imaging are performed with subcutaneously growing human UM-UC-3luc2 cells in BALB/c-nude mice. The translational potential is confirmed in a metastasising UM-UC-3luc2 orthotopic mouse model. Infliximab-IRDye800CW and rituximab-IRDye800CW are used as controls. Results: UCCs show prominent uPAR expression at the tumour-stroma interface and EpCAM on epithelial cells. uPAR and EpCAM are expressed by 6/7 and 4/7 UCC cell lines, respectively. In vitro, MNPR-101-IRDye800CW has a picomolar affinity for domain 2-3 of uPAR. In vivo fluorescence imaging with MNPR-101-IRDye800CW, specifically delineates both subcutaneous and orthotopic tumours with tumour-to-background ratios reaching as high as 6.8, differing significantly from controls (p &lt; 0.0001). Photoacoustic 3D in depth imaging confirms the homogenous distribution of MNPR-101-IRDye800CW through the tumour. Conclusions: MNPR-101-IRDye800CW is suitable for multimodal imaging of UCC, awaiting clinical translation