Search

Pancreatic ductal adenocarcinoma (PDAC) remains a deadly malignancy with no efficient therapy available up‐to‐date. Glycolysis is the main provider of energetic substrates to sustain cancer dissemination of PDAC. Accordingly, altering the glycolytic pathway is foreseen as a sound approach to trigger pancreatic cancer regression. Here, we show for the first time that high transforming growth factor beta‐induced (TGFBI) expression in PDAC patients is associated with a poor outcome. We demonstrate that, although usually secreted by stromal cells, PDAC cells synthesize and secrete TGFBI in quantity correlated with their migratory capacity. Mechanistically, we show that TGFBI activates focal adhesion kinase signaling pathway through its binding to integrin αVβ5, leading to a significant enhancement of glycolysis and to the acquisition of an invasive phenotype. Finally, we show that TGFBI silencing significantly inhibits PDAC tumor development in a chick chorioallantoic membrane assay model. Our study highlights TGFBI as an oncogenic extracellular matrix interacting protein that bears the potential to serve as a target for new anti‐PDAC therapeutic strategies. What's new? While pancreatic ductal adenocarcinoma (PDAC) continues to have exceedingly low 5‐year survival rates, there is hope that the discovery of reliable biomarkers and therapeutic targets can improve early diagnosis and treatment outcomes. To that end, the authors of this study identify the extracellular matrix protein transforming growth factor beta‐induced (TGFBI) as a promising PDAC target. In PDAC patients, high TGFBI expression was associated with poor outcome. In PDAC cells, tumor aggressiveness was associated with greater TGFBI synthesis and secretion. Mechanistic analyses show that TGFBI activates FAK signaling via integrin αVβ5 binding, enhancing glycolysis and invasiveness in PDAC cells. [ABSTRACT FROM AUTHOR]

Our study investigates the biochemical and functional impact of selective histone deacetylase 6 (HDAC6) inhibitors, a promising class of novel therapeutics, in several cancer models. Selective HDAC6 inhibitors (Tubathian A, Tubastatin A, Tubacin and Ricolinostat) and a non‐selective HDAC inhibitor (Vorinostat) were evaluated on cancer cell lines derived from multiple tumour types in both an in vitro and in vivo setting as potential cancer therapeutics. Selective HDAC6 inhibitors resulted in α‐tubulin acetylation with no impact on histone acetylation but failed to show any anti‐cancer properties. Only the use of high concentrations of selective HDAC6 inhibitors resulted in co‐inhibition of other HDAC enzymes and consequently in reduced growth, migratory and/or invasive activity of cancer cells in vitro as well as in vivo. The specificity of HDAC6 inhibition was confirmed using a CRISPR/Cas9 knockout cell line. Our results suggest that selective HDAC6 inhibitors may fall short as potential single agent anti‐cancer drugs and prove that many previous data regarding this promising class of compounds need to be interpreted with great care due to their use in high concentrations resulting in low selectivity and potential off‐target effects. What's new? Histone deacetylase 6 (HDAC6) helps regulate cell growth, metastasis, and invasion in tumour cells, making it an appealing target for anti‐cancer therapies. This study shows that in cancer cells, selective HDAC6 inhibitors increase acetylation levels of α‐tubulin, an HDAC6 substrate, without affecting histone acetylation. HDAC6 inhibition, however, did not reduce tumour cell growth or block tumour cell migration or invasion. Moreover, HDAC6 inhibitors exhibited anti‐tumour activity in vitro and in vivo only when administered in high concentrations, which also produced off‐target effects. The findings suggest that further study is needed to clarify data on anti‐cancer effects of selective HDAC6 inhibitors. [ABSTRACT FROM AUTHOR]

This work reports on the development of amide bond bioconjugation for the production of -NOTA and -NODAGA PRGD2 using batch strategy and microfluidic reactor technology. The final radiolabelling step was fully optimized using Design of Experiments and Design Space approaches, hence targeting robust labelling yields in routine. Optimal labelling conditions were defined in sodium acetate buffer as 168 μg/mL peptide concentration, 4.9 pH, 47.5°C temperature, and 12.5-minute reaction time. Upon optimization, the Gallium-68 radiolabelling was fully automated. All the work was designed to be compliant to the GMP environment and to support the pharmaceutical scale-up. [ABSTRACT FROM AUTHOR]

The present work focuses on the semisynthesis of halogenated and glycosylated derivatives of pheophorbide a (Ph a). Because of the low reaction yields enocuntered en route to halogenated derivatives, we then focused only on the semisynthesis of glycosylated derivatives of Ph a with the aim at enhancing the Ph a specificity for cancer cells by introducing β-galactose moieties expected to bind gal-1. We applied LC-SPE-NMR/MS, to facilitate the direct identification of glycosylated derivatives. The transposition of these analytical methods to a preparative scale facilitated the isolation of glycosylated compounds in quantities sufficient to evaluate in vitro photodynamic efficacies. The in vitro growth inhibitory activity of semisynthesized compounds was then evaluated using the MTT colorimetric assay in the presence and absence of light. However, this pharmacological evaluation method seems to be unable to efficiently yield information about carbohydrate effects in relation to possible compound specificities for gal-1 overexpressed by B16F10 cancer cells. [ABSTRACT FROM AUTHOR]

ABSTRACT Introduction As a complement to the classic metabolomics biofluid studies, the visualisation of the metabolites contained in cells or tissues could be a very powerful tool to understand how the local metabolism and biochemical pathways could be affected by external or internal stimuli or pathologies. Therefore, extraction and/or lysis is necessary to obtain samples adapted for use with the current analytical tools (liquid NMR and MS). These extraction or lysis work-ups are often the most labour-intensive and rate-limiting steps in metabolomics, as they require accuracy and repeatability as well as robustness. Many of the procedures described in the literature appear to be very time-consuming and not easily amenable to automation. Objective To find a fast, simplified procedure that allows release of the metabolites from cells and tissues in a way that is compatible with NMR analysis. Methods We assessed the use of sonication to disrupt cell membranes or tissue structures. Both a vibrating probe and an automated bath sonicator were explored. Results The application of sonication as the disruption procedure led to reproducible NMR spectral data compatible with metabolomics studies. This method requires only a small biological tissue or cell sample, and a rapid, reduced work-up was applied before analysis. The spectral patterns obtained are comparable with previous, well-described extraction protocols. Conclusion The rapidity and the simplicity of this approach could represent a suitable alternative to the other protocols. Additionally, this approach could be favourable for high- throughput applications in intracellular and intratissular metabolite measurements. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Introduction - The genus Echinacea ( Asteraceae) comprises about 10 species originally distributed in North America. Three species are very well known as they are used worldwide as medicinal plants: Echinacea purpurea, E. pallida, E. angustifolia. Objective - To discriminate between these three Echinacea species and E. simulata by 1H NMR-based metabolomics. Methodology - 1H NMR and multivariate analysis techniques were applied to diverse Echinacea plants including roots and aerial parts, authentic plants, commercial plants and commercial dry extracts. Results - Using the 1H NMR metabolomics, it was possible, without previous evaporation or separation steps, to obtain a metabolic fingerprint to distinguish between species. Conclusion - A clear distinction between the three pharmaceutical species was possible and some useful metabolites were identified. Copyright © 2009 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]