Biological evidence of cancer stem-like cells and recurrent disease in osteosarcoma

Sarcomas are a large family of cancers originating in the mesenchyme. Composed of more than 100 histological subtypes, soft tissue and bone sarcomas remain clinically challenging, particularly in children and adolescents in whom sarcomas are the second most common malignant entities. Osteosarcoma is the main primary bone tumor in adolescents and young adults and is characterized by a high propensity to induce distant metastatic foci and become multi-drug resistant. The innate and acquired resistance of osteosarcoma can be explained by high histological heterogeneity and genetic/molecular diversity. In the last decade, the notion of cancer stem-like cells (CSCs) has emerged. This subset of cancer cells has been linked to drug resistance properties, recurrence of the disease, and therapeutic failure. Although CSCs remain controversial, many elements are in favor of them playing a role in the development of the drug resistance profile. The present review gives a brief overview of the most recent biological evidence of the presence of CSCs in osteosarcomas and their role in the drug resistance profile of these rare oncological entities. Their use as promising therapeutic targets is discussed

Technical report: liquid overlay technique allows the generation of homogeneous osteosarcoma, glioblastoma, lung and prostate adenocarcinoma spheroids that can be used for drug cytotoxicity measurements

Introduction: The mechanisms involved in cancer initiation, progression, drug resistance, and disease recurrence are traditionally investigated through in vitro adherent monolayer (2D) cell models. However, solid malignant tumor growth is characterized by progression in three dimensions (3D), and an increasing amount of evidence suggests that 3D culture models, such as spheroids, are suitable for mimicking cancer development. The aim of this report was to reaffirm the relevance of simpler 3D culture methods to produce highly reproducible spheroids, especially in the context of drug cytotoxicity measurements. Methods: Human A549 lung adenocarcinoma, LnCaP prostate adenocarcinoma, MNNG/HOS osteosarcoma and U251 glioblastoma cell lines were grown into spheroids for 20 days using either Liquid Overlay Technique (LOT) or Hanging Drop (HD) in various culture plates. Their morphology was examined by microscopy. Sensitivity to doxorubicin was compared between MNNG/HOS cells grown in 2D and 3D. Results: For all cell lines studied, the morphology of spheroids generated in round-bottom multiwell plates was more repeatable than that of those generated in flat-bottom multiwell plates. HD had no significant advantage over LOT when the spheroids were cultured in round-bottom plates. Finally, the IC50 of doxorubicin on MNNG/HOS cultured in 3D was 18.8 times higher than in 2D cultures (3D IC50 = 15.07 ± 0.3 µM; 2D IC50 = 0.8 ± 0.4 µM; *p < 0.05). Discussion: In conclusion, we propose that the LOT method, despite and because of its simplicity, is a relevant 3D model for drug response measurements that could be scaled up for high throughput screening

Inhibiting endothelin receptors with macitentan strengthens the bone protective action of RANKL inhibition and reduces metastatic dissemination in osteosarcoma

Current treatments for osteosarcoma, combining conventional polychemotherapy and surgery, make it possible to attain a five-year survival rate of 70% in affected individuals. The presence of chemoresistance and metastases significantly shorten the patient’s lifespan, making identification of new therapeutic tools essential. Inhibiting bone resorption has been shown to be an efficient adjuvant strategy impacting the metastatic dissemination of osteosarcoma, tumor growth, and associated bone destruction. Unfortunately, over-apposition of mineralized matrix by normal and tumoral osteoblasts was associated with this inhibition. Endothelin signaling is implicated in the functional differentiation of osteoblasts, raising the question of the potential value of inhibiting it alone, or in combination with bone resorption repression. Using mouse models of osteosarcoma, the impact of macitentan, an endothelin receptor inhibitor, was evaluated regarding tumor growth, metastatic dissemination, matrix over-apposition secondary to RANKL blockade, and safety when combined with chemotherapy. The results showed that macitentan has no impact on tumor growth or sensitivity to ifosfamide, but significantly reduces tumoral osteoid tissue formation and the metastatic capacity of the osteosarcoma. To conclude, macitentan appears to be a promising therapeutic adjuvant for osteosarcoma alone or associated with bone resorption inhibitors

Three-dimensional in vitro culture models in oncology research

Cancer is a multifactorial disease that is responsible for 10 million deaths per year. The intra- and inter-heterogeneity of malignant tumors make it difficult to develop single targeted approaches. Similarly, their diversity requires various models to investigate the mechanisms involved in cancer initiation, progression, drug resistance and recurrence. Of the in vitro cell-based models, monolayer adherent (also known as 2D culture) cell cultures have been used for the longest time. However, it appears that they are often less appropriate than the three-dimensional (3D) cell culture approach for mimicking the biological behavior of tumor cells, in particular the mechanisms leading to therapeutic escape and drug resistance. Multicellular tumor spheroids are widely used to study cancers in 3D, and can be generated by a multiplicity of techniques, such as liquid-based and scaffold-based 3D cultures, microfluidics and bioprinting. Organoids are more complex 3D models than multicellular tumor spheroids because they are generated from stem cells isolated from patients and are considered as powerful tools to reproduce the disease development in vitro. The present review provides an overview of the various 3D culture models that have been set up to study cancer development and drug response. The advantages of 3D models compared to 2D cell cultures, the limitations, and the fields of application of these models and their techniques of production are also discussed

Comparaison du nivellement direct, du positionnement GPS bifréquence et de l'altimétrie satellitaire radar pour le nivellement des stations limnimétriques du bassin Amazonien

Three levelling techniques were compared for the levelling of hydrometric stations along Amazonian rivers : spirit levelling, bi-frequency GPS positioning and satellite radar altimetry (Topex/Poseidon T/P. Results show a good agreement between GPS positioning and satellite radar altimetry techniques, the two techniques having been compared on 23 hydrometric stations with an average root mean square error of 0.05 +/- 0.64m. Comparison with spirit levelling allowed the identification of discrepancies in direct geometric land levelling results from Brazilian and Peruvian networks over the Amazon Basin. Levelling hydrometric stations through satellite radar altimetry required a specific methodology : maximum annual water levels (referred to the geoid, using EGM96 geopotential model) are quantified at intersections between the satellite ground tracks and the river network. Maximum annual water levels at hydrometric stations are then derived through longitudinal interpolation. Comparison with maximum annual readings at gauges allows the determination of local orthometric heights at these stations. Altimetric levelling from Topex/Poseidon measurements has been realized for 97 hydrometric stations along 27 740 km of the Amazon hydrographic network. Resulting river surface longitudinal profiles proved to be fully consistent in terms of hydrodynamics

Etablissement d'un référentiel altimétrique sur le bassin amazonien par altimétrie satellitaire radar (Topex Poseidon)

Satellite radar altimetry (Topex/Poseidon T/P) was used to establish a consistent altimetric reference network over the Amazon basin. A methodology was developed to quantify maximum annual water levels (referred to the geoid, using EGM96 geoid model) at intersections between the satellite ground tracks and the river network. Maximum annual water levels (referred to the geoid) at hydrometric stations where then derived using longitudinal interpolation. Comparison with maximum annual readings at gauges allowed the determination of local orthometric heights at these stations. Altimetric leveling from Topex/Poseidon measurements has been realized for 97 hydrometric stations along 27 740 km of the Amazon hydrographic network. Validation has been realized both by checking the overall hydraulic consistency of longitudinal river profiles at low and high river stages and by comparing, for 23 hydrometric stations, orthometric heights obtained from T/P measurements with values obtained from bi-frequency GPS positioning. Results show a good agreement between the two techniques (average root mean square error of 0.05 +/- 0.64m). This levelling method allowed the identification of discrepancies in direct geometric land leveling results from Brazilian and Peruvian networks. These results are of major importance for the study of Amazon river flow dynamics and sediment transport

Investigation of humoral and cellular immunity of dairy cattle after one or two year of vaccination with a phase I Coxiella vaccine

AbstractQ fever is a worldwide zoonosis that may cause reproductive disorders such as abortion, endometritis or infertility in livestock. The implementation of a vaccination with a phase I vaccine is the nowadays the most relevant way to control the spread of the infection within herds. Annual boosters are recommended for ruminants by the manufacturer whereas in humans, to prevent side effects, no booster must be done before 5 years and the lack of humoral and cellular immunity has to be confirmed before any additional vaccination. The aim of this study was to investigate, in dairy cattle, the interest of such annual booster by assessing the level of different immune markers among 142 animals (from infected and uninfected herds) vaccinated either 2 year (i.e. 2 times) or 1 year before with an efficient commercial phase I Q fever vaccine. One year after vaccination, more than 80 % of the vaccinated cows had still immune markers, whereas 68 % of the heifers from uninfected herd did not. These data suggested that an annual booster would not be necessary for all vaccinated animals within a herd. In order to detect the immune animals and then to optimize the number of animals needing a boost, the skin test method, performed at least 3 days before the vaccination could be used