9 results on '"Ivan Adanja"'
Search Results
2. VEGFR1 and VEGFR2 involvement in extracellular galectin-1- and galectin-3-induced angiogenesis.
- Author
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Nicky D'Haene, Sébastien Sauvage, Calliope Maris, Ivan Adanja, Marie Le Mercier, Christine Decaestecker, Linda Baum, and Isabelle Salmon
- Subjects
Medicine ,Science - Abstract
AIM: Accumulating evidence suggests that extracellular galectin-1 and galectin-3 promote angiogenesis. Increased expression of galectin-1 and/or galectin-3 has been reported to be associated with tumour progression. Thus, it is critical to identify their influence on angiogenesis. METHODS: We examined the individual and combined effects of galectin-1 and galectin-3 on endothelial cell (EC) growth and tube formation using two EC lines, EA.hy926 and HUVEC. The activation of vascular endothelial growth factor receptors (VEGFR1 and VEGFR2) was determined by ELISA and Western blots. We evaluated the VEGFR1 and VEGFR2 levels in endosomes by proximity ligation assay. RESULTS: We observed different responses to exogenous galectins depending on the EC line. An enhanced effect on EA.hy926 cell growth and tube formation was observed when both galectins were added together. Focusing on this enhanced effect, we observed that together galectins induced the phosphorylation of both VEGFR1 and VEGFR2, whereas galectin-1 and -3 alone induced VEGFR2 phosphorylation only. In the same way, the addition of a blocking VEGFR1 antibody completely abolished the increase in tube formation induced by the combined addition of both galectins. In contrast, the addition of a blocking VEGFR2 antibody only partially inhibited this effect. Finally, the addition of both galectins induced a decrease in the VEGFR1 and VEGFR2 endocytic pools, with a significantly enhanced effect on the VEGFR1 endocytic pool. These results suggest that the combined action of galectin-1 and galectin-3 has an enhanced effect on angiogenesis via VEGFR1 activation, which could be related to a decrease in receptor endocytosis.
- Published
- 2013
- Full Text
- View/download PDF
3. A new method to address unmet needs for extracting individual cell migration features from a large number of cells embedded in 3D volumes.
- Author
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Ivan Adanja, Véronique Megalizzi, Olivier Debeir, and Christine Decaestecker
- Subjects
Medicine ,Science - Abstract
BACKGROUND: In vitro cell observation has been widely used by biologists and pharmacologists for screening molecule-induced effects on cancer cells. Computer-assisted time-lapse microscopy enables automated live cell imaging in vitro, enabling cell behavior characterization through image analysis, in particular regarding cell migration. In this context, 3D cell assays in transparent matrix gels have been developed to provide more realistic in vitro 3D environments for monitoring cell migration (fundamentally different from cell motility behavior observed in 2D), which is related to the spread of cancer and metastases. METHODOLOGY/PRINCIPAL FINDINGS: In this paper we propose an improved automated tracking method that is designed to robustly and individually follow a large number of unlabeled cells observed under phase-contrast microscopy in 3D gels. The method automatically detects and tracks individual cells across a sequence of acquired volumes, using a template matching filtering method that in turn allows for robust detection and mean-shift tracking. The robustness of the method results from detecting and managing the cases where two cell (mean-shift) trackers converge to the same point. The resulting trajectories quantify cell migration through statistical analysis of 3D trajectory descriptors. We manually validated the method and observed efficient cell detection and a low tracking error rate (6%). We also applied the method in a real biological experiment where the pro-migratory effects of hyaluronic acid (HA) were analyzed on brain cancer cells. Using collagen gels with increased HA proportions, we were able to evidence a dose-response effect on cell migration abilities. CONCLUSIONS/SIGNIFICANCE: The developed method enables biomedical researchers to automatically and robustly quantify the pro- or anti-migratory effects of different experimental conditions on unlabeled cell cultures in a 3D environment.
- Published
- 2011
- Full Text
- View/download PDF
4. Phase contrast image segmentation by weak watershed transform assembly.
- Author
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Olivier Debeir, Ivan Adanja, Nadine Warzée, Philippe Van Ham, and Christine Decaestecker
- Published
- 2008
- Full Text
- View/download PDF
5. Quantitative Analysis of Melanocyte Migrationin vitroBased on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions
- Author
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Françoise Xavier, Stuart J. Gallagher, Sylvain Fouliard, Olivier Debeir, Véronique Letort, Gaëlle Letort, Lionel Larue, Ivan Adanja, Mayuko Y. Kumasaka, Mathématiques Appliquées aux Systèmes - EA 4037 (MAS), Ecole Centrale Paris, Laboratory of Image Synthesis and Analysis (LISA), Université libre de Bruxelles (ULB), Institut Curie [Paris], Régulations cellulaires et oncogenèse (RCO), and Institut Curie [Paris]-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Cell division ,Integrin ,Cell ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Melanocyte migration ,Extracellular matrix ,03 medical and health sciences ,0302 clinical medicine ,[MATH.MATH-ST]Mathematics [math]/Statistics [math.ST] ,medicine ,Cell migration ,Cell-matrix interaction ,Fibronectin ,030304 developmental biology ,Automated cell tracking ,0303 health sciences ,biology ,Applied Mathematics ,Contact inhibition ,[STAT.TH]Statistics [stat]/Statistics Theory [stat.TH] ,Cell biology ,medicine.anatomical_structure ,Beta-catenin ,Modeling and Simulation ,Video microscopy ,biology.protein ,Melanocytes ,030217 neurology & neurosurgery - Abstract
International audience; The aim of this study was to describe and analyze the regulation and spatio-temporal dynamics of melanocyte migration in vitro and its coupling to cell division and interaction with the matrix. The melanocyte lineage is particularly interesting because it is involved in both embryonic development and oncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocyte cell lines established from wild-type and beta-catenin-transgenic mice (bcat*). The multi-functional beta-catenin molecule is known to be able to regulate the transcription of various genes involved in cell proliferation and migration, particularly in the melanocyte lineage. We also investigated fibronectin, an extra-cellular matrix protein that binds integrins, thereby providing adhesion points for cells and encouraging migration. As the migration of individual cells were followed, automated methods were required for processing the large amount of data generated by the time-lapse video-microscopy. A model-based approach for automated cell tracking was evaluated on a sample by comparison with manual tracking. This method was found reliable in studying overall cell behaviour. Its application to all the observed sequences provided insight into the factors affecting melanocyte migration in vitro: melanocytes of mutated form of beta-catenin showed higher division rates and no contact inhibition of growth was induced by the resulting increase in cell density. However, cell density limited the amplitude of cell displacements, although their motility was less affected. The high fibronectin concentration bound to substratum promoted cell migration and motility, the effect being more intense for wild-type cells than for cells with beta-catenin over-expression. During the division process, cell migration speed increased rapidly then decreased slowly. Analyses of such data is expected to lead both to biological answers and to a framework for a better modeling processes in the future.
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- 2010
6. VEGFR1 and VEGFR2 Involvement in Extracellular Galectin-1- and Galectin-3-Induced Angiogenesis
- Author
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Christine Decaestecker, Marie Le Mercier, Linda G. Baum, Isabelle Salmon, Ivan Adanja, Nicky D'Haene, Calliope Maris, and Sébastien Sauvage
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Galectin 1 ,Angiogenesis ,Galectin 3 ,Endocytic cycle ,Glycobiology ,lcsh:Medicine ,Cardiovascular ,Biochemistry ,chemistry.chemical_compound ,Molecular Cell Biology ,Basic Cancer Research ,Phosphorylation ,lcsh:Science ,Cells, Cultured ,Tube formation ,Extracellular Matrix Proteins ,Multidisciplinary ,Protein Kinase Signaling Cascade ,Neovascularization, Pathologic ,Sciences bio-médicales et agricoles ,Signaling Cascades ,Endocytosis ,Cell biology ,Endothelial stem cell ,Vascular endothelial growth factor ,Oncology ,Galectin-3 ,Galectin-1 ,cardiovascular system ,Medicine ,Cellular Types ,Research Article ,Signal Transduction ,animal structures ,Blotting, Western ,Enzyme-Linked Immunosorbent Assay ,Biology ,Cell Growth ,Vascular Biology ,otorhinolaryngologic diseases ,Humans ,Galectin ,Vascular Endothelial Growth Factor Receptor-1 ,lcsh:R ,Proteins ,Endothelial Cells ,Molecular biology ,Vascular Endothelial Growth Factor Receptor-2 ,stomatognathic diseases ,chemistry ,lcsh:Q - Abstract
Aim:Accumulating evidence suggests that extracellular galectin-1 and galectin-3 promote angiogenesis. Increased expression of galectin-1 and/or galectin-3 has been reported to be associated with tumour progression. Thus, it is critical to identify their influence on angiogenesis.Methods:We examined the individual and combined effects of galectin-1 and galectin-3 on endothelial cell (EC) growth and tube formation using two EC lines, EA.hy926 and HUVEC. The activation of vascular endothelial growth factor receptors (VEGFR1 and VEGFR2) was determined by ELISA and Western blots. We evaluated the VEGFR1 and VEGFR2 levels in endosomes by proximity ligation assay.Results:We observed different responses to exogenous galectins depending on the EC line. An enhanced effect on EA.hy926 cell growth and tube formation was observed when both galectins were added together. Focusing on this enhanced effect, we observed that together galectins induced the phosphorylation of both VEGFR1 and VEGFR2, whereas galectin-1 and -3 alone induced VEGFR2 phosphorylation only. In the same way, the addition of a blocking VEGFR1 antibody completely abolished the increase in tube formation induced by the combined addition of both galectins. In contrast, the addition of a blocking VEGFR2 antibody only partially inhibited this effect. Finally, the addition of both galectins induced a decrease in the VEGFR1 and VEGFR2 endocytic pools, with a significantly enhanced effect on the VEGFR1 endocytic pool. These results suggest that the combined action of galectin-1 and galectin-3 has an enhanced effect on angiogenesis via VEGFR1 activation, which could be related to a decrease in receptor endocytosis. © 2013 D'Haene et al., SCOPUS: ar.j, info:eu-repo/semantics/published
- Published
- 2013
7. In vivo assessment of temozolomide local delivery for lung cancer inhalation therapy
- Author
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Sven Saussez, Robert Kiss, Véronique Mathieu, Christine Decaestecker, Philippe Deleuze, Julien Hecq, Karim Amighi, Isabelle Roland, Olivier Debeir, Ivan Adanja, and Nathalie Wauthoz
- Subjects
Pathology ,medicine.medical_specialty ,Lung Neoplasms ,Melanoma, Experimental ,Pharmaceutical Science ,Antineoplastic Agents ,Pharmacology ,Lung pseudo-metastases ,NSCLC ,Mice ,In vivo ,Cell Line, Tumor ,Administration, Inhalation ,medicine ,Temozolomide ,Distribution (pharmacology) ,Animals ,Particle Size ,Lung cancer ,Fluorescent microparticles ,Chromatography, High Pressure Liquid ,Fluorescent Dyes ,Aerosol therapy ,Inhalation ,business.industry ,Melanoma ,Cancer ,Reproducibility of Results ,Sciences bio-médicales et agricoles ,medicine.disease ,Dacarbazine ,Drug delivery ,Female ,Mouse B16F10 melanoma ,business ,medicine.drug - Abstract
The aim of this study was to compare the efficacy of local drug delivery by inhalation to intravenous delivery in a B16F10 melanoma metastatic lung model. Temozolomide was formulated as a suspension, which was elaborated and evaluated in terms of particle size, shape and agglomeration. An endotracheal administration device was used to aerosolise the suspension. This mode of delivery was evaluated at different temozolomide concentrations and was optimized for the uniformity of delivered dose, the droplet size distribution and the distribution of droplets in vivo. Of the particles in the stabilised suspension, 79% were compatible with the human respirable size range, and this formulation retained 100% in vitro anticancer activity as compared to temozolomide alone in three distinct cancer cell lines. The pulmonary delivery device provided good reproducibility in terms of both the dose delivered and the droplet size distribution. Most of the lung tissues that were exposed to aerosol droplets contained the particles, as revealed by fluorescent microscopy techniques. The global in vivo antitumour activity of the inhaled temozolomide provided a median survival period similar to that for intravenous temozolomide delivery, and three out of 27 mice (11%) survived with almost complete eradication of the lung tumours. The present study thus shows that inhalation of a simple liquid formulation is well tolerated and active against a very biologically aggressive mouse melanoma pulmonary pseudo-metastatic model. This inhalation delivery could be used to deliver other types of anticancer drugs., JOURNAL ARTICLE, info:eu-repo/semantics/published
- Published
- 2010
8. Automated tracking of unmarked cells migrating in three-dimensional matrices applied to anti-cancer drug screening
- Author
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Nadine Warzée, Christine Decaestecker, Olivier Debeir, Robert Kiss, Ivan Adanja, and Véronique Megalizzi
- Subjects
Cytochalasin D ,Template matching ,Cell Culture Techniques ,Reproducibility of Results ,Context (language use) ,Cell migration ,Cell Biology ,Biology ,Tracking (particle physics) ,Automation ,Transformation (function) ,Robustness (computer science) ,Cell Movement ,Cell Line, Tumor ,Cancer cell ,Image Processing, Computer-Assisted ,Preprocessor ,Humans ,Microscopy, Phase-Contrast ,Collagen ,Drug Screening Assays, Antitumor ,Biological system ,Cell Migration Assays ,Gels ,Algorithms - Abstract
In oncology, combating the spread of tumor cells is a clinical need which currently remains unsatisfied. Identifying anti-migratory compounds usually requires in vitro screening of a large number of molecules. Efficient and realistic (i.e., preferably 3D) in vitro tests are thus required in order to quantify the anti-migratory effects of anti-cancer drugs. To remain compatible with high-throughput screening, we focus on assays where unlabeled cells are migrating in 3D transparent gels and are observed under time-lapse 3D phase-contrast microscopy. In this context, we present a method for automatically tracking cells that combines a template matching preprocessing step with a mean-shift process. The preprocessing step consists in performing a correlation of a cell template with each observed volume in order to provide a phase-contrast artifact-free volume where the cells appear as correlation peaks surrounded by smooth gradients. This transformation enables the cells to be efficiently tracked by a mean-shift process. Robustness and efficiency of this approach are qualitatively and quantitatively shown in various experiments. Finally, we successfully applied our method to the quantitative characterization of the anti-migratory impact of cytochalasin-D on cancer cells. In conclusion, our method can efficiently be used for drug screening aiming to evidence drug-induced effects on cell migration in 3D transparent environments, such as matrix gels.
- Published
- 2009
9. Phase contrast image segmentation by weak watershed transform assembly
- Author
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Nadine Warzée, Christine Decaestecker, Olivier Debeir, Ivan Adanja, and P. Van Ham
- Subjects
Watershed ,Pixel ,business.industry ,Segmentation-based object categorization ,Computer science ,Scale-space segmentation ,Pattern recognition ,Image segmentation ,Mathematical morphology ,Thresholding ,Computer vision ,Segmentation ,Artificial intelligence ,business - Abstract
We present here a method giving a robust segmentation for in vitro cells observed under standard phase-contrast microscopy. We tackle the problem using the watershed transform. Watershed transform is known for its ability to generate closed contours and its extreme sensitivity to image borders. One main drawback of this method is over- segmentation. In order to circumvent this, marked watershed based on the "modified gradient" method has been developed. However, the choice of the watershed mark locations is critical and their inadequacy may cause wrong results. Similarly to randomization and combination procedures used in the machine learning field, the present paper promotes the use of an assembly of marked watershed transforms, in order to increase the segmentation robustness. This results in the definition of candidate segmentations margins (expressed in terms of object border confidence) from which final segmentation can be chosen by means of thresholding.
- Published
- 2008
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