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tel.:+32 2 650 22 95
fax:+32 2 650 22 98

ULB - SLN CP165/57
50, Av. F.Roosevelt
1050 Bruxelles


Development of methods and tools for image analysis and automatic feature extraction in phase-contrast cell images (project page:Cellular imaging). Specifically, the goal of my research is to develop methods for 3D cell tracking and morphological features extraction.



N. D'Haene, S. Sauvage, C. Maris, I. Adanja, M. Le Mercier, C. Decaestecker, L. Baum, I. Salmon,
VEGFR1 and VEGFR2 Involvement in Extracellular Galectin-1- and Galectin-3-Induced Angiogenesis
PloS one, Vol. 8, 6, 2013
Bibtex : info:hdl:2013/148927
Note : SCOPUS: ar.j
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.

I. Adanja, V. Megalizzi, O. Debeir, C. Decaestecker,
A New Method to Address Unmet Needs for Extracting Individual Cell Migration Features from a Large Number of Cells Embedded in 3D Volumes
PloS one, Vol. 6, 7, 2011
Bibtex : info:hdl:2013/94206
Note : SCOPUS: ar.j
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. © 2011 Adanja et al.

I. Adanja, O. Debeir, V. Megalizzi, R. Kiss, N. Warzée, C. Decaestecker,
Automated tracking of unmarked cells migrating in three-dimensional matrices applied to anti-cancer drug screening.
Experimental cell research, Vol. 316, 2, pp. 181-193, 2010
Bibtex : info:hdl:2013/52535
Note : Journal Article
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.

N. Wauthoz, P. Deleuze, J. Hecq, I. Roland, S. Saussez, I. Adanja, O. Debeir, C. Decaestecker, V. Mathieu, R. Kiss, K. Amighi,
In vivo assessment of temozolomide local delivery for lung cancer inhalation therapy.
European journal of pharmaceutical sciences, Vol. 39, pp. 402-411, 2010
Bibtex : info:hdl:2013/51846
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.


O. Debeir, I. Adanja, N. Warzée, P. Van Ham, C. Decaestecker,
Phase contrast image segmentation by weak watershed transform assembly
Proceedings of 5th IEEE International Symposium on Biomedical Imaging : From Nano to Macro (ISBI), pp. 724-727, ISBI)(Paris, France, 2008
Bibtex : info:hdl:2013/53020
Note : Language of publication: en

O. Debeir, R. Van Ginckel, I. Adanja, F. Darro, R. Kiss, C. Decaestecker,
High throughput characterization of anticancer compounds by means of cellular imaging and automatic image analysis.
Conference: (April 14-18: Los Angeles - USA), 2007
Bibtex : info:hdl:2013/71810
Note : Présentation orale (abstract 4965)

O. Debeir, I. Adanja, R. Kiss, C. Decaestecker,
Automatic in vitro cell division characterization by automatic image sequence analysis
Conference: Annual Symposium of the IEEE/EMBS Benelux Chapter(I: 7-8 December 2006: Brussels), 2006
Bibtex : info:hdl:2013/158307
Note : Language of publication: en
Abstract : In vitro cell observation has been widely used by biologists for years and covers a wide range of applications: cancer and cell proliferation, cell migration or chemotactism, embryology, drug testing. Nowadays the use of computer assisted-microscopy (or time-laps microscopy) allows the handling of considerably large amount of image data, which is acquired during experiments possibly lasting from several hours to several days. If cell cultures are observed for long periods of time (2-4 days), it becomes possible to detect less frequent cell events, such as cell division or cell death. We describe in this work a semi-automatic approach of studying the cell division event itself - starting from the cell exhibiting a round shaped form to the final daughter cells separation.

Books chapters

O. Debeir, I. Adanja, R. Kiss, C. Decaestecker,
Models of cancer cell migration and cellular imaging and analysis
Anja Lambrechts, Christophe Ampe (Eds). The Motile Actin System in Health and Disease, pp. 123-156, 2008
Bibtex : info:hdl:2013/99600
Note : Language of publication: en