Engineering and Imaging multicellular tumour spheroids to study 3D cell proliferation – SPH-IM-3D

Adaptive optics
Phoromanipulation
Signal treatment and analysis

The results obtained relate to the introduction of the instruments and imaging techniques sheet of light on the development of a new wavefront analyzer , the appropriate biological engineering models , and the development of mathematical tools to analyze and use the data obtained .
The engineering spheroids expressing biosensors or fluorescent markers allows us to perform imaging studies of dynamic events at the cellular level occurring at depth. These studies are made possible by combining the skills of partners , particularly those mathematicians who develop software tools specifically adapted . Important results have already been obtained and . They focus on the characterization of the effects of stress on the development of the various phases of mitosis. For example, we demonstrated the impact of mechanical stress on the progression in prometaphase and involvement of actomyosin cytoskeleton in this response.

This work opens up many opportunities, particularly for the use of models and technologies developed in the field of identification of new therapeutic targets and evaluation of the effectiveness of anti-tumor agents.

After a first phase mainly dedicated to the development of tools, instruments and biological models, the project begins to enter an operational phase and recovery.
This translates communications in many specialized seminars, both in imaging, cancer biology, in applied mathematics.
Several papers are in preparation for next submission in international journals with referees. An example of such work is presented below. It is dedicated to study the characterization of the progress of the division in different situations and impact on tumor growth. This study would not have been possible without the technical and mathematical developments made ??within the SPH-IM-3D consortium.

Engineering and imaging multicellular tumour spheroids to study 3D cell proliferation.
SPH-IM-3D stands for SPHeroids and IMaging in 3D, that are the three keywords associated with this project aiming at developing imaging approaches and at engineering original multicellular tumor spheroid models to explore cancer cell proliferation dynamics in 3D.
This project is organized into three complementary objectives. First, we will aim at the development of a new 3D imaging technology, the SPIM (Single Plane Illumination Microscope), a non-commercial device which there are only a few prototypes in the world. We will conduct innovative developments on the use of adaptive optics, on the implementation of photomanipulation devices, and on signal and image processing. In parallel, we will develop new and original models of 3D growth in which cellular tumor heterogeneity will be considered. Multicellular tumor spheroids (MCTS) will be genetically engineered to express fluorescent biosensors and markers specific to a function or a parameter of interest. Finally, we will demonstrate that these complementary technological and biological innovative tools allow monitoring the exploration of the biology of cell division and tumor proliferation in integrated 3D models, giving much attention to the spatial regulation of mitosis and the response to genotoxic stress. This work opens up various perspectives, in particularly in the use of the models and technologies developed in this project for the identification of new putative therapeutic targets and for the assessment of anti-tumor efficacy of new compounds.
This project is supported by two academic groups with complementary skills and expertise in cancer biology, cell imaging, optics and mathematics and by a SME specialized in adaptive optics. It is coordinated by the imaging group of the newly created multidisciplinary Institute for Advanced Technologies in the Life Sciences (ITAV – Centre Pierre Poitier).