Microscopy of living cancer cells at physiological oxygen levels: the MICROX platform

Description du projet

Microscopie à des niveaux d’oxygène physiologiques

La recherche et le développement de médicaments remplacent les tests de lecture de plaques standard par un dépistage par microscopie. Combiné à des tests fonctionnels innovants, il est possible de surveiller les événements dynamiques dans les cellules vivantes. Le MICROX, financé par l’UE, travaillera sur des dosages microscopiques semblables aux niveaux réels d’oxygène rencontrés par nos cellules. Les protocoles conventionnels maintiennent les cellules à 21 % d’oxygène, alors que physiologiquement la plupart des cellules sont exposées à 2 à 5 % d’oxygène et les cellules cancéreuses à des conditions hypoxiques. La plateforme de microscopie MICROX résultante permettra d’ajuster les conditions atmosphériques en fonction du type de cellule et, surtout, facilitera la découverte et le criblage de médicaments anticancéreux.

Objectif

Functional evaluation of bioactive compounds using cell-based assays is key in discovering new and improved drugs to address our societies growing medical needs. In a large number of academic and private R&D facilities around the world, hi-content microscopy screening is used to complement and often outperform the conventional luminescence and fluorescence plate-reader assays. The combination of fluorescent-probe technology, modern optical microscopes and innovative functional assays allows monitoring highly dynamic events in living cells with exquisite temporal and spatial resolution.
Remarkably, drug candidates and treatment regimens are commonly tested on living cells maintained at atmospheric oxygenation levels (i.e. at 21% O2) while in reality, cells in our bodies never experience such high oxygen levels. Rather, most cells experience 2-5% of O2 and cancer cells in solid tumours are generally hypoxic, i.e. they function at < 1% of O2. It has become evident that oxygenation of the preparation dramatically affects efficacy of our drugs, in particular of cancer treatments. Thus, there is an urgent need to establish functional microscopy assays for (cancer)drug efficacy at hypoxic conditions.
However, it is not trivial to enclose the necessary high-end microscopes in O2-tight incubators while maintaining good access to the cells. In this project, experienced biochemist and pharmacologist dr. Olga Mazina aims to team up with the advanced microscopy group of prof. Kees Jalink to establish an innovative MICROX microscopy platform with fully adjustable atmospheric conditions, along with a panel of biosensors optimized for use at hypoxia. MICROX will be developed in close collaboration with leading European manufacturers and validated in studies addressing cancer cell signalling and sensitivity to cancer drugs at various O2 levels.