6533b85dfe1ef96bd12bdb66

RESEARCH PRODUCT

L'hyperthermie provoque l'agrégation de FLIP et restaure l'apoptose induite par TRAIL

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The TNF-family member TRAIL (TNF-related apoptosis inducing ligand) is a cytokine involved in the immune anti-tumour surveillance. TRAIL is a promising agent currently under investigation for its anti-cancer properties with limited side effects on healthy cells. However, the use of TRAIL in oncology has been limited due to its lack of efficiency, mainly associated with cell resistance to apoptosis. The aim of this project was to study the interest of hyperthermia (or heat shock - HS) as an adjuvant for TRAIL therapy and the mechanisms involved in this sensitization.We have first evaluated the significance of this combination in a large variety of cancer cell lines known to be sensitive or resistant to TRAIL. We could demonstrate that hyperthermia was able to efficiently sensitize resistant cancer cells to TRAIl-induced apoptosis in almost every cell lines tested.We next, focused our work on the molecular mechanisms responsible for the sensitization, during hyperthermia. Analyses of the DISC (Death-Inducing Signaling Complex) revealed a lack of recruitment of FLIP in the DISC, the main inhibitor of the extrinsic pathway, and a delay in the formation of the complex under hyperthermic conditions. Inhibition of FLIP recruitment was associated with enhanced initiator caspases activation when cells were reincubated at 37°C after the HS.The absence of FLIP within the TRAIL DISC was due to its aggregation during HS and was independent of post-translational modifications. Inhibition of FLIP aggregation by glycerol, which stabilizes denaturerated proteins, restored FLIP recruitment within the TRAIL DISC and consequently inhibited TRAIL-induced cell death. Taken together, these results highlight the interest of combining TRAIL with hyperthermia and highlight new mechanisms explaining its efficiency.