Informace o projektu

Anotace je dostupná pouze v anglickém jazyce.

Fibroblast growth factor receptor (FGFR)/ERK MAP kinase signaling pathway is often deregulated in human disperse and therefore constitutes a major target for therapeutic intervention. While several small compound inhibitors exist to target FGFR/ERK signaling, they show only a limited use in clinic, warranting development of novel treatment strategies. Cell-based high-throughput screening (HTS) of chemical libraries represents one effective approach for identification of novel compounds affecting the signaling pathways. Compared to biochemical assays based on the protein-protein or protein-chemical interaction, the cell-based approach offers an advantage to identify modulators of entire pathway in intact cell invironment, complete with regulatory networks and feedback control mechanisms. The major disadvantage of the assays based on cell growth is high false-positive rate, i.e. toxic compounds as well as those targeting other pro-growth pathways will be picked up. We propose to overcome the latter by utilizing cell growth-arrest instead of proliferation as a reporter for HTS screening. In our previous work we characterized, in detail, the molecular features of FGFR/ERK-mediated growth arrest in rat chondrosarcoma (RCS) cells, including the proof-of-concept experiments demonstrating the potential of RCS growth-arrest assay for discovery of novel FGFR/ERK inhibitors. Here, we propose to develop and successfully apply the RCS growth-arrest assay for image-based screening of large chemical libraries aimed on discovery and further characterization of novel, potentially therapeutic inhibitors of pathological FGFR/ERK signaling. Subsequently, we will combine modern methods of molecular biology, genetics and biological approaches to identify the targeted proteins and characterize the molecular mechanism of inhibition. Finally, we will combine several in vivo approaches to determine the basic pharmacological properties of the lead compounds identified in screenings, and to validate their therapeutic potential in preclinical models to pathological FGFR/ERK signaling.