Description du sujet de thèse

Development of Fluorescent Biosensors of Kinases involved in inflammation for applications in juvenile arthritis

Contexte de travail

Protein Kinases (PKs) play important roles in promoting inflammation and immune response and PK dysregulation is well documented in inflammatory pathologies [8-13]. Moreover, several PKs constitute attractive pharmacological targets for therapeutic purposes, as exemplified by the Janus family of kinases (JAKs) targeted by Tofacitinib both in rheumatoid arthritis in adults and in JIA [9-10]. PKs involved in upregulation of inflammation and immune response signalling pathways include JAKs, Tyk2, IRAKs, GRKs and RIPK kinases, Syk, Src, NIK, Lck, ERK, Lyn and Hck [8-13]. However, despite the wealth of literature concerning these PKs and the arsenal of PK inhibitors available for clinical therapies or in development in the drug discovery pipeline [14], there are currently no diagnostic approaches based on detection and quantification of PK activities. Although the relative abundance of PKs can be determined through standard antigenic approaches such as Western blotting or ELISA, there is no direct correlation between PK levels and their functional activity. Development of technologies to profile PK activities that are dysregulated in inflammatory pathologies could provide a solid basis for development of predictive diagnostics, subsequently guiding towards personalized treatments.

This thesis project will aim to develop a toolbox of fluorescent peptide biosensors that report on the activity of PKs involved in inflammation, thus contributing to the development of a biosensing platform for profiling PK signatures involved in inflammation, in particular in patients with juvenile arthritis.

Specifically, the thesis project will consist of (1) developing and characterizing a family of specific peptide substrates for the PKs of interest (PKs of the JAK, GRK, IRAK, RIPK and ERK/MAPK families) (2) developing and optimizing fluorescent peptide biosensors from the identified substrates and characterizing the selectivity and sensitivity of their response to recombinant and endogenous PK activities, in cell extracts stimulated or treated with inhibitors (3) exploiting these fluorescent peptide biosensors to develop a platform for profiling several PK activities simultaneously in the synovial fluid of patients with arthritis.
This project is part of the research topics developed by Dr. M.C. Morris over the past ten years aiming to target and detection protein kinases thanks to fluorescent biosensor technologies.

Le poste se situe dans un secteur relevant de la protection du potentiel scientifique et technique (PPST), et nécessite donc, conformément à la réglementation, que votre arrivée soit autorisée par l'autorité compétente du MESR.

Contraintes et risques

none