Project aims:

• Design of MMP-2 and -9 covalent inhibitors. The covalent inhibitors will be conceived to target specific Lysine residues present within the S₃’ subsite of MMP2/9 and absent in the one of other MMPs. Their selectivity profile will be finely tuned by exploring several key parameters including i) binding affinity and selectivity of the recognition motif directed toward MMP catalytic cleft, ii) reactivity of the electrophilic warhead used to target Lysine, and iii) subtle orientation of this reactive motif within the S₃’ subsite of MMP.
• Test the effect of MMP-2/9 inhibitors in combination with irradiation in in vitro 3D GBM invasion models, comprising also tumour microenvironment.

Project will be implemented in two work packages.

WP1: Design, screening and selection of covalent inhibitors targeting specifically MMP-2 and MMP-9 (MMP-2/9), partner CEA

Task 1. Design, synthesis and characterization of a focused library

Task 2. In vitro evaluation of covalent inhibitors on a set of MMPs through enzyme assays enabling to determine their inhibitory/selectivity properties

Task 3. For the best candidates, optimization of their reactivity in complex proteomes, consisting in modulating their structure to limit their unspecific reactivity toward abundant proteins

WP2: Test the effect of MMP2/9 inhibitors and irradiation in in vitro GBM models of invasion and organoid invasion model, partner NIB

Task 4. Analyse protein levels and activity of MMP-2/-9 in GBM cell model lysates

Task 5. Determine cytotoxicity of MMP2/9 covalent inhibitors and irradiation in GBM cellular models

Task 6. Perform GBM cell invasion assays with MMP2/9 covalent inhibitors and irradiation

Task 7. Perform organoid invasion assay