Date : 06/04/2010

Laboratory
Institut Cochin
INSERM U1016, CNRS UMR8104,
Université Paris Descartes
22 rue Méchain 75014 Paris
Director : Pierre-Olivier Couraud

Team
Insulin signaling, glucose sensing and glucotoxicity

PhD Supervisors
Tarik ISSAD / Anne-Françoise BURNOL
email : Cet e-mail est protégé contre les robots collecteurs de mails, votre navigateur doit accepter le Javascript pour le voir
phone : +33 1 40 51 64 09

Subjects / Tools-Methodologies

1 : Insulin signaling/Protein-protein interactions - BRET methodology
2 : Structural bioinformatics/Chemoinformatics - Virtual screening
3 : Pathophysiology of insulin action/animal models of diabetes and obesity

Summary of lab's interests

Groups involved:
Tarik ISSAD (BRET, Insulin signaling)
Anne-Françoise BURNOL (Pathophysiology of insulin action)
Bruno VILLOUTREIX (Bioinformatics, Chemoinformatics)

The group of Tarik Issad has a long standing experience in insulin signaling and its regulation in normal and pathological conditions. This lab has developed a recognized expertise on the BRET technique for the study of therapeutic targets, particularly in the field of insulin and IGF1 receptors.
The group of Anne-Françoise Burnol has cloned and studied the inhibitor of insulin signaling Grb14. This lab has a recognized expertise in the use of animal models for the study of insulin action.
The group of Bruno Villoutreix (Inserm-Paris Diderot UMR-S 973) has been working in the field of structural bioinformatics, chemoinformatics and drug design for many years, both in term of methodological developments and applications to relevant therapeutic targets. The key axes in this group with regard to drug design are around in silico prediction of ADME/Tox properties and rational analysis of the chemical space to identify protein-protein interaction inhibitors.

Summary of project

Insulin binding to its receptor (IR) stimulates its tyrosine kinase activity, which in turns activates intracellular signalling pathway. Grb14 is an adaptor that binds to and inhibits the tyrosine kinase activity of the IR. Its expression is increased in some tissues of type 2 diabetic patients. Therefore, small molecule inhibitors of the IR-Grb14 interaction may help improve insulin sensitivity in diabetic patients. In a previous work, using structured-based virtual screening, we have selected 1000 molecules (out of a compound collection containing over 300000 molecules) with high-score for potential inhibition of the IR-Grb14 interaction. These compounds were then purchased and experimentally evaluated for inhibition of IR-Grb14 interaction using an in vitro BRET assay. Several inhibitors were detected, among which one compound (C8) appears to be particularly efficient, both in BRET and co-immunoprecipitation experiments.
The aim of the present project will be:
1. to further characterize the inhibitory activity of C8 on IR-Grb14 interaction, using BRET as well as biochemical techniques
2. to evaluate the effect of C8 on insulin signalling (PI-3 kinase, Erk Kinase) and insulin biological effects (glucose uptake, cell proliferation and survival) in cultured cells and in animals
3. to develop, on the basis of C8 structure, optimized inhibitors of IR-Grb14 interaction using ligand-based and structure-based virtual docking, as well as other chemoinformatic tools (e.g., QSAR, ADME/Tox prediction...)
4. to study the inhibitory effect of new C8-derived molecules on BRET, on cell signalling in cultured cells and on insulin sensitivity in animal models. The strength of the project relies on the complementarities of the three participating groups, including bioinformatics, chemoinformatics, biophysical techniques and physiological approaches. This multidisciplinary approach has already proven to be fruitful, as shown by the discovery of C8 compound. The use of C8 as a lead compound should now allow developing new molecules with high therapeutic interest for the treatment of diabetes.

Publications :
Goenaga, D., Hampe, C., Carré, N., Cailliau, K., Browaeys-Poly, E., Perdereau, D., Holt, L.J., Daly, R.J., Girard, J., Broutin, I., Issad, T., Burnol, A-F. Molecular determinants of Grb14-mediated inhibition of insulin signaling (2009) Mol. Endocrinol. 23(7): 1043ñ1051

Dual effect of the adapter growth factor receptor-bound protein 14 (grb14) on insulin action in primary hepatocytes.CarrÈ N, Ca‚zac M, Girard J, Burnol AF. Endocrinology. 2008 149(6):3109-17.

Nouaille S, Blanquart C, Zilberfarb V, Boute N, Perdereau D, Roix J, Burnol AF, Issad T. (2006) Interaction with Grb14 results in site-specific regulation of tyrosine phosphorylation of the insulin receptor. EMBO Reports. 7: 512-518.

Nouaille S, Blanquart C, Zilberfarb V, Boute N, Perdereau D, Burnol AF, Issad T. Interaction between the insulin receptor and Grb14: A dynamic study in living cells using BRET. (2006) Biochem Pharmacol. 72:1355-1366

Issad, T., Blanquart, C. and Gonzalez-Yanes, C. The use of Bioluminescence Resonance Energy Transfer (BRET) for the study of therapeutic targets: application to tyrosine kinase receptors (2007) Expert. Opin. Ther. Targets 11: 541-556

Reynès C, Host H, Camproux AC, Laconde G, Leroux F, Mazars A, Deprez B, Fahraeus R, Villoutreix BO, Sperandio O. Designing focused chemical libraries enriched in protein-protein interaction inhibitors using machine-learning methods. PLoS Comput Biol. 2010 Mar 5;6(3):e1000695.

Sperandio O, Reynès CH, Camproux AC, Villoutreix BO. Rationalizing the chemical space of protein-protein interaction inhibitors. Drug Discov Today. 2010 Mar;15(5-6):220-9.

Basse N, Montes M, Maréchal X, Qin L, Bouvier-Durand M, Genin E, Vidal J, Villoutreix BO, Reboud-Ravaux M. Novel organic proteasome inhibitors identified by virtual and in vitro screening. J Med Chem. 2010 Jan 14;53(1):509-13.

Lagorce D, Sperandio O, Galons H, Miteva MA, Villoutreix BO. FAF-Drugs2: free ADME/tox filtering tool to assist drug discovery and chemical biology projects. BMC Bioinformatics. 2008 Sep 24;9:396.

Sperandio O, Mouawad L, Pinto E, Villoutreix BO, Perahia D, Miteva MA. How to choose relevant multiple receptor conformations for virtual screening: a test case of Cdk2 and normal mode analysis. Eur Biophys J. 2010 Mar 18.