Physiologie de la Reproduction et des Comportements, UMR 7247, équipe “Biologie et Bioinformatique des Systèmes de Signalisation” http://bios.tours.inra.fr

Topic of the thesis: Intracellular antibodies to explore the relationships between conformations

and activity of the FSH receptor, and applications in reverse pharmacology

G protein-coupled receptors (GPCRs) are membrane receptors involved in most physiological functions. The follicle-stimulating hormone receptor (FSHR) has been studied for many years by the host laboratory. Given its key role in reproduction, this receptor is the main target of medically-assisted therapies in human. In addition, molecules with FSH activity are injected to breeding animals to induce ovulation in synchronized females, prior to artificial insemination. As expected from studies on other GPCRs, it should be possible to “freeze” one or the other activated conformations of the FSHR with intracellular antibodies (intrabodies, Ibs), especially with single chain antibody fragments. Hence, the present thesis project consists in the identification and functional characterization of anti-activated FSHR Ibs. First, these Ibs will permit to address some exciting scientific questions in the field of GPCRs: they are expected i/ to correlate the dynamics of the FSHR activation to the dynamics of the signaling network engaged, ii/ to follow in real-time the FSHR intracellular trafficking, that is poorly known in gonadal cells, iii/ to address the question of endosomal signaling, as reported for other GPCRs recently, and iv/ to provide a precious tool for solving the crystal structure of the complete FSHR (in collaboration). In addition, in woman or in animals, administration of FSH leads to deleterious responses, hence there is a need for pharmacological substitutes to FSH action. This is why, in the second part of the thesis, the Ibs isolated in the first part will be used as tools for a reverse pharmacology approach, for screening ligands of FSHR active conformations of interest. A proof of concept in female rat will be attempted at the end of the research program. The PhD student is expected to have a background in cell biology and to exhibit interest in cell signaling. He/she will benefit of an interdisciplinary scientific environment of biologists, bioinformaticians and biomathematicians. He/she will be educated to many different methodologies that are routinely used in our group: phage display, epitope mapping, pharmacological characterization, functional assays for cell signaling, physiology of the reproductive track. Expected start: October 2019 Contact : Pascale Crépieux, DR CNRS, Co-responsable de l'équipe BIOS Physiologie de la Reproduction et des Comportements, UMR 7247 INRA- CNRS- Univ. Tours Centre de Recherches Val de Loire, 37380 Nouzilly, France Pascale.Crepieux@inra.fr tél: 33 2 47 42 75 14

http://bios.tours.inra.fr/