Fiche Projet - Ecole Doctorale Complexité du vivant

Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS
Nom et prénom du directeur de thèse (et si besoin du co-directeur) : Weil Robert
Le directeur de thèse et le co-directeur doivent impérativement être habilités à diriger les recherches (HDR)
Coordonnées Laboratoire de Signalisation et Pathogenèse, UMR 3691-CNRS Institut Pasteur
25 rue du Docteur Roux, 75724 Paris Cedex 15
Tel : 01-40-61-30-38
e-mail : [email protected]
Nom et prénom du co-encadrant (non HdR ) (s’il y a lieu) : Pierre Génin
Coordonnées Laboratoire de Signalisation et Pathogenèse, UMR 3691-CNRS Institut Pasteur
25 rue du Docteur Roux, 75724 Paris Cedex 15
Tel : 01-40-61-30-38
e-mail : [email protected]
Nom et prénom du responsable de l’équipe : Robert Weil (équipe 1), Génin Pierre (équipe 2)
Nombre de chercheurs et enseignants-chercheurs statutaires de l’équipe titulaires d’une HDR : 2
Nom et prénom du responsable du laboratoire : Weil Robert
Intitulé du laboratoire et N° d’unité : Signalisation et Pathogenèse – UMR 3691
Spécialité : Immunologie, Biologie Moléculaire, Biologie Cellulaire
Titre du projet de thèse :
Project 1 : E-Syt2 dysfunction in lymphoproliferative diseases, a therapeutic target ?
Dysfonctionnement d’E-Syt2 dans les maladies lymphoprolifératives, une cible thérapeutique ?
Direction : Robert Weil
Project 2 : Regulation of the mitophagic function by the protein Optineurin
Régulation de la fonction mitophagique par la protéine Optineurine
Direction : Robert Weil, Responsable : Pierre Génin
Résumé du projet de thèse (1 page maximum, en anglais)
Project 1 : E-Syt2 dysfunction in lymphoproliferative diseases, a therapeutic target ?
My laboratory is working mainly on the mechanism of activation of NF-B transcription factor, a master regulator of T cell
responses and perturbations of its activity are linked to immune and inflammatory diseases. An important activator of the
TCR-induced NF-B pathway is the CBM complex, composed of CARMA1, BCL10 and MALT1, that promotes the
activation of downstream signaling. The localization of the CBM complex at the immune synapse (IS), between the Antigen
Presenting Cell (APC) and the T cell, is critical for the initiation of the immune response. We identified, Extended
Synaptotagmin-Like Protein 2 (E-Syt2) as a CARMA1-interacting protein essential for positioning signaling effectors at the
IS. To further assess the physiological importance of this protein, we developed the classical knockout of E-Syt2 gene and
its conditional knockout version in the thymus of mice (manuscript submitted). We observed that E-Syt2-deficient mice
present defects in T lymphocytes homeostasis, impaired TCR-mediated cytokine production, NFsurvival. In addition, these mice develop severe age-dependent inflammatory skin disorder and splenic defects.
Since loss- and gain-of-function mutations within the CBM complex have been associated respectively with severe human
combined immunodeficiencies (SCID) and diffuse large B-cell lymphomas (DLBCL-ABC), we seek to determine whether
E-Syt2 contributes to the survival of these lymphomas using retroviruses to express E-Syt2 shRNA in ABC-DLBCL cell
lines. We will also set up a high-throughput screening (HTS) inhibition assay to identify drugs able to dissociate the
interaction between CARMA1 and E-Syt2. Their effectiveness on DLBCL will be tested by biochemical and imaging
techniques. The aim of this project is also to search for loss-of-function mutations of E-Syt2 gene responsible for human
immunodeficiencies in a cohort of immunodeficient patients
Project 2 : Regulation of the mitophagic function by the protein Optineurin
Optineurin (Optn), homolog of NEMO, the central regulator of the NF-B pathway, is an extremely versatile protein
involved in distinct cellular processes such as Trans-Golgi network organization, membrane secretion, antiviral immune
response and bacterial-induced autophagy (1). Optn gene has been linked to diverse pathological processes such as open
angle glaucoma, amyotrophic lateral sclerosis and Paget’s disease of bone, although the links between Optn functions and
associated pathologies have not yet been discovered. Recently, Optn was found to participate to the mitophagic process –
i.e. degradation of damaged mitochondria – by linking the autophagosomal marker LC3 to the damaged mitochondria.
However, the mechanism by which Optn exerts this function is still unclear and especially the kinase and phosphorylation
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Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS
events that control Optn-LC3 interaction in this case are unknown. In our group, it was previously shown that the protein
kinase D (PKD) or also called PKC mu, specifically interacts with and phosphorylates a specific residue of Optn (serine
342) when activated. As PKD is activated in response to oxidative stress, a signal for mitophagy, we sought to determine
whether phosphorylation of Optn by PKD could constitute the missing link in the mitophagic process. In addition,
mitophagy is considered as a tumor suppressor mechanism through elimination of damaged mitochondria, thereby limiting
the tumor-promoting effects of Reactive Oxygen Species accumulation by oxidative stress. Since decreased mitophagy has
been widely observed in human malignancies (4) and as Optn deficiency is associated to lung cancer development (5), we
also sought to assess the association between cancer and downregulation of the PKD-Optn axis using tumorigenic lung
epithelial and breast lineages as a cellular model. This project should therefore reveal a new association between human
malignancies and deficiency of the PKD-Optn axis and lead to a model of tumor-suppression by this axis based on the
mitophagy induction events.
Bibliography:
1. Kachaner, D., Genin, P., Laplantine, E. & Weil, R. (2012). Toward an integrative view of Optineurin functions. Cell
Cycle 11, 2808-2818.
2. LaValle CR, George KM, Sharlow ER, Lazo JS, Wipf P, et al. (2010) Protein kinase D as a potential new target for
cancer therapy. Biochimica et biophysica acta 1806: 183-192.
3. Liu Z, Chen P, Gao H, Gu Y, Yang J, et al. (2014) Ubiquitylation of autophagy receptor Optineurin by HACE1 activates
selective autophagy for tumor suppression. Cancer cell 26: 106-120.
Thèses actuellement en cours dans l’équipe
Nom et Prénom du doctorant
Nom du directeur de thèse
Année de 1ere
inscription et
Ecole Doctorale
Financement pendant la thèse
Trois publications récentes du directeur de thèse (du co-directeur ou du co-encadrant s’il y a lieu).Mettre en gras le nom
du directeur de thèse.
N. Messali, H. Soares, N. Satoh, A. Trivellas, A. Alcover, J. P. Di Santo, G. Jouvion, F. Langa-Vives and R. Weil. (2016) E-Syt2 is
essential for TCR-mediated NF-B signaling and immune responses. Manuscript submitted for publication.
Génin P, Cuvelier F, Lambin S, Côrte-Real Filipe J, Autrusseau E, Laurent C, Laplantine E, Weil R. (2015) Optineurin regulates the
interferon response in a cell cycle-dependent manner. PLoS Pathogens 11, e1004877.
Messali N., Génin P., Weil R. (2013) Dysregulation of the antigen-induced NF-κB signaling pathway in the development of human Bcells lymphomas. Journal of Leukemia 1: 105.
Docteurs encadrés par le directeur de thèse ayant soutenu après septembre 2010 et publications relatives à leur sujet de thèse.
Mettre en gras le nom du directeur de thèse et celui du docteur.
Nom Prénom : Kachaner David
Date de soutenance : 24 Septembre 2012
Durée de thèse (en mois): 38
Ecole Doctorale : Complexité du vivant
Publications :
D. Kachaner, J. Filipe, E. Laplantine, A. Bauch, K. L. Bennett, G. Superti-Furga, A. Israël, and R. Weil. Plk1dependent phosphorylation of Optineurin provides a negative feedback mechanism for mitotic progression (2012) Mol.
Cell, 45: 553-566.
D. Kachaner, E. Laplantine, P. Génin and R. Weil. Optineurin: a new vision on the regulation of cell division (2012)
Cell cycle, 11:1481-1482.
D. Kachaner, P. Génin, E. Laplantine and R. Weil. Towards an integrative view of Optineurin functions (2012) Cell
cycle, 11:2808-2818.
Nom Prénom : Messali Nassima (co-encadrée)
Date de soutenance : 24 Septembre 2014
Durée de thèse (en mois): 48
Ecole Doctorale : Biologie et Biotechnologie
Publications :
N. Messali, P. Génin and R. Weil. Deregulated antigen-induced NF-κB signalling pathway in the development of
human B-cells lymphomas (2013) Journal of leukemia, 1, 105.
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