Scientific contact information: Ignacio Anegon or Arnaud Nicot

Transcription

1 Transplantation and Immunology Nantes, France "Progreffe 2012" Inserm UMRS 1064/ITUN [ in Nantes (France) is launching its International doctoral program by opening a competitive call for one PhD fellowship starting in October This Program financed by the Progreffe Foundation [ aims to recruit a highly motivated and successful student of any nationality with a Master 2 degree. The PhD position will cover 3 years of salary for research activity (~16,000 tax-free/year) and will provide training in a strong research environment and participation to local seminars and international meetings. Applicants are asked to choose one project from the ones proposed by each team (see in the attached document). Pre-selected candidates will be interviewed on site or by conference call for final selection. The academic degree of the applicant should meet the requirements of the University of Nantes to be engaged in a PhD program (Master degree "M2" or equivalent required & obtained at the time of recruitment) [ defaultstructureksup]. Thus, the call is open immediately till May 1st, The selected candidate will be informed in July. Scientific Research at the ITUN Institute is organized in 5 research teams, with strong interfaces with clinical departments. Topics span many areas of research related to transplantation and autoimmune diseases, including Molecular & Cell Biology, Immunology, Neuroimmunology, Infectious Diseases, Regenerative Medicine Stem Cells, and Gene Therapy. Located within the Nantes University Hospital, the ITUN includes the research unit 1064 from INSERM (French National Institute for Health and Biomedical Research) and the clinical unit that performs among the highest number of kidney and pancreas transplantations in Europe. The unit 1064 comprises laboratories and animal facilities (~3,000 m 2 ) staffed by ~150 people and houses several core facilities (Q-PCR, high-speed cell sorting, microsurgery, protein and antibody purification, rat transgenesis, lentiviral vector production). The institute is part of a local research bringing together other INSERM units with more than 750 faculty staff and several core facilities (e.g. confocal imaging, microarrays, production of viral gene vectors). The city of Nantes offers an active cultural environment, and is conveniently located within 45 min drive (or train) from beautiful sea resorts and 2 hours train from Paris. For foreigners, the association «Chercheurs Etrangers à Nantes» will provide help to settle down in Nantes. The application should be sent electronically to Valerie.Pecqueret@univnantes.fr (with "Progreffe 2012" in subject) and should include: a letter (1-2 pages) with 1) a short description of achievements and career planning, and 2) your specific interest, motivation and qualifications for the project in question. a CV including lab experience, personal contact information & two references with and phone number proof of engagement in a Master degree Scientific contact information: Ignacio Anegon (ianegon@nantes.inserm.fr) or Arnaud Nicot (arnaud.nicot@inserm.fr) PhD reasearch projects proposed by Team 1: Dr E. Chiffoleau; Team 2: Drs L. Caron & I. Anegon; Team 3: Drs B. Vanhove & G. Blancho; Team 4: Dr N. Degauque; Team 5: Dr L. Le Berre.

2 Team 1 PhD project: Role of CLEC-1 in dendritic cells Supervisor: Dr E. Chiffoleau The aim of the project is to decipher the function of CLEC-1, a C-type lectin-like receptor, that we identified as being over-expressed in an experimental model of allograft tolerance (1,2). Clecl-1 belongs to a subfamily of lectin-like receptor genes located in the NK gene complex that encodes numerous receptors with important immune functions in monocytes, dendritic (DC) or endothelial cells (EC). CLEC-1 is an orphan receptor that is expressed by DCs and ECs. We have recently showed that CLEC-1 expression in rat DCs is increased by CD4 + CD25 + regulatory T cells and that CLEC-1 inhibits effector CD4 + T cell activation in vitro (3). These results suggest that CLEC-1 may act as an inhibitory receptor in DCs, in the activated T cells (expressing presumably the ligand) or in both. The expression and the role of CLEC-1 in DCs will be further explored by using innovative tools and relevant in vitro and in vivo assays. We will study the regulation of CLEC-1 expression in rat and human DCs and ECs with newly developped antibodies. Moreover, to block in vivo and in vitro interaction of CLEC-1 with its ligand, we have generated a fusion protein containing the extracellular domain of rat CLEC-1 and a mutated murine immunoglobulin Fc fragment. This fusion protein will also allow us to generate an antibody directed against the extra-cellular domain of CLEC-1 that will serve to analyze the role of CLEC-1 in immunity. In addition, we will use gene silencing in DCs using stealth sirnas to study the role of CLEC-1 in T cell differentiation of allogeneic or transgenic CD4 + T cells in vitro. In parallel, we will test the direct effect of the recombinant CLEC-1 protein on CD4 + T cell activation and differentiation. Moreover, we will identify the CLEC-1 interacting partners that co-immunoprecipitate with anti-clec-1 antibodies. Importantly, CLEC-1 KO rats, currently under generation in our laboratory, will be used to determine the role of CLEC-1 in different cell subset development and function, in transplantation and in various immune responses. The lab has all the expertises to develop these approaches during this 3-year project. In the long-term, CLEC-1 may represent an important therapeutic tool to modulate the immune response in transplantation, auto-immunity or cancer. 1. Heslan J.M. et al. (2006) New evidence for a role of allograft accommodation in long-term tolerance. Transplantation 82: Thebault P. et al. (2007) Role of IFNgamma in allograft tolerance mediated by CD4+CD25+ regulatory T cells by induction of IDO in endothelial cells. Am J Transplant 7: Thebault P., et al. (2009) The C-type lectin-like receptor CLEC-1, expressed by myeloid cells and endothelial cells, is up-regulated by immunoregulatory mediators and moderates T cell activation. J Immunol 183:

3 Team 2 PhD Project: Role of interleukin 34 in mechanisms of heart tolerance induction in allotransplantation Supervisor: Dr L. Caron, with Dr I. Anegon We are investigating the immune mechanisms of tolerance in rodent models of allotransplantation with the aim to prevent chronic rejection and to translate to human. The team has recently shown that treatment of rats with a fusion molecule blocking the CD40-CD40L interaction provides a drastic extension of graft survival through induction of regulatory CD8 + T lymphocytes (1,2). Such CD8+ T cells are also present in humans. The objectives are now to define the mechanisms governing the generation and the suppressive functions of CD40Ig-induced Tregs in the rat (3). Our unpublished studies show that the cytokine IL-34 is over-expressed in these induced Tregs and that it plays a role in the immunosuppression via dendritic cells. This recently discovered cytokine share some characteristics with M-CSF as it binds to the same receptor (M-CSFR), though functional studies have suggested different signalling pathways. The key and specific role of IL-34 in immune responses is still ill defined. In order to elucidate the specific role of IL-34, IL-34 KO rats are currently under generation using ZFNs as previously described (4,5) and specific AAV vectors encoding IL-34 have been developped in our lab. The PhD candidate will use these new tools to address the role of IL-34 in immune responses, particularly in transplantation tolerance. We will take particular attention to the effects of IL-34 vs. M-CSF in different populations of dendritic cells and macrophages. This should help to understand the specific role of IL-34 in immunity and tissue homeostasis. 1. Guillonneau C, et al. (2007) CD40Ig treatment results in allograft acceptance mediated by CD8CD45RC T cells, IFN-gamma, and indoleamine 2,3-dioxygenase. J Clin Invest 117: Guillonneau C, et al. (2010) CD8+ regulatory T cells in solid organ transplantation. Curr Opin Organ Transplant.[Epub ahead of print] 3. Li XL, et al. (2010) Mechanism and localization of CD8 regulatory T cells in a heart transplant model of tolerance. J Immunol 185: Geurts A, et al (2009) Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases. Science 24: Ménoret et al. (2010) Characterization of immunoglobulin heavy chain knockout rats. Eur J Immunol. 40:

4 Team 3 PhD Project: Controlling immune responses by modulating expression of cystathionine-γ-lyase in T lymphocytes Supervisors: Pr G. Blancho & Dr B. Vanhove Cystathionine-γ-lyase (CSE) is a rate-limiting enzyme of the trans-sulfuration pathway which converts methionine and cystathionine into cysteine and H2S. T cells being deficient in CSE and cysteine import, they are metabolically dependent on accessory cells for cysteine supply and growth. We demonstrated that immune tolerance after organ transplantation is associated with a repression of CSE in dendritic cells, resulting in a modification of T cell responses (1). This suppressive mechanism of action in transplantation is shared with solid tumors where infiltrating Treg cells and myeloidderived suppressor cells (MDSC) prevent rejection in part by blocking cysteine supply to CD8+ T Cells specific for tumor antigens. Our working hypothesis is based on the concept that ectopic expression of CSE in tumor antigen-specific T cells might prevent their functional inactivation by Treg and MDSC and confer a potential therapeutical benefit. Proof of concept will be assessed by using a thymoma model in mice infused with tumor-specific CD8+ T cells expressing CSE. This research opens a new avenue in immunointervention to control T cell reactivity in cancer and immunosuppression research, two major topics of INSERM UMR-S 1064-Team Vuillefroy de Silly R. et al. (2012) Transplant tolerance is associated with reduced expression of cystathionine-γ-lyase that controls interleukin-12 production by dendritic cells and TH-1 immune responses. Blood Jan FrancoSalinas G. et al. (2011) TNF blockade abrogates the induction of T cell-dependent humoral responses in an allotransplantation model. J Leukoc Biol : Poirier N. et al. (2010) Inducing CTLA-4-dependent immune regulation by selective CD28 blockade promotes regulatory T cells in organ transplantation. Sci Transl Med. 2(17):17ra Dugast AS. et al. (2008) Myeloid-derived suppressor cells accumulate in kidney allograft tolerance and specifically suppress effector T cell expansion. J Immunol. 180: Jovanovic V. et al. (2008) Implication of matrix metalloproteinase 7 and the noncanonical wingless-type signaling pathway in a model of kidney allograft tolerance induced by the administration of anti-donor class II antibodies. J Immunol. 180:

5 Team 4 PhD project: Role of alloreactive CD8 T+ cells in the evolution of kidney graft Supervisor: Dr N. Degauque Organ transplantation is the treatment of choice for life-sustaining terminal organ deficiencies. Survival of kidney transplantation has been greatly improved over the last decade. Yet, all patients are facing a late graft loss and nearly half of them will lose their kidney graft 10 years post-transplantation. Prevention of rejection requires life-long nonspecific immunosuppression, which not only increases the risk of complications related to immunosuppression such as infections and malignancies, but also has specific side effects such as renal toxicity. Identifying early biomarkers of graft dysfunction and understanding the cause of graft dysfunction are two major challenges for improving the graft survival. The role of CD8 T cells in chronic rejection had been poorly characterized despite the fact that CD8 T cells are potent effector cells. Indeed, recent lines of evidence indicate that CD8 T cells play an active role in the physiopathology of chronic rejection. In order to define the role of recipient alloreactive CD8 T cells in kidney graft dysfunction, the PhD candidate will first setup an ex-vivo model of recipient immune cells donor endothelial cell culture and then use it to screen a large cohort of kidney-transplanted patients. For this project, the PhD student will use a broad spectrum of modern state-ofart immunological techniques (molecular biology, cellular biology, multicolor flow cytometry, cellular imaging). In collaboration with a biostatical team, we will correlate the CD8 T cell characteristics with the kidney transplant recipient outcome. Thanks to a close collaboration with the clinical team and a unique biocollection of human recipient and donor cells, the successful applicant will have the opportunity to address a major challenge in transplant immunology. Selected relevant publications of the team: 1. Degauque N, et al. (2011) The blood of healthy individuals exhibits CD8 T cells with a highly altered TCR Vb repertoire but with an unmodified phenotype. PLoS One 6: e Pallier A, et al. (2010) Patients with drug-free long-term graft function display increased numbers of peripheral B cells with a memory and inhibitory phenotype. Kidney international 78: Ashton-Chess J, et al. (2009) Regulatory, effector, and cytotoxic T cell profiles in long-term kidney transplant patients. J Am Soc Nephrol 20: Brouard S, et al. (2007) Identification of a peripheral blood transcriptional biomarker panel associated with operational renal allograft tolerance. Proc Natl Acad Sci U S A 104: Lair D, et al. (2007) Functional compartmentalization following induction of long-term graft survival with pregraft donor-specific transfusion. Am J Transplant 7:

6 Team 5 PhD project: Exploring a novel immunotherapy for a recurring renal pathology in a preclinical model of Idiopathic Nephrotic Syndrome Supervisor: Dr. L. Le Berre, with Pr. J. Dantal Our research group is working on a specific human renal disease: the idiopathic nephrotic syndrome (INS) and particularly its form with recurrence after renal transplantation. Our main goal is the characterization of immunological factors responsible of this glomerulonephritis in order to propose new avenues for immune therapeutic intervention. This nephropathy is characterized by a selective proteinuria and glomerular lesions, reversible after steroid therapy in 80% of cases. Resistance to other immunosuppressive treatments leads to a chronic impairment of renal function requiring hemodialysis and/or renal transplantation. Until now, mechanisms of this orphan disease remain unknown. Thanks to an original animal model very closed to the human disease, the Buffalo/Mna rat, we are able to explore physiopathological hypotheses and to propose new therapeutic approaches for INS. We have recently shown that LF , a derivative of deoxyspergualin, has a major and long-lasting beneficial effect on the proteinuria and glomerular lesions of the Buff/Mna rat and that LF induced regulatory T cells transferred into nephrotic Buff/Mna rats alleviate their proteinuria (1). The PhD candidate will investigate in vitro and in vivo the mechanisms of action of this immunomodulatory compound that improves nephropathy of our rat model. The protective role of induced regulatory T cells will be characterized in this context. The candidate should have an excellent knowledge and keen interests in immunology or renal physiology, and should be willing to work on rats. Supervision will be ensured by a clinician and a scientist, providing a good opportunity to work in translational research. 1. Le Berre L, et al (2009) Induction of T regulatory cells attenuates idiopathic nephrotic syndrome. JASN 20: Le Berre L, et al. (2002) Extrarenal effects on the pathogenesis and relapse of idiopathic nephrotic syndrome in Buffalo/Mna rats. J. Clin. Invest. 109: Le Berre L, et al. (2005) Renal macrophage activation and Th2-polarisation precedes the development of nephrotic syndrome in Buff/Mna rats. Kidney Int 68: Bruneau S, et al.. (2009) High levels of soluble ST2 protein after idiopathic nephrotic syndrome recurrence following kidney transplantation. Am J Kidney Dis. 54: Le Berre L, et al. (2010) Development of initial INS and post-transplantation recurrence: evidence of a same biological entity. Nephrol. Dial. Transplant. 26: