Fondation MEDIC

www.fondation-medic.ch

Identifying high-avidity and high-quality individual T-cells in cancer patients
[24.10.2016]
Project leader – Nathalie Rufer
Nathalie Rufer

Dr. Nathalie Rufer received her PhD from the University of Geneva in 1996. She carried out her post-doctoral work at the Terry Fox Laboratory in Vancouver (Canada) before joining in 2001, the Swiss Institute for Cancer Research (ISREC) as an associate scientist within the Lausanne Oncology program. Since 2009, she has been appointed by the Lausanne University Hospital Center (CHUV), and affiliated to the Ludwig Center for Cancer Research. In 2012, Nathalie Rufer finished her full education in clinical medicine at the University of Lausanne and joined the newly formed Department of Oncology led by Prof. George Coukos. Since January 2013, she is extending her research activities together with her clinical education in internal medicine, in oncology and currently in hematology. Her research interests are focused on human T lymphocyte responses against tumor antigens in cancer patients and the development of T cell-based therapies.

Hôpital orthopédique, niveau 5, lab. 1532
Avenue Pierre Decker 4
CH-1011 Lausanne

nathalie.rufer@unil.ch

Group members
 

Michael Hebeisen, PhD

project leader

Mathilde Allard, PhD

post-doctoral fellow

Minh Ngoc Duong, PhD

postdoctoral fellow

Laura Carretero, PhD

postdoctoral fellow

Efe Erdes

PhD student

Barbara Couturaud

PhD student

Project description
 

During recent years, immunotherapy has taken a leading role in innovative oncology. Immune modulation can be achieved by vaccination, adoptive T cell transfer or by checkpoint blockade. However, therapeutic efficacy mostly relies on the identification of high-quality tumor antigen-specific cytotoxic CD8 T-lymphocytes (T-cells).

The efficiency of a T-cell response critically depends on how well a T-cell receptor (TCR) binds to a stimulating peptide-MHC (pMHC) complex (also defined as TCR affinity/avidity) with stronger interactions believed to trigger more vigorous T-cell activation than weaker interactions. This is of particular interest for immunotherapy based on adoptive T-cell transfer, aiming to convey enhanced immune reactivity against tumor-associated antigens, for which endogenous T-cell responses are usually weak. Many tumor antigens are in fact self-antigens that are expressed in the thymus, and accordingly most tumor-reactive T-cells of high avidity are deleted during thymic maturation, in contrast to pathogen-specific T-cells. At present, a major challenge of cancer immunotherapy lies in the identification and activation of those rare tumor-specific CD8 T-cells that escape thymic selection, offering maximal functional avidity against malignant cells (Figure 1).

TCR-pMHC avidity has yet to be considered as a determining factor in the selection of antigen-specific T-cells for clinical protocols. However, technical limitations are the major reasons why T-cell selection for immunotherapy is currently based on assays measuring antigen specificity, proliferation and cytokine (e.g. IFNγ) production. Furthermore, the precise impact of TCR-pMHC avidity for various T-cell functions (e.g. target cell killing, cytokine secretion, and proliferation) as well as on the selection, homing to and function in metastatic lesions remains to be directly evaluated. Because TCR-pMHC avidity is one of the most important correlates of immune protection, it is greatly justified to further understand its impact on clinically and biologically relevant parameters with the aim to identify CD8 T-cells with best suitable TCRs and high functional qualities against tumor cells, and with limited toxicity (Figure 1).

The overall aim of this project is to determine the TCR avidities from vaccinated melanoma patients, together with other clinically and biologically relevant parameters using in vitro and in vivo experimental settings. We hypothesize that rare anti-tumor TCRs of high avidity and high functional quality are present in those melanoma patients and that the TCR-pMHC avidity is a major determinant allowing identifying and isolating such cells.

Specifically, we will use the recently developed NTAmer technology, which allows for the direct quantification of TCR-pMHC dissociation kinetics on living antigen-specific CD8 T-cells. The analysis of TCR affinity/avidity will be combined with various in vitro functional assays (e.g. protein signaling, target-cell-killing, cytokine release and proliferation) to clarify the causality between TCR affinity, T-cell activation and signaling, differentiation and functionality. Once identified, we will further assess the impact of structural TCR avidity on in vivo T-cell migration to tumor lesions and potency to recognize and control tumor growth following adoptive transfer in immunodeficient NSG mice. Overall, these studies directly support the development of more effective T-cell mediated therapies, such as adoptive T-cell transfer or vaccination.

figure 1

Figure 1. General outline for identifying and selecting tumor-specific CD8 T-cells offering maximal functional capacities for adoptive cell transfer in cancer patients. A large repertoire of tumor-specific T-cells can be generated following immunotherapeutic vaccination (a) and various biological attributes may be used for the identification, selection and activation of the most potent anti-cancer T-cells (b), before short-term in vitro expansion and reinfusion into patients (so-called “ adoptive T-cell transfer” therapy) (c).

Publications
 
2016 / 10.1158/1078-0432.CCR-15-1212
Legat A, Maby-El Hajjami H, Baumgaertner P, Cagnon L, Abed Maillard S, Geldhof C, Iancu EM, Lebon L, Guillaume P, Dojcinovic D, Michielin O, Romano E, Berthod G, Rimoldi D, Triebel F, Luescher I, Rufer N, Speiser DE. Vaccination with LAG-3Ig (IMP321) and Peptides Induces Specific CD4 and CD8 T-Cell Responses in Metastatic Melanoma Patients--Report of a Phase I/IIa Clinical Trial. Clin Cancer Res. 2016;22:1330-40
» PubMed
2015 / 10.1158/0008-5472.CAN-14-3516
Hebeisen M, Schmidt J, Guillaume P, Baumgaertner P, Speiser DE, Luescher I, Rufer N. Identification of Rare High-Avidity, Tumor-Reactive CD8+ T Cells by Monomeric TCR-Ligand Off-Rates Measurements on Living Cells. Cancer Res. 2015;75:1983-91
» PubMed
2015 / 10.1080/2162402X.2015.1073882
Bron S, Henry L, Faes-van’t Hull E, Turrini R, Vanhecke D, Guex N, Ifticene-Treboux A, Majo F, Nicolas M, Iancu EM, Semilietof M, Rufer N, Lehr HA, Xenarios I, Coukos G, Delaloye J.-F., Doucey M-A. TIE-2-expressing monocytes are lymphangiogenic and incorporate specifically into lymphatics of human breast cancer. Oncoimmunology
» PubMed
2015 / 10.4049/jimmunol.1403145
Gannon PO, Wieckowski S, Baumgaertner P, Hebeisen M, Allard M, Speiser DE, Rufer N. Quantitative TCR:pMHC Dissociation Rate Assessment by NTAmers Reveals Antimelanoma T Cell Repertoires Enriched for High Functional Competence. J Immunol. 2015;195:356-66
» PubMed
2015 / 10.1126/scitranslmed.aaa3700
Fuertes Marraco SA, Soneson C, Cagnon L, Gannon PO, Allard M, Maillard SA, Montandon N, Rufer N, Waldvogel S, Delorenzi M, Speiser DE. Long-lasting stem cell-like memory CD8+ T cells with a naïve-like profile upon yellow fever vaccination. Sci Transl Med. 2015 Apr 8;7(282):282ra48
» PubMed
2015 / 10.3389/fimmu.2015.00582
Hebeisen M, Allard M, Gannon PO, Schmidt J, Speiser DE, Rufer N. Identifying Individual T Cell Receptors of Optimal Avidity for Tumor Antigens. Front Immunol. 2015;6:582
» PubMed
2013 / 10.1172/JCI65325
Hebeisen M, Baitsch L, Presotto D, Baumgaertner P, Romero P, Michielin O, Speiser DE, Rufer N. SHP-1 phosphatase activity counteracts increased T cell receptor affinity. J Clin Invest. 2013 Mar;123(3):1044-56
» PubMed
2013 / 10.1016/j.immuni.2012.12.006
Dudda JC, Salaun B, Ji Y, Palmer DC, Monnot GC, Merck E, Boudousquie C, Utzschneider DT, Escobar TM, Perret R, Muljo SA, Hebeisen M, Rufer N, Zehn D, Donda A, Restifo NP, Held W, Gattinoni L, Romero P. MicroRNA-155 is required for effector CD8+ T cell responses to virus infection and cancer. Immunity. 2013 Apr 18;38(4):742-53
» PubMed
2013 / 10.3389/fimmu.2013.00154
Hebeisen M, Oberle SG, Presotto D, Speiser DE, Zehn D, Rufer N. Molecular insights for optimizing T cell receptor specificity against cancer. Front Immunol. 2013;4:154
» PubMed
2013 / 10.1371/journal.pone.0078686
Iancu EM, Gannon PO, Laurent J, Gupta B, Romero P, Michielin O, Romano E, Speiser DE, Rufer N. Persistence of EBV antigen-specific CD8 T cell clonotypes during homeostatic immune reconstitution in cancer patients. PLoS One. 2013 Oct 25;8(10):e78686
» PubMed
2012 / 10.1002/ijc.26297
Baumgaertner P, Jandus C, Rivals JP, Derré L, Lövgren T, Baitsch L, Guillaume P, Luescher IF, Berthod G, Matter M, Rufer N, Michielin O, Speiser DE. Vaccination-induced functional competence of circulating human tumor-specific CD8 T-cells. Int J Cancer. 2012;130:2607-17
» PubMed