CD - Modulation du microenvironnement tumoral par les cellules iNKT pour améliorer la thérapie cellulaire CAR dans le glioblastome

Offre de thèse

Date limite de candidature

09-06-2026

Date de début de contrat

01-10-2026

Directeur de thèse

REPPEL Loïc

Encadrement

The PhD candidate will benefit from close supervision thanks to the availability of a principal supervisor and a co-supervisor working in the same laboratory. The candidate's progress will be monitored through regular support meetings : • A thesis kick-off meeting in the presence of the principal supervisor, the co-supervisor, team members, and PhD students (with opportunities for discussion), as well as the managers of the relevant technical platforms. • A weekly meeting in the presence of the principal supervisor and/or co-supervisor. • A bi-monthly progress review meeting attended by the principal supervisor, the co-supervisor, and two team members involved in the PhD project. During these meetings, the candidate's progress, results obtained, and any difficulties encountered will be discussed. • A meeting every two months with the partners involved in the project. • A presentation of their work every six months during team meetings. PhD training : • PhD candidates in our team take part in both disciplinary and interdisciplinary training courses offered by the University of Lorraine, the CNRS, and other organizations. • PhD candidates also attend the “Doctoriales” organized once a year by the doctoral schools. • If they wish, our PhD candidates may gain teaching experience through a temporary teaching and research position (e.g., ATER) or receive support from PEEL (Pôle Entrepreneuriat Étudiant de Lorraine). Technical and methodological supervision : The PhD candidate will be trained in all the techniques required to carry out their research project. The candidate's well-being will be a constant priority. Particular attention will be paid to the development of their technical, intellectual, and psychological maturity to ensure they have a positive experience within partner laboratories. The supervisory team will encourage early thesis writing and the valorization of research outcomes. If a patent application can be considered, the necessary steps will be initiated, and dissemination through publications and conference presentations will be encouraged and supported. Sufficient time will be dedicated throughout the PhD to ensure that the candidate meets the objectives defined by the doctoral school. Career prospects after the PhD will naturally depend on the candidate's own aspirations. However, should they express interest, we will discuss the possibility of a postdoctoral position in Professor Dotti's laboratory or with other partners involved in the project. The collaborative nature of this project provides valuable opportunities to interact with numerous researchers, enabling the candidate to refine their future professional goals.

Type de contrat

Concours pour un contrat doctoral

Candidater à cette offre

école doctorale

BioSE - Biologie Santé Environnement

équipe

Equipe 6 : Cell-engineering and Immunomodulation of Inflammatoryand Neoplastic Disorders (CImIND

contexte

Survival rates for patients with glioblastoma (GBM) remain very low, with minimal progress made over the past decade. More than two-thirds of patients diagnosed with GBM, the most common malignant brain tumor, die within two years. Even among the 5% of patients surviving beyond five years, many suffer from long-term consequences due to medical interventions. GBM is considered a high-priority area for cancer research, as it is a cancer of unmet medical need. Several challenges make GBM difficult to treat: (i) it exhibits high resistance to both chemotherapy and radiotherapy, with tumors often evolving into more aggressive, treatment-resistant phenotypes; (ii) its location in the central nervous system (CNS) often precludes complete surgical resection, and the blood-brain barrier limits the exposure of systemic therapies, enabling resistant clones to emerge1. GBM is characterized by a hostile and immunosuppressive tumor microenvironment (TME). GBM is categorized within the ‘cold tumors” with few effector T and NK cells contrasting with many immunosuppressive cells including M2 tumor-associated macrophages (TAMs), myeloid suppressive cells (MDSCs) and regulatory T cells (Tregs)1. Single cell transcriptomic characterization of grade IV GBM microenvironment confirmed a predominant infiltration by MDSCs capable of supporting tumor stem cells and inhibiting T cell functions2. Given the immense unmet need of GBM, there is great interest in developing new immunotherapy strategies that could overcome tumor escape mechanisms. CAR-T therapies have proven successful in certain hematological malignancies. However, their efficacy has been limited in solid tumors. In GBM, recent studies have shown promising clinical signs in patients treated with CAR-T cells targeting antigens such as EGFRvIII, HER2, and IL-13Rα23. Nevertheless, the responses are often short-lived, and patients frequently relapse3. Two additional targets of interest in GBM are GD2 and B7-H3, both of which are highly expressed in this cancer1. Preclinical studies have shown that anti-GD2 CAR-T cells have the potential to target GBM cells effectively in vitro and in patient-derived xenograft (PDX) mouse models4. B7-H3 (CD276) plays a role in immune evasion by inhibiting T cell activity, contributing to tumor escape mechanisms, particularly in brain tumors5. Initial clinical trials are now underway, further exploring these therapeutic approaches in GBM3. Even though recent clinical trials of CAR-T cells in GBM have shown positive signs and are ongoing in lung cancers, the CAR-T strategy needs to be rethought with approaches that enhance migration, persistence, effectiveness and modulation of tumor immunosuppressive microenvironment in solid tumors like GBM. Invariant NKT (iNKT) cells offer a promising alternative to conventional CAR-T cells. These innate immune cells express a unique TCR (V&#945;24-J&#945;18/V&#946;11 in humans) that recognizes glycolipids presented by CD1d on antigen-presenting and cancer cells, leading to their rapid activation and cytokine secretion6. They directly kill tumor cells and modulate the immune response by converting the TME to a proinflammatory state, suppressing MDSCs, and promoting T and NK cell activation7. An additional advantage of iNKT cells is their lack of alloreactivity, as demonstrated by the Rubio team8,9, positioning them as promising candidates for off-the-shelf therapies. These attributes xmake iNKT cells excellent candidates for CAR-based therapies10. Importantly, CAR-iNKT cells could target tumor cells not only through their CAR but also via their iTCR, particularly in tumors expressing CD1d, such as GBM11. Few studies have compared CAR-iNKT to CAR-T cells. In a preclinical lymphoma model, anti-CD19 CAR-iNKT cells showed superior efficacy to CAR-T cells, particularly in homing to brain lymphoma localization12. In a neuroblastoma pre-clinical model, anti-GD2 CAR-iNKT cells migrate more actively to tumor sites than CAR-T cells13. Engineered hematopoietic stem cells to express NKT invariant TCR (CSH-iNKT) and an anti-BCMA CAR demonstrated enhanced control of myeloma by clearing CD1d-expressing immunosuppressive MDSCs, macrophages, and dendritic cells from the TME14,15. Several clinical trials are exploring CAR-iNKT cells in hematological malignancies10. In solid tumors, anti-GD2 CAR iNKT cells have shown good tolerance profile and some efficacy in phase I clinical trials in neuroblastoma16. Even if iNKT cells represent a new and promising cellular platform for chimeric antigen receptor-therapies, we still have an incomplete understanding of how these cells exert their antitumoral effects in solid tumors, especially in the presence of an intact TME. Our project aims to investigate the mechanisms of action of CAR-iNKT cells and compare them to CAR-T cells targeting GD2 and B7-H3 in GBM models. The originality of our approach lies in utilizing two distinct, innovative preclinical models that preserve the integrity of the TME. These include human tumor explants from GBM, maintained under survival conditions, as well as a syngeneic mouse model of GBM. These models will enable a more accurate replication of the complexities of the TME, allowing us to assess the efficacy of CAR-iNKT cells in real-time, and compare their interactions with both the tumor and surrounding microenvironment to those of CAR-T cells. We hypothesize that CAR-iNKT cells will outperform CAR-T cells in terms of migration and immune synapse formation, which will be monitored through dynamic fluorescence imaging. This advantage may arise from their iTCR, which enables recognition of CD1d-expressing tumor cells. Given that the TME presents significant barriers to immune activation, we will examine these obstacles by correlating CAR-iNKT and CAR-T functional responses with the TME composition through baseline transcriptomic studies. Additionally, due to the known ability of iNKT cells to produce substantial levels of IFN&#947;, a key cytokine in reshaping the TME, and their potential to target immunosuppressive CD1d-positive myeloid cells, we hypothesize that CAR-iNKT cells will demonstrate superior capacity for TME remodeling. These properties will be evaluated in our models, with the goal of optimizing this innovative immune therapy approach.

spécialité

Sciences de la Vie et de la Santé - BioSE

laboratoire

IMoPA - Ingénierie Moléculaire et Physiopathologie Articulaire

Mots clés

Récepteur chimérique d'antigène, immunothérapie allogénique, cellules iNKT, microenvironnement, glioblastome

Détail de l'offre

Le glioblastome (GBM) est une tumeur solide agressive associée à un mauvais pronostic pour les patients, principalement en raison de son microenvironnement tumoral (TME) immunosuppresseur et de sa résistance aux thérapies actuelles. Des défis tels qu'une faible infiltration tumorale, l'évasion immunitaire et l'immunosuppression au sein du TME entravent leur 'efficacité. Les iNKT modifiées par CAR (CAR-iNKT) présentent des avantages uniques, notamment leur capacité à moduler le TME et leur indépendance vis-à-vis de l'alloréactivité, ce qui en fait une alternative prometteuse. Notre projet vise à décrypter les mécanismes d'action des cellules CAR-iNKT et à comparer leur efficacité à celles des cellules CAR-T ciblant les antigènes associés aux tumeurs GD2 et B7-H3 dans des modèles précliniques de GBM. À l'aide d'explants tumoraux humains frais et de modèles murins syngéniques, nous évaluerons des paramètres clés tels que la migration cellulaire, la formation de synapses immunologiques et la cytotoxicité au sein de microenvironnements tumoraux intacts. De plus, des analyses transcriptomiques dynamiques et des tests fonctionnels seront utilisés afin d'explorer comment les cellules CAR-iNKT et CAR-T sont influencées par le TME immunosuppresseur et comment, en retour, elles peuvent reprogrammer cet environnement pour renforcer les réponses antitumorales dans le GBM.

Keywords

Chimeric Antigen Receptor, Allogeneic Immunotherapy, iNKT cells, Microenvironment, Glioblastoma

Subject details

Glioblastoma (GBM) is aggressive solid tumor with poor patient prognoses, primarily due to their immunosuppressive tumor microenvironments (TME) and resistance to current therapies. While chimeric antigen receptor (CAR) T-cell therapies have achieved remarkable success in hematological malignancies, their efficacy in solid tumors has been limited. Challenges such as poor tumor infiltration, immune evasion, and suppression within the TME hinder the effectiveness of CAR-T cells. In contrast, CAR invariant natural killer T (CAR-iNKT) cells offer unique advantages, including their ability to modulate the TME and their independence from alloreactivity, positioning them as a promising alternative. Our project aims to dissect the mechanisms by which CAR-iNKT cells act and compare their efficacy to CAR-T cells targeting the tumor-associated antigens GD2 and B7-H3 in preclinical models of GBM. Through the use of fresh human tumor explants and syngeneic mouse models, we will evaluate key parameters such as cell migration, immune synapse formation, and cytotoxicity within intact TMEs. Moreover, dynamic transcriptomic and functional assays will be employed to explore how CAR-iNKT and CAR-T cells are influenced by the immunosuppressive TME and how they, in turn, may reprogram this environment to enhance anti-tumor responses in GBM.

Profil du candidat

Profil recherché: Étudiant niveau M2 en biologie cellulaire, immunologie

Compétences: recherche bibliographique, analyse et interprétation de résultats, culture cellulaire (primaire et lignées), transduction cellulaire, cytométrie en flux, tests de cytotoxicité

Candidate profile

Required profile: Master's student in cell biology, immunology

Skills: bibliographic research, analysis and interpretation of results, cell culture (primary and cell lines), flow cytometry, cell transduction, cytotoxicity assays

Référence biblio

1. Yuan, B., Wang, G., Tang, X., Tong, A. & Zhou, L. Immunotherapy of glioblastoma: Recent advances and future prospects. Hum Vaccin Immunother 18, 2055417, doi:10.1080/21645515.2022.2055417 (2022).
2. Jackson, C. et al. Distinct Myeloid Derived Suppressor Cell Populations Promote Tumor Aggression in Glioblastoma. bioRxiv, doi:10.1101/2023.03.26.534192 (2023).
3. Agosti, E. et al. CAR-T Cells Therapy in Glioblastoma: A Systematic Review on Molecular Targets and Treatment Strategies. Int J Mol Sci 25, doi:10.3390/ijms25137174 (2024).
4. Gargett, T. et al. GD2-targeting CAR-T cells enhanced by transgenic IL-15 expression are an effective and clinically feasible therapy for glioblastoma. J Immunother Cancer 10, doi:10.1136/jitc-2022-005187 (2022).
5. Guo, X., Chang, M., Wang, Y., Xing, B. & Ma, W. B7-H3 in Brain Malignancies: Immunology and Immunotherapy. Int J Biol Sci 19, 3762-3780, doi:10.7150/ijbs.85813 (2023).
6. Lantz, O. & Bendelac, A. An invariant T cell receptor alpha chain is used by a unique subset of major histocompatibility complex class I-specific CD4+ and CD4-8- T cells in mice and humans. J Exp Med 180, 1097-1106, doi:10.1084/jem.180.3.1097 (1994).
7. Nelson, A., Lukacs, J. D. & Johnston, B. The Current Landscape of NKT Cell Immunotherapy and the Hills Ahead. Cancers (Basel) 13, doi:10.3390/cancers13205174 (2021).
8. Rubio, M. T. et al. Early posttransplantation donor-derived invariant natural killer T-cell recovery predicts the occurrence of acute graft-versus-host disease and overall survival. Blood 120, 2144-2154, doi:10.1182/blood-2012-01-404673 (2012).
9. Rubio, M. T. et al. Pre-transplant donor CD4(-) invariant NKT cell expansion capacity predicts the occurrence of acute graft-versus-host disease. Leukemia 31, 903-912, doi:10.1038/leu.2016.281 (2017).
10. O'Neal, J., Mavers, M., Jayasinghe, R. G. & DiPersio, J. F. Traversing the bench to bedside journey for iNKT cell therapies. Front Immunol 15, 1436968, doi:10.3389/fimmu.2024.1436968 (2024).
11. Hara, A. et al. CD1d expression in glioblastoma is a promising target for NKT cell-based cancer immunotherapy. Cancer Immunol Immunother 70, 1239-1254, doi:10.1007/s00262-020-02742-1 (2021).
12. Rotolo, A. et al. Enhanced Anti-lymphoma Activity of CAR19-iNKT Cells Underpinned by Dual CD19 and CD1d Targeting. Cancer Cell 34, 596-610 e511, doi:10.1016/j.ccell.2018.08.017 (2018).
13. Heczey, A. et al. Invariant NKT cells with chimeric antigen receptor provide a novel platform for safe and effective cancer immunotherapy. Blood 124, 2824-2833, doi:10.1182/blood-2013-11-541235 (2014).
14. Li, Y. R. et al. Generation of allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells using a clinically guided culture method. Nat Biotechnol, doi:10.1038/s41587-024-02226-y (2024).
15. Li, Y. R. et al. Development of allogeneic HSC-engineered iNKT cells for off-the-shelf cancer immunotherapy. Cell Rep Med 2, 100449, doi:10.1016/j.xcrm.2021.100449 (2021).
16. Heczey, A. et al. Anti-GD2 CAR-NKT cells in relapsed or refractory neuroblastoma: updated phase 1 trial interim results. Nat Med 29, 1379-1388, doi:10.1038/s41591-023-02363-y (2023).
17. Coman, T. et al. Human CD4- invariant NKT lymphocytes regulate graft versus host disease. Oncoimmunology 7, e1470735, doi:10.1080/2162402X.2018.1470735 (2018).
18. Reppel, L. et al. Targeting disialoganglioside GD2 with chimeric antigen receptor-redirected T cells in lung cancer. J Immunother Cancer 10, doi:10.1136/jitc-2021-003897 (2022).
19. Durand, M. et al. Radiosensitization with Gadolinium Chelate-Coated Gold Nanoparticles Prevents Aggressiveness and Invasiveness in Glioblastoma. Int J Nanomedicine 18, 243-261, doi:10.2147/IJN.S375918 (2023).
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21. Simula, L. et al. Mitochondrial metabolism sustains CD8(+) T cell migration for an efficient infiltration into solid tumors. Nat Commun 15, 2203, doi:10.1038/s41467-024-46377-7 (2024).
22. Landoni, E. et al. IL-12 reprograms CAR-expressing natural killer T cells to long-lived Th1-polarized cells with potent antitumor activity. Nat Commun 15, 89, doi:10.1038/s41467-023-44310-y (2024).