Innate lymphoid cells (ILCs) are lymphocytes lacking antigen-specific receptors, and as the innate counterpart of lymphoid cells, they have been identified as playing a major role in controlling the immune response and constitute promising therapeutic targets. Type 2 ILCs (ILC2) accumulate in the tumor microenvironment (TME) and blood of patients with hepatocellular carcinoma (HCC)1,2. Single-cell analyses reveal two subsets of ILC2 in HCC, one being an ICOS⁺ KLRG1⁻ IL 13⁺ population specifically enriched in tumors and associated with poor prognosis3
. The molecular and cellular basis of their protumor activity remains unknown. During this internship you will participate to: Identify the most relevant pre-clinical model to study ILC2 in HCC You will work with two established mouse models of HCC available in the lab: a genetically induced model (c-Myc/NRas-driven)1 and an orthotopic Hepa1.6 model. You will compare immune infiltration patterns across both models using spectral flow cytometry to identify which best mirrors the ILC2-enriched TME observed in human HCC samples.
Evaluate how ILC2 deficiency reshapes the tumor microenvironment (TME) in HCC Using IL7RCreRORαf/f conditional KO-mice, which lack ILC24,5, you will assess the impact of ILC2 deficiency on tumor progression (via bioluminescence imaging) and TME composition (via spectral flow cytometry and single-cell RNA sequencing).
Characterize the crosstalk between ILC2 and myeloid cells in-vivo and ex-vivo. Given that tumor-enriched ILC2 are key producers of IL-131,3, a cytokine known to drive the polarization of tumor-associated macrophages6,7 and myeloid-derived suppressor cells8 toward immunosuppressive phenotypes, you will investigate how ILC2 influence the myeloid compartment. In vivo, you will use spectral flow cytometry to analyze the recruitment and activation status of myeloid cells in the presence or absence of ILC2. Ex vivo, you will co-culture ILC2 isolated from the tumor environment with bone marrowderived macrophages to assess the ability of ILC2 to induce a pro-tumoral polarization.
Assess the role of Notch signaling in reprogramming tumor specific ILC2 toward a KLRG1⁺ state. Notch signaling has been shown to up-regulate KLRG1 while dampening IL-13 production in ILC2 following IL33 stimulation in lung inflammation models9 . Moreover, in pancreatic cancer, KLRG1⁺ ILC2 accumulation correlates with improved prognosis10. To elucidate the relevance of this pathway in HCC, you will activate Notch ex vivo and profile cytokine output from tumor-derived ILC2 after IL-33 stimulation, and employ IL7RCreNotch2f/f conditional-KO mice (available in the lab)11 to examine how Notch2 in ILC2 affects HCC development in vivo.
Methodology & training You will be exposed to state-of-the-art technologies and gain skills in:
• Tumor immunology
• Biology of innate lymphoid cells, myeloid cells, and T cells
• Mouse models of HCC and conditional-KO.
• High-parameter flow & spectral cytometry, cell sorting and ex-vivo functional assays.
The project offers full immersion in tumor immunology, data generation and analysis, and teamwork in a multidisciplinary environment.

Bibliography
1.Xu, X. et al. Hepatology 74, 2526–2543 (2021)
2.Heinrich, B. et al. Gut 71, 1161–1175 (2022)
3.He, Y. et al. Hepatology 76, 1013–1029 (2022)
4.Wong, S. H. et al. Nat Immunol 13, 229–236 (2012)
5.Oliphant, C. J. et al. Immunity 41, 283–295 (2014)
6.Bronte, V. et al. J Immunol 170, 270–278 (2003)
7. Mantovani, A., Sica, A., Allavena, P., Garlanda, C. & Locati, M. Hum Immunol 70, 325–330 (2009)
8. Gabrilovich, D.I., Ostrand-Rosenberg, S. & Bronte, V. Nat Rev Immunol 12, 253–268 (2012)
9. Zhang, K. et al. J Immunol 198, 1798–1803 (2017)
10. Amisaki, M. et al. Nature 638, 1076–1084 (2025)
11. Golub, R. Biomed J 44, 133–143 (2021)

Dernières Publications en lien avec le projet :
1. Bourayou, E. & Golub, R. Signaling Pathways Tuning Innate Lymphoid Cell Response to Hepatocellular Carcinoma. Front. Immunol. 13, 846923 (2022).
2. Perchet T, Petit M, Banchi EG, Meunier S, Cumano A, Golub R (2018) The Notch Signaling Pathway Is Balancing Type 1 Innate Lymphoid Cell Immune Functions. Front Immunol. 9:1252.
3. Xu W, Cherrier D, Chea S, Vosshenrich C, Serafini N, Petit M, Liu P, Golub R, Di Santo J (2019) A novel Id2RFP reporter mouse strain redefines ILC precursor potentials. Immunity 50:1054.
4. Chea S, Perchet T, Petit M, Verrier T, Guy-Grand D, Banchi EG, Vosshenrich CAJ, Di Santo JP, Cumano A. & Golub R (2016) Notch signaling in group 3 ILC modulates their plasticity. Sci Signal. 9(426):ra45.
5. Chea S, Schmutz S, Berthault C, Burlen-Defranoux O, Goldrath A, Rodewald HR, Cumano A & Golub R (2016) Single gene expression analyses reveal heteregeneous responsiveness of innate lymphoid progenitors to Notch signaling. Cell Rep. 2016 Feb 16;14(6):1500-16.

Ce projet s’inscrit dans la perspective d’une thèse
Type de financement prévu : Concours ED
Ecole Doctorale de rattachement : BioSPC