Keywords: STING, Autophagy, Non-canonical autophagy(CASM), Immune responses, Cancer Internship Duration: 30/11/-1 - 30/11/-1
Head of the hosting team: Pr Christian POÜS
Address of the host laboratory: Inserm UMR-S 1193 Team Inserm UMR-S 1193 Team 5 17 Av. des Sciences 91400 Orsay France
Supervisor: Dr Elise JACQUINE-mail: elise.jacquin@universite-paris-saclay.fr Phone: +33 01.80.00.63.16
The Stimulator of Interferon Gene (STING) has emerged as a promising target to modulate antitumor immunity. Following STING activation in response cytosolic DNA detection, proinflammatory molecules and type I Interferons (IFN) are released and stimulate T cell activation in the tumor microenvironment (TME) thus favoring adaptive antitumor responses1. T cell-intrinsic activation of STING can also directly stimulate CD4 T cell antitumor activity2. A deeper molecular understanding of the STING pathway and its regulation in the TME is now needed to improve STING-based anticancer therapies. Autophagy has been proposed to regulate the STING pathway. While STING activation drives LC3 lipidation, a key marker of autophagy, the underlying mechanisms and consequences of STING-driven autophagy are still unclear and context-dependent. STING agonists have been proposed to activate both macroautophagy and non-canonical autophagy, which is the alternative Conjugation of Atg8 proteins to Single Membranes (CASM)3-5. However, the functional consequences of STING-associated marcoautophagy and CASM in STING-relevant immune cells remain unexplored. Using STING-relevant primary myeloid and lymphoid cells, we found that STING agonists can trigger CASM and macroautophagy in parallel. We are currently exploring the functional consequences of these STING-driven autophagy pathways by measuring the conventional responses to STING activation and by exploring other unknown responses using unbiased approaches. We are looking for a highly motivated candidate to characterize the response to STING activation of cell models of macroautophagy or CASM deficiency. RAW 264.7 macrophages genetically engineered by CRIPSR-Cas9 to be incompetent for macroautophagy or CASM will be cultured and stimulated with STING agonists in vitro. Primary immune cells from mice will also be used ex vivo. The candidate will then analyze LC3 lipidation and assess the production of immune mediators following STING activation. She/he will thus allow us to understand the impact of STING-driven autophagies on immune responses and the potential consequences on tumor growth control. Previous technical experience in cell biology will be much appreciated.
This project will use various techniques including cell culture and transfection, immunofluorescence and imaging, Western-blotting, ELISA, cytokine arrays, and RT-qPCR.
1. Zhu Y, et al. Mol Cancer. 2019. PMID: 31679519 2. Benoit-Lizon I, Jacquin E, et al. J Immunother Cancer. 2022. PMID: 35091453 3. Durgan J, Florey O. Sci Adv. 2022. PMID: 36288315 4. Hooper KM, Jacquin E, et al. J Cell Biol. 2022. PMID: 35511089 5. Fischer TD, et al. J Cell Biol. 2020. PMID: 33201170
