Development of cellular models for evaluating active compounds in cancer therapy

The INSERM U1242 laboratory is a translational research structure bringing together biologists, chemists and clinicians to better understand oncogenesis in brain and gynecological cancers. By associating biologists working on cancer stress signaling pathways, chemists developing molecular tools and cancer clinicians, the laboratory aims at deciphering how tumor cells cope with stresses to resist to death and how they could be sensitized to treatments. The candidate will join an exciting and multidisciplinary scientific environment with on-site access to cutting-edge facilities in biochemistry, biophysics, molecular biology, cell biology and structural biology. The INSERM U1242 laboratory Is affiliated to the Université of Rennes, a top French university that offers a dynamic and lively international campus with an ideal location less than 2 hours from Paris and 1 hour from Saint Malo and the beautiful Britany coast and its beaches. 

Supervisors: Dr X. Guillory (PI) Remuneration: 580€ net/month; Duration: 6 months between January and July 2025

Scientific background – Cancer cells undergo endoplasmic reticulum stress due to unfavorable environmental factors, such as hypoxia or nutrient shortage, high metabolic demand associated in particular with the production of misfolded proteins. To cope with this situation, cancer cells use IRE1 signaling as an adaptive mechanism and, as such, IRE1 plays a crucial role as a rheostatic regulator of cell fate, controlling both cell survival and death. Proof-of-concept of IRE1 inhibition as a novel therapeutic modality has been demonstrated in various cancer models [1], and we are currently developing small molecules inhibitors as adjuvant for Glioblastoma treatments [2]. In parallel to this approach, we are developing a targeted protein degradation (TPD)[3] approach that would induce IRE1 degradation, thus neutralizing its catalytic and interactional functions (PPI, scaffolding) and leading to a durable control of IRE1 activity level in vivo.

As part of our efforts to develop these IRE1 degraders, your goal will be to genetically known-in the HiBit tag using a CRISPR approach to establish a stable cell-line with endogenous expression of tagged IRE1, allowing you to quantify protein level in real-time. Once the cell line established, you will develop the assay protocol, automate it on a pipetting robot, and validate its reliability and robustness. Finally, you will evaluate our existing library of degraders in order to determine their potency. If time allows, you will also work on establishing a stable XBP1s-nLuc reporter cell lines using lentiviral transduction.

Main activities and techniques involved –

Cell line culture, cell treatment, generation of knock-out and knock-in cells using the CRISPR/Cas9 approach, transfection (DNA and siRNA), DNA cloning and primers design, cell viability testing, Western Blot, enzymatic assays.

This internship will take place in the context of a collaboration between medicinal chemists (CNRS UMR 6226, team COrInt) and biologists (Inserm U1242, team PROSAC) and is taking place on the Villejean health campus of the University of Rennes.

References:

[1] D. P. Raymundo, D. Doultsinos, X. Guillory, A. Carlesso, L. A. Eriksson, E. Chevet, Trends Cancer 2020, 6, 1018.

[2] D. P. Raymundo et al., iScience 2023, 26, 5, 106687.

[3] M. Békés, D. R. Langley, C. M. Crews, Nat Rev Drug Discov 2022, 21, 181.

Applicant profile – We encourage applications from highly motivated students from the national and international community in their final year of Master or Engineering school with a background in cell biology, molecular biology and/or biochemistry with experience in cell culture. Python or scripting skills represent a plus but are not required.