PhD Defense of Eden HASEEB

HASEEB Eden, PhLAM Laboratory - UMR8523 - Team DYSCO 

The defence will take place on Wednesday, 11th June 2025 at 14h30, at the Pierre Glorieux Amphitheater, CERLA.

Title : ‘Targeting Kinases: A Strategic Approach to Mitigate TNF-Induced Necroptosis’.

Jury : Franck RIQUET (Supervisor), Pierre Vincent (Rapporteur), Benjamin PFEUTY (Supervisor), Christel VANBESIEN (Examiner), May MORRIS (Examiner), Giulia BERTOLIN (Rapporteur). 

Abstract :

Necroptosis is a genetically regulated form of cell death that is characterized by the release of cytokines, chemokines, and damage-associated molecular patterns, leading to immunogenic consequences. This type of cell death is involved in various pathological disorders, including neurodegenerative and cardiovascular diseases. Among various modulators of Tumor Necrosis Factor (TNF)-induced necroptosis is Receptor-Interacting serine/threonine Protein kinase 1 (RIPK1). Despite numerous clinical trials to inhibit RIPK1, no effective necroptosis inhibitors have been successfully translated into clinical practice. Therefore, we reasoned that additional molecular targets might exist and drug repurposing may provide alternative therapeutic strategies for necroptosis management. We conducted a high-throughput screening of 2800 FDA-approved compounds to evaluate their modulatory effects on TNF-induced necroptosis in L929 cells. Secondary screening validated 151 compounds that modulated TNF-induced necroptosis significantly, of which Pyrvinium Pamoate (PyrPam), an FDA-approved drug, spiked our interest. PyrPam is a Casein Kinase 1 (CK1) activator that promoted necroptosis significantly compared to the TNF-treated alone, which led us to concentrate on the role of CK1 in necroptosis. We found that prior studies have indicated that CK1 inhibition can attenuate necroptosis in murine and human cellular models by inhibiting the phosphorylation of RIPK3 at Serine 227, which blocks necroptosis. However, the dose-response study revealed PyrPam to reduce necroptosis whilst Epiblastin A (inhibitor of CK1) reduced necroptosis as well in L929 cells. We resorted to studying the signaling dynamics of CK1 at the single-cell level through a FRET-based biosensor, which revealed CK1 activity to be differential across cell survival and necroptosis. The early and sharp increase in CK1 activity potentially suggests its involvement in TNF-induced necroptosis. Importantly, it highlighted that CK1 activity could be dispensable for L929 cells, however, further research is needed to delineate the precise mechanisms through which CK1 activity is being compensated (if any) through other pathways, which are sustaining its viability in this context.