INFORMAZIONI SU
AGOSTINI Francesca
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Supervisori: Prof.ssa Rapozzi e Dr. Di Giorgio
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| In pancreatic and lung cancer, the transcription factor NRF2 is implicated in KRAS signaling by sustaining antioxidant responses and aerobic glycolysis. Recently, it has been demonstrated in the lab that the depletion of NRF2 causes the metabolic reprogramming of cancer cells which become highly dependent on aminoacids, in particular arginine, and accumulate high levels of nitric oxide. Here we aim to identify the metabolic shortcircuits which are activated following the depletion of NRF2 and the KRAS shutdown achieved by its PROTAC mediated degradation. In particular, the work will focus on the investigation of the anabolic and catabolic pathways modulated by KRAS/NRF2 axis which control the conversion of arginine to citrulline and nitric oxide, studying their impact on ATP production and oncogenesis. Two classes of enzymes mediate arginine to citrulline conversion: nitric oxide synthases (NOS) and peptidyl arginine deiminases (PADI), working respectively on free arginine and conjugated arginine. CRISPR/Cas9 technology will be adopted to achieve their knock-out in healthy and malignant PDAC cells. Label-free mass-spec quantification methods will be used to quantify metabolites. ChIP-seq, ATAC-seq and Hi-C will be used to quantify the impact of histone citrullination on chromatin remodeling. The transcriptome of NOS and PADI depleted cells will be obtained by means of RNA-seq. The integration of epigenomic, proteomic and biochemical analysis will allow to clarify the role of these metabolic short-circuits in oncogenesis. |
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