Alterations of metabolic pathways in tumours

Cancer progression is shaped by extensive metabolic rewiring and multilayered regulatory mechanisms. This thesis integrates transcriptomic, proteomic, and regulatory network analyses to characterise metabolic reprogramming across multiple tumour types, with a particular focus on glycolysis and the multifunctional enzyme phosphoglycerate kinase 1 (PGK1). Motivated by the growing recognition of metabolic rewiring as a hallmark of cancer, this work addressed key knowledge gaps concerning the regulatory roles of metabolic RBPs and the poorly understood RNA-binding activity of PGK1. An integrated computational–experimental strategy was employed. Differential expression analyses using transcriptomic and proteomic datasets revealed a widespread metabolic activation across tumours and marked transcript–protein discordance within glycolysis. PGK1 and GAPDH were identified as the only glycolytic genes consistently upregulated across all the solid tumours analysed, although protein levels do not always mirror transcript abundance, suggesting extensive post-transcriptional regulation and moonlighting functions. Computational predictions indicated that PGK1 may interact with both coding and non-coding RNAs, including VEGF and metabolic transcripts such as ALDH7A1. Experimental assays revealed a weak or context dependent binding between the PGK1 protein and the VEGF RNA, although further experimental evidences are needed to confirm the interaction. Collectively, this study offers a framework for identifying metabolic vulnerabilities and novel strategies to disrupt tumour-promoting networks.