Functional characterization of the role of glucose metabolism heterogeneity on an in-vitro 3D model of colorectal cancer (CRC)

Colorectal cancer (CRC) is the third most common type of cancer and the second most common cause of cancer death. CRC is a very heterogeneous disease that evolves from the interaction of genetic and environmental factors. Genetic heterogeneity is a key feature of cancer that leads to the formation of diverse subpopulations of cancer cells displaying different mutational landscapes and cellular phenotypes, thus contributing to disease progression and resistance to common therapies. However, little is known about how non-genetic events contribute to this phenotypic heterogeneity. In a recent study a subpopulation of highly glycolytic cells within colorectal tumors characterized by high pyruvate dehydrogenase kinase (PDK) activity has been described. These cells are quiescent and exhibit robust tumor-initiating potential. The aim of this thesis is to further investigate this metabolic heterogeneity and its role on tumor progression and resistance to chemotherapies. To do this, we set up an in-vitro 3D model of CRC expressing unique genetically-encoded metabolic reporters, which, in combination with imaging, cell-sorting, transcriptomic and metabolomic experiments allowed us to track and characterize cells with different glycolytic activity at single cell level.