Fig 7A and 7B show the network pathway for intracellular compounds that were significantly associated with extracellular compounds, including cysteine, glutamine, and homoserine. Taken together, we conclude that the depletion determined by CD44 altered extra- and intra-cellular metabolic profiles of CCA cells, leading to increased levels of ROS, particularly due to the alteration of amino acids such as cysteine, glutamate, and glutamine, that is driven by CD44-mediated Akt activation and its downstream targets contributing to CCA progression (Fig 8). This evidence concerns the gene AKT1 and cholangiocarcinoma.