Supporting data for Combination therapies to enhance efficacy of immune checkpoint inhibitors in hepatocellular carcinoma

My studies explore new therapeutic strategies to combine with immune checkpoint inhibitors in hepatocellular carcinoma (HCC). Inhibition of FSP1 induced ferroptosis, a newr form of immunogenic cell death that elicited immune responses. I studied the regulation of FSP1 by NRF2/KEAP1 pathway. On the other hand, I studied how hypoxia promoted the accumulation of adenosine in the tumor microenvironment (TME) in HCC. HIF is the key transcription factor mediating hypoxia-induced adenosine accumulation. I studied the HIF regulation of ADK, an enzyme that removes adenosine intracellularly, and ENT4, an adenosine transporter that pump out adenosine to the TME. <br> qRT-PCR analysis was performed to study mRNA expression of taregeted genes to investigate their regulation by NRF2/KEAP1 or HIF pathway. ChIP assay was used to study the binding of NRF2 or HIF to targeted genes. Luciferase reporter assay was also performed to study the regulation of targeted gene by NRF2.