Loss-of-function genetic screening identifies ALDOA as an essential driver for hepatocellular carcinoma cell growth under hypoxia

Hypoxia is a common feature of tumor microenvironment (TME), which promotes tumor progression, metastasis and therapeutic drug resistance via a myriad of cell activities in tumor and stroma cells. While targeting hypoxic TME is emerging as a promising strategy for treating solid tumors, preclinical studies with this approach are lacking in hepatocellular carcinoma (HCC). From a genome-wide CRISPR/Cas9 gene knockout screening, we identified aldolase A (ALDOA), a key enzyme in glycolysis and gluconeogenesis, as a master driver for HCC cell growth under hypoxia. To delineate the functional implications of ALDOA in HCC, transcriptome sequencing is performed to interrogate the differential gene expression in ALDOA-knockdown HepG2 cells under hypoxia. Overall design: The HCC cells are cultured in 20% O2 and 1% O2 for 3 days to mimic normoxia and hypoxia, genome-wide CRISPR/Cas9 knockout library screening and sequencing to identify essential genes responding to hypoxia in HCC tumor cells. RNA sequencing is performed to interrogate the differential gene expression in ALDOA-knockdown HepG2 cells under hypoxia.