CRISPR/Cas9 Screens Reveal that Hexokinase 2 Enhances Cancer Stemness and Tumorigenicity by Activating the ACSL4‐Fatty Acid β‐Oxidation Pathway

Abstract Metabolic reprogramming is often observed in carcinogenesis, but little is known about the aberrant metabolic genes involved in the tumorigenicity and maintenance of stemness in cancer cells. Sixty‐seven oncogenic metabolism‐related genes in liver cancer by in vivo CRISPR/Cas9 screening are identified. Among them, acetyl‐CoA carboxylase 1 (ACC1), aldolase fructose‐bisphosphate A (ALDOA), fatty acid binding protein 5 (FABP5), and hexokinase 2 (HK2) are strongly associated with stem cell properties. HK2 further facilitates the maintenance and self‐renewal of liver cancer stem cells. Moreover, HK2 enhances the accumulation of acetyl‐CoA and epigenetically activates the transcription of acyl‐CoA synthetase long‐chain family member 4 (ACSL4), leading to an increase in fatty acid β ‐oxidation activity. Blocking HK2 or ACSL4 effectively inhibits liver cancer growth, and GalNac‐siHK2 administration specifically targets the growth of orthotopic tumor xenografts. These results suggest a promising therapeutic strategy for the treatment of liver cancer.