USP29-mediated HIF1α stabilization is associated with Sorafenib resistance of hepatocellular carcinoma cells by upregulating glycolysis

Abstract

Understanding the mechanisms underlying evasive resistance in cancer is an unmet medical need to improve the efficacy of current therapies. In hepatocellular carcinoma (HCC), aberrant expression of hypoxia-inducible factor 1 α (HIF1α) and increased aerobic glycolysis metabolism are drivers of resistance to therapy with the multi-kinase inhibitor Sorafenib. However, it has remained unknown how HIF1α is activated and how its activity and the subsequent induction of aerobic glycolysis promote Sorafenib resistance in HCC. Here, we report the ubiquitin-specific peptidase USP29 as a new regulator of HIF1α and of aerobic glycolysis during the development of Sorafenib resistance in HCC. In particular, we identified USP29 as a critical deubiquitylase (DUB) of HIF1α, which directly deubiquitylates and stabilizes HIF1α and, thus, promotes its transcriptional activity. Among the transcriptional targets of HIF1α is the gene encoding hexokinase 2 (HK2), a key enzyme of the glycolytic pathway. The absence of USP29, and thus of HIF1α transcriptional activity, reduces the levels of aerobic glycolysis and restores sensitivity to Sorafenib in Sorafenib-resistant HCC cells in vitro and in xenograft transplantation mouse models in vivo. Notably, the absence of USP29 and high HK2 expression levels correlate with the response of HCC patients to Sorafenib therapy. Together, the data demonstrate that, as a DUB of HIF1α, USP29 promotes Sorafenib resistance in HCC cells, in parts by upregulating glycolysis, thereby opening new avenues for therapeutically targeting Sorafenib-resistant HCC in patients.

YAP/TAZ and ATF4 drive resistance to Sorafenib in hepatocellular carcinoma by preventing ferroptosis

“…To identify the molecular mechanisms underlying therapy resistance in HCC, we had previously established Sorafenib‐resistant HCC cell lines, called Huh7‐IR and Huh7‐CR (IC50 of 10.7 and 10.8 µM, respectively) and Hep3B‐IR and Hep3B‐CR (IC50 of 7.2 and 8.3 µM, respectively), as compared to their Sorafenib‐sensitive parental cell lines Huh7 and Hep3B (IC50 of 1.7 and 3.0 µM, respectively) (Appendix Fig S1A–C ) (Tang et al , 2019 ; Gao et al , 2021 ). Global transcriptomic analysis between the Sorafenib‐sensitive parental cells and their Sorafenib‐resistant derivatives revealed that, in addition to changes in various signaling pathways, cellular metabolism pathways had dynamically shifted in HCC cells upon the establishment of Sorafenib resistance, including amino acid metabolism (Appendix Fig S1D ; Dataset [Link] , [Link] ).…”

The emerging role of deubiquitylating enzymes as therapeutic targets in cancer metabolism

“…Moreover, systematic knockout of Usp29 in MYC-driven animal models markedly decreased the expression of intratumoral MYC, HIF1α, and their key downstream metabolic targets [ 33 ]. Consistently, another group recently reported that USP29 promotes aerobic glycolysis via stabilizing HIF1α to mediate sorafenib resistance in HCC cell lines, suggesting that USP29 may play a key role in the regulation of aerobic glycolysis in different cancer types [ 34 ]. In NSCLC, OTUB2 (OTU deubiquitinase, ubiquitin aldehyde binding 2) was significantly upregulated in primary tissues and associated with tumor malignancy [ 35 ].…”

A genome‐wide CRISPR activation screen reveals Hexokinase 1 as a critical factor in promoting resistance to multi‐kinase inhibitors in hepatocellular carcinoma cells

“…These results are generally in line with other studies linking sorafenib resistance to enhanced glycolysis. [40][41][42] Interestingly, Gao et al 43 recently reported yellowing of the cell media in sorafenib-resistant Huh7 cells, indicating increased acidification caused by increased glycolysis and lactate excretion. Furthermore, this study showed an induction of HK2, but not HK1, in these cells at the RNA level.…”