Genome-wide CRISPR screen reveals SGOL1 as a druggable target of sorafenib-treated hepatocellular carcinoma

Abstract

The genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screen is a powerful tool used to identify therapeutic targets that can be harnessed for cancer treatment. This study aimed to assess the efficacy of genome-wide CRISPR screening to identify druggable genes associated with sorafenib-treated hepatocellular carcinoma (HCC). A genome-scale CRISPR knockout (GeCKO v2) library containing 123,411 single guide RNAs (sgRNAs) was used to identify loss-of-function mutations conferring sorafenib resistance upon HCC cells. Resistance gene screens identified SGOL1 as an indicator of prognosis of patients treated with sorafenib. Of the 19,050 genes tested, the knockout screen identified inhibition of SGOL1 expression as the most-effective genetic suppressor of sorafenib activity. Analysis of the survival of 210 patients with HCC after hepatic resection revealed that high SGOL1 expression shortened overall survival (P = 0.021). Further, matched pairs analysis of the TCGA database revealed that SGOL1 is differentially expressed. When we used a lentivirus Cas9 vector to determine the effect of targeting SGOL1 with a specific sgRNA in HCC cells, we found that SGOL1 expression was efficiently inhibited and that loss of SGOL1 was associated with sorafenib resistance. Further, loss of SGOL1 from HCC cell decreased the cytotoxicity of sorafenib in vivo. We conclude that the CRISPR screen is a powerful tool for therapeutic target analysis of sorafenib treatment and that SGOL1 serves as a druggable target for HCC treated with sorafenib and an indicator of prognosis.

CRISPR/Cas9 for overcoming drug resistance in solid tumors

“…A genome wide CRISPR screening study revealed that loss of SGOL1, a protein involved in mitosis, is the main cause of resistance to sorafenib in HuH7 and SMMC-7721 HCC cells. Further in vivo studies showed decreased sorafenib cytotoxicity and increased tumor size supporting contribution of SGOL1 to sorafenib resistance [46]. These findings suggest that SGOL1 might be a suitable therapeutic target for HCC patients and individuals with SGOL1 overexpression might be appropriate candidates to be treated with sorafenib.…”

CRISPR/Cas: From Tumor Gene Editing to T Cell-Based Immunotherapy of Cancer

“…sgRNAs in the cells affect their competence for viability during the proliferation. The enrichment or depletion of these sgRNAs thereupon reveals the genes responsible for the cognate phenotype (82)(83)(84)(85)(86)(87)(88).…”

Delivery Approaches for Therapeutic Genome Editing and Challenges

“…The discovery of the host factors and receptor for Norovirus, the leading cause of gastroenteritis, was the result of CRISPR screens using LVs for the delivery of the sgRNA library [ 155 ]. Similarly, Sun et al identified new drug targets conferring sorafenib resistance in the treatment of hepatocellular carcinoma [ 156 ].…”