Abstract
The p53 mutation and altered Pten expression are two most common genetic events in Hepatitis B virus (HBV) infection related hepatocellular carcinoma (HCC). To confirm the causative role of p53 and Pten somatic mutation in HCC development, we established CRISPR/Cas9-mediated somatic gene disruption via hydrodynamic tail vein injection, allowing for in vivo targeting p53 and Pten simultaneously in adult HBV transgenic mice. Here we demonstrated that the utility of this approach resulted in macroscopic liver tumors as early as 4 months’ post injection and most tumors harbored both p53 and Pten loss-of-function alterations. Immunohistochemical (IHC) and histopathology analysis demonstrated that the tumors were positive for Glutamine synthetase (GS), a marker of HCC and accompanied with prominent lipid accumulation. The study here indicated that CRISPR/Cas9-mediated p53 and Pten somatic mutation accelerated hepatocarcinogenesis in adult HBV transgenic mice. This method also provides a fast and convenient system for generating mouse model of HCC with HBV infection characteristics.
Highlights
Hepatocellular carcinoma (HCC) is one of the most common liver cancer and becoming the second leading cause of cancer-related deaths in the world[1]
Compared with tumors associated with other risk factors including hepatitis C virus (HCV) infection, exposition to aflatoxin B1, alcohol consumption and metabolic diseases, Hepatitis B virus (HBV)-related liver tumors have a higher rate (32–47%) of p53 inactive mutations[4,5,6,7,8]
We showed that CRISPR/Cas[9] system delivered by hydrodynamic tail vein injection can be used to somatically induce p53 and phosphatase and tensin homolog (Pten) mutation in the liver of HBV-transgenic mice, leading to the induction of hepatocellular carcinoma (HCC) as early as 4 months’ post injection
Summary
Hepatocellular carcinoma (HCC) is one of the most common liver cancer and becoming the second leading cause of cancer-related deaths in the world[1]. Xue et al Showed that hydrodynamic injection of the CRISPR/ Cas[9] system to target p53 and Pten genes in adult mice could yield cancer-related phenotypes in liver after carbon tetrachloride treatment[22]. They showed that hydrodynamic injection can deliver plasmid DNA to only about 20% of hepatocytes in mice, the CRISPR/Cas9–mediated genome editing is sufficient to induce multifocal tumors in adult mouse liver. We showed that CRISPR/Cas[9] system delivered by hydrodynamic tail vein injection can be used to somatically induce p53 and Pten mutation in the liver of HBV-transgenic mice, leading to the induction of HCC as early as 4 months’ post injection. This study provided a fast and convenient method for generating mouse model of liver cancer with HBV infection characteristics
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