A conditional transposon-based insertional mutagenesis screen for genes associated with mouse hepatocellular carcinoma

Keng, Vincent W.; Villanueva, Augusto; Chiang, Derek Y.; Dupuy, Adam J.; Ryan, Barbara J.; Matise, Ilze; Silverstein, Kevin A.T.; Sarver, Aaron L.; Starr, Timothy K.; Akagi, Keiko; Tessarollo, Lino; Collier, Lara S.; Powers, Scott; Lowe, Scott W.; Jenkins, Nancy A.; Copeland, Neal G.; Llovet, Josep M.; Largaespada, David A.

doi:10.1038/nbt.1526

Cited by 194 publications

(260 citation statements)

References 51 publications

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“…Recently, this approach has been modified through the conditional activation of the SB transposase in specific tissues using Cre/LoxP technology (5,6). In one of these studies, an SB screen identified 19 genes that accelerate liver cancer induced by expression of a dominant-negative Trp53 transgene.…”

mentioning

confidence: 99%

A Sleeping Beauty mutagenesis screen reveals a tumor suppressor role for Ncoa2/Src-2 in liver cancer

O’Donnell

Keng

York

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The Sleeping Beauty (SB) transposon mutagenesis system is a powerful tool that facilitates the discovery of mutations that accelerate tumorigenesis. In this study, we sought to identify mutations that cooperate with MYC, one of the most commonly dysregulated genes in human malignancy. We performed a forward genetic screen with a mouse model of MYC-induced liver cancer using SBmediated mutagenesis. We sequenced insertions in 63 liver tumor nodules and identified at least 16 genes/loci that contribute to accelerated tumor development. RNAi-mediated knockdown in a liver progenitor cell line further validate three of these genes, Ncoa2/ Src-2, Zfx, and Dtnb, as tumor suppressors in liver cancer. Moreover, deletion of Ncoa2/Src-2 in mice predisposes to diethylnitrosamineinduced liver tumorigenesis. These findings reveal genes and pathways that functionally restrain MYC-mediated liver tumorigenesis and therefore may provide targets for cancer therapy. T ransposable elements (TEs) are powerful genetic tools that are widely used in insertional mutagenesis screens because of the facile identification of transposon-induced mutations. The application of transposon-based approaches to cancer gene identification provides opportunities to study the consequences of specific mutations in the context of tumor cell initiation, progression, and maintenance in well-defined, genetically engineered mouse models. Sleeping Beauty (SB), a member of the Tc1/mariner superfamily of DNA transposons, is highly active in mammalian cells (1). A growing body of evidence has demonstrated that SB is an efficient tool for cancer gene discovery when used in forward genetic screens in mice (2, 3).The SB system is based on the use of two transgenic mouse lines, one harboring a transposase and the other with a concatemerized mutagenic transposon containing sequences that can disrupt gene function through either gain-of-function or loss-offunction mechanisms. The transposase binds to the transposon ends and catalyzes its mobilization to new sites. The effect of ubiquitous transposition in mice is the development of T-cell leukemia and brain tumors (3, 4). In contrast, restricted expression of SB transposase accelerates fibrosarcoma development in p19 Arf −/− mice (2).Recently, this approach has been modified through the conditional activation of the SB transposase in specific tissues using Cre/LoxP technology (5, 6). In one of these studies, an SB screen identified 19 genes that accelerate liver cancer induced by expression of a dominant-negative Trp53 transgene. However, the SB system has not been used previously to identify genes that cooperate with oncogenes that initiate liver tumorigenesis.Hepatocellular carcinoma (HCC) is the fifth most common solid tumor worldwide and is the third leading cause of death from cancer (7,8). In the majority of cases, it occurs in a setting of chronic inflammation or cirrhosis. Although numerous genomic alterations have been documented in liver cancer, the key genetic alterations that drive hepatocellular transforma...

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mentioning

confidence: 99%

A Sleeping Beauty mutagenesis screen reveals a tumor suppressor role for Ncoa2/Src-2 in liver cancer

O’Donnell

Keng

York

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Indeed, transposons can be remobilized from chromosomally resident loci and reintegrated somewhere else in the target genome by providing the transposase transiently (Geurts et al, 2006;Keng et al, 2009). SB transposon has been successfully mobilized in mouse somatic cells at frequencies high enough to induce tumors .…”

Section: Discussionmentioning

confidence: 99%

Effective Expression-Independent Gene Trapping and Mutagenesis Mediated by Sleeping Beauty Transposon

Song

Long

Hackett

2012

Journal of Genetics and Genomics

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“…Much is known about the signals and mechanisms of regeneration, both derived from the ligant/ receiver system, and from the whole body, including circulatory events and signals from other organs. Hepatocellular carcinoma study suggested that there may be more mechanisms contributing to the regulation of hepatocyte growth than those currently known 11 .…”

Section: Introductionmentioning

confidence: 92%

The ileum positively regulates hepatic regeneration in rats

et al. 2014

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PURPOSE:To test the hypothesis that liver regeneration after partial hepatectomy can be influenced by the ileum. METHODS:Eighteen Wistar rats were distributed into groups of six animals: 1 -ileum resection+ hepatectomy 2/3; 2 -hepatectomy 2/3, and 3 -sham. Anesthesia with ketamine and xylazine i.p., aseptic technique, analgesia with meperidine (10mg/kg s.c.). On day 6, serum ALT, AST, alkaline phosphatase (AP) and albumin were measured. Liver regeneration and hepatocyte mitosis were quantified.Statistical analysis with ANOVA and Tukey tests, with significance p<0.05. RESULTS:In group hepatectomy+ileal resection, ALT, AST and AP were 180.6±24.9, 58.6±3.1 and 254.6±46.6 respectively. They were significantly higher than in the hepatectomy group, whose values were 126.0±16.5, 44.1±3.9 and 163.5±8.6, respectively (p<0.001).Albumin levels were not significantly different among groups. Liver regeneration in hepatectomy group (94.17%) was statistically higher (p<0.001) than in ileal resection+hepatectomy group (55.96%). In the latter group the mitosis of hepatocytes were significantly less frequent than in the hepatectomy group. CONCLUSION:The data confirm that the ileum positively influence on liver regeneration in rats undergoing hepatectomy.

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A conditional transposon-based insertional mutagenesis screen for genes associated with mouse hepatocellular carcinoma

Cited by 194 publications

References 51 publications

A Sleeping Beauty mutagenesis screen reveals a tumor suppressor role for Ncoa2/Src-2 in liver cancer

A Sleeping Beauty mutagenesis screen reveals a tumor suppressor role for Ncoa2/Src-2 in liver cancer

Effective Expression-Independent Gene Trapping and Mutagenesis Mediated by Sleeping Beauty Transposon

The ileum positively regulates hepatic regeneration in rats

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