Whole-Body <i>Sleeping Beauty</i> Mutagenesis Can Cause Penetrant Leukemia/Lymphoma and Rare High-Grade Glioma without Associated Embryonic Lethality

Collier, Lara S.; Adams, David J.; Hackett, Christopher S.; Bendzick, Laura; Akagi, Keiko; Davies, Michael N.; Diers, Miechaleen D.; Rodríguez, Fausto J.; Bender, Aaron M.; Tieu, Christina; Matise, Ilze; Dupuy, Adam J.; Copeland, Neal G.; Jenkins, Nancy A.; Hodgson, J. Graeme; Weiss, William A.; Jenkins, Robert B.; Largaespada, David A.

doi:10.1158/0008-5472.can-09-1760

Cited by 71 publications

(75 citation statements)

References 34 publications

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“…Despite the small number of lymphomas analyzed, two of the genes with SB insertions, Myb and Erg, had been found previously as CISs that contribute to leukemias/lymphomas in a larger cohort of 58 T2/Onc; Rosa26-SB11 animals (4). In contrast, none of the lymphoma CIS genes from the Collier et al study (4) overlapped with liver CIS genes identified in the present study. Interestingly, the only CIS that overlapped between our lymphoma-and liver tumor-sequencing datasets was Ncoa2 (Fig.…”

Section: Sb Insertions In Lymphomas Are Distinct From Liver Tumor Inscontrasting

confidence: 97%

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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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Section: Sb Insertions In Lymphomas Are Distinct From Liver Tumor Inscontrasting

confidence: 97%

“…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).…”

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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“…Functional genomic strategies have been routinely used to identify genes important in driving cancer initiation, progression, and therapeutic resistance, which ultimately complements large-scale sequencing strategies. Implementation of gene insertion strategies, including the sleeping beauty transposon method, have led to the identification of several novel cancer genes (6)(7)(8). We hypothesize that nontransformed astrocytes harboring alterations in relevant glioblastoma pathways, including the Ras or p53 signaling, can be transformed using random retroviral insertions to inactivate genes involved in initiation and or progression.…”

Section: Introductionmentioning

confidence: 99%

PINK1 Is a Negative Regulator of Growth and the Warburg Effect in Glioblastoma

et al. 2016

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Proliferating cancer cells are characterized by high rates of glycolysis, lactate production, and altered mitochondrial metabolism. This metabolic reprogramming provides important metabolites for proliferation of tumor cells, including glioblastoma. These biological processes, however, generate oxidative stress that must be balanced through detoxification of reactive oxygen species (ROS). Using an unbiased retroviral loss-of-function screen in nontransformed human astrocytes, we demonstrate that mitochondrial PTEN-induced kinase 1 (PINK1) is a regulator of the Warburg effect and negative regulator of glioblastoma growth. We report that loss of PINK1 contributes to the Warburg effect through ROS-dependent stabilization of hypoxia-inducible factor-1A and reduced pyruvate kinase muscle isozyme 2 activity, both key regulators of aerobic glycolysis. Mechanistically, PINK1 suppresses ROS and tumor growth through FOXO3a, a master regulator of oxidative stress and superoxide dismutase 2. These findings highlight the importance of PINK1 and ROS balance in normal and tumor cells. PINK1 loss was observed in a significant number of human brain tumors including glioblastoma (n > 900) and correlated with poor patient survival. PINK1 overexpression attenuates in vivo glioblastoma growth in orthotopic mouse xenograft models and a transgenic glioblastoma model in Drosophila.

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“…Transposon-mediated mutagenesis has emerged as a powerful methodology for functionally annotating cancer genomes. A number of studies have utilized transposon-based insertional mutagenesis screens in mice to identify and validate a number of genes relevant to tumorigenesis, including T-cell leukemia, hepatocellular carcinoma, medulloblastoma, and nerve sheath tumors (14)(15)(16)(17)(18)(19)(20)(21). To date, a vast majority of studies have relied upon the mobilization of mutagenic DNA transposons such as Sleeping Beauty (SB) or piggyBac in transgenic mouse models.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Ex Vivo Transposon Mutagenesis Identifies Genes That Promote Growth Factor Independence and Leukemogenesis

et al. 2016

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Aberrant signaling through cytokine receptors and their downstream signaling pathways is a major oncogenic mechanism underlying hematopoietic malignancies. To better understand how these pathways become pathologically activated and to potentially identify new drivers of hematopoietic cancers, we developed a high-throughput functional screening approach using ex vivo mutagenesis with the Sleeping Beauty transposon. We analyzed over 1,100 transposon-mutagenized pools of Ba/F3 cells, an IL3-dependent pro-B-cell line, which acquired cytokine independence and tumor-forming ability. Recurrent transposon insertions could be mapped to genes in the JAK/ STAT and MAPK pathways, confirming the ability of this strategy to identify known oncogenic components of cytokine signaling pathways. In addition, recurrent insertions were identified in a large set of genes that have been found to be mutated in leukemia or associated with survival, but were not previously linked to the JAK/STAT or MAPK pathways nor shown to functionally contribute to leukemogenesis. Forced expression of these novel genes resulted in IL3-independent growth in vitro and tumorigenesis in vivo, validating this mutagenesis-based approach for identifying new genes that promote cytokine signaling and leukemogenesis. Therefore, our findings provide a broadly applicable approach for classifying functionally relevant genes in diverse malignancies and offer new insights into the impact of cytokine signaling on leukemia development.

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Whole-Body Sleeping Beauty Mutagenesis Can Cause Penetrant Leukemia/Lymphoma and Rare High-Grade Glioma without Associated Embryonic Lethality

Cited by 71 publications

References 34 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

PINK1 Is a Negative Regulator of Growth and the Warburg Effect in Glioblastoma

Comprehensive Ex Vivo Transposon Mutagenesis Identifies Genes That Promote Growth Factor Independence and Leukemogenesis

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