CaRLO — Cancer Research Library Online

Hits per page

hits 1 - 5 | 5 hits

Sorting

Unknown

A conditional piggyBac transposition system for genetic screening in mice identifies oncogenic networks in pancreatic cancer

Rad R ; Rad L ; Wang W ; [et al.]

Nature Genetics 47 (1), 47-56

add to mindlist on the mindlist

Details

Keywords: GENOME ; T-CELLS ; MOUSE ; DROSOPHILA-MELANOGASTER ; EMBRYONIC STEM-CELLS ; SOMATIC MUTAGENESIS ; CORONARY-ARTERY-DISEASE ; next-generation ; SLEEPING-BEAUTY ; CHROMOSOMAL TRANSPOSITION

Abstract: Here we describe a conditional piggyBac transposition system in mice and report the discovery of large sets of new cancer genes through a pancreatic insertional mutagenesis screen. We identify Foxp1 as an oncogenic transcription factor that drives pancreatic cancer invasion and spread in a mouse model and correlates with lymph node metastasis in human patients with pancreatic cancer. The propensity of piggyBac for open chromatin also enabled genome-wide screening for cancer-relevant noncoding DNA, which pinpointed a Cdkn2a cis-regulatory region. Histologically, we observed different tumor subentities and discovered associated genetic events, including Fign insertions in hepatoid pancreatic cancer. Our studies demonstrate the power of genetic screening to discover cancer drivers that are difficult to identify by other approaches to cancer genome analysis, such as downstream targets of commonly mutated human cancer genes. These piggyBac resources are universally applicable in any tissue context and provide unique experimental access to the genetic complexity of cancer.

DOI: 10.1038/ng.3164

Type of Publication: Journal article published

PubMed ID: 25485836

Deep Link: http://www.dkfz.de/cgi-bin/sel?http://www.dkfz.de/PublicationManager/Show/ShowJournal.aspx%3fpublishedId=59324

Permalink

Signatur	Availability

Others were also interested in ...

DKFZ PUBLICATION

Unknown

Genome-wide transposon screening and quantitative insertion site sequencing for cancer gene discovery in mice

Friedrich MJ ; Rad L ; Bronner IF ; [et al.]

Nature Protocols 12 (2), 289-309

add to mindlist on the mindlist

Details

Abstract: Transposon-mediated forward genetics screening in mice has emerged as a powerful tool for cancer gene discovery. It pinpoints cancer drivers that are difficult to find with other approaches, thus complementing the sequencing-based census of human cancer genes. We describe here a large series of mouse lines for insertional mutagenesis that are compatible with two transposon systems, PiggyBac and Sleeping Beauty, and give guidance on the use of different engineered transposon variants for constitutive or tissue-specific cancer gene discovery screening. We also describe a method for semiquantitative transposon insertion site sequencing (QiSeq). The QiSeq library preparation protocol exploits acoustic DNA fragmentation to reduce bias inherent to widely used restriction-digestion-based approaches for ligation-mediated insertion site amplification. Extensive multiplexing in combination with next-generation sequencing allows affordable ultra-deep transposon insertion site recovery in high-throughput formats within 1 week. Finally, we describe principles of data analysis and interpretation for obtaining insights into cancer gene function and genetic tumor evolution.

Type of Publication: Journal article published

PubMed ID: 28079877

Deep Link: http://www.dkfz.de/cgi-bin/sel?http://www.dkfz.de/PublicationManager/Show/ShowJournal.aspx%3fpublishedId=78327

Permalink

Signatur	Availability

Others were also interested in ...

DKFZ PUBLICATION

Unknown

A single-copy Sleeping Beauty transposon mutagenesis screen identifies new PTEN-cooperating tumor suppressor genes

de la Rosa J ; Weber J ; Friedrich MJ ; [et al.]

Nature Genetics 49 (5), 730-741

add to mindlist on the mindlist

Details

Abstract: The overwhelming number of genetic alterations identified through cancer genome sequencing requires complementary approaches to interpret their significance and interactions. Here we developed a novel whole-body insertional mutagenesis screen in mice, which was designed for the discovery of Pten-cooperating tumor suppressors. Toward this aim, we coupled mobilization of a single-copy inactivating Sleeping Beauty transposon to Pten disruption within the same genome. The analysis of 278 transposition-induced prostate, breast and skin tumors detected tissue-specific and shared data sets of known and candidate genes involved in cancer. We validated ZBTB20, CELF2, PARD3, AKAP13 and WAC, which were identified by our screens in multiple cancer types, as new tumor suppressor genes in prostate cancer. We demonstrated their synergy with PTEN in preventing invasion in vitro and confirmed their clinical relevance. Further characterization of Wac in vivo showed obligate haploinsufficiency for this gene (which encodes an autophagy-regulating factor) in a Pten-deficient context. Our study identified complex PTEN-cooperating tumor suppressor networks in different cancer types, with potential clinical implications.

Type of Publication: Journal article published

PubMed ID: 28319090

Deep Link: http://www.dkfz.de/cgi-bin/sel?http://www.dkfz.de/PublicationManager/Show/ShowJournal.aspx%3fpublishedId=79209

Permalink

Signatur	Availability

Others were also interested in ...

DKFZ PUBLICATION

Unknown

Selective Requirement of PI3K/PDK1 Signaling for Kras Oncogene-Driven Pancreatic Cell Plasticity and Cancer

Eser S ; Reiff N ; Messer M ; [et al.]

Cancer Cell 23 (3), 406-420

add to mindlist on the mindlist

Details

Keywords: IN-VIVO ; MICE ; RAS ONCOGENE ; EGF RECEPTOR ; MOUSE MODEL ; TUMOR-SUPPRESSOR ; DUCTAL ADENOCARCINOMA ; TUMORIGENESIS ; PI3K ; C-RAF

Abstract: Oncogenic Kras activates a plethora of signaling pathways, but our understanding of critical Ras effectors is still very limited. We show that cell-autonomous phosphoinositide 3-kinase (PI3K) and 3-phosphoinositide-dependent protein kinase 1 (PDK1), but not Craf, are key effectors of oncogenic Kras in the pancreas, mediating cell plasticity, acinar-to-ductal metaplasia (ADM), and pancreatic ductal adenocarcinoma (PDAC) formation. This contrasts with Kras-driven non-small cell lung cancer, where signaling via Craf, but not PDK1, is an essential tumor-initiating event. These in vivo genetic studies together with pharmacologic treatment studies in models of human ADM and PDAC demonstrate tissue-specific differences of oncogenic Kras signaling and define PI3K/PDK1 as a suitable target for therapeutic intervention specifically in PDAC.

DOI: 10.1016/j.ccr.2013.01.023

Type of Publication: Journal article published

PubMed ID: 23453624

Deep Link: http://www.dkfz.de/cgi-bin/sel?http://www.dkfz.de/PublicationManager/Show/ShowJournal.aspx%3fpublishedId=39843

Permalink

Signatur	Availability

Others were also interested in ...

DKFZ PUBLICATION

Unknown

A Genetic Progression Model of Braf(V600E)-Induced Intestinal Tumorigenesis Reveals Targets for Therapeutic Intervention

Rad R ; Cadinanos J ; Rad L ; [et al.]

Cancer Cell 24 (1), 15-29

add to mindlist on the mindlist

Details

Keywords: COLORECTAL-CANCER ; COLON-CANCER ; KRAS MUTATIONS ; HYPERPLASTIC POLYPS ; ONCOGENIC K-RAS ; MEK INHIBITION ; EARLY NEOPLASTIC PROGRESSION ; SESSILE SERRATED ADENOMAS ; SENESCENCE IN-VIVO ; HIGH-GRADE TUMORS

Abstract: We show that BRAF(V600E) initiates an alternative pathway to colorectal cancer (CRC), which progresses through a hyperplasia/adenoma/carcinoma sequence. This pathway underlies significant subsets of CRCs with distinctive pathomorphologic/genetic/epidemiologic/clinical characteristics. Genetic and functional analyses in mice revealed a series of stage-specific molecular alterations driving different phases of tumor evolution and uncovered mechanisms underlying this stage specificity. We further demonstrate dose-dependent effects of oncogenic signaling, with physiologic Braf(V600E) expression being sufficient for hyperplasia induction, but later stage intensified Mapk-signaling driving both tumor progression and activation of intrinsic tumor suppression. Such phenomena explain, for example, the inability of p53 to restrain tumor initiation as well as its importance in invasiveness control, and the late stage specificity of its somatic mutation. Finally, systematic drug screening revealed sensitivity of this CRC subtype to targeted therapeutics, including Mek or combinatorial PI3K/Braf inhibition.

DOI: 10.1016/j.ccr.2013.05.014

Type of Publication: Journal article published

PubMed ID: 23845441

Deep Link: http://www.dkfz.de/cgi-bin/sel?http://www.dkfz.de/PublicationManager/Show/ShowJournal.aspx%3fpublishedId=41723

Permalink

Signatur	Availability

Others were also interested in ...

DKFZ PUBLICATION

hits 1 - 5 | 5 hits