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

Roland Rad; Lena Rad; Wei Wang; Alexander Strong; Hannes Ponstingl; Iraad F Bronner; Matthew Mayho; Katja Steiger; Julia Weber; Maren Hieber; Christian Veltkamp; Stefan Eser; Ulf Geumann; Rupert Öllinger; Magdalena Zukowska; Maxim Barenboim; Roman Maresch; Juan Cadiñanos; Mathias Friedrich; Ignacio Varela; Fernando Constantino-Casas; Aaron Sarver; Jelle Ten Hoeve; Haydn Prosser; Barbara Seidler; Judith Bauer; Mathias Heikenwälder; Emmanouil Metzakopian; Anne Krug; Ursula Ehmer; Günter Schneider; Thomas Knösel; Petra Rümmele; Daniela Aust; Robert Grützmann; Christian Pilarsky; Zemin Ning; Lodewyk Wessels; Roland M Schmid; Michael A Quail; George Vassiliou; Irene Esposito; Pentao Liu; Dieter Saur; Allan Bradley

doi:10.1038/ng.3164

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

Nat Genet. 2015 Jan;47(1):47-56. doi: 10.1038/ng.3164. Epub 2014 Dec 8.

Authors

Roland Rad¹, Lena Rad², Wei Wang², Alexander Strong², Hannes Ponstingl², Iraad F Bronner², Matthew Mayho², Katja Steiger³, Julia Weber⁴, Maren Hieber⁵, Christian Veltkamp⁵, Stefan Eser⁵, Ulf Geumann⁴, Rupert Öllinger⁵, Magdalena Zukowska⁵, Maxim Barenboim⁴, Roman Maresch⁴, Juan Cadiñanos⁶, Mathias Friedrich², Ignacio Varela⁷, Fernando Constantino-Casas⁸, Aaron Sarver⁹, Jelle Ten Hoeve¹⁰, Haydn Prosser², Barbara Seidler⁵, Judith Bauer¹¹, Mathias Heikenwälder¹¹, Emmanouil Metzakopian², Anne Krug⁵, Ursula Ehmer⁵, Günter Schneider⁵, Thomas Knösel¹², Petra Rümmele¹³, Daniela Aust¹⁴, Robert Grützmann¹⁵, Christian Pilarsky¹⁵, Zemin Ning², Lodewyk Wessels¹⁰, Roland M Schmid⁵, Michael A Quail², George Vassiliou², Irene Esposito¹⁶, Pentao Liu², Dieter Saur⁴, Allan Bradley²

Affiliations

¹ 1] Department of Medicine II, Klinikum Rechts der Isar, Technische Universität München, München, Germany. [2] German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany. [3] The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridgeshire, UK.
² The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridgeshire, UK.
³ Department of Pathology, Klinikum Rechts der Isar, Technische Universität München, München, Germany.
⁴ 1] Department of Medicine II, Klinikum Rechts der Isar, Technische Universität München, München, Germany. [2] German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany.
⁵ Department of Medicine II, Klinikum Rechts der Isar, Technische Universität München, München, Germany.
⁶ Instituto de Medicina Oncológica y Molecular de Asturias (IMOMA), Oviedo, Spain.
⁷ Instituto de Biomedicina y Biotecnología de Cantabria (UC-CSIC-SODERCAN), Santander, Spain.
⁸ Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
⁹ Biostatistics and Bioinformatics Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA.
¹⁰ Bioinformatics and Statistics, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
¹¹ Institute of Virology, Technische Universität München, Munich, Germany.
¹² Institute of Pathology, Ludwig Maximilians Universität München, München, Germany.
¹³ Institute of Pathology, Universität Regensburg, Regensburg, Germany.
¹⁴ Institute of Pathology, Technische Universität Dresden, Dresden, Germany.
¹⁵ Department of Surgery, Technische Universität Dresden, Dresden, Germany.
¹⁶ Institute of Pathology, Medizinische Universität Insbruck, Insbruck, Austria.

PMID: 25485836
DOI: 10.1038/ng.3164

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Cell Transformation, Neoplastic / genetics*
DNA Transposable Elements / genetics*
Forkhead Transcription Factors / analysis
Forkhead Transcription Factors / antagonists & inhibitors
Forkhead Transcription Factors / genetics
Gene Expression Profiling
Gene Expression Regulation
Gene Knock-In Techniques
Gene Regulatory Networks*
Genes, Synthetic
Genes, p16
Humans
Mice
Mice, Transgenic
Molecular Sequence Data
Moths / genetics
Mutagenesis, Insertional*
Neoplasm Proteins / antagonists & inhibitors
Neoplasm Proteins / genetics
Pancreatic Neoplasms / chemistry
Pancreatic Neoplasms / genetics*
Pancreatic Neoplasms / pathology
Proton-Translocating ATPases / genetics
RNA, Small Interfering / pharmacology
Repressor Proteins / analysis
Repressor Proteins / antagonists & inhibitors
Repressor Proteins / genetics
Transgenes
Transposases / genetics
Transposases / physiology

Substances

DNA Transposable Elements
FOXP1 protein, human
Forkhead Transcription Factors
Foxp1 protein, mouse
Foxp2 protein, mouse
Neoplasm Proteins
RNA, Small Interfering
Repressor Proteins
Transposases
Proton-Translocating ATPases

Associated data

BioProject/PRJNA266573
SRA/SRP049610

Grants and funding

095663/Wellcome Trust/United Kingdom