title: Human somatic cell mutagenesis creates genetically tractable sarcomas.
abstract: Creating spontaneous yet genetically tractable human tumors from normal cells presents a fundamental challenge. Here we combined retroviral and transposon insertional mutagenesis to enable cancer gene discovery starting with human primary cells. We used lentiviruses to seed gain- and loss-of-function gene disruption elements, which were further deployed by Sleeping Beauty transposons throughout the genome of human bone explant mesenchymal cells. De novo tumors generated rapidly in this context were high-grade myxofibrosarcomas. Tumor insertion sites were enriched in recurrent somatic copy-number aberration regions from multiple cancer types and could be used to pinpoint new driver genes that sustain somatic alterations in patients. We identified HDLBP, which encodes the RNA-binding protein vigilin, as a candidate tumor suppressor deleted at 2q37.3 in greater than one out of ten tumors across multiple tissues of origin. Hybrid viral-transposon systems may accelerate the functional annotation of cancer genomes by enabling insertional mutagenesis screens in higher eukaryotes that are not amenable to germline transgenesis.
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Sam D
authors.initials: SD
authors.lastname: Molyneux
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Waterhouse
authors.affiliation: Digital-biology Center, Bio-Rad Laboratories, Pleasanton, California, USA.
authors.forename: Dawne
authors.initials: D
authors.lastname: Shelton
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Yang W
authors.initials: YW
authors.lastname: Shao
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Watling
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Qing-Lian
authors.initials: QL
authors.lastname: Tang
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Isaac S
authors.initials: IS
authors.lastname: Harris
authors.affiliation: Department of Pathology and Division of Orthopaedic Surgery, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: Brendan C
authors.initials: BC
authors.lastname: Dickson
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Pirashaanthy
authors.initials: P
authors.lastname: Tharmapalan
authors.affiliation: Department of Informatics, University of Oslo, Blindern, Oslo, Norway.
authors.forename: Geir K
authors.identifier: 0000000249591409
authors.initials: GK
authors.lastname: Sandve
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Swneke D
authors.initials: SD
authors.lastname: Bailey
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Hal
authors.initials: H
authors.lastname: Berman
authors.affiliation: Department of Pathology and Division of Orthopaedic Surgery, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: Jay S
authors.initials: JS
authors.lastname: Wunder
authors.affiliation: Max-Delbruck-Center for Molecular Medicine, Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: Max-Delbruck-Center for Molecular Medicine, Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Iszvak
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Lupien
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Tak W
authors.initials: TW
authors.lastname: Mak
authors.affiliation: Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Rama
authors.initials: R
authors.lastname: Khokha
mesh_terms: D002460:Cell Line; D004251:DNA Transposable Elements; D005822:Genetic Vectors / Q000235:genetics; D015894:Genome, Human; D057809:HEK293 Cells; D006801:Humans; D016254:Mutagenesis, Insertional; D016601:RNA-Binding Proteins / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D012509:Sarcoma / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; D016601:RNA-Binding Proteins; C075197:high density lipoprotein binding protein