title: Comprehensive Ex Vivo Transposon Mutagenesis Identifies Genes That Promote Growth Factor Independence and Leukemogenesis.
abstract: 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.
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Yabin
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Barrett L
authors.initials: BL
authors.lastname: Updegraff
authors.affiliation: Department of Clinical Sciences, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Sunho
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Deniz
authors.initials: D
authors.lastname: Durakoglugil
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Victoria H
authors.initials: VH
authors.lastname: Cruz
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Sarah
authors.initials: S
authors.lastname: Maddux
authors.affiliation: Department of Clinical Sciences, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Tae Hyun
authors.initials: TH
authors.lastname: Hwang
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas. Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas. Kathryn.ODonnell@UTSouthwestern.edu.
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: O'Donnell
mesh_terms: D000818:Animals; D063646:Carcinogenesis / Q000235:genetics*; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000473:pathology; D051379:Mice; D016296:Mutagenesis; D015398:Signal Transduction