Signal transducer and activator of transcription 5 (STAT5) activation occurs frequently in human progenitor B-cell acute lymphoblastic leukemia (B-ALL). To identify gene alterations that cooperate with STAT5 activation to initiate leukemia, we crossed mice expressing a constitutively active form of STAT5 (Stat5b-CA) with mice in which a mutagenic Sleeping Beauty transposon (T2/Onc) was mobilized only in B cells. Stat5b-CA mice typically do not develop B-ALL (<2% penetrance); in contrast, 89% of Stat5b-CA mice in which the T2/Onc transposon had been mobilized died of B-ALL by 3 months of age. High-throughput sequencing approaches were used to identify genes frequently targeted by the T2/Onc transposon; these included Sos1 (74%), Kdm2a (35%), Jak1 (26%), Bmi1 (19%), Prdm14 or Ncoa2 (13%), Cdkn2a (10%), Ikzf1 (8%), Caap1 (6%) and Klf3 (6%). Collectively, these mutations target three major cellular processes: (i) the Janus kinase/STAT5 pathway (ii) progenitor B-cell differentiation and (iii) the CDKN2A tumor-suppressor pathway. Transposon insertions typically resulted in altered expression of these genes, as well as downstream pathways including STAT5, extracellular signal-regulated kinase (Erk) and p38. Importantly, expression of Sos1 and Kdm2a, and activation of p38, correlated with survival, further underscoring the role these genes and associated pathways have in B-ALL.
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We thank Peter Schoettler, Chris Reis, Alyssa Kne, Amy Mack and Emilea Sykes for assistance with animal husbandry, Maya Raghunandan for technical assistance, Paul Champoux for assistance with Flow cytometry and Dr Richard Williams for helpful comments. This work was supported by a Brainstorm grant from the University of Minnesota Masonic Cancer Center (MAF and DAL), NIH grants R01 CA154998 and CA151845 to MAF, NIH grant 5R00CA151672-04 to TKS, NIH R01 CA113636 to DAL, an NIH Institutional Shared Resource grant to the Masonic Cancer Center P30-CA77598 and a Leukemia and Lymphoma Society Scholar award (MAF).
LMH-H, DAL and MAF designed research; LMH-H, JDL, TKS and GKH performed experiments and analyzed data; ALS and MAF analyzed data, LMH-H and MAF wrote the paper; and all authors critically reviewed and edited the paper.
DAL has ownership interest (including patents) in Discovery Genomics, Inc., and NeoClone Biotechnologies International and is also a consultant/advisory board member of Discovery Genomics, Inc., and NeoClone Biotechnologies International.
Supplementary Information accompanies this paper on the Oncogene website
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Heltemes-Harris, L., Larson, J., Starr, T. et al. Sleeping Beauty transposon screen identifies signaling modules that cooperate with STAT5 activation to induce B-cell acute lymphoblastic leukemia. Oncogene 35, 3454–3464 (2016). https://doi.org/10.1038/onc.2015.405
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