Sequencing a mouse acute promyelocytic leukemia genome reveals genetic events relevant for disease progression
Lukas D. Wartman, … , Richard K. Wilson, Timothy J. Ley
Lukas D. Wartman, … , Richard K. Wilson, Timothy J. Ley
Published March 23, 2011
Citation Information: J Clin Invest. 2011;121(4):1445-1455. https://doi.org/10.1172/JCI45284.
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Sequencing a mouse acute promyelocytic leukemia genome reveals genetic events relevant for disease progression

Abstract

Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML). It is characterized by the t(15;17)(q22;q11.2) chromosomal translocation that creates the promyelocytic leukemia–retinoic acid receptor α (PML-RARA) fusion oncogene. Although this fusion oncogene is known to initiate APL in mice, other cooperating mutations, as yet ill defined, are important for disease pathogenesis. To identify these, we used a mouse model of APL, whereby PML-RARA expressed in myeloid cells leads to a myeloproliferative disease that ultimately evolves into APL. Sequencing of a mouse APL genome revealed 3 somatic, nonsynonymous mutations relevant to APL pathogenesis, of which 1 (Jak1 V657F) was found to be recurrent in other affected mice. This mutation was identical to the JAK1 V658F mutation previously found in human APL and acute lymphoblastic leukemia samples. Further analysis showed that JAK1 V658F cooperated in vivo with PML-RARA, causing a rapidly fatal leukemia in mice. We also discovered a somatic 150-kb deletion involving the lysine (K)-specific demethylase 6A (Kdm6a, also known as Utx) gene, in the mouse APL genome. Similar deletions were observed in 3 out of 14 additional mouse APL samples and 1 out of 150 human AML samples. In conclusion, whole genome sequencing of mouse cancer genomes can provide an unbiased and comprehensive approach for discovering functionally relevant mutations that are also present in human leukemias.

Authors

Lukas D. Wartman, David E. Larson, Zhifu Xiang, Li Ding, Ken Chen, Ling Lin, Patrick Cahan, Jeffery M. Klco, John S. Welch, Cheng Li, Jacqueline E. Payton, Geoffrey L. Uy, Nobish Varghese, Rhonda E. Ries, Mieke Hoock, Daniel C. Koboldt, Michael D. McLellan, Heather Schmidt, Robert S. Fulton, Rachel M. Abbott, Lisa Cook, Sean D. McGrath, Xian Fan, Adam F. Dukes, Tammi Vickery, Joelle Kalicki, Tamara L. Lamprecht, Timothy A. Graubert, Michael H. Tomasson, Elaine R. Mardis, Richard K. Wilson, Timothy J. Ley

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Figure 3

Functional validation of the recurrent Jak1 mutation identified in the sequenced mouse APL genome.

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Functional validation of the recurrent Jak1 mutation identified in the s...
(A) The mouse Jak1 V657F (mJAK1) mutation is orthologous to the human JAK1 V658F (hJAK1) and human JAK2 V617F (hJAK2) mutations. B41, band 4.1 domain; SH2, src-homology domain; JH2, pseudokinase domain; JH1, kinase domain. (B) Mouse APL Jak1 V657F mutation allele frequency by 454 sequencing. The variant allele frequencies of the V657F mutation from tumor 9500 and the 6 other tumors with V657F mutations are shown. On the far right are the read counts for the controls of a WT 129/SvJ DNA pool (n = 6) and a WT B6/T DNA pool (n = 6). F, female; M, male. (C) Functional validation with human JAK1 cDNA MSCV retroviral constructs. IRES-GFP (WT/GFP) alone versus JAK1 WT-IRES-GFP (WT/JAK1 WT) versus JAK1 V658F (WT/JAK1 V658F) mutant-IRES-GFP retroviral constructs were transduced into WT B6/T or mCG-PR bone marrow from 6-week-old mice. The transduced marrow was transplanted into lethally irradiated WT B6/T mice. The JAK1 V658F mutant cooperates with PML-RARA (PR) in a mouse model of APL and causes a fatal leukemia; a Kaplan-Meier overall survival plot is shown. This result was replicated in 2 independent experiments. (D) Serial flow cytometric analysis of peripheral blood shows a massive expansion of GFP+/Gr-1+ cells in the mCG-PR/JAK1 V658F cohort compared with that in the other experimental cohorts and controls. Error bars are mean ± SEM. (E) Flow cytometric analysis of peripheral blood, bone marrow, and spleen at the end of 1 JAK1 functional validation experiment showed no significant expansion of GFP+/Gr-1+ cells in cohorts other than mCG-PR/JAK1 V658F mice. Each symbol represents an individual mouse. Error bars are mean ± SEM.