Abstract
Mutations in genes encoding splicing factors (which we refer to as spliceosomal genes) are commonly found in patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML)1,2,3. These mutations recurrently affect specific amino acid residues, leading to perturbed normal splice site and exon recognition4,5,6. Spliceosomal gene mutations are always heterozygous and rarely occur together with one another, suggesting that cells may tolerate only a partial deviation from normal splicing activity. To test this hypothesis, we engineered mice to express a mutated allele of serine/arginine-rich splicing factor 2 (Srsf2P95H)—which commonly occurs in individuals with MDS and AML—in an inducible, hemizygous manner in hematopoietic cells. These mice rapidly succumbed to fatal bone marrow failure, demonstrating that Srsf2-mutated cells depend on the wild-type Srsf2 allele for survival. In the context of leukemia, treatment with the spliceosome inhibitor E7107 (refs. 7,8) resulted in substantial reductions in leukemic burden, specifically in isogenic mouse leukemias and patient-derived xenograft AMLs carrying spliceosomal mutations. Whereas E7107 treatment of mice resulted in widespread intron retention and cassette exon skipping in leukemic cells regardless of Srsf2 genotype, the magnitude of splicing inhibition following E7107 treatment was greater in Srsf2-mutated than in Srsf2-wild-type leukemia, consistent with the differential effect of E7107 on survival. Collectively, these data provide genetic and pharmacologic evidence that leukemias with spliceosomal gene mutations are preferentially susceptible to additional splicing perturbations in vivo as compared to leukemias without such mutations. Modulation of spliceosome function may thus provide a new therapeutic avenue in genetically defined subsets of individuals with MDS or AML.
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Change history
11 May 2016
In the version of this article initially published online, the graphs in Figure 3b–d were laid out incorrectly and were not consistent with the figure legend and text. The error has been corrected for the print, PDF and HTML versions of this article.
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Acknowledgements
This work was supported by the Leukemia and Lymphoma Society (S.C.-W.L. and O.A.-W.), the US Department of Defense Breast Cancer Research Program grant W81XWH-14-1-0044 (H.D.), the US Department of Defense Bone Marrow Failure Research Program grants BM150092 (O.A.-W.) and W81XWH-12-1-0041 (R.K.B. and O.A.-W.), the Worldwide Cancer Research Fund (E.K.), the Fondation de France (J.-B.M.), the American Society of Hematology (B.H.D. and O.A.-W.), the Edward P. Evans Foundation (R.K.B. and O.A.-W.), the US National Institutes of Health (NIH)-NHLBI grant R01 HL128239 (R.K.B. and O.A.-W.), the NIH–NCI grant 1K08CA160647-01 (O.A.-W.), the Ellison Medical Foundation grant AG-NS-1030-13 (R.K.B.), the Damon Runyon Foundation (R.K.B. and O.A.-W.), the NIH–NIDDK grant R01 DK103854 (R.K.B.), the Starr Foundation grant I8-A8-075 (O.A.-W.), the Josie Robertson Investigator Program (O.A.-W.), the Mr. William H. Goodwin and Mrs. Alice Goodwin Commonwealth Foundation for Cancer Research (O.A.-W.), and the Experimental Therapeutics Center of MSKCC (O.A.-W.).
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S.C.-W.L., H.D., E.K., R.K.B., and O.A.-W. designed the study; S.C.-W.L., E.K., H.C., Y.R.C., J.-B.M., B.H.D., A.Y., Y.J.K., C.-W.C., A.P., J.T., X.W., A.K., S.A.A., and O.A.-W. performed experiments; H.D. and R.K.B. performed RNA-seq analysis; M.T., J.P., S.B., and P.G.S. provided E7107 and advice with drug-dosing experiments; and S.C.-W.L., H.D., E.K., C.L., R.K.B., and O.A.-W. prepared the manuscript with help from all co-authors.
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M.T., J.P., S.B., and P.G.S. are employees of H3 Biomedicine, Inc.
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Supplementary Text and Figures
Supplementary Figures 1–6 and Supplementary Tables 2 and 4 (PDF 40706 kb)
Supplementary Table 1
Median normalized gene expression values across replicates in each of the murine mRNA sequencing experiments shown in Figures 1-3. (XLSX 2160 kb)
Supplementary Table 3
Median exon inclusion rates (PSI (percent spliced in) values (ψ)) for cassette exons across replicates in each of the murine mRNA sequencing experiments shown in Figures 1-3. (XLSX 1831 kb)
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Lee, SW., Dvinge, H., Kim, E. et al. Modulation of splicing catalysis for therapeutic targeting of leukemia with mutations in genes encoding spliceosomal proteins. Nat Med 22, 672–678 (2016). https://doi.org/10.1038/nm.4097
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DOI: https://doi.org/10.1038/nm.4097
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