American Association for Cancer Research
21598290cd130419-sup-supp_methods_and_figs.pdf (601.21 kB)

Supplementary Methods and Figures from NUP98–PHF23 Is a Chromatin-Modifying Oncoprotein That Causes a Wide Array of Leukemias Sensitive to Inhibition of PHD Histone Reader Function

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journal contribution
posted on 2023-04-03, 20:46 authored by Sheryl M. Gough, Fan Lee, Fan Yang, Robert L. Walker, Yeulin J. Zhu, Marbin Pineda, Masahiro Onozawa, Yang Jo Chung, Sven Bilke, Elise K. Wagner, John M. Denu, Yi Ning, Bowen Xu, Gang Greg Wang, Paul S. Meltzer, Peter D. Aplan

PDF file 601K, Supplementary Methods. Figure S1. The NUP98-PHF23 (NP23) transgene and peripheral blood (PB) characteristics of NP23 mice. Figure S2. NP23 AML and pre-T LBL. Figure S3. NP23 bi-clonal AML/pre-T LBL, B-ALL and erythroid leukemias. Figure S4.Impaired hematopoietic development in NP23 progenitor cell populations. Figure S5. Validation of differential gene expression in NP23 tissues. Figure S6. NP23 cells are sensitive to disulfiram, an inhibitor of PHD domain H3K4me3 binding



In this report, we show that expression of a NUP98–PHF23 (NP23) fusion, associated with acute myeloid leukemia (AML) in humans, leads to myeloid, erythroid, T-cell, and B-cell leukemia in mice. The leukemic and preleukemic tissues display a stem cell–like expression signature, including Hoxa, Hoxb, and Meis1 genes. The PHF23 plant homeodomain (PHD) motif is known to bind to H3K4me3 residues, and chromatin immunoprecipitation experiments demonstrated that the NP23 protein binds to chromatin at a specific subset of H3K4me3 sites, including at Hoxa, Hoxb, and Meis1. Treatment of NP23 cells with disulfiram, which inhibits the binding of PHD motifs to H3K4me3, rapidly and selectively killed NP23-expressing myeloblasts; cell death was preceded by decreased expression of Hoxa, Hoxb, and Meis1. Furthermore, AML driven by a related fusion gene, NUP98–JARID1A (NJL), was also sensitive to disulfiram. Thus, the NP23 mouse provides a platform to evaluate compounds that disrupt binding of oncogenic PHD proteins to H3K4me3.Significance: NP23 and NJL belong to a subset of chromatin-modifying fusion oncoproteins that cause leukemia characterized by overexpression of Hoxa and Meis1 genes. Inhibition of NP23 binding to H3K4me3 at Hoxa and Meis1 loci by disulfiram, a U.S. Food and Drug Administration–approved drug, leads to leukemic cell death, demonstrating the feasibility of targeting this subset of oncoproteins. Cancer Discov; 4(5); 564–77. ©2014 AACR.This article is highlighted in the In This Issue feature, p. 495