Genetic regulation of the RUNX transcription factor family has antitumor effects
Ken Morita, Kensho Suzuki, Shintaro Maeda, Akihiko Matsuo, Yoshihide Mitsuda, Chieko Tokushige, Gengo Kashiwazaki, Junichi Taniguchi, Rina Maeda, Mina Noura, Masahiro Hirata, Tatsuki Kataoka, Ayaka Yano, Yoshimi Yamada, Hiroki Kiyose, Mayu Tokumasu, Hidemasa Matsuo, Sunao Tanaka, Yasushi Okuno, Manabu Muto, Kazuhito Naka, Kosei Ito, Toshio Kitamura, Yasufumi Kaneda, Paul P. Liu, Toshikazu Bando, Souichi Adachi, Hiroshi Sugiyama, Yasuhiko Kamikubo
Ken Morita, Kensho Suzuki, Shintaro Maeda, Akihiko Matsuo, Yoshihide Mitsuda, Chieko Tokushige, Gengo Kashiwazaki, Junichi Taniguchi, Rina Maeda, Mina Noura, Masahiro Hirata, Tatsuki Kataoka, Ayaka Yano, Yoshimi Yamada, Hiroki Kiyose, Mayu Tokumasu, Hidemasa Matsuo, Sunao Tanaka, Yasushi Okuno, Manabu Muto, Kazuhito Naka, Kosei Ito, Toshio Kitamura, Yasufumi Kaneda, Paul P. Liu, Toshikazu Bando, Souichi Adachi, Hiroshi Sugiyama, Yasuhiko Kamikubo
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Research Article Hematology Oncology

Genetic regulation of the RUNX transcription factor family has antitumor effects

Abstract

Runt-related transcription factor 1 (RUNX1) is generally considered to function as a tumor suppressor in the development of leukemia, but a growing body of evidence suggests that it has pro-oncogenic properties in acute myeloid leukemia (AML). Here we have demonstrated that the antileukemic effect mediated by RUNX1 depletion is highly dependent on a functional p53-mediated cell death pathway. Increased expression of other RUNX family members, including RUNX2 and RUNX3, compensated for the antitumor effect elicited by RUNX1 silencing, and simultaneous attenuation of all RUNX family members as a cluster led to a much stronger antitumor effect relative to suppression of individual RUNX members. Switching off the RUNX cluster using alkylating agent–conjugated pyrrole-imidazole (PI) polyamides, which were designed to specifically bind to consensus RUNX-binding sequences, was highly effective against AML cells and against several poor-prognosis solid tumors in a xenograft mouse model of AML without notable adverse events. Taken together, these results identify a crucial role for the RUNX cluster in the maintenance and progression of cancer cells and suggest that modulation of the RUNX cluster using the PI polyamide gene-switch technology is a potential strategy to control malignancies.

Authors

Ken Morita, Kensho Suzuki, Shintaro Maeda, Akihiko Matsuo, Yoshihide Mitsuda, Chieko Tokushige, Gengo Kashiwazaki, Junichi Taniguchi, Rina Maeda, Mina Noura, Masahiro Hirata, Tatsuki Kataoka, Ayaka Yano, Yoshimi Yamada, Hiroki Kiyose, Mayu Tokumasu, Hidemasa Matsuo, Sunao Tanaka, Yasushi Okuno, Manabu Muto, Kazuhito Naka, Kosei Ito, Toshio Kitamura, Yasufumi Kaneda, Paul P. Liu, Toshikazu Bando, Souichi Adachi, Hiroshi Sugiyama, Yasuhiko Kamikubo

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Figure 2

RUNX1 inhibition activates the p53 pathway by transcriptionally controlling BCL11A and TRIM24 expression.

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RUNX1 inhibition activates the p53 pathway by transcriptionally controll...
(A) Identification of genes under the control of RUNX1. One hundred and thirty-two genes were commonly upregulated in 4 independent AML patients with a higher RUNX1 expression (GSE22845, GSE67936, GSE21261, and GSE19577). Among them, RUNX1 bound to 66 gene regulatory regions (GSE22178 and GSE31221). See Supplemental Tables 1 and 2 for the list of the indicated genes. (B) Expression levels of BCL11A and TRIM24 in MV4-11 cells transduced with control or RUNX1 shRNAs. Cells were treated with 3 μM doxycycline for 48 hours. Values are normalized to that of control vector–transduced cells (n = 3). (C) Downregulation of BCL11A and TRIM24 in RUNX1-silenced MV4-11 cells. Nondepleted and RUNX1-depleted MV4-11 cells were treated as in B. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D and E) Rescue of cell proliferation rate by forced expression of BCL11A (D) or TRIM24 (E) in RUNX1-depleted MV4-11 cells. The indicated MV4-11 cells were cultured in the presence of 3 μM doxycycline (n = 3). (F) p53-dependent growth suppression of RUNX1-depleted cells. Nondepleted and RUNX1-depleted MV4-11 (left panel) and MOLM-13 cells (right panel) were transduced with lentivirus vector encoding shRNA targeting p53 (sh_p53 #1 or sh_p53 #2) (n = 3). Data are the mean ± SEM values. *P < 0.05, by 2-tailed Student’s t test.