Selective cytotoxic mechanism of GTP-14564, a novel tyrosine kinase inhibitor in leukemia cells expressing a constitutively active Fms-like tyrosine kinase 3 (FLT3)

Ken Murata, Hidetoshi Kumagai, Toshiyuki Kawashima, Kaori Tamitsu, Mariko Irie, Hideaki Nakajima, Shinya Suzu, Masabumi Shibuya, Shimeru Kamihira, Tetsuya Nosaka, Shigetaka Asano, Toshio Kitamura

Research output: Contribution to journalArticle

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Abstract

The receptor tyrosine kinase FLT3 is constitutively activated by an internal tandem duplication (ITD) mutation within the juxtamembrane domain in 20-30% of patients with acute myeloid leukemia. In this stcudy, we identified GTP-14564 as a specific kinase inhibitor for ITD-FLT3 and investigated the molecular basis of its specificity. GTP-14564 inhibited the growth of interleukin-3-independent Ba/F3 expressing ITD-FLT3 at 1 μM, whereas a 30-fold higher concentration of GTP-14564 was required to inhibit FLT3 ligand-dependent growth of Ba/F3 expressing wild type FLT3 (wt-FLT3). However, this inhibitor suppressed the kinase activities of wt-FLT3 and ITD-FLT3 equally, suggesting that the signaling pathways for proliferation differ between wt-FLT3 and ITD-FLT3. Analysis of downstream targets of FLT3 using GTP-14564 revealed STAT5 activation to be essential for growth signaling of ITD-FLT3. In contrast, wt-FLT3 appeared to mainly use the MAPK pathway rather than the STAT5 pathway to transmit a proliferative signal. Further analysis demonstrated that the first two tyrosines in an ITD were critical for STAT5 activation and growth induction but that all of the tyrosines in the juxtamembrane region were dispensable in terms of the proliferation signals of wt-FLT3. These results indicate that an ITD mutation in FLT3 elicits an aberrant STAT5 activation that results in increased sensitivity to GTP-14564. Thus, FLT3-targeted inhibition is an attractive approach, with the potential for selective cytotoxicity, to the treatment of ITD-FLT3-positive acute myeloid leukemia.

Original languageEnglish (US)
Pages (from-to)32892-32898
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number35
DOIs
StatePublished - Aug 29 2003

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fms-Like Tyrosine Kinase 3
Protein-Tyrosine Kinases
Leukemia
Cells
Chemical activation
Growth
Acute Myeloid Leukemia
Tyrosine
GTP 14564
Phosphotransferases
Mutation
Interleukin-3
Receptor Protein-Tyrosine Kinases
Cytotoxicity

ASJC Scopus subject areas

  • Biochemistry

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Selective cytotoxic mechanism of GTP-14564, a novel tyrosine kinase inhibitor in leukemia cells expressing a constitutively active Fms-like tyrosine kinase 3 (FLT3). / Murata, Ken; Kumagai, Hidetoshi; Kawashima, Toshiyuki; Tamitsu, Kaori; Irie, Mariko; Nakajima, Hideaki; Suzu, Shinya; Shibuya, Masabumi; Kamihira, Shimeru; Nosaka, Tetsuya; Asano, Shigetaka; Kitamura, Toshio.

In: Journal of Biological Chemistry, Vol. 278, No. 35, 29.08.2003, p. 32892-32898.

Research output: Contribution to journalArticle

Murata, K, Kumagai, H, Kawashima, T, Tamitsu, K, Irie, M, Nakajima, H, Suzu, S, Shibuya, M, Kamihira, S, Nosaka, T, Asano, S & Kitamura, T 2003, 'Selective cytotoxic mechanism of GTP-14564, a novel tyrosine kinase inhibitor in leukemia cells expressing a constitutively active Fms-like tyrosine kinase 3 (FLT3)', Journal of Biological Chemistry, vol. 278, no. 35, pp. 32892-32898. https://doi.org/10.1074/jbc.M210405200
Murata, Ken ; Kumagai, Hidetoshi ; Kawashima, Toshiyuki ; Tamitsu, Kaori ; Irie, Mariko ; Nakajima, Hideaki ; Suzu, Shinya ; Shibuya, Masabumi ; Kamihira, Shimeru ; Nosaka, Tetsuya ; Asano, Shigetaka ; Kitamura, Toshio. / Selective cytotoxic mechanism of GTP-14564, a novel tyrosine kinase inhibitor in leukemia cells expressing a constitutively active Fms-like tyrosine kinase 3 (FLT3). In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 35. pp. 32892-32898.
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AU - Murata, Ken

AU - Kumagai, Hidetoshi

AU - Kawashima, Toshiyuki

AU - Tamitsu, Kaori

AU - Irie, Mariko

AU - Nakajima, Hideaki

AU - Suzu, Shinya

AU - Shibuya, Masabumi

AU - Kamihira, Shimeru

AU - Nosaka, Tetsuya

AU - Asano, Shigetaka

AU - Kitamura, Toshio

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N2 - The receptor tyrosine kinase FLT3 is constitutively activated by an internal tandem duplication (ITD) mutation within the juxtamembrane domain in 20-30% of patients with acute myeloid leukemia. In this stcudy, we identified GTP-14564 as a specific kinase inhibitor for ITD-FLT3 and investigated the molecular basis of its specificity. GTP-14564 inhibited the growth of interleukin-3-independent Ba/F3 expressing ITD-FLT3 at 1 μM, whereas a 30-fold higher concentration of GTP-14564 was required to inhibit FLT3 ligand-dependent growth of Ba/F3 expressing wild type FLT3 (wt-FLT3). However, this inhibitor suppressed the kinase activities of wt-FLT3 and ITD-FLT3 equally, suggesting that the signaling pathways for proliferation differ between wt-FLT3 and ITD-FLT3. Analysis of downstream targets of FLT3 using GTP-14564 revealed STAT5 activation to be essential for growth signaling of ITD-FLT3. In contrast, wt-FLT3 appeared to mainly use the MAPK pathway rather than the STAT5 pathway to transmit a proliferative signal. Further analysis demonstrated that the first two tyrosines in an ITD were critical for STAT5 activation and growth induction but that all of the tyrosines in the juxtamembrane region were dispensable in terms of the proliferation signals of wt-FLT3. These results indicate that an ITD mutation in FLT3 elicits an aberrant STAT5 activation that results in increased sensitivity to GTP-14564. Thus, FLT3-targeted inhibition is an attractive approach, with the potential for selective cytotoxicity, to the treatment of ITD-FLT3-positive acute myeloid leukemia.

AB - The receptor tyrosine kinase FLT3 is constitutively activated by an internal tandem duplication (ITD) mutation within the juxtamembrane domain in 20-30% of patients with acute myeloid leukemia. In this stcudy, we identified GTP-14564 as a specific kinase inhibitor for ITD-FLT3 and investigated the molecular basis of its specificity. GTP-14564 inhibited the growth of interleukin-3-independent Ba/F3 expressing ITD-FLT3 at 1 μM, whereas a 30-fold higher concentration of GTP-14564 was required to inhibit FLT3 ligand-dependent growth of Ba/F3 expressing wild type FLT3 (wt-FLT3). However, this inhibitor suppressed the kinase activities of wt-FLT3 and ITD-FLT3 equally, suggesting that the signaling pathways for proliferation differ between wt-FLT3 and ITD-FLT3. Analysis of downstream targets of FLT3 using GTP-14564 revealed STAT5 activation to be essential for growth signaling of ITD-FLT3. In contrast, wt-FLT3 appeared to mainly use the MAPK pathway rather than the STAT5 pathway to transmit a proliferative signal. Further analysis demonstrated that the first two tyrosines in an ITD were critical for STAT5 activation and growth induction but that all of the tyrosines in the juxtamembrane region were dispensable in terms of the proliferation signals of wt-FLT3. These results indicate that an ITD mutation in FLT3 elicits an aberrant STAT5 activation that results in increased sensitivity to GTP-14564. Thus, FLT3-targeted inhibition is an attractive approach, with the potential for selective cytotoxicity, to the treatment of ITD-FLT3-positive acute myeloid leukemia.

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