Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro

Benjamin M F Mow, Joya Chandra, Phyllis A. Svingen, Christopher G. Hallgren, Ellen Weisberg, Timothy J. Kottke, Ven L. Narayanan, Mark R Litzow, James D. Griffin, Edward A. Sausville, Ayalew Tefferi, Scott H Kaufmann

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Abstract

The adenosine triphosphate binding-site-directed agent STI571 and the tyrphostin adaphostin are undergoing evaluation as bcr/abl kinase inhibitors. The current study compared the effects of these agents on the survival of K562 cells, bcr/abl-transduced FDC-P1 cells, and myeloid progenitors from patients with chronic myelogenous leukemia (CML) compared with healthy donors. Treatment of K562 cells with 10 μM adaphostin resulted in decreased p210bcr/abl polypeptide levels in the first 6 hours, followed by caspase activation and accumulation of apoptotic cells in less than 12 hours. By 24 hours, 90% of the cells were apoptotic and unable to form colonies, In contrast, 20 μM STI571 caused rapid inhibition of bcr/abl autophosphorylation without p210bcr/abl degradation. Although this was followed by the inhibition of Stat5 phosphorylation and the down-regulation of Bcl-XL and Mcl-1, only 7% ± 3% and 25% ± 9% of cells were apoptotic at 16 and 24 hours, respectively. Instead, the cytotoxic effects of STI571 became more pronounced with prolonged exposure, with IC90 values greater than 20 μM and 1.0 ± 0.6 μM after 24 and 48 hours, respectively. Consistent with these results, 24-hour adaphostin exposure inhibited CML granulocyte colony-forming units (CFU-G) (median IC50, 12 μM) but not normal CFU-G (median IC50, greater than 20 μM), whereas 24-hour STI571 treatment had no effect on CML or normal CFU-G. Additional experiments revealed that STI571-resistant K562 cells remained sensitive to adaphostin. Moreover, the combination of STI571 + adaphostin induced more cytotoxicity in K562 cells and in CML CFU-G than either agent alone did. Collectively, these results identify adaphostin as a mechanistically distinct CML-selective agent that retains activity in STI571-resistant cell lines.

Original languageEnglish (US)
Pages (from-to)664-671
Number of pages8
JournalBlood
Volume99
Issue number2
DOIs
StatePublished - Jan 15 2002

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NSC 680410
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Phosphotransferases
K562 Cells
Inhibitory Concentration 50
Cells
Tyrphostins
Myeloid Progenitor Cells
Phosphorylation
Cytotoxicity
Caspases
In Vitro Techniques
Imatinib Mesylate
Granulocytes
Stem Cells
Down-Regulation
Adenosine Triphosphate
Chemical activation

ASJC Scopus subject areas

  • Hematology

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Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro. / Mow, Benjamin M F; Chandra, Joya; Svingen, Phyllis A.; Hallgren, Christopher G.; Weisberg, Ellen; Kottke, Timothy J.; Narayanan, Ven L.; Litzow, Mark R; Griffin, James D.; Sausville, Edward A.; Tefferi, Ayalew; Kaufmann, Scott H.

In: Blood, Vol. 99, No. 2, 15.01.2002, p. 664-671.

Research output: Contribution to journalArticle

Mow, BMF, Chandra, J, Svingen, PA, Hallgren, CG, Weisberg, E, Kottke, TJ, Narayanan, VL, Litzow, MR, Griffin, JD, Sausville, EA, Tefferi, A & Kaufmann, SH 2002, 'Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro', Blood, vol. 99, no. 2, pp. 664-671. https://doi.org/10.1182/blood.V99.2.664
Mow, Benjamin M F ; Chandra, Joya ; Svingen, Phyllis A. ; Hallgren, Christopher G. ; Weisberg, Ellen ; Kottke, Timothy J. ; Narayanan, Ven L. ; Litzow, Mark R ; Griffin, James D. ; Sausville, Edward A. ; Tefferi, Ayalew ; Kaufmann, Scott H. / Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro. In: Blood. 2002 ; Vol. 99, No. 2. pp. 664-671.
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abstract = "The adenosine triphosphate binding-site-directed agent STI571 and the tyrphostin adaphostin are undergoing evaluation as bcr/abl kinase inhibitors. The current study compared the effects of these agents on the survival of K562 cells, bcr/abl-transduced FDC-P1 cells, and myeloid progenitors from patients with chronic myelogenous leukemia (CML) compared with healthy donors. Treatment of K562 cells with 10 μM adaphostin resulted in decreased p210bcr/abl polypeptide levels in the first 6 hours, followed by caspase activation and accumulation of apoptotic cells in less than 12 hours. By 24 hours, 90{\%} of the cells were apoptotic and unable to form colonies, In contrast, 20 μM STI571 caused rapid inhibition of bcr/abl autophosphorylation without p210bcr/abl degradation. Although this was followed by the inhibition of Stat5 phosphorylation and the down-regulation of Bcl-XL and Mcl-1, only 7{\%} ± 3{\%} and 25{\%} ± 9{\%} of cells were apoptotic at 16 and 24 hours, respectively. Instead, the cytotoxic effects of STI571 became more pronounced with prolonged exposure, with IC90 values greater than 20 μM and 1.0 ± 0.6 μM after 24 and 48 hours, respectively. Consistent with these results, 24-hour adaphostin exposure inhibited CML granulocyte colony-forming units (CFU-G) (median IC50, 12 μM) but not normal CFU-G (median IC50, greater than 20 μM), whereas 24-hour STI571 treatment had no effect on CML or normal CFU-G. Additional experiments revealed that STI571-resistant K562 cells remained sensitive to adaphostin. Moreover, the combination of STI571 + adaphostin induced more cytotoxicity in K562 cells and in CML CFU-G than either agent alone did. Collectively, these results identify adaphostin as a mechanistically distinct CML-selective agent that retains activity in STI571-resistant cell lines.",
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T1 - Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro

AU - Mow, Benjamin M F

AU - Chandra, Joya

AU - Svingen, Phyllis A.

AU - Hallgren, Christopher G.

AU - Weisberg, Ellen

AU - Kottke, Timothy J.

AU - Narayanan, Ven L.

AU - Litzow, Mark R

AU - Griffin, James D.

AU - Sausville, Edward A.

AU - Tefferi, Ayalew

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