FLT3/ITD mutation signaling includes suppression of SHP-1

Peili Chen, Mark J Levis, Patrick A Brown, Kyu Tae Kim, Jeffrey Allebach, Donald Small

Research output: Contribution to journalArticle

Abstract

Mutations in the FLT3 gene are the most common genetic alteration found in AMDL patients. FLT3 internal tandem duplication (ITD) mutations result in constitutive activation of FLT3 tyrosine kinase activity. The consequences of this activation are an increase in total phosphotyrosine content, persistent downstream signaling, and ultimately transformation of hematopoietic cells to factor-independent growth. The Src homology (SH)2 domain-containing protein-tyrosine phosphatase (SHP)-1 is involved in the down-regulation of a broad range of growth, factor and cytokine-driven signaling cascades. Loss-of-function or deficiency of SHP-1 activity results in a hyperproliferative response of myelomonocytic cell populations to growth factor stimulation. In this study, we examined the possible role of SHP-1 in regulating FLT3 signaling. We found that transformation of TF-1 cells with FLT3/ITD mutations suppressed the activity of SHP-1 by ∼3-fold. Suppression was caused by decreased SHP-1 protein expression, as analyzed at both the protein and RNA levels. In contrast, protein levels of SHP-2, a phosphatase that plays a stimulatory role in signaling through a variety of receptors, did not change significantly in FLT3 mutant cells. Suppressed SHP-1 protein levels in TF-1/ITD cells were partially overcome after cells were exposed to CEP-701, a selective FLT3 inhibitor. SHP-1 protein levels also increased in naturally occurring FLT3/ITD expressing AML cell lines and in primary FLT3/ITD AML samples after CEP-701 treatment. Furthermore, a small but reproducible growth/survival advantage was observed in both TF-1 and TF-1/ITD cells when SHP-1 expression was knocked down by RNAi. Taken together, these data provide the first evidence that suppression of SHP-1 by FLT3/ITD signaling may be another mechanism contributing to the transformation by FLT3/ITD mutations.

Original languageEnglish (US)
Pages (from-to)5361-5369
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number7
DOIs
StatePublished - Feb 18 2005

Fingerprint

Mutation
Cells
Proteins
Intercellular Signaling Peptides and Proteins
Non-Receptor Type 6 Protein Tyrosine Phosphatase
Chemical activation
Phosphotyrosine
SH2 Domain-Containing Protein Tyrosine Phosphatases
Phosphoric Monoester Hydrolases
Protein-Tyrosine Kinases
Protein Tyrosine Phosphatases
Population Growth
Thermodynamic properties
Genes
RNA Interference
RNA
Cytokines
Down-Regulation
Cell Line
Survival

ASJC Scopus subject areas

  • Biochemistry

Cite this

FLT3/ITD mutation signaling includes suppression of SHP-1. / Chen, Peili; Levis, Mark J; Brown, Patrick A; Kim, Kyu Tae; Allebach, Jeffrey; Small, Donald.

In: Journal of Biological Chemistry, Vol. 280, No. 7, 18.02.2005, p. 5361-5369.

Research output: Contribution to journalArticle

Chen, Peili ; Levis, Mark J ; Brown, Patrick A ; Kim, Kyu Tae ; Allebach, Jeffrey ; Small, Donald. / FLT3/ITD mutation signaling includes suppression of SHP-1. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 7. pp. 5361-5369.
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abstract = "Mutations in the FLT3 gene are the most common genetic alteration found in AMDL patients. FLT3 internal tandem duplication (ITD) mutations result in constitutive activation of FLT3 tyrosine kinase activity. The consequences of this activation are an increase in total phosphotyrosine content, persistent downstream signaling, and ultimately transformation of hematopoietic cells to factor-independent growth. The Src homology (SH)2 domain-containing protein-tyrosine phosphatase (SHP)-1 is involved in the down-regulation of a broad range of growth, factor and cytokine-driven signaling cascades. Loss-of-function or deficiency of SHP-1 activity results in a hyperproliferative response of myelomonocytic cell populations to growth factor stimulation. In this study, we examined the possible role of SHP-1 in regulating FLT3 signaling. We found that transformation of TF-1 cells with FLT3/ITD mutations suppressed the activity of SHP-1 by ∼3-fold. Suppression was caused by decreased SHP-1 protein expression, as analyzed at both the protein and RNA levels. In contrast, protein levels of SHP-2, a phosphatase that plays a stimulatory role in signaling through a variety of receptors, did not change significantly in FLT3 mutant cells. Suppressed SHP-1 protein levels in TF-1/ITD cells were partially overcome after cells were exposed to CEP-701, a selective FLT3 inhibitor. SHP-1 protein levels also increased in naturally occurring FLT3/ITD expressing AML cell lines and in primary FLT3/ITD AML samples after CEP-701 treatment. Furthermore, a small but reproducible growth/survival advantage was observed in both TF-1 and TF-1/ITD cells when SHP-1 expression was knocked down by RNAi. Taken together, these data provide the first evidence that suppression of SHP-1 by FLT3/ITD signaling may be another mechanism contributing to the transformation by FLT3/ITD mutations.",
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