Limits...
The conformational control inhibitor of tyrosine kinases DCC-2036 is effective for imatinib-resistant cells expressing T674I FIP1L1-PDGFRα.

Shen Y, Shi X, Pan J - PLoS ONE (2013)

Bottom Line: This study evaluated the effect of DCC-2036 on FIP1L1-PDGFRα-positive cells, including the wild type (WT) and the T674I mutant.We found that DCC-2036 decreased the phosphorylated levels of PDGFRα and its downstream targets without apparent effects on total protein levels.DCC-2036 may be a potential compound to treat imatinib-resistant HES.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, PR China.

ABSTRACT
The cells expressing the T674I point mutant of FIP1-like-1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRα) in hypereosinophilics syndrome (HES) are resistant to imatinib and some second-generation tyrosine kinase inhibitors (TKIs). There is a desperate need to develop therapy to combat this acquired drug resistance. DCC-2036 has been synthesized as a third-generation TKI to combat especially the Bcr-Abl T315I mutant in chronic myeloid leukemia. This study evaluated the effect of DCC-2036 on FIP1L1-PDGFRα-positive cells, including the wild type (WT) and the T674I mutant. The in vitro effects of DCC-2036 on the PDGFRα signal pathways, proliferation, cell cycling and apoptosis of FIP1L1-PDGFRα-positive cells were investigated, and a nude mouse xenograft model was employed to assess the in vivo antitumor activity. We found that DCC-2036 decreased the phosphorylated levels of PDGFRα and its downstream targets without apparent effects on total protein levels. DCC-2036 inhibited proliferation, and induced apoptosis with MEK-dependent up-regulation of the pro-apoptotic protein Bim in FIP1L1-PDGFRα-positive cells. DCC-2036 also exhibited in vivo antineoplastic activity against cells with T674I FIP1L1-PDGFRα. In summary, FIP1L1-PDGFRα-positive cells are sensitive to DCC-2036 regardless of their sensitivity to imatinib. DCC-2036 may be a potential compound to treat imatinib-resistant HES.

Show MeSH

Related in: MedlinePlus

DCC-2036 inhibits the phosphorylated-PDGFRα level and its downstream targets.A, EOL-1 cells and BaF3 cells harboring WT or T674I FIP1L1-PDGFRα were incubated with indicated concentrations of DCC-2036 for 24 h (EOL-1) or 36 h (BaF3-WT and -T674I mutant), the phosphorylated and total levels of PDGFRα, STAT3, Akt and Erk1/2 were analyzed by immunoblotting. B, EOL-1 cells and BaF3 cells expressing WT or T674I FIP1L1-PDGFRα were treated with DCC-2036 for indicated durations with different concentrations (6 nM for EOL-1 cells, 400 nM for BaF3 cells), the phosphorylated and total levels of indicated proteins were analyzed by immunoblotting. C, BaF3-T674I cells were exposed to the indicated concentrations of DCC-2036, imatinib or sorafenib for 36 h, the phosphorylated PDGFRα and total PDGFRα were analyzed by immunoblotting. Imatinib was used for comparison.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3756952&req=5

pone-0073059-g001: DCC-2036 inhibits the phosphorylated-PDGFRα level and its downstream targets.A, EOL-1 cells and BaF3 cells harboring WT or T674I FIP1L1-PDGFRα were incubated with indicated concentrations of DCC-2036 for 24 h (EOL-1) or 36 h (BaF3-WT and -T674I mutant), the phosphorylated and total levels of PDGFRα, STAT3, Akt and Erk1/2 were analyzed by immunoblotting. B, EOL-1 cells and BaF3 cells expressing WT or T674I FIP1L1-PDGFRα were treated with DCC-2036 for indicated durations with different concentrations (6 nM for EOL-1 cells, 400 nM for BaF3 cells), the phosphorylated and total levels of indicated proteins were analyzed by immunoblotting. C, BaF3-T674I cells were exposed to the indicated concentrations of DCC-2036, imatinib or sorafenib for 36 h, the phosphorylated PDGFRα and total PDGFRα were analyzed by immunoblotting. Imatinib was used for comparison.

Mentions: Conceived and designed the experiments: YS JP. Performed the experiments: YS XS. Analyzed the data: YS. Contributed reagents/materials/analysis tools: JP. Wrote the paper: YS XS. Other: Figure 1, Figure 2A, Figure 3, 4, 5, S1 for two BaF3 cells, Figure 6ABDE, Figure 7ABCD: YS. Figure 2B, Figure 3, 4, 5, S1 for EOL-1 cells, Figure 6C, Figure 7AB: XS.


The conformational control inhibitor of tyrosine kinases DCC-2036 is effective for imatinib-resistant cells expressing T674I FIP1L1-PDGFRα.

Shen Y, Shi X, Pan J - PLoS ONE (2013)

DCC-2036 inhibits the phosphorylated-PDGFRα level and its downstream targets.A, EOL-1 cells and BaF3 cells harboring WT or T674I FIP1L1-PDGFRα were incubated with indicated concentrations of DCC-2036 for 24 h (EOL-1) or 36 h (BaF3-WT and -T674I mutant), the phosphorylated and total levels of PDGFRα, STAT3, Akt and Erk1/2 were analyzed by immunoblotting. B, EOL-1 cells and BaF3 cells expressing WT or T674I FIP1L1-PDGFRα were treated with DCC-2036 for indicated durations with different concentrations (6 nM for EOL-1 cells, 400 nM for BaF3 cells), the phosphorylated and total levels of indicated proteins were analyzed by immunoblotting. C, BaF3-T674I cells were exposed to the indicated concentrations of DCC-2036, imatinib or sorafenib for 36 h, the phosphorylated PDGFRα and total PDGFRα were analyzed by immunoblotting. Imatinib was used for comparison.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756952&req=5

pone-0073059-g001: DCC-2036 inhibits the phosphorylated-PDGFRα level and its downstream targets.A, EOL-1 cells and BaF3 cells harboring WT or T674I FIP1L1-PDGFRα were incubated with indicated concentrations of DCC-2036 for 24 h (EOL-1) or 36 h (BaF3-WT and -T674I mutant), the phosphorylated and total levels of PDGFRα, STAT3, Akt and Erk1/2 were analyzed by immunoblotting. B, EOL-1 cells and BaF3 cells expressing WT or T674I FIP1L1-PDGFRα were treated with DCC-2036 for indicated durations with different concentrations (6 nM for EOL-1 cells, 400 nM for BaF3 cells), the phosphorylated and total levels of indicated proteins were analyzed by immunoblotting. C, BaF3-T674I cells were exposed to the indicated concentrations of DCC-2036, imatinib or sorafenib for 36 h, the phosphorylated PDGFRα and total PDGFRα were analyzed by immunoblotting. Imatinib was used for comparison.
Mentions: Conceived and designed the experiments: YS JP. Performed the experiments: YS XS. Analyzed the data: YS. Contributed reagents/materials/analysis tools: JP. Wrote the paper: YS XS. Other: Figure 1, Figure 2A, Figure 3, 4, 5, S1 for two BaF3 cells, Figure 6ABDE, Figure 7ABCD: YS. Figure 2B, Figure 3, 4, 5, S1 for EOL-1 cells, Figure 6C, Figure 7AB: XS.

Bottom Line: This study evaluated the effect of DCC-2036 on FIP1L1-PDGFRα-positive cells, including the wild type (WT) and the T674I mutant.We found that DCC-2036 decreased the phosphorylated levels of PDGFRα and its downstream targets without apparent effects on total protein levels.DCC-2036 may be a potential compound to treat imatinib-resistant HES.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, PR China.

ABSTRACT
The cells expressing the T674I point mutant of FIP1-like-1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRα) in hypereosinophilics syndrome (HES) are resistant to imatinib and some second-generation tyrosine kinase inhibitors (TKIs). There is a desperate need to develop therapy to combat this acquired drug resistance. DCC-2036 has been synthesized as a third-generation TKI to combat especially the Bcr-Abl T315I mutant in chronic myeloid leukemia. This study evaluated the effect of DCC-2036 on FIP1L1-PDGFRα-positive cells, including the wild type (WT) and the T674I mutant. The in vitro effects of DCC-2036 on the PDGFRα signal pathways, proliferation, cell cycling and apoptosis of FIP1L1-PDGFRα-positive cells were investigated, and a nude mouse xenograft model was employed to assess the in vivo antitumor activity. We found that DCC-2036 decreased the phosphorylated levels of PDGFRα and its downstream targets without apparent effects on total protein levels. DCC-2036 inhibited proliferation, and induced apoptosis with MEK-dependent up-regulation of the pro-apoptotic protein Bim in FIP1L1-PDGFRα-positive cells. DCC-2036 also exhibited in vivo antineoplastic activity against cells with T674I FIP1L1-PDGFRα. In summary, FIP1L1-PDGFRα-positive cells are sensitive to DCC-2036 regardless of their sensitivity to imatinib. DCC-2036 may be a potential compound to treat imatinib-resistant HES.

Show MeSH
Related in: MedlinePlus