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Proteomic analysis reveals heat shock protein 70 has a key role in polycythemia Vera.

Gallardo M, Barrio S, Fernandez M, Paradela A, Arenas A, Toldos O, Ayala R, Albizua E, Jimenez A, Redondo S, Garcia-Martin RM, Gilsanz F, Albar JP, Martinez-Lopez J - Mol. Cancer (2013)

Bottom Line: Immunohistochemistry of 46 MPN bone marrow samples confirmed HSP70 expression.In an ex vivo model KNK437 was used as an inhibition model assay of HSP70, showed dose-dependent inhibition of cell growth and burst formation unit erythroid (BFU-E) in PV and ET, increased apoptosis in the erythroid lineage, and decreased pJAK2 signaling, as well as a specific siRNA for HSP70.These data suggest a key role for HSP70 in proliferation and survival of the erythroid lineage in PV, and may represent a potential therapeutic target in MPN, especially in PV.

View Article: PubMed Central - HTML - PubMed

Affiliation: Hematology Service, Hospital Universitario 12 de Octubre, Avenida, Córdoba, s/n, 28041, Madrid, Spain. miguelgallardodelgado@gmail.com.

ABSTRACT
JAK-STAT signaling through the JAK2V617F mutation is central to the pathogenesis of myeloproliferative neoplasms (MPN). However, other events could precede the JAK2 mutation. The aim of this study is to analyze the phenotypic divergence between polycytemia vera (PV) and essential thrombocytemia (ET) to find novel therapeutics targets by a proteomic and functional approach to identify alternative routes to JAK2 activation. Through 2D-DIGE and mass spectrometry of granulocyte protein from 20 MPN samples, showed differential expression of HSP70 in PV and ET besides other 60 proteins. Immunohistochemistry of 46 MPN bone marrow samples confirmed HSP70 expression. The median of positive granulocytes was 80% in PV (SD 35%) vs. 23% in ET (SD 34.25%). In an ex vivo model KNK437 was used as an inhibition model assay of HSP70, showed dose-dependent inhibition of cell growth and burst formation unit erythroid (BFU-E) in PV and ET, increased apoptosis in the erythroid lineage, and decreased pJAK2 signaling, as well as a specific siRNA for HSP70. These data suggest a key role for HSP70 in proliferation and survival of the erythroid lineage in PV, and may represent a potential therapeutic target in MPN, especially in PV.

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Related in: MedlinePlus

Flow chart describing the three-steps to proteomic screening for differentially expressed proteins in polycythemia vera (PV) and essential thrombocythemia (ET). The training set includes two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS); the protein validation set includes immunohistochemistry (IHC); and the functional validation set includes burst formation unit-erythroid (BFU-E) and stem cells studies.
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Figure 1: Flow chart describing the three-steps to proteomic screening for differentially expressed proteins in polycythemia vera (PV) and essential thrombocythemia (ET). The training set includes two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS); the protein validation set includes immunohistochemistry (IHC); and the functional validation set includes burst formation unit-erythroid (BFU-E) and stem cells studies.

Mentions: The study was approved by the 12 Octubre Hospital ethics committee and written informed consent was obtained from all patients, according to the Declaration of Helsinki. A flow diagram of the patients is shown in Figure 1.


Proteomic analysis reveals heat shock protein 70 has a key role in polycythemia Vera.

Gallardo M, Barrio S, Fernandez M, Paradela A, Arenas A, Toldos O, Ayala R, Albizua E, Jimenez A, Redondo S, Garcia-Martin RM, Gilsanz F, Albar JP, Martinez-Lopez J - Mol. Cancer (2013)

Flow chart describing the three-steps to proteomic screening for differentially expressed proteins in polycythemia vera (PV) and essential thrombocythemia (ET). The training set includes two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS); the protein validation set includes immunohistochemistry (IHC); and the functional validation set includes burst formation unit-erythroid (BFU-E) and stem cells studies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4225507&req=5

Figure 1: Flow chart describing the three-steps to proteomic screening for differentially expressed proteins in polycythemia vera (PV) and essential thrombocythemia (ET). The training set includes two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS); the protein validation set includes immunohistochemistry (IHC); and the functional validation set includes burst formation unit-erythroid (BFU-E) and stem cells studies.
Mentions: The study was approved by the 12 Octubre Hospital ethics committee and written informed consent was obtained from all patients, according to the Declaration of Helsinki. A flow diagram of the patients is shown in Figure 1.

Bottom Line: Immunohistochemistry of 46 MPN bone marrow samples confirmed HSP70 expression.In an ex vivo model KNK437 was used as an inhibition model assay of HSP70, showed dose-dependent inhibition of cell growth and burst formation unit erythroid (BFU-E) in PV and ET, increased apoptosis in the erythroid lineage, and decreased pJAK2 signaling, as well as a specific siRNA for HSP70.These data suggest a key role for HSP70 in proliferation and survival of the erythroid lineage in PV, and may represent a potential therapeutic target in MPN, especially in PV.

View Article: PubMed Central - HTML - PubMed

Affiliation: Hematology Service, Hospital Universitario 12 de Octubre, Avenida, Córdoba, s/n, 28041, Madrid, Spain. miguelgallardodelgado@gmail.com.

ABSTRACT
JAK-STAT signaling through the JAK2V617F mutation is central to the pathogenesis of myeloproliferative neoplasms (MPN). However, other events could precede the JAK2 mutation. The aim of this study is to analyze the phenotypic divergence between polycytemia vera (PV) and essential thrombocytemia (ET) to find novel therapeutics targets by a proteomic and functional approach to identify alternative routes to JAK2 activation. Through 2D-DIGE and mass spectrometry of granulocyte protein from 20 MPN samples, showed differential expression of HSP70 in PV and ET besides other 60 proteins. Immunohistochemistry of 46 MPN bone marrow samples confirmed HSP70 expression. The median of positive granulocytes was 80% in PV (SD 35%) vs. 23% in ET (SD 34.25%). In an ex vivo model KNK437 was used as an inhibition model assay of HSP70, showed dose-dependent inhibition of cell growth and burst formation unit erythroid (BFU-E) in PV and ET, increased apoptosis in the erythroid lineage, and decreased pJAK2 signaling, as well as a specific siRNA for HSP70. These data suggest a key role for HSP70 in proliferation and survival of the erythroid lineage in PV, and may represent a potential therapeutic target in MPN, especially in PV.

Show MeSH
Related in: MedlinePlus