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Shp-2 has a positive regulatory role in ES cell differentiation and proliferation
1Department of Biochemistry and Molecular Biology, Walther Oncology Center, Indiana University School of Medicine and Walther Cancer Institute, Indianapolis 46202-5121, USA.
Abstract:
Shp-2 is a ubiquitously expressed tyrosine phosphatase with two SH2 domains. Homozygous mutant mice with a targeted deletion of 65 amino acid residues in the N-terminal SH2 domain of Shp-2 die in utero at mid-gestation, with multiple defects in mesodermal patterning. To surpass the embryonic lethality in dissecting the Shp-2 function in cell growth and differentiation, we established homozygous Shp-2 mutant embryonic stem (ES) cell lines. Our previous data showed a severe suppression of hematopoietic cell differentiation from Shp-2 mutant ES cells. Here we demonstrate that development of cardiac muscle cells was dramatically delayed and impaired in embryoid bodies (EBs) of Shp-2 mutant origin. Shp-2 mutant ES cells failed to differentiate into epithelial and fibroblast cells in vitro. However, higher efficiency of secondary EB formation was observed from the mutant than the wild-type ES cells. Further, mutant ES cells were more sensitive than wild-type cells to the differentiation suppressing effect of leukemia inhibitory factor (LIF). In addition, mutant ES cells showed a reduced growth rate compared to wild-type cells. These results suggest that the Shp-2 tyrosine phosphatase is a positive regulator for both cell differentiation and proliferation, in contrast to the Src-family kinases which promote cell growth but block differentiation.
Insights
Shp-2 tyrosine phosphatase is crucial for embryonic development, regulating cell differentiation and proliferation. Mutant cells show impaired cardiac muscle development and reduced growth, highlighting Shp-2
Area of Science:
- Molecular Biology
- Developmental Biology
- Cell Signaling
Background:
- Shp-2 is a ubiquitously expressed tyrosine phosphatase with essential roles in development.
- Homozygous Shp-2 mutant mice exhibit embryonic lethality due to mesodermal patterning defects.
- Previous studies indicated suppressed hematopoietic cell differentiation in Shp-2 mutant ES cells.
Purpose of the Study:
- To investigate the function of Shp-2 in cell differentiation and proliferation beyond embryonic lethality.
- To analyze the impact of Shp-2 deficiency on cardiac muscle cell development in vitro.
- To assess Shp-2's role in ES cell differentiation into epithelial and fibroblast lineages.
Main Methods:
- Generation and characterization of homozygous Shp-2 mutant embryonic stem (ES) cell lines.
- Differentiation of ES cells into embryoid bodies (EBs) to study cardiac muscle cell development.
- In vitro differentiation assays for epithelial and fibroblast cell lineages.
- Assessment of ES cell sensitivity to leukemia inhibitory factor (LIF).
Main Results:
- Cardiac muscle cell development was significantly delayed and impaired in Shp-2 mutant EBs.
- Shp-2 mutant ES cells failed to differentiate into epithelial and fibroblast cells.
- Mutant ES cells exhibited increased secondary EB formation and heightened sensitivity to LIF.
- A reduced growth rate was observed in Shp-2 mutant ES cells compared to wild-type.
Conclusions:
- Shp-2 tyrosine phosphatase acts as a positive regulator of both cell differentiation and proliferation.
- Shp-2 is essential for normal cardiac muscle, epithelial, and fibroblast cell development.
- Unlike Src-family kinases, Shp-2 promotes both growth and differentiation, suggesting a distinct regulatory role.