Time-restricted activation of Protein Kinase D2 directs vasculogenesis during mouse embryonic stem cell differentiation
| Autor: | J Schroeer, N Azoitei, F Genze, Anett Illing, Alexander Kleger, Martin Müller, Stefan Liebau, Thomas Seufferlein |
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| Rok vydání: | 2014 |
| Předmět: | |
| Zdroj: | Zeitschrift für Gastroenterologie. 52 |
| ISSN: | 1439-7803 0044-2771 |
| DOI: | 10.1055/s-0034-1386212 |
| Popis: | The protein kinase D (PKD) isoenzymes PKD1, -2, and -3, are prominent downstream targets of PKCs and phospholipase D in various biological systems. Recent data from our laboratory identified PKD isoforms as novel, essential mediators of tumour cell-endothelial cell communication but also as regulators of tumour cell motility and metastasis formation. The role of PKD isoforms during vascular development remains elusive. In the current study, we aimed to dissect this contribution using mouse embryonic stem (ES) cells as bona fide tool. First, we identified Protein Kinase D2 as the predominant isoform in undifferentiated ES cells leading us to particularly focus on this isoform. Time-restricted PKD2 activation using an inducible knock-in allele in differentiating mouse ES cells prevented cardiac mesoderm but activated a vascular differentiation program as shown by gene and protein regulation. Interestingly, the proliferative capacity is strongly diminished as a consequence of forced PKD2 expression. Finally, we aimed to underpin our findings in two independent in vivo models: First, embryoid bodies were transplanted on the chorioallantois membrane (CAM) of fertilised chicken eggs, a widely used model to study pro- and anti-angiogenesis. In line, with our in vitro data pronounced vessel formation was evident in the tumour-like structures arising at day 4 of the CAM assay. Second, we used the teratoma assay and induced PKD2 in immunodeficient mice during teratoma formation. While there was no difference in teratoma weight or size, a strong increase of CD31 expression as an indicator of vasculogenesis was observed in teratoma lysates. Thus, our data obtained in ES cells demonstrate that PKD2 contributes to the regulation of vasculogenesis during early development and ascribes a vascular fate in two independent embryonic tumorgenesis models. |
| Databáze: | OpenAIRE |
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