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Multiple actions of lysophosphatidic acid on fibroblasts revealed by transcriptional profiling.

Stortelers C, Kerkhoven R, Moolenaar WH - BMC Genomics (2008)

Bottom Line: The subset of LPA-specific genes was enriched for those associated with cytoskeletal remodeling, in keeping with LPA's ability to regulate cell shape and motility.This study highlights the importance of LPA in programming fibroblasts not only to proliferate and migrate but also to produce many paracrine mediators of tissue remodeling, angiogenesis, inflammation and tumor progression.Furthermore, our results show that G protein-coupled receptors and receptor tyrosine kinases can signal independently to regulate broadly overlapping sets of genes in the same cell type.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Cellular Biochemistry and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. c.stortelers@gmail.com

ABSTRACT

Background: Lysophosphatidic acid (LPA) is a lipid mediator that acts through specific G protein-coupled receptors to stimulate the proliferation, migration and survival of many cell types. LPA signaling has been implicated in development, wound healing and cancer. While LPA signaling pathways have been studied extensively, it remains unknown how LPA affects global gene expression in its target cells.

Results: We have examined the temporal program of global gene expression in quiescent mouse embryonic fibroblasts stimulated with LPA using 32 k oligonucleotide microarrays. In addition to genes involved in growth stimulation and cytoskeletal reorganization, LPA induced many genes that encode secreted factors, including chemokines, growth factors, cytokines, pro-angiogenic and pro-fibrotic factors, components of the plasminogen activator system and metalloproteases. Strikingly, epidermal growth factor induced a broadly overlapping expression pattern, but some 7% of the genes (105 out of 1508 transcripts) showed differential regulation by LPA. The subset of LPA-specific genes was enriched for those associated with cytoskeletal remodeling, in keeping with LPA's ability to regulate cell shape and motility.

Conclusion: This study highlights the importance of LPA in programming fibroblasts not only to proliferate and migrate but also to produce many paracrine mediators of tissue remodeling, angiogenesis, inflammation and tumor progression. Furthermore, our results show that G protein-coupled receptors and receptor tyrosine kinases can signal independently to regulate broadly overlapping sets of genes in the same cell type.

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Differential regulation of selected genes by LPA (black) and EGF (red) validated by qPCR. qPCR data were normalized to HPRT mRNA concentration and plotted relative to the level at time zero. Data are presented as means ± SD of duplicate experiments.
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Figure 9: Differential regulation of selected genes by LPA (black) and EGF (red) validated by qPCR. qPCR data were normalized to HPRT mRNA concentration and plotted relative to the level at time zero. Data are presented as means ± SD of duplicate experiments.

Mentions: Fibroblasts have long been used as a model to study peptide growth factor signaling. When stimulated by distinct peptide growth factors (EGF, FGF, PDGF), fibroblasts show a strongly conserved gene-expression signature [31]. This is not too surprising since the cognate receptor tyrosine kinases (RTKs) all use the same signaling principle. To our knowledge, however, it is unknown to what extent the transcriptional response to GPCR stimulation bears comparison with that to RTK stimulation in the same cell type. We therefore compared the temporal gene expression programs of LPA and EGF in MEFs at five different time points (0.5–6 hrs). We found that EGF (20 ng/ml) induced many of the same genes as LPA (5 μM), although LPA stimulation often led to a higher level of induction and/or more prolonged kinetics (Figure 8A,B). For example, LPA caused a much more prolonged upregulation of the immediate early genes Fos, Dusp1 and Cxcl1 than did EGF (Figure 8B; see additional file 8: cluster 1). LPA was also more efficacious in inducing genes that encode paracrine factors (Ccl2, Ereg, Il1RL1, Ctgf, Vegfa) and components of the plasminogen activator system (Plaur, Pai-1) (Figure 8B). Quantitative PCR analysis confirmed the differential regulation of selected genes by LPA and EGF (Figure 9). A complete list of the differentially regulated genes is shown in additional file 9. To what extent these quantitative differences reflect different expression levels of the respective receptors is currently unknown.


Multiple actions of lysophosphatidic acid on fibroblasts revealed by transcriptional profiling.

Stortelers C, Kerkhoven R, Moolenaar WH - BMC Genomics (2008)

Differential regulation of selected genes by LPA (black) and EGF (red) validated by qPCR. qPCR data were normalized to HPRT mRNA concentration and plotted relative to the level at time zero. Data are presented as means ± SD of duplicate experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2536681&req=5

Figure 9: Differential regulation of selected genes by LPA (black) and EGF (red) validated by qPCR. qPCR data were normalized to HPRT mRNA concentration and plotted relative to the level at time zero. Data are presented as means ± SD of duplicate experiments.
Mentions: Fibroblasts have long been used as a model to study peptide growth factor signaling. When stimulated by distinct peptide growth factors (EGF, FGF, PDGF), fibroblasts show a strongly conserved gene-expression signature [31]. This is not too surprising since the cognate receptor tyrosine kinases (RTKs) all use the same signaling principle. To our knowledge, however, it is unknown to what extent the transcriptional response to GPCR stimulation bears comparison with that to RTK stimulation in the same cell type. We therefore compared the temporal gene expression programs of LPA and EGF in MEFs at five different time points (0.5–6 hrs). We found that EGF (20 ng/ml) induced many of the same genes as LPA (5 μM), although LPA stimulation often led to a higher level of induction and/or more prolonged kinetics (Figure 8A,B). For example, LPA caused a much more prolonged upregulation of the immediate early genes Fos, Dusp1 and Cxcl1 than did EGF (Figure 8B; see additional file 8: cluster 1). LPA was also more efficacious in inducing genes that encode paracrine factors (Ccl2, Ereg, Il1RL1, Ctgf, Vegfa) and components of the plasminogen activator system (Plaur, Pai-1) (Figure 8B). Quantitative PCR analysis confirmed the differential regulation of selected genes by LPA and EGF (Figure 9). A complete list of the differentially regulated genes is shown in additional file 9. To what extent these quantitative differences reflect different expression levels of the respective receptors is currently unknown.

Bottom Line: The subset of LPA-specific genes was enriched for those associated with cytoskeletal remodeling, in keeping with LPA's ability to regulate cell shape and motility.This study highlights the importance of LPA in programming fibroblasts not only to proliferate and migrate but also to produce many paracrine mediators of tissue remodeling, angiogenesis, inflammation and tumor progression.Furthermore, our results show that G protein-coupled receptors and receptor tyrosine kinases can signal independently to regulate broadly overlapping sets of genes in the same cell type.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Cellular Biochemistry and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. c.stortelers@gmail.com

ABSTRACT

Background: Lysophosphatidic acid (LPA) is a lipid mediator that acts through specific G protein-coupled receptors to stimulate the proliferation, migration and survival of many cell types. LPA signaling has been implicated in development, wound healing and cancer. While LPA signaling pathways have been studied extensively, it remains unknown how LPA affects global gene expression in its target cells.

Results: We have examined the temporal program of global gene expression in quiescent mouse embryonic fibroblasts stimulated with LPA using 32 k oligonucleotide microarrays. In addition to genes involved in growth stimulation and cytoskeletal reorganization, LPA induced many genes that encode secreted factors, including chemokines, growth factors, cytokines, pro-angiogenic and pro-fibrotic factors, components of the plasminogen activator system and metalloproteases. Strikingly, epidermal growth factor induced a broadly overlapping expression pattern, but some 7% of the genes (105 out of 1508 transcripts) showed differential regulation by LPA. The subset of LPA-specific genes was enriched for those associated with cytoskeletal remodeling, in keeping with LPA's ability to regulate cell shape and motility.

Conclusion: This study highlights the importance of LPA in programming fibroblasts not only to proliferate and migrate but also to produce many paracrine mediators of tissue remodeling, angiogenesis, inflammation and tumor progression. Furthermore, our results show that G protein-coupled receptors and receptor tyrosine kinases can signal independently to regulate broadly overlapping sets of genes in the same cell type.

Show MeSH
Related in: MedlinePlus