Limits...
Pigment epithelium-derived factor (PEDF): a novel trophoblast-derived factor limiting feto-placental angiogenesis in late pregnancy.

Loegl J, Nussbaumer E, Hiden U, Majali-Martinez A, Ghaffari-Tabrizi-Wizy N, Cvitic S, Lang I, Desoye G, Huppertz B - Angiogenesis (2016)

Bottom Line: Notably, human recombinant PEDF reduced network formation only in combination with VEGF.Analysis of phosphorylation of ERK1/2 and FAK, two key players in VEGF-induced proliferation and migration, revealed that PEDF altered VEGF signaling, while PEDF alone did not affect phosphorylation of ERK1/2 and FAK.These data suggest that the trophoblast-derived anti-angiogenic molecule PEDF is involved in restricting growth and expansion of the feto-placental endothelium predominantly in late pregnancy and targets to modulate the intracellular effect of VEGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Graz, Austria.

ABSTRACT
The rapidly expanding feto-placental vasculature needs tight control by paracrine and endocrine mechanisms. Here, we focused on paracrine influence by trophoblast, the placental epithelium. We aimed to identify differences in regulation of feto-placental angiogenesis in early versus late pregnancy. To this end, the effect of conditioned media (CM) from early and late pregnancy human trophoblast was tested on network formation, migration and proliferation of human feto-placental endothelial cells. Only CM of late pregnancy trophoblast reduced network formation and migration. Screening of trophoblast transcriptome for anti-angiogenic candidates identified pigment epithelium-derived factor (PEDF) with higher expression and protein secretion in late pregnancy trophoblast. Addition of a PEDF-neutralizing antibody restored the anti-angiogenic effect of CM from late pregnancy trophoblast. Notably, human recombinant PEDF reduced network formation only in combination with VEGF. Also in the CAM assay, the combination of PEDF with VEGF reduced branching of vessels below control levels. Analysis of phosphorylation of ERK1/2 and FAK, two key players in VEGF-induced proliferation and migration, revealed that PEDF altered VEGF signaling, while PEDF alone did not affect phosphorylation of ERK1/2 and FAK. These data suggest that the trophoblast-derived anti-angiogenic molecule PEDF is involved in restricting growth and expansion of the feto-placental endothelium predominantly in late pregnancy and targets to modulate the intracellular effect of VEGF.

No MeSH data available.


Related in: MedlinePlus

Effect of first (FTB) and third (TTB) trimester trophoblast-conditioned medium (CM) on network formation, migration, proliferation and survival of feto-placental endothelial cells. a Representative phase contrast light microscopic images show network formation of feto-placental endothelial cells after 12 h on growth factor-reduced Matrigel. CM of peripheral blood monocyte cells (PBMC) used as positive control. b Quantitative analysis of networks depicts the inhibitory effect of TTB CM on feto-placental endothelial cells (n = 6). c CM of FTB and TTB reduced migration of feto-placental endothelial cells compared to control (n = 6). Endothelial basal medium (EBM) supplemented with FBS and growth factors (EBM++) was used as positive control. d Proliferation was unchanged between early and late trophoblast CM (n = 4). EBM supplemented with FCS and growth factors (EBM++) was used as positive control. e FTB and TTB CM did not increase LDH release of feto-placental endothelial cells (n = 4). Data are given as mean ± SEM. Statistical analysis used the mean of the triplicate of the number of individual biological replicates. Control medium (C) = DMEM/EBM + 7.5 % FCS for 48 h at 37 °C without cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4930480&req=5

Fig1: Effect of first (FTB) and third (TTB) trimester trophoblast-conditioned medium (CM) on network formation, migration, proliferation and survival of feto-placental endothelial cells. a Representative phase contrast light microscopic images show network formation of feto-placental endothelial cells after 12 h on growth factor-reduced Matrigel. CM of peripheral blood monocyte cells (PBMC) used as positive control. b Quantitative analysis of networks depicts the inhibitory effect of TTB CM on feto-placental endothelial cells (n = 6). c CM of FTB and TTB reduced migration of feto-placental endothelial cells compared to control (n = 6). Endothelial basal medium (EBM) supplemented with FBS and growth factors (EBM++) was used as positive control. d Proliferation was unchanged between early and late trophoblast CM (n = 4). EBM supplemented with FCS and growth factors (EBM++) was used as positive control. e FTB and TTB CM did not increase LDH release of feto-placental endothelial cells (n = 4). Data are given as mean ± SEM. Statistical analysis used the mean of the triplicate of the number of individual biological replicates. Control medium (C) = DMEM/EBM + 7.5 % FCS for 48 h at 37 °C without cells

Mentions: To investigate the paracrine effect of trophoblast from early versus late pregnancy on angiogenesis, the influence of CM on network formation, migration, proliferation and survival of feto-placental endothelial cells was analyzed (Fig. 1). CM of trophoblast from late pregnancy (term trophoblasts; TTB) reduced network formation of feto-placental endothelial cells by 30 ± 14 % (p = 0.024) (Fig. 1a, b), while CM of trophoblast from early pregnancy (first trimester trophoblasts; FTB) had no effect. CM of peripheral blood monocyte cells (PBMC) used as positive control [18] stimulated network formation by 21 ± 10 % (p = 0.049). Similar results were obtained with CM that had been ultracentrifuged prior to use (not shown), indicating that soluble factors secreted by trophoblast induce the anti-angiogenic effects.Fig. 1


Pigment epithelium-derived factor (PEDF): a novel trophoblast-derived factor limiting feto-placental angiogenesis in late pregnancy.

Loegl J, Nussbaumer E, Hiden U, Majali-Martinez A, Ghaffari-Tabrizi-Wizy N, Cvitic S, Lang I, Desoye G, Huppertz B - Angiogenesis (2016)

Effect of first (FTB) and third (TTB) trimester trophoblast-conditioned medium (CM) on network formation, migration, proliferation and survival of feto-placental endothelial cells. a Representative phase contrast light microscopic images show network formation of feto-placental endothelial cells after 12 h on growth factor-reduced Matrigel. CM of peripheral blood monocyte cells (PBMC) used as positive control. b Quantitative analysis of networks depicts the inhibitory effect of TTB CM on feto-placental endothelial cells (n = 6). c CM of FTB and TTB reduced migration of feto-placental endothelial cells compared to control (n = 6). Endothelial basal medium (EBM) supplemented with FBS and growth factors (EBM++) was used as positive control. d Proliferation was unchanged between early and late trophoblast CM (n = 4). EBM supplemented with FCS and growth factors (EBM++) was used as positive control. e FTB and TTB CM did not increase LDH release of feto-placental endothelial cells (n = 4). Data are given as mean ± SEM. Statistical analysis used the mean of the triplicate of the number of individual biological replicates. Control medium (C) = DMEM/EBM + 7.5 % FCS for 48 h at 37 °C without cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4930480&req=5

Fig1: Effect of first (FTB) and third (TTB) trimester trophoblast-conditioned medium (CM) on network formation, migration, proliferation and survival of feto-placental endothelial cells. a Representative phase contrast light microscopic images show network formation of feto-placental endothelial cells after 12 h on growth factor-reduced Matrigel. CM of peripheral blood monocyte cells (PBMC) used as positive control. b Quantitative analysis of networks depicts the inhibitory effect of TTB CM on feto-placental endothelial cells (n = 6). c CM of FTB and TTB reduced migration of feto-placental endothelial cells compared to control (n = 6). Endothelial basal medium (EBM) supplemented with FBS and growth factors (EBM++) was used as positive control. d Proliferation was unchanged between early and late trophoblast CM (n = 4). EBM supplemented with FCS and growth factors (EBM++) was used as positive control. e FTB and TTB CM did not increase LDH release of feto-placental endothelial cells (n = 4). Data are given as mean ± SEM. Statistical analysis used the mean of the triplicate of the number of individual biological replicates. Control medium (C) = DMEM/EBM + 7.5 % FCS for 48 h at 37 °C without cells
Mentions: To investigate the paracrine effect of trophoblast from early versus late pregnancy on angiogenesis, the influence of CM on network formation, migration, proliferation and survival of feto-placental endothelial cells was analyzed (Fig. 1). CM of trophoblast from late pregnancy (term trophoblasts; TTB) reduced network formation of feto-placental endothelial cells by 30 ± 14 % (p = 0.024) (Fig. 1a, b), while CM of trophoblast from early pregnancy (first trimester trophoblasts; FTB) had no effect. CM of peripheral blood monocyte cells (PBMC) used as positive control [18] stimulated network formation by 21 ± 10 % (p = 0.049). Similar results were obtained with CM that had been ultracentrifuged prior to use (not shown), indicating that soluble factors secreted by trophoblast induce the anti-angiogenic effects.Fig. 1

Bottom Line: Notably, human recombinant PEDF reduced network formation only in combination with VEGF.Analysis of phosphorylation of ERK1/2 and FAK, two key players in VEGF-induced proliferation and migration, revealed that PEDF altered VEGF signaling, while PEDF alone did not affect phosphorylation of ERK1/2 and FAK.These data suggest that the trophoblast-derived anti-angiogenic molecule PEDF is involved in restricting growth and expansion of the feto-placental endothelium predominantly in late pregnancy and targets to modulate the intracellular effect of VEGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Graz, Austria.

ABSTRACT
The rapidly expanding feto-placental vasculature needs tight control by paracrine and endocrine mechanisms. Here, we focused on paracrine influence by trophoblast, the placental epithelium. We aimed to identify differences in regulation of feto-placental angiogenesis in early versus late pregnancy. To this end, the effect of conditioned media (CM) from early and late pregnancy human trophoblast was tested on network formation, migration and proliferation of human feto-placental endothelial cells. Only CM of late pregnancy trophoblast reduced network formation and migration. Screening of trophoblast transcriptome for anti-angiogenic candidates identified pigment epithelium-derived factor (PEDF) with higher expression and protein secretion in late pregnancy trophoblast. Addition of a PEDF-neutralizing antibody restored the anti-angiogenic effect of CM from late pregnancy trophoblast. Notably, human recombinant PEDF reduced network formation only in combination with VEGF. Also in the CAM assay, the combination of PEDF with VEGF reduced branching of vessels below control levels. Analysis of phosphorylation of ERK1/2 and FAK, two key players in VEGF-induced proliferation and migration, revealed that PEDF altered VEGF signaling, while PEDF alone did not affect phosphorylation of ERK1/2 and FAK. These data suggest that the trophoblast-derived anti-angiogenic molecule PEDF is involved in restricting growth and expansion of the feto-placental endothelium predominantly in late pregnancy and targets to modulate the intracellular effect of VEGF.

No MeSH data available.


Related in: MedlinePlus