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Epidermal growth factor-like domain 7 (Egfl7) is a gene that encodes a partially secreted protein and whose expression is largely restricted to the endothelia. We recently reported that EGFL7 is also expressed by trophoblast cells in mouse and human placentas. Here, we investigated the molecular pathways that are regulated by EGFL7 in trophoblast cells. Stable EGFL7 overexpression in a Jeg3 human choriocarcinoma cell line resulted in significantly increased cell migration and invasiveness, while cell proliferation was unaffected. Analysis of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways showed that EGFL7 promotes Jeg3 cell motility by activating both pathways. We show that EGFL7 activates the epidermal growth factor receptor (EGFR) in Jeg3 cells, resulting in downstream activation of extracellular regulated kinases (ERKs). In addition, we provide evidence that EGFL7-triggered migration of Jeg3 cells involves activation of NOTCH signaling. EGFL7 and NOTCH1 are co-expressed in Jeg3 cells, and blocking of NOTCH activation abrogates enhanced migration of Jeg3 cells overexpressing EGFL7. We also demonstrate that signaling through EGFR and NOTCH converged to mediate EGFL7 effects. Reduction of endogenous EGFL7 expression in Jeg3 cells significantly decreased cell migration. We further confirmed that EGFL7 stimulates cell migration by using primary human first trimester trophoblast (PTB) cells overexpressing EGFL7. In conclusion, our data suggest that in trophoblast cells, EGFL7 regulates cell migration and invasion by activating multiple signaling pathways. Our results provide a possible explanation for the correlation between reduced expression of EGFL7 and inadequate trophoblast invasion observed in placentopathies.

Background and Purpose: The lectin-like oxidised LDL receptor-1 (OLR1) gene encodes a scavenger receptor implicated in the pathogenesis of atherosclerosis. Although functional roles have been suggested for two variants, epidemiological studies on OLR1 have been inconsistent. Methods - We tested the association between the non-synonymous substitution K167N (rs11053646) and intima media thickness of the common carotid artery (CCA-IMT) in 2,141 samples from the Progression of Lesions in the Intima of the Carotid (PLIC) study (a prospective population-based study). Results: Significantly increased IMT was observed in male carriers of the minor C (N) allele compared to GC and GG (KN and KK) genotype. Functional analysis on macrophages suggested a decreased association to Ox-LDL in NN carriers compared to KN and KK carriers which is also associated with a reduced OLR1 mRNA expression. Macrophages from NN carriers present also a specific inflammatory gene expression pattern compared to cells from KN and KK carriers. Conclusions: These data suggest that the 167N variant of LOX-1 receptor affects the atherogenic process in the carotid artery prior to evidence of disease through an inflammatory process. © 2012 Predazzi et al.

Objectives: Implantation and placental development are essential for the patterning of the embryo, for its growth and survival upon implantation, as well as for preserving the health of the fetus and mother. Inadequate invasion of trophoblast cells into the maternal decidua and the deficient growth of fetal blood vessels are actually recognized as possible pathogenetic factors of preeclampsia (PE). To examine molecular mechanisms that control these events, we are focusing in our studies on the endothelial gene Egfl7. Our preliminary studies show that Egfl7 has a critical role in molecular processes that control human vascular development and proper placental implantation: in particular, Egfl7 is expressed in the trophectoderm and in the fetal placental vasculature. To assess the involvement of Egfl7 in the development of PE, we investigate the EGFL7 expression patterns between PE and control placentas. Materials and Methods: Two study groups are considered: (1) a first of women with pregnancies complicated by early onset PE ; (2) a second study group of healthy women. The placental biopsies were obtained immediately after caesarean section from both groups, the PE patients and the normal controls. Real-time PCR and immunofluorescence analysis of EGFL7 expression in normal and preeclamptic human placental tissue were performed. Results: Real-time PCR analysis revealed that EGFL7 is strongly down regulated in the villi of PE placentas when compared to normal samples. In agreement with published data, VEGF expression was also decreased in PE placentas, although to a lesser extent compared to EGFL7. At variance, expression of the pan-endothelial marker CD31 was not significantly altered in PE as compared to normal placentas. These results were confirmed by immunofluorescence analysis. Conclusion: Our results suggest a role for Egfl7 in the pathogenesis of PE. We hypothesize that in PE placental villi the total number of endothelial cells is not significantly decreased (

Objectives: Implantation and placental development are essential for the patterning of the embryo, for its growth and survival upon implantation, as well as for preserving the health of the fetus and mother. Inadequate invasion of trophoblast cells into the maternal decidua and the deficient growth of fetal blood vessels are actually recognized as possible pathogenetic factors of preeclampsia (PE). To examine molecular mechanisms that control these events, we are focusing in our studies on the endothelial gene Egfl7. Our preliminary studies show that Egfl7 has a critical role in molecular processes that control human vascular development and proper placental implantation: in particular, Egfl7 is expressed in the trophectoderm and in the fetal placental vasculature. To assess the involvement of Egfl7 in the development of PE, we investigate the EGFL7 expression patterns between PE and control placentas. Materials and Methods: Two study groups are considered: (1) a first of women with pregnancies complicated by early onset PE ; (2) a second study group of healthy women. The placental biopsies were obtained immediately after caesarean section from both groups, the PE patients and the normal controls. Real-time PCR and immunofluorescence analysis of EGFL7 expression in normal and preeclamptic human placental tissue were performed. Results: Real-time PCR analysis revealed that EGFL7 is strongly down regulated in the villi of PE placentas when compared to normal samples. In agreement with published data, VEGF expression was also decreased in PE placentas, although to a lesser extent compared to EGFL7. At variance, expression of the pan-endothelial marker CD31 was not significantly altered in PE as compared to normal placentas. These results were confirmed by immunofluorescence analysis. Conclusion: Our results suggest a role for Egfl7 in the pathogenesis of PE. We hypothesize that in PE placental villi the total number of endothelial cells is not significantly decreased (

Inhibition of bone formation after surgery to correct craniosynostosis would alleviate the need for secondary surgeries and decrease morbidity and mortality. This study used a single dose of Noggin protein to prevent resynostosis and improve postoperative outcomes in a rabbit model of craniosynostosis. Introduction: Craniosynostosis is defined as the premature fusion of one or more of the cranial sutures, which causes secondary deformations of the cranial vault, cranial base, and brain. Current surgical intervention involves extirpation of the fused suture to allow unrestricted brain growth. However, resynostosis of the extirpated regions often occurs. Several bone morphogenetic proteins (BMPs), well-described inducers of ossification, are involved in bone healing. This study tested the hypothesis that a postoperative treatment with Noggin, an extracellular BMP inhibitor, can inhibit resynostosis in a rabbit model of human familial nonsyndromic craniosynostosis. Materials and Methods: Thirty-one New Zealand white rabbits with bilateral coronal suture synostosis were divided into three groups: (1) suturectomy controls (n = 13); (2) suturectomy with BSA in a slow-resorbing collagen vehicle, (n = 8); and (3) suturectomy with Noggin in a slow-resorbing collagen vehicle (n = 10). At 10 days of age, a 3 x 15-mm coronal suturectomy was performed. The sites in groups 2 and 3 were immediately filled with BSA-loaded gel or Noggin-loaded gel, respectively. Serial 3D-CT scan reconstructions of the defects and standard radiographs were obtained at 10, 25, 42, and 84 days of age, and the sutures were harvested for histological analysis. Results: Radiographic analysis revealed that Noggin-treated animals had significantly greater coronal suture marker separation by 25 days and significantly greater craniofacial length at 84 days of age compared with controls. 3D-CT analysis revealed that Noggin treatment led to significantly greater defect areas through 84 days and to increased in

Inhibition of bone formation after surgery to correct craniosynostosis would alleviate the need for secondary surgeries and decrease morbidity and mortality. This study used a single dose of Noggin protein to prevent resynostosis and improve postoperative outcomes in a rabbit model of craniosynostosis. Introduction: Craniosynostosis is defined as the premature fusion of one or more of the cranial sutures, which causes secondary deformations of the cranial vault, cranial base, and brain. Current surgical intervention involves extirpation of the fused suture to allow unrestricted brain growth. However, resynostosis of the extirpated regions often occurs. Several bone morphogenetic proteins (BMPs), well-described inducers of ossification, are involved in bone healing. This study tested the hypothesis that a postoperative treatment with Noggin, an extracellular BMP inhibitor, can inhibit resynostosis in a rabbit model of human familial nonsyndromic craniosynostosis. Materials and Methods: Thirty-one New Zealand white rabbits with bilateral coronal suture synostosis were divided into three groups: (1) suturectomy controls (n = 13); (2) suturectomy with BSA in a slow-resorbing collagen vehicle, (n = 8); and (3) suturectomy with Noggin in a slow-resorbing collagen vehicle (n = 10). At 10 days of age, a 3 x 15-mm coronal suturectomy was performed. The sites in groups 2 and 3 were immediately filled with BSA-loaded gel or Noggin-loaded gel, respectively. Serial 3D-CT scan reconstructions of the defects and standard radiographs were obtained at 10, 25, 42, and 84 days of age, and the sutures were harvested for histological analysis. Results: Radiographic analysis revealed that Noggin-treated animals had significantly greater coronal suture marker separation by 25 days and significantly greater craniofacial length at 84 days of age compared with controls. 3D-CT analysis revealed that Noggin treatment led to significantly greater defect areas through 84 days and to increased in