Regulation of the Matricellular Proteins CYR61 (CCN1) and NOV (CCN3) by Hypoxia-Inducible Factor-1α and Transforming-Growth Factor-β3 in the Human Trophoblast

Abstract

It is known that a hypoxic environment is critical for trophoblast migration and invasion and is fundamental for appropriate placental perfusion. Because cysteine-rich 61 (CYR61, CCN1) and nephroblastoma overexpressed (NOV, CCN3) are expressed in the extravillous trophoblast and expression levels are deregulated in preeclampsia, we investigated their regulation properties in first-trimester placental explants and in JEG3 choriocarcinoma cells upon a physiological low oxygen tension of 1-3%. In placental explants, both proteins were expressed in the extravillous trophoblast cells and were increased upon hypoxia. JEG3 cells revealed a significant up-regulation of CYR61 and NOV intracellular as well as secreted protein upon hypoxic treatment accompanied by the stabilization of the hypoxia-inducible factor-1alpha (HIF-1alpha). Treatment with dimethyloxalylglycine to mimic hypoxia and silencing of HIF-1alpha using small interfering RNA revealed that only the increase in intracellular protein expression seems to be dependent on HIF-1alpha but obviously not the secretion process. Moreover, recombinant TGF-beta3 was able to further enhance the amount of intracellular CCN proteins as well as secreted CYR61 levels under hypoxia. These results indicate that low oxygen levels trigger elevation of intracellular as well as secreted CYR61 and NOV protein probably in two independent pathways. Addition of recombinant CYR61 and NOV proteins increases migration as well as invasion properties of JEG3 trophoblast cells, which strengthen their role in supporting trophoblast migration invasion properties. In summary, CYR61 and NOV are regulated by HIF-1alpha and TGF-beta3 in the trophoblast cell line JEG3, and their enhanced secretion could be implicated in appropriate placental invasion.

CCN1 (CYR61) and CCN3 (NOV) signaling drives human trophoblast cells into senescence and stimulates migration properties

“…Comparing the effect of CCNs on migration properties in both cell lines showed that Jeg3 trophoblast cells and SGHPL-5 cells are mostly stimulated by non-glycosylated CCN1 and CCN3. [12][13][14] Thus, the regulation properties of CCNs on proliferation differ between the malignant and the benign trophoblast cell lines, and migration seems to be similarly regulated.…”

“…[12][13][14][15] CCN proteins are known to regulate pivotal cellular processes, such as differentiation, proliferation, migration, and angiogenesis. 16,17 Downstream signaling events are mediated by integrins, bone morphogenetic proteins (BMPs), vascular endothelial growth factor (VEGF), Wnt proteins, and Notch.…”

“…23 We have already demonstrated that in the malignant trophoblast cell line Jeg3, a model of invasive EVT, CCN3 reduces cell proliferation and enhances migration properties. [12][13][14] These studies showed that CCN3 acts by inducing the mitogen-activated protein kinase/extracellular signal-related kinase (MAPK/ERK), phosphatidylinositol 3-kinase/protein kinase B (PI3K/ Akt), and Notch/p21 pathways, mediating these separate functions for proliferation and migration/invasion in Jeg3 cells. 14 In the study reported here, we investigated the proliferation control of CCN1 and CCN3 in benign SGHPL-5 trophoblast cells, which are more similar to the in vivo situation than previous models.…”

Cysteine-rich matricellular protein improves callus regenerate in a rabbit trauma model

“…Moreover, the experimentally induced soft-tissue trauma in our experiment and with it the collapse of vascular volumes can lead to local hypoxia [24], resulting in a callus deficiency [9], which explains the smaller callus diaphsis index in the control group but a higher one after local CYR61 treatment. In turn, hypoxia stabilises and activates the transcription factor HIF-1α, one of the key regulators of CYR61 next to TGF-β3 [25]. The HIF and CYR61 pathway is likely to be critically important for the interplay of angio-and osteogenesis and thus for fracture healing and therefore for bone volume.…”

Hypoxia-Inducible Factor 1 Alpha Contributes to Cardiac Healing in Mesenchymal Stem Cells-Mediated Cardiac Repair

“…We next investigated the impact of HIF-1a on MSC gene expression. Gain-of-function studies in MSC showed that HIF-1a coordinately regulated the expression of genes encoding proteins and growth factors implicated in wound repair, angiogenesis, and extracellular matrix production, most of them previously described as HIF targets [30][31][32][33][34][35]. Proangiogenic factors including angiopoietin (ANGPT1), apelin (APLN), and the matricellular protein CYR61 (CNN1) were upregulated in MSC-HIF.…”