G Protein α Subunit 14 Mediates Fibroblast Growth Factor 2‐Induced Cellular Responses in Human Endothelial Cells

Abstract

During pregnancy, a tremendous increase in fetoplacental angiogenesis is associated with elevated blood flow. Aberrant fetoplacental vascular function may lead to pregnancy complications including pre‐eclampsia. Fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor A (VEGFA) are crucial regulators of fetoplacental endothelial function. G protein α subunit 14 (GNA14), a member of Gαq/11 subfamily is involved in mediating hypertensive diseases and tumor vascularization. However, little is known about roles of GNA14 in mediating the FGF2‐ and VEGFA‐induced fetoplacental endothelial function. Using human umbilical vein endothelial cells (HUVECs) cultured under physiological chronic low oxygen (3% O2) as a cell model, we show that transfecting cells with adenovirus carrying GNA14 complementary DNA (cDNA; Ad‐GNA14) increases (p < 0.05) protein expression of GNA14. GNA14 overexpression blocks (p < 0.05) FGF2‐stimulated endothelial migration, whereas it enhances (p < 0.05) endothelial monolayer integrity (maximum increase of ~35% over the control at 24 hr) in response to FGF2. In contrast, GNA14 overexpression does not significantly alter VEGFA‐stimulated cell migration, VEGFA‐weakened cell monolayer integrity, and intracellular Ca++ mobilization in response to adenosine triphosphate (ATP), FGF2, and VEGFA. GNA14 overexpression does not alter either FGF2‐ or VEGFA‐induced phosphorylation of ERK1/2. However, GNA14 overexpression time‐dependently elevates (p < 0.05) phosphorylation of phospholipase C‐β3 (PLCβ3) at S1105 in response to FGF2, but not VEGFA. These data suggest that GNA14 distinctively mediates fetoplacental endothelial cell migration and permeability in response to FGF2 and VEGFA, possibly in part by altering activation of PLCβ3 under physiological chronic low oxygen.

Endothelial Ca2+ Signaling, Angiogenesis and Vasculogenesis: Just What It Takes to Make a Blood Vessel

“…It has been reported that Gα q monomers activate PLCβ isotypes according to the following rank order: PLCβ1 ≥ PLCβ3 > PLCβ2, while PLCβ4 recruitment is heavily limited by ribonucleotides, such as GTP-γ-S [60,61]. The role played by the distinct PLCβ isotypes in the endothelial Ca 2+ response to G q/11 PCR has not been carefully dissected, but preliminary evidence suggested the involvement of PLCβ1 [62] and PLCβ3 [63]. In addition, PLCβ1-3, but not PLCβ4, are also sensitive to Gβγ dimers, although they display high affinity only towards PLCβ2 [60].…”

“…As expected by their ability to bind to RTK coupled to the PLCγ1/InsP 3 signaling axis, many other growth factors can induce pro-angiogenic Ca 2+ signals in vascular endothelial cells [41,199,200]. For instance, bFGF, also known as FGF-2, triggers an increase in [Ca 2+ ] i in a variety of vascular endothelial cells, including HUVEC [63,199,201], SV40-transfected human corneal endothelial cells (SV40-HCEC) [202], and BAEC [192,200]. The Ca 2+ response to bFGF is initiated by FGF receptor-1 (FGFR-1) [179,203], requires PLCγ1 activation [201], although the contribution of PLCβ3 has also been reported [63], and is essentially mediated by extracellular Ca 2+ entry [200].…”

Sexual Dimorphisms of Preeclampsia-Dysregulated Transcriptomic Profiles and Cell Function in Fetal Endothelial Cells

“…Cell monolayer integrity was determined using the ECIS Zθ+96-well array station and 96W10idf plates (Applied BioPhysics, NY) 10 . Cell proliferation was assessed using the CCK-8 kit (Dojindo Molecular Technologies, MD) 33 . Cell migration was assessed using a transwell system (Corning, NY) 33,34 .…”

“…Cell proliferation was assessed using the CCK-8 kit (Dojindo Molecular Technologies, MD) 33 . Cell migration was assessed using a transwell system (Corning, NY) 33,34 .…”

Tufted angioma with associated Kasabach‐Merritt phenomenon caused by somatic mutation in GNA14

“…GNA14 , a member of the alpha q subfamily of G proteins, is known to regulate Ras activity and modulate endothelial cell permeability and migration in response to FGF2 and VEGFA . Indeed, activating mutations in GNA14 and the Ras subfamily have been found in the same class of vascular tumors, and GNA14 p.Q205L mutation is common in anastomosing hemangiomas and cherry hemangiomas .…”