3,200 research outputs found
Role of endothelial cell extracellular signal-regulated kinase1/2 in urokinase-type plasminogen activator upregulation and in vitro angiogenesis by fibroblast growth factor-2.
No full textDownstream signaling triggered by the binding of fibroblast growth factor-2 (FGF2) to its tyrosine-kinase receptors involves the activation of mitogen-activated protein kinase kinase (MEK) with consequent phosphorylation of extracellular signal-regulated kinases (ERKs). Here we demonstrate that FGF2 induces ERK(1/2) activation in bovine aortic endothelial (BAE) cells and that the continuous presence of the growth factor is required for sustained ERK(1/2) phosphorylation. This is prevented by the MEK inhibitors PD 098059 and U0126, which also inhibit FGF2-mediated upregulation of urokinase-type plasminogen activator (uPA) and in vitro formation of capillary-like structures in three-dimensional type I collagen gel. Various FGF2 mutants originated by deletion or substitution of basic amino acid residues in the amino terminus or in the carboxyl terminus of FGF2 retained the capacity to induce a long-lasting activation of ERK(1/2) in BAE cells. Among them, K128Q/R129Q-FGF2 was also able to stimulate uPA production and morphogenesis whereas R129Q/K134Q-FGF2 caused uPA upregulation only. In contrast, K27,30Q/R31Q-FGF2, K128Q/K138Q-FGF2 and R118,129Q/K119,128Q-FGF2 exerted a significant uPA-inducing and morphogenic activity in an ERK(1/2)-dependent manner only in the presence of heparin. Furthermore, no uPA upregulation and morphogenesis was observed in BAE cells treated with the deletion mutant Δ27-32-FGF2 even in the presence of soluble heparin. Thus, mutational analysis of FGF2 dissociates the capacity of the growth factor to induce a persistent activation of ERK(1/2) from its ability to stimulate uPA upregulation and/or in vitro angiogenesis. In conclusion, the data indicate that ERK(1/2) phosphorylation is a key step in the signal transduction pathway switched on by FGF2 in endothelial cells. Nevertheless, a sustained ERK(1/2) activation is not sufficient to trigger uPA upregulation and morphogenesis. FGF2 mutants may represent useful tools to dissect the signal transduction pathway(s) mediating the complex response elicited by an angiogenic stimulus in endothelial cells
The interaction of basic fibroblast growth factor (bFGF) with heparan sulfate proteoglycans: biochemical bases and biological implications
No full textImpact of fibroblast growth factor-2 (FGF2) on tumor growth and vascularization: Establishing a "FGF2-dependent" tumor model
No full textVariations on the Author
No full text“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Fibroblast growth factor modulates mast cell recruitment in a murine model of prostate cancer
No full textMast cells are important modifiers of prostate tumor microenvironment. The fibroblast growth factor/fibroblast growth factor receptor (FGF/FGFR) system plays a non-redundant autocrine/paracrine role in the growth, vascularization and progression of prostate tumors. Accordingly, the FGF antagonist long pentraxin-3 (PTX3) and the PTX3-derived small molecule FGF-trap NSC12 have been shown to inhibit the growth and vascularization of different FGF-dependent tumor types, including prostate cancer. In this study, we show that recombinant FGF2 is able to cause mast cell recruitment in vivo in the Matrigel plug assay. Conversely, PTX3 overexpression in transgenic mice or treatment with the FGF inhibitor NSC12 result in a significant inhibition of the growth and vascularization of TRAMP-C2 tumor grafts, a murine model of prostate cancer, that were paralleled by a decrease of mast cell infiltrate into the lesion. These data confirm and extend previous observations about the capacity of mast cells to respond chemotactically to FGF2 stimulation and provide evidence about a relationship among mast cell recruitment, angiogenesis, and tumor growth in human prostate adenocarcinom
β3 Integrin promotes long-lasting activation and polarization of vascular endothelial growth factor receptor 2 by immobilized ligand
No full textOBJECTIVE:
During neovessel formation, angiogenic growth factors associate with the extracellular matrix. These immobilized factors represent a persistent stimulus for the otherwise quiescent endothelial cells (ECs), driving directional EC migration and proliferation and leading to new blood vessel growth. Vascular endothelial growth factor receptor 2 (VEGFR2) is the main mediator of angiogenesis. Although VEGFR2 signaling has been deeply characterized, little is known about its subcellular localization during neovessel formation. Aim of this study was the characterization and molecular determinants of activated VEGFR2 localization in ECs during neovessel formation in response to matrix-immobilized ligand.
APPROACH AND RESULTS:
Here we demonstrate that ECs stimulated by extracellular matrix-associated gremlin, a noncanonical VEGFR2 ligand, are polarized and relocate the receptor in close contact with the angiogenic factor-enriched matrix both in vitro and in vivo. GM1 (monosialotetrahexosylganglioside)-positive planar lipid rafts, β3 integrin receptors, and the intracellular signaling transducers focal adhesion kinase and RhoA (Ras homolog gene family, member A) cooperate to promote VEGFR2 long-term polarization and activation.
CONCLUSIONS:
A ligand anchored to the extracellular matrix induces VEGFR2 polarization in ECs. Long-lasting VEGFR2 relocation is closely dependent on lipid raft integrity and activation of β3 integrin pathway. The study of the endothelial responses to immobilized growth factors may offer insights into the angiogenic process in physiological and pathological conditions, including cancer, and for a better engineering of synthetic tissue scaffolds to blend with the host vasculature
Long Pentraxin-3 Inhibits FGF8b-Dependent Angiogenesis and Growth of Steroid Hormone-Regulated Tumors.
No full textFibroblast growth factor-8b (FGF8b) exerts nonredundant autocrine/paracrine functions in steroid hormone-regulated tumors. Previous observations had shown that the soluble pattern recognition receptor long pentraxin-3 (PTX3) is a natural selective antagonist for a restricted number of FGF family members, inhibiting FGF2 but not FGF1 and FGF4 activity. Here, we assessed the capacity of PTX3 to antagonize FGF8b and to inhibit the vascularization and growth of steroid hormone-regulated tumors. Surface plasmon resonance analysis shows that PTX3 binds FGF8b with high affinity (K d = 30-90 nmol/L). As a consequence, PTX3 prevents the binding of FGF8b to its receptors, inhibits FGF8b-driven ERK1/2 activation, cell proliferation, and chemotaxis in endothelial cells, and suppresses FGF8b-induced neovascularization in vivo. Also, PTX3 inhibits dihydrotestosterone (DHT)- and FGF8b-driven proliferation of androgen-regulated Shionogi 115 (S115) mouse breast tumor cells. Furthermore, DHT-treated, PTX3 overexpressing hPTX3-S115 cell transfectants show a reduced proliferation rate in vitro and a limited angiogenic activity in the chick embryo chorioallantoic membrane and murine s.c. Matrigel plug assays. Accordingly, hPTX3-S115 cells show a dramatic decrease of their tumorigenic activity when grafted in immunodeficient male mice. These results identify PTX3 as a novel FGF8b antagonist endowed with antiangiogenic and antineoplastic activity with possible implications for the therapy of hormonal tumor
Imparare le lingue per abbracciare il mondo
No full textL’autrice che si colloca, come Daniela Zorzi, tra i fautori di una linguistica “impegnata”, propone in ricordo della studiosa amica, una riflessione sul ruolo cruciale del multilinguismo e delle lingue- culture, per il successo professionale e il benessere sociale, nella società contemporanea. Il multilinguismo appare come un’urgenza educativa che interpella studiosi e formatori a tutti i livelli al fine di promuovere nei giovani un apprendimento consapevole efficace ed autonomo.Multilingualism constitutes an urgent objective for education, which needs to involve teachers and researchers at all levels if we are to promote effective and autonomous learning in the young. Like Daniela Zorzi, the author considers herself a socially engaged linguist, and in memory of her friend’s work, offers a reflection on the key roles of languages and cultures and of multilingualism for professional success and social welfare today
Basic fibroblast growth factor expression in endothelial cells: an autocrine role in angiogenesis?
No full textBiochemical bases of the interaction of the human basic fibroblast growth factor with glycosaminoglycans: new insights from trypsin digestion studies
No full textHeparins from bovine mucosa and lung, and chemically modified
heparins were assayed for their capacity to: (i) protect human
recombinant basic fibroblast growth factor (bFGF) from tryptic
cleavage; (ii) prevent 1251-bFGF binding to heparan sulphate
proteoglycans present in the extracellular matrix and on the cell
surface of fetal bovine aortic endothelial GM 7373 cell cultures;
(iii) affect 1251-bFGF binding to high-affinity tyrosine kinase
FGF receptors present on the cell membrane of GM 7373 cells;
(iv) inhibit the mitogenic activity exerted by bFGF in the same
cells. The results demonstrate thatthe potency shown by mucosal
heparins in the different assays is a direct function of size, verylow-
molecular-mass heparin (2.0 kDa) being significantly less
effective on a molar basis than unfractionated heparin (13.6 kDa).
Increased flexibility of the backbone structure, as observed in
reduced/oxidized heparins of different size, does not affect the
capacity of the polysaccharide to interact with bFGF. In contrast,
selective 2-O-desulphation, but not 6-O-desulphation, drastically
reduced the capacity of heparin to protect bFGF from proteolytic
cleavage, to affect its interaction with low- and high-affinity sites,
and to inhibit its mitogenic activity. Two preparations of bovine
lung heparin, differing in molecular mass, were as effective as
mucosal heparin in the bFGF-tryptic-digestion assay and the
endothelial-cell proteoglycan-binding assay, but they were highly
inefficient at inhibiting the capacity of bFGF to interact with its
tyrosine kinase receptors. Bovine lung heparins were also less
effective than mucosal heparin as bFGF antagonists in GM
7373-cell-proliferation assays. N-Desulphated/N-acetylated
bovine lung heparin retained only a significant capacity to
protect bFGF from tryptic cleavage. The results demonstrate
that different chemical features of the heparin molecule, including
decrease in molecular mass, selective desulphation, disaccharide
composition and clustering, affect differently the capacity of the
glycosaminoglycan to interact with bFGF and to influence its
biological behaviour in different assays in vitro and in endothelial
cell cultures. Our findings should aid the design of synthetic
oligosaccharides aimed at improving the. bioavailability of bFGF
when administered in vivo as a therapeutic agent
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