Glycated fibroblast growth factor-2 is quickly produced in vitro upon low-millimolar glucose treatment and detected in vivo in diabetic mice

Francesco Facchiano, Daniela D'Arcangelo, Katia Russo, Vincenzo Fogliano, Carmela Mennella, Raffaele Ragone, Giovanna Zambruno, Virginia Carbone, Domenico Ribatti, Cesare Peschle, Maurizio C. Capogrossi, Antonio Facchiano

Research output: Contribution to journalArticle

Abstract

Angiogenesis impairment in hyperglycemic patients represents a leading cause of severe vascular complications of both type-1 and -2 diabetes mellitus (DM). Angiogenesis dysfunction in DM is related to glycemic control; however, molecular mechanisms involved are still unclear. Fibroblast growth factor-2 (FGF-2) is a potent angiogenic factor and, according to previous evidence, may represent a key target of molecular modifications triggered by high-sugar exposure. Therefore, the purpose of this study was to investigate whether short incubation with hyperglycemic levels of glucose affected FGF-2 and whether glucose-modified FGF-2 was detectable in vivo. Biochemical analyses carried out with SDS-PAGE, fluorescence emission, mass-spectrometry, immunoblot, and competitive ELISA experiments demonstrated that human FGF-2 undergoes a rapid and specific glycation upon 12.5-50 mM glucose exposure. In addition, FGF-2 exposed for 30 min to 12.5 mM glucose lost mitogenic and chemotactic activity in a time- and dose-dependent manner. Under similar conditions, binding affinity to FGF receptor 1 was dramatically reduced by 20-fold, as well as FGF receptor 1 and ERK-1/2 phosphorylation, and FGF-2 lost about 45% of angiogenic activity in two different in vivo angiogenic (Matrigel and chorioallantoic-membrane) assays. Such glucose-induced modification was specific, because other angiogenic growth factors, namely platelet-derived growth factor BB and placental-derived growth factor were not significantly or markedly less modified. Finally, for the first time, glycated-FGF-2 was detected in vivo, in tissues from hyperglycemic nonobese diabetic mice, in significantly higher amounts than in normoglycemic mice. In conclusion, hyperglycemic levels of glucose may strongly affect FGF-2 structure and impair its angiogenic features, and endogenous glycated-FGF-2 is present in diabetic mice, indicating a novel pathogenetic mechanism underlying angiogenesis defects in DM.

Original languageEnglish
Pages (from-to)2806-2818
Number of pages13
JournalMolecular Endocrinology
Volume20
Issue number11
DOIs
Publication statusPublished - Nov 2006

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Fibroblast Growth Factor 2
Glucose
Fibroblast Growth Factor 1
Fibroblast Growth Factor Receptors
Therapeutics
Angiogenesis Inducing Agents
Intercellular Signaling Peptides and Proteins
Diabetes Mellitus
In Vitro Techniques
Chorioallantoic Membrane
Obese Mice
Inbred NOD Mouse
Type 1 Diabetes Mellitus
Type 2 Diabetes Mellitus
Blood Vessels
Polyacrylamide Gel Electrophoresis
Mass Spectrometry
Fluorescence
Enzyme-Linked Immunosorbent Assay
Phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Glycated fibroblast growth factor-2 is quickly produced in vitro upon low-millimolar glucose treatment and detected in vivo in diabetic mice. / Facchiano, Francesco; D'Arcangelo, Daniela; Russo, Katia; Fogliano, Vincenzo; Mennella, Carmela; Ragone, Raffaele; Zambruno, Giovanna; Carbone, Virginia; Ribatti, Domenico; Peschle, Cesare; Capogrossi, Maurizio C.; Facchiano, Antonio.

In: Molecular Endocrinology, Vol. 20, No. 11, 11.2006, p. 2806-2818.

Research output: Contribution to journalArticle

Facchiano, Francesco ; D'Arcangelo, Daniela ; Russo, Katia ; Fogliano, Vincenzo ; Mennella, Carmela ; Ragone, Raffaele ; Zambruno, Giovanna ; Carbone, Virginia ; Ribatti, Domenico ; Peschle, Cesare ; Capogrossi, Maurizio C. ; Facchiano, Antonio. / Glycated fibroblast growth factor-2 is quickly produced in vitro upon low-millimolar glucose treatment and detected in vivo in diabetic mice. In: Molecular Endocrinology. 2006 ; Vol. 20, No. 11. pp. 2806-2818.
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abstract = "Angiogenesis impairment in hyperglycemic patients represents a leading cause of severe vascular complications of both type-1 and -2 diabetes mellitus (DM). Angiogenesis dysfunction in DM is related to glycemic control; however, molecular mechanisms involved are still unclear. Fibroblast growth factor-2 (FGF-2) is a potent angiogenic factor and, according to previous evidence, may represent a key target of molecular modifications triggered by high-sugar exposure. Therefore, the purpose of this study was to investigate whether short incubation with hyperglycemic levels of glucose affected FGF-2 and whether glucose-modified FGF-2 was detectable in vivo. Biochemical analyses carried out with SDS-PAGE, fluorescence emission, mass-spectrometry, immunoblot, and competitive ELISA experiments demonstrated that human FGF-2 undergoes a rapid and specific glycation upon 12.5-50 mM glucose exposure. In addition, FGF-2 exposed for 30 min to 12.5 mM glucose lost mitogenic and chemotactic activity in a time- and dose-dependent manner. Under similar conditions, binding affinity to FGF receptor 1 was dramatically reduced by 20-fold, as well as FGF receptor 1 and ERK-1/2 phosphorylation, and FGF-2 lost about 45{\%} of angiogenic activity in two different in vivo angiogenic (Matrigel and chorioallantoic-membrane) assays. Such glucose-induced modification was specific, because other angiogenic growth factors, namely platelet-derived growth factor BB and placental-derived growth factor were not significantly or markedly less modified. Finally, for the first time, glycated-FGF-2 was detected in vivo, in tissues from hyperglycemic nonobese diabetic mice, in significantly higher amounts than in normoglycemic mice. In conclusion, hyperglycemic levels of glucose may strongly affect FGF-2 structure and impair its angiogenic features, and endogenous glycated-FGF-2 is present in diabetic mice, indicating a novel pathogenetic mechanism underlying angiogenesis defects in DM.",
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AU - D'Arcangelo, Daniela

AU - Russo, Katia

AU - Fogliano, Vincenzo

AU - Mennella, Carmela

AU - Ragone, Raffaele

AU - Zambruno, Giovanna

AU - Carbone, Virginia

AU - Ribatti, Domenico

AU - Peschle, Cesare

AU - Capogrossi, Maurizio C.

AU - Facchiano, Antonio

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AB - Angiogenesis impairment in hyperglycemic patients represents a leading cause of severe vascular complications of both type-1 and -2 diabetes mellitus (DM). Angiogenesis dysfunction in DM is related to glycemic control; however, molecular mechanisms involved are still unclear. Fibroblast growth factor-2 (FGF-2) is a potent angiogenic factor and, according to previous evidence, may represent a key target of molecular modifications triggered by high-sugar exposure. Therefore, the purpose of this study was to investigate whether short incubation with hyperglycemic levels of glucose affected FGF-2 and whether glucose-modified FGF-2 was detectable in vivo. Biochemical analyses carried out with SDS-PAGE, fluorescence emission, mass-spectrometry, immunoblot, and competitive ELISA experiments demonstrated that human FGF-2 undergoes a rapid and specific glycation upon 12.5-50 mM glucose exposure. In addition, FGF-2 exposed for 30 min to 12.5 mM glucose lost mitogenic and chemotactic activity in a time- and dose-dependent manner. Under similar conditions, binding affinity to FGF receptor 1 was dramatically reduced by 20-fold, as well as FGF receptor 1 and ERK-1/2 phosphorylation, and FGF-2 lost about 45% of angiogenic activity in two different in vivo angiogenic (Matrigel and chorioallantoic-membrane) assays. Such glucose-induced modification was specific, because other angiogenic growth factors, namely platelet-derived growth factor BB and placental-derived growth factor were not significantly or markedly less modified. Finally, for the first time, glycated-FGF-2 was detected in vivo, in tissues from hyperglycemic nonobese diabetic mice, in significantly higher amounts than in normoglycemic mice. In conclusion, hyperglycemic levels of glucose may strongly affect FGF-2 structure and impair its angiogenic features, and endogenous glycated-FGF-2 is present in diabetic mice, indicating a novel pathogenetic mechanism underlying angiogenesis defects in DM.

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