Fibroblast Growth Factor 23 Stimulates Cardiac Fibroblast Activity through Phospholipase C-Mediated Calcium Signaling

Fibroblast growth factor (FGF)-23 induces hypertrophy and calcium (Ca2+) dysregulation in cardiomyocytes, leading to cardiac arrhythmia and heart failure. However, knowledge regarding the effects of FGF-23 on cardiac fibrogenesis remains limited. This study investigated whether FGF-23 modulates cardiac fibroblast activity and explored its underlying mechanisms. We performed MTS analysis, 5-ethynyl-2’-deoxyuridine assay, and wound-healing assay in cultured human atrial fibroblasts without and with FGF-23 (1, 5 and 25 ng/mL for 48 h) to analyze cell proliferation and migration. We found that FGF-23 (25 ng/mL, but not 1 or 5 ng/mL) increased proliferative and migratory abilities of human atrial fibroblasts. Compared to control cells, FGF-23 (25 ng/mL)-treated fibroblasts had a significantly higher Ca2+ entry and intracellular inositol 1,4,5-trisphosphate (IP3) level (assessed by fura-2 ratiometric Ca2+ imaging and enzyme-linked immunosorbent assay). Western blot analysis showed that FGF-23 (25 ng/mL)-treated cardiac fibroblasts had higher expression levels of calcium release-activated calcium channel protein 1 (Orai1) and transient receptor potential canonical (TRPC) 1 channel, but similar expression levels of α-smooth muscle actin, collagen type IA1, collagen type Ⅲ, stromal interaction molecule 1, TRPC 3, TRPC6 and phosphorylated-calcium/calmodulin-dependent protein kinase II when compared with control fibroblasts. In the presence of ethylene glycol tetra-acetic acid (a free Ca2+ chelator, 1 mM) or U73122 (an inhibitor of phospholipase C, 1 μM), control and FGF-23-treated fibroblasts exhibited similar proliferative and migratory abilities. Moreover, polymerase chain reaction analysis revealed that atrial fibroblasts abundantly expressed FGF receptor 1 but lacked expressions of FGF receptors 2-4. FGF-23 significantly increased the phosphorylation of FGF receptor 1. Treatment with PD166866 (an antagonist of FGF receptor 1, 1 μM) attenuated the effects of FGF-23 on cardiac fibroblast activity. In conclusion, FGF-23 may activate FGF receptor 1 and subsequently phospholipase C/IP3 signaling pathway, leading to an upregulation of Orai1 and/or TRPC1-mediated Ca2+ entry and thus enhancing human atrial fibroblast activity.

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The association of circulating ferritin with serum concentrations of fibroblast growth factor-23 measured by three commercial assays

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/000456307781646102 ◽

2007 ◽

Vol 44 (5) ◽

pp. 463-466 ◽

Cited By ~ 51

Author(s):

Brian H Durham ◽

Frank Joseph ◽

Lisa M Bailey ◽

William D Fraser

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Serum Ferritin ◽

Fibroblast Growth Factor 23 ◽

Inverse Correlation ◽

Biologically Active ◽

Enzyme Linked Immunosorbent Assay ◽

Fibroblast Growth ◽

Serum Samples ◽

Fgf 23

Background: The measurement of the serum concentration of fibroblast growth factor-23 (FGF-23) is beginning to be used as a diagnostic tool in renal phosphate wasting disorders. Having observed an increased serum FGF-23 in three subjects with low circulating ferritin concentrations we investigated the association between low ferritin and raised serum FGF-23. Methods: We measured FGF-23 in 150 random anonymized serum samples with ferritin concentrations between <5 and 50 µg/L using three commercially available enzyme-linked immunosorbent assay (ELISA) kits. One kit, Human FGF-23[C-term] (Immutopics Inc, USA) measures total FGF-23 whereas the other two kits, Immutopics intact and FGF-23 ELISA (Kainos, Japan) are reported to measure only the biologically active intact molecule. Results: We have detected a significant inverse correlation of -0.565 ( P<0.0001) between serum ferritin when <50 µg/L and FGF-23 using the C-terminal assay. This relationship is also shown with the Immutopics intact assay but is not demonstrated with the Kainos intact assay. Conclusion: The measurement of FGF-23 by both Immutopics assays is altered in the presence of low circulating concentrations of serum ferritin whereas with the Kainos intact assay this effect was not demonstrated. Serum ferritin should be measured when an elevated FGF-23 is obtained using the Immutopics C-terminal or intact FGF-23 assay to prevent misdiagnosis of the cause of this abnormality.

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Fibroblast Growth Factor (FGF) 23 Regulates the Plasma Levels of Parathyroid Hormone In Vivo Through the FGF Receptor in Normocalcemia, But Not in Hypocalcemia

Calcified Tissue International ◽

10.1007/s00223-017-0333-9 ◽

2017 ◽

Vol 102 (1) ◽

pp. 85-92 ◽

Cited By ~ 17

Author(s):

Maria L. Mace ◽

Eva Gravesen ◽

Anders Nordholm ◽

Klaus Olgaard ◽

Ewa Lewin

Keyword(s):

Parathyroid Hormone ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Plasma Levels ◽

Fibroblast Growth ◽

Fgf Receptor ◽

Fgf 23

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Direct activation of phospholipase C-gamma by fibroblast growth factor receptor is not required for mesoderm induction in Xenopus animal caps.

Molecular and Cellular Biology ◽

10.1128/mcb.14.5.3006 ◽

1994 ◽

Vol 14 (5) ◽

pp. 3006-3012 ◽

Cited By ~ 22

Author(s):

A J Muslin ◽

K G Peters ◽

L T Williams

Keyword(s):

Growth Factor ◽

Phospholipase C ◽

Fibroblast Growth Factor ◽

Mammalian Cells ◽

Platelet Derived Growth Factor ◽

Mesoderm Induction ◽

Fibroblast Growth ◽

Gamma Activation ◽

Fgf Receptor ◽

Phospholipase C Gamma

Members of the fibroblast growth factor (FGF) family induce mesoderm formation in explants of Xenopus embryonic ectoderm (animal caps). Recent studies have been directed at determining signaling pathways downstream of the FGF receptor that are important in mesoderm induction. We have recently shown that a point mutation in the FGF receptor changing tyrosine 766 to phenylalanine (Y/F mutation) abolishes phospholipase C-gamma (PLC-gamma) activation in mammalian cells. To explore the role of PLC-gamma activation in FGF-stimulated mesoderm induction, we constructed two chimeric receptors, each consisting of the extracellular portion of the platelet-derived growth factor beta receptor, with one having the transmembrane and intracellular portions of the wild-type FGF receptor 1 (PR-FR wt) and the other having the corresponding region of the Y/F766 mutant FGF receptor 1 (PR-FR Y/F766). When expressed in Xenopus oocytes, only PR-FR wt was able to mediate PLC gamma phosphorylation, inositol-1,4,5-trisphosphate accumulation, and calcium efflux in response to platelet-derived growth factor stimulation. However, both receptors mediated mesoderm induction in Xenopus animal caps as measured by cap elongation, muscle-specific actin mRNA induction, and skeletal muscle formation. These results demonstrate that PLC gamma activation by the FGF receptor is not required for FGF-stimulated mesoderm induction.

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Faculty Opinions recommendation of Pharmacological inhibition of fibroblast growth factor (FGF) receptor signaling ameliorates FGF23-mediated hypophosphatemic rickets.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717993579.793474216 ◽

2013 ◽

Author(s):

Murat Bastepe ◽

Serap Turan

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Hypophosphatemic Rickets ◽

Fibroblast Growth ◽

Receptor Signaling ◽

Fgf Receptor ◽

Pharmacological Inhibition

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Low Serum Levels of Fibroblast Growth Factor 2 in Gunn Rats: A Hyperbilirubinemia Animal Model of Schizophrenic Symptoms

CNS & Neurological Disorders - Drug Targets ◽

10.2174/1871527319999200729153907 ◽

2020 ◽

Vol 19 (7) ◽

pp. 503-508

Author(s):

Maiko Hayashida ◽

Sadayuki Hashioka ◽

Kenji Hayashida ◽

Shoko Miura ◽

Keiko Tsuchie ◽

...

Keyword(s):

Animal Model ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Wistar Rats ◽

Serum Levels ◽

Significant Negative Correlation ◽

Enzyme Linked Immunosorbent Assay ◽

Normal Strain ◽

Fibroblast Growth ◽

Gunn Rats

Background: Fibroblast growth factor (FGF) 2 (also referred to as basic FGF) is a multifunctional growth factor that plays a pivotal role in the pro-survival, pro-migration and pro-differentiation of neurons. Method: Because alterations in FGF2 levels are suggested to contribute to the pathogenesis schizophrenia, we investigated serum levels of FGF2 in the Gunn rat, a hyperbilirubinemia animal model of schizophrenic symptoms. Results: The enzyme-linked immunosorbent assay showed that the serum levels of FGF2 in Gunn rats were 5.09 ± 0.236 pg/mL, while those in the normal strain Wistar rats were 11.90 ± 2.142 pg/mL. The serum FGF2 levels in Gunn rats were significantly lower than those in Wistar rats. We also measured serum levels of unconjugated bilirubin (UCB) and found a significant negative correlation between UCB and FGF2 at serum levels in all the rats studied. Conclusion: Since it is known that FGF2 regulates dopaminergic neurons and have anti-neuroinflammatory effects, our finding suggests that low FGF2 levels may contribute to the pathogenesis of schizophrenia, in which disbalanced dopamin-ergic signaling and neuroinflammation are supposed to play certain roles.

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Half-life modeling of basic fibroblast growth factor released from growth factor-eluting polyelectrolyte multilayers

Scientific Reports ◽

10.1038/s41598-021-89229-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ivan Ding ◽

Amy M. Peterson

Keyword(s):

Cell Culture ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Basic Fibroblast Growth Factor ◽

Half Life ◽

Cell Types ◽

Polyelectrolyte Multilayers ◽

Enzyme Linked Immunosorbent Assay ◽

Fibroblast Growth ◽

Cell Culture Study

AbstractGrowth factor-eluting polymer systems have been widely reported to improve cell and tissue outcomes; however, measurements of actual growth factor concentration in cell culture conditions are limited. The problem is compounded by a lack of knowledge of growth factor half-lives, which impedes efforts to determine real-time growth factor concentrations. In this work, the half-life of basic fibroblast growth factor (FGF2) was determined using enzyme linked immunosorbent assay (ELISA). FGF2 release from polyelectrolyte multilayers (PEMs) was measured and the data was fit to a simple degradation model, allowing for the determination of FGF2 concentrations between 2 and 4 days of culture time. After the first hour, the FGF2 concentration for PEMs assembled at pH = 4 ranged from 2.67 ng/mL to 5.76 ng/mL, while for PEMs assembled at pH = 5, the concentration ranged from 0.62 ng/mL to 2.12 ng/mL. CRL-2352 fibroblasts were cultured on PEMs assembled at pH = 4 and pH = 5. After 2 days, the FGF2-eluting PEM conditions showed improved cell count and spreading. After 4 days, only the pH = 4 assembly condition had higher cells counts, while the PEM assembled at pH = 5 and PEM with no FGF2 showed increased spreading. Overall, the half-life model and cell culture study provide optimal concentration ranges for fibroblast proliferation and a framework for understanding how temporal FGF2 concentration may affect other cell types.

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Increased Levels of sRAGE in Diabetic CKD-G5D Patients: A Potential Protective Mechanism against AGE-Related Upregulation of Fibroblast Growth Factor 23 and Inflammation

Mediators of Inflammation ◽

10.1155/2017/9845175 ◽

2017 ◽

Vol 2017 ◽

pp. 1-9 ◽

Cited By ~ 6

Author(s):

Elena Dozio ◽

Valentina Corradi ◽

Elena Vianello ◽

Elisa Scalzotto ◽

Massimo de Cal ◽

...

Keyword(s):

Cardiovascular Risk ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Cardiac Remodeling ◽

Glycated Albumin ◽

Fibroblast Growth Factor 23 ◽

Protective Mechanism ◽

Fibroblast Growth ◽

Increased Risk ◽

Fgf 23

Advanced glycation end products (AGEs) may induce cardiac remodeling in kidney disease by promoting fibroblast growth factor 23 (FGF-23) expression. Since AGEs are increased in diabetes mellitus (DM), our first aim was to evaluate the existence of any potential association between AGEs, FGF-23, inflammation, and increased cardiovascular risk in DM patients on dialysis (CKD-G5D). Secondarily, we explored the potential role of the soluble receptor for AGEs (sRAGE) as a marker of heart failure. Levels of glycated albumin (GA), sRAGE, c-terminal FGF-23 (cFGF-23), brain natriuretic peptide (BNP), and inflammatory mediators were compared between DM and non-DM CKD-G5D patients. The levels of sRAGE, cFGF-23, BNP, and proinflammatory markers were over the ranges of normality in both DM and non-DM groups. Only GA and sRAGE levels were increased in DM compared to non-DM patients. Plasma levels of sRAGE and CRP were the only independent predictors of BNP concentration. In conclusion, in DM CKD-G5D patients, sRAGE appeared to be a marker of cardiac remodeling. Indeed, its increase could be a potential protective mechanism against the increased risk of cardiovascular complications related to AGEs and inflammation. The causal relationship between sRAGE and cardiovascular risk in these patients needs to be further confirmed by mechanistic studies.

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