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. 2013 May 10;6(1):10.
doi: 10.1186/1755-1536-6-10.

Platelet-derived growth factor alpha and beta receptors have overlapping functional activities towards fibroblasts

Johanna Donovan  1 , Xu Shiwen, Jill Norman, David Abraham
Affiliations

Platelet-derived growth factor alpha and beta receptors have overlapping functional activities towards fibroblasts

Johanna Donovan et al. Fibrogenesis Tissue Repair. .

Abstract

Background: Platelet-derived growth factor (PDGF) signalling is essential for many key cellular processes in mesenchymal cells. As there is redundancy in signalling between the five PDGF ligand isoforms and three PDGF receptor isoforms, and deletion of either of the receptors in vivo produces an embryonic lethal phenotype, it is not know which ligand and receptor combinations mediate specific cellular functions. Fibroblasts are key mediators in wound healing and tissues repair. Recent clinical trials using broad spectrum tyrosine kinase inhibitors in fibrotic diseases have highlighted the need to further examine the specific cellular roles each of the tyrosine kinases plays in fibrotic processes. In this study, we used PDGFR-specific neutralising antibodies to dissect out receptor-specific signalling events in fibroblasts in vitro, to further understand key cellular processes involved in wound healing and tissue repair.

Results: Neutralising antibodies against PDGFRs were shown to block signalling through PDGFRα and PDGFRβ receptors, reduce human PDGF-AA and PDGF-BB-induced collagen gel remodelling in dermal fibroblasts, and reduce migration stimulated by all PDGF ligands in human dermal and lung fibroblasts.

Conclusions: PDGFRα and PDGFRβ neutralising antibodies can be a useful tool in studying PDGFR isoform-specific cellular events.

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Figures

Figure 1
Figure 1
Schematic diagram showing PDGFR-PDGF interactions in vitro: PDGF ligand dimers bind either PDGFRα or PDGFRβ homodimers or the α/β heterodimer.Arrows show proven in vitro ligand-receptor interactions. Each PDGF receptor has five extracellular immunoglobulin-like domains and two intracellular tyrosine kinase domains. PDGFR chains are shown in blue (PDGFRα) and red (PDGFRβ). PDGF ligands are shown in green (PDGF-A), yellow (PDGF-B), orange (PDGF-C) and purple (PDGF-D).
Figure 2
Figure 2
Phosphorylation of PDGFRα and PDGFRβ in response to PDGF ligands. Dermal fibroblasts (a) and lung fibroblasts (b) were grown in 10% FCS and serum-starved overnight or kept in 10% FCS. Cells were then stimulated with either 10% FCS, 0% FCS, PDGF-AA, PDGF-BB, PDGF-DD or Imatinib for 15 min. Whole cell lysate were Western blotted using antibodies against phospho-PDGFRα, PDGFRα, phospho-PDGFRβ, PDGFRβ and GAPDH (loading control).
Figure 3
Figure 3
Phosphorylation of PDGFRα and PDGFRβ in response different doses of PDGF-AA and PDGF-CC ligands. Dermal fibroblasts were grown in 10% FCS and serum-starved overnight. Cells were stimulated with 0% FCS, PDGF-BB, PDGF-AA (5-200ng/ml) and PDGF-CC (5-200ng/ml) for 15 min. Whole cell lysates were Western blotted using antibodies against phospho-PDGFRα, phospho-PDGFRβ and GAPDH (loading control).
Figure 4
Figure 4
Western blot showing effects of PDGFRα and PDGFRβ neutralising antibodies. (a) Dermal fibroblasts were treated with neutralising antibodies to anit-PDGFRα, anti-PDGFRβ or vehicle for 1 h at room temperature and then stimulated with either PDGF-AA or PDGF-BB or vehicle for 15 min. Cells were then washed in ice-cold PBS and lysed. Cell lysates were Western blotted for PDGFRα, pPDGFRα, PDGFRβ, pPDGFRβ, ERK, pERK and GAPDH. The relative amount of pPDGFRα, pPDGFRβ, as measure by densitometry, is shown in (b) and (c). (d) Lung fibroblasts were treated and analysed in the same manner (e) and (f).
Figure 5
Figure 5
The effect of collagen gel contraction of dermal fibroblasts after treatment with PDGFRα and PDGFRβ neutralising antibodies. Dermal fibroblasts were treated with neutralising antibodies to anti-PDGFRα, anti-PDGFRβ or blank for 1 h at room temperature and were added to a collagen gel. The gels were then grown in media containing either PDGF-AA or PDGF-BB. After 24 h the gels were weighed to assess contraction. Error bars are standard error of the mean. Two-sample T-test statistical analysis was performed compared to compared gel weights compared to the 0% FCS control p = 0.004 PDGF-AA, p = 0.032 PDGF-BB, p = 0.82 anti-PDGFR-α., p = 0.62 anti-PDGFR-α.+ PDGF-AA, p = 0.03 anti-PDGFR-α.+ PDGF-BB, p = 0.77 anti-PDGFR-β, p = 0.035 anti-PDGFR-β + PDGF-AA, p = 0.0007 anti-PDGFR-β + PDGF-BB. *p > 0.05.
Figure 6
Figure 6
The effect on migration of dermal and lung fibroblasts after treatment with PDGFRα and PDGFRβ neutralising antibodies. Dermal fibroblasts were grown in 10% serum. A scratch was made in the cell layer and cells were treated with mitomycin C with neutralising antibodies to either anti-PDGFRα, anti-PDGFRβ, anti-PDGFRα and anti-PDGFRβ, control IgG, Imatinib or 10% FCS for 1 h at room temperature and then stimulated with PDGF-AA, PDGF-BB, PDGF-CC or PDGF-DD. (a). Percentage migration was calculated by measuring the average density of cells migrated into the scratched area after 24 h (b-f). Error bars are standard errors of the mean. Two-sample T-test was performed - 10% FCS vs. Media + mitomycin C, 10% FCS vs. IgG treated, anti-PDGFRα + anti-PDGFRβ vs. anti-PDGFRα + anti-PDGFRβ + PDGF-BB, Imatinib vs. Imatinib + PDGF-BB, p = 0.032, 0.035, 0.92, and 0.1 respectively. Anti-PDGFRα + PDGF-AA vs. PDGF-AA, anti-PDGFRβ + PDGF-AA vs. PDGF-AA, anti-PDGFRβ + PDGF-BB vs. PDGF-BB, anti-PDGFRα + PDGF-BB vs. PDGF-BB, anti-PDGFRβ + PDGF-CC vs. PDGF-CC, anti-PDGFRα + PDGF-CC vs. PDGF-CC, anti-PDGFRβ + PDGF-DD vs. PDGF-DD, anti-PDGFRα + PDGF-DD vs. PDGF-DD —p = 0.24, 0.93, 0.006, 0.24, 0.06, 0.34, 0.38 and 0.64 respectively. Lung fibroblasts were treated and analysed in identical fashion. (g-k). Two-sample T-test was performed - FCS vs. Media + mitomycin C, 10% FCS vs. IgG treated, anti-PDGFRα + anti-PDGFRβ vs. anti-PDGFRα + anti-PDGFRβ + PDGF-BB, Imatinib vs. Imatinib + PDGF-BB —p = 0.585, 0.18, 0.64 and 0.21 respectively. Anti-PDGFRα + PDGF-AA vs. PDGF-AA, anti-PDGFRβ + PDGF-AA vs. PDGF-AA, anti-PDGFRβ + PDGF-BB vs. PDGF-BB, anti-PDGFRα + PDGF-BB vs. PDGF-BB, anti-PDGFRβ + PDGF-CC vs. PDGF-CC, anti-PDGFRα + PDGF-CC vs. PDGF-CC, anti-PDGFRβ + PDGF-DD vs. PDGF-DD, anti-PDGFRα + PDGF-DD vs. PDGF-DD —p = 0.25, 0.75, 0.05, 0.64, 0.82, 0.69, 0.1 and 0.23 respectively.
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References

    1. Alvarez RH, Kantarjian HM, Cortes JE. Biology of platelet-derived growth factor and its involvement in disease. Mayo Clin Proc. 2006;81(9):1241–57. doi: 10.4065/81.9.1241. - DOI - PubMed
    1. Andrae J, Gallini R, Betsholtz C. Role of platelet-derived growth factors in physiology and medicine. Genes Dev. 2008;22(10):1276–312. doi: 10.1101/gad.1653708. - DOI - PMC - PubMed
    1. Grotendorst GR. Platelet-derived growth factor is a chemoattractant for vascular smooth muscle cells. J Cell Physiol. 1982;113(2):261–6. doi: 10.1002/jcp.1041130213. - DOI - PubMed
    1. Seppa H. Platelet-derived growth factor in chemotactic for fibroblasts. J Cell Biol. 1982;92(2):584–8. doi: 10.1083/jcb.92.2.584. - DOI - PMC - PubMed
    1. Claesson-Welsh L, Ronnstrand L, Heldin CH. Biosynthesis and intracellular transport of the receptor for platelet-derived growth factor. Proc Natl Acad Sci U S A. 1987;84(24):8796–800. doi: 10.1073/pnas.84.24.8796. - DOI - PMC - PubMed