TGFβ modulates cell-cell communication in early epithelial-to-mesenchymal transition
Hills, Claire E., Siamantouras, Eleftherios, Smith, S. W., Cockwell, Paul, Liu, Kuo-Kang and Squires, Paul E.. (2012) TGFβ modulates cell-cell communication in early epithelial-to-mesenchymal transition. Diabetologia, Vol.55 (No.3). pp. 812-824. ISSN 0012-186X
WRAP_Squires_manuscript_final.pdf - Accepted Version - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
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Official URL: http://dx.doi.org/10.1007/s00125-011-2409-9
Aims/hypothesis A key pathology in diabetic nephropathy is tubulointerstitial fibrosis.
Characterized by increased deposition of the extracellular matrix, fibrotic scar formation and
declining renal function, the prosclerotic cytokine TGF-β1 mediates many of these catastrophic
changes. The current study investigates a role for TGF-β1 induced epithelial to mesencyhmal
transition (EMT) in alterations in cell-adhesion, cell coupling and cell communication in the
human renal proximal tubule.
Methods: Whole-cell and cell-compartment expression of E-cadherin, N-cadherin, Snail,
vimentin, β-catenin and connexin-43 (Cx43) was determined in HK2 and human proximal tubule
cells (hPTC) +/-TGF-β1, using western blotting and immunocytochemistry, and quantified using
densitometry. The contribution of Cx43 in PT cell communication was quantified using small
interfering RNA knockdown, whilst dye-transfer assessed gap-junction mediated intercellular
communication (GJIC). Functional tethering was assessed by single-cell force spectroscopy +/-
TGF-β1 or immunoneutralization of cadherin ligation.
Results: High glucose (25mmol/l) increased the secretion of TGF-β1 from HK2 cells. Analysis
confirmed early TGF-β1 induced morphological and phenotypic changes of EMT with altered
expression of adhesion and adheren junction proteins. These changes correlated with impaired cell
adhesion and decreased tethering between coupled cells. Impaired E-cadherin mediated adhesion
induced a loss of Cx43 expression and GJIC, an effect mimicked by neutralizing E-cadherin
ligation. Up-regulation of N-cadherin failed to restore adhesion or Cx43 mediated GJIC.
Conclusions: Our study provides compelling evidence that TGF-β1 induced EMT instigates a loss
in E-cadherin, cell adhesion and ultimately connexin-mediated cell communication in the
proximal tubule under diabetic conditions ahead of overt signs of renal damage.
|Item Type:||Journal Article|
|Subjects:||Q Science > QP Physiology|
|Divisions:||Faculty of Science > Engineering
Faculty of Science > Life Sciences (2010- )
|Library of Congress Subject Headings (LCSH):||Diabetic nephropathies -- Pathogenesis, Kidneys -- Fibrosis, Cytokines|
|Journal or Publication Title:||Diabetologia|
|Official Date:||4 March 2012|
|Number of Pages:||13|
|Page Range:||pp. 812-824|
|Access rights to Published version:||Restricted or Subscription Access|
|Funder:||Diabetes UK, Diabetes Research & Wellness Foundation (DRWF), European Foundation for the Study of Diabetes (EFSD), Janssen Pharmaceutical Ltd., University of Warwick Research Development Fund, Wellcome Trust (London, England)|
|Grant number:||BDA:11/0004215 (DUK)|
1. Eddy AA (1996) Molecular insights into renal interstitial fibrosis. J Am Soc Nephrol 12:
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