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

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Abstract

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
Publisher:	Springer
ISSN:	0012-186X
Date:	4 March 2012
Volume:	Vol.55
Number:	No.3
Number of Pages:	13
Page Range:	pp. 812-824
Identification Number:	10.1007/s00125-011-2409-9
Status:	Peer Reviewed
Publication Status:	Published
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)
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URI:	http://wrap.warwick.ac.uk/id/eprint/40468

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