The TGF-β profibrotic cascade targets ecto-5'-nucleotidase gene in proximal tubule epithelial cells and is a traceable marker of progressive diabetic kidney disease

Biochim Biophys Acta Mol Basis Dis. 2020 Jul 1;1866(7):165796. doi: 10.1016/j.bbadis.2020.165796. Epub 2020 Apr 11.


Progressive diabetic nephropathy (DN) and loss of renal function correlate with kidney fibrosis. Crosstalk between TGF-β and adenosinergic signaling contributes to the phenotypic transition of cells and to renal fibrosis in DN models. We evaluated the role of TGF-β on NT5E gene expression coding for the ecto-5`-nucleotidase CD73, the limiting enzyme in extracellular adenosine production. We showed that high d-glucose may predispose HK-2 cells towards active transcription of the proximal promoter region of the NT5E gene while additional TGF-β results in full activation. The epigenetic landscape of the NT5E gene promoter was modified by concurrent TGF-β with occupancy by the p300 co-activator and the phosphorylated forms of the Smad2/3 complex and RNA Pol II. Transcriptional induction at NT5E in response to TGF-β was earlier compared to the classic responsiveness genes PAI-1 and Fn1. CD73 levels and AMPase activity were concomitantly increased by TGF-β in HK-2 cells. Interestingly, we found increased CD73 content in urinary extracellular vesicles only in diabetic patients with renal repercussions. Further, CD73-mediated AMPase activity was increased in the urinary sediment of DN patients. We conclude that the NT5E gene is a target of the profibrotic TGF-β cascade and is a traceable marker of progressive DN.

Keywords: CD73; Diabetic nephropathy; Histone modification; NT5E gene regulation; TGF-β.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / genetics*
  • Adenosine / biosynthesis
  • Biomarkers / metabolism
  • Cell Line
  • Diabetic Nephropathies / genetics*
  • Diabetic Nephropathies / pathology
  • E1A-Associated p300 Protein / genetics
  • Epigenesis, Genetic / genetics
  • Epithelial Cells / metabolism
  • Epithelial Cells / pathology
  • Fibrosis / genetics*
  • Fibrosis / pathology
  • GPI-Linked Proteins / genetics
  • Gene Expression Regulation
  • Humans
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Tubules, Proximal / metabolism
  • Kidney Tubules, Proximal / pathology
  • Nucleotidases / genetics
  • Promoter Regions, Genetic / genetics
  • RNA Polymerase II / genetics
  • Transforming Growth Factor beta / genetics*


  • Biomarkers
  • GPI-Linked Proteins
  • Transforming Growth Factor beta
  • E1A-Associated p300 Protein
  • EP300 protein, human
  • RNA Polymerase II
  • Nucleotidases
  • adenosine monophosphatase
  • 5'-Nucleotidase
  • NT5E protein, human
  • Adenosine