PKC-alpha modulates TGF-beta signaling and impairs podocyte survival

Cell Physiol Biochem. 2009;24(5-6):627-34. doi: 10.1159/000257518. Epub 2009 Nov 4.

Abstract

Background: Progressive loss of podocytes has been documented as an early lesion in the development of glomerular disease. In a variety of glomerular diseases, including diabetic nephropathy the activation of transforming growth factor-beta (TGF-beta) has been demonstrated to promote podocyte death and the development of glomerulosclerosis. In this manuscript we analyzed the role of PKC-alpha (PKCalpha) on TGF-beta1 induced apoptosis in podocytes.

Methods: To accomplish this we generated stable murine PKCalpha deficient podocyte cell lines and examined survival- and pro-apoptotic signaling signatures as well as caspase activation after stimulation with TGF-beta.

Results: After stimulation with TGF-beta we can demonstrate an enhanced and prolonged activation of PI3K/AKT and ERK1/2 in PKCalpha-knockout (PKCalpha-/-) podocytes compared to PKCalpha-wildtype (PKCalpha+/ +) podocytes, whereas proapoptotic signaling via p38MAPK is significantly reduced. Interestingly, activation of the Smad-pathway is also prolonged in the PKCalpha-/-podocytes. When we analyzed the underlying mechanisms we found a TGF-beta inducible interaction of PKCalpha with the TGF-beta-type-I-receptor (TGFbetaRI). Moreover, endocytosis assays showed that the TGFbetaRI is less internalized in PKCalpha-/- podocytes.

Conclusion: Since we can demonstrate a key role for PKCalpha in the signaling response after stimulation with TGF-beta we conclude that PKCalpha might be an interesting target molecule as a "podocyte protective" therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Cell Line
  • Endocytosis
  • Gene Knockout Techniques
  • Mice
  • Mice, Knockout
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism
  • Podocytes / enzymology*
  • Protein Kinase C-alpha / deficiency
  • Protein Kinase C-alpha / genetics
  • Protein Kinase C-alpha / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Receptors, Transforming Growth Factor beta / metabolism
  • Signal Transduction
  • Transforming Growth Factor beta1 / metabolism*
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Receptors, Transforming Growth Factor beta
  • Transforming Growth Factor beta1
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • Protein Kinase C-alpha
  • Mitogen-Activated Protein Kinase 1
  • p38 Mitogen-Activated Protein Kinases