TY - JOUR

T1 - Activation of Rac-1 and RhoA Contributes to Podocyte Injury in Chronic Kidney Disease

AU - Babelova, Andrea

AU - Jansen, Felix

AU - Sander, Kerstin

AU - Löhn, Matthias

AU - Schäfer, Liliana

AU - Fork, Christian

AU - Ruetten, Hartmut

AU - Plettenburg, Oliver

AU - Stark, Holger

AU - Daniel, Christoph

AU - Amann, Kerstin

AU - Pavenstädt, Hermann

AU - Jung, Oliver

AU - Brandes, Ralf P.

N1 - Funding Information: The authors have read the journal's policy and have the following conflicts of financial interest: Hartmut Rütten, Oliver Pletternburg and Matthias Löhn are employees to Sanofi-Aventis, which developed SAR407899 as a novel Rho kinase inhibitor for the treatment of disease associated with Rho kinase activation. The project was supported by a grant from Sanofi-Aventis (50.000 €) to Ralf Brandes. Co-authors Oliver Plettenurh and Matthias Loehn are patent holders for the following patent/patent applications: WO 2009156100 Preparation of substituted isoquinolines and isoquinolinones as rho kinase inhibitors for therapy. WO 2009156099 6-Substituted isoquinolines and isoquinolinones as Rho, PKA and PKG kinase inhibitors and their preparation, pharmaceutical compositions and use in the treatment of diseases. WO 2009156092 Preparation of bi- and polycyclic substituted isoquinoline and isoquinolinone compounds as Rho-kinase inhibitors for therapy. WO 2008077555 New substituted isoquinoline and isoquinolinone derivatives. WO 2008077551 Cycloalkylamine substituted isoquinolone derivatives. WO 2008077554 Cycloalkylamine substituted isoquinoline derivatives. WO 2008077553 Cycloalkylamine substituted isoquinolone and isoquinolinone derivatives. WO 2008077552 Substituted isoquinoline and isoquinolinone derivatives. WO 2008077550 Substituted isoquinoline and isoquinolinone derivatives as inhibitors of rho-kinase. WO 2008077556 Cycloalkylamine substituted isoquinoline and isoquinolinone derivatives. WO 2007012421 Preparation of piperidinyl isoquinolone derivatives as Rho-kinase inhibitors. WO 2007012422 Preparation of cyclohexylamine substituted isoquinolones as Rho-kinase inhibitors. WO 2007000240 Preparation of piperidinyl substituted isoquinoline derivatives as inhibitors of Rho-kinase. EHT1846 has become commercially available at Sigma-Adrich. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. The authors Andrea Babelova PhD and Ralf P. Brandes MD, on behalf of Sanofi and Dr. Matthias Loehn confirm that researchers wishing to replicate the results presented here will be provided access to SAR407899 (ie. that the tech transfer department at Sanofi is willing to share SAR407899 for non-commercial research purposes).

PY - 2013/11/7

Y1 - 2013/11/7

N2 - Rho-family GTPases like RhoA and Rac-1 are potent regulators of cellular signaling that control gene expression, migration and inflammation. Activation of Rho-GTPases has been linked to podocyte dysfunction, a feature of chronic kidney diseases (CKD). We investigated the effect of Rac-1 and Rho kinase (ROCK) inhibition on progressive renal failure in mice and studied the underlying mechanisms in podocytes. SV129 mice were subjected to 5/6-nephrectomy which resulted in arterial hypertension and albuminuria. Subgroups of animals were treated with the Rac-1 inhibitor EHT1846, the ROCK inhibitor SAR407899 and the ACE inhibitor Ramipril. Only Ramipril reduced hypertension. In contrast, all inhibitors markedly attenuated albumin excretion as well as glomerular and tubulo-interstitial damage. The combination of SAR407899 and Ramipril was more effective in preventing albuminuria than Ramipril alone. To study the involved mechanisms, podocytes were cultured from SV129 mice and exposed to static stretch in the Flexcell device. This activated RhoA and Rac-1 and led via TGFβ to apoptosis and a switch of the cells into a more mesenchymal phenotype, as evident from loss of WT-1 and nephrin and induction of α-SMA and fibronectin expression. Rac-1 and ROCK inhibition as well as blockade of TGFβ dramatically attenuated all these responses. This suggests that Rac-1 and RhoA are mediators of podocyte dysfunction in CKD. Inhibition of Rho-GTPases may be a novel approach for the treatment of CKD.

AB - Rho-family GTPases like RhoA and Rac-1 are potent regulators of cellular signaling that control gene expression, migration and inflammation. Activation of Rho-GTPases has been linked to podocyte dysfunction, a feature of chronic kidney diseases (CKD). We investigated the effect of Rac-1 and Rho kinase (ROCK) inhibition on progressive renal failure in mice and studied the underlying mechanisms in podocytes. SV129 mice were subjected to 5/6-nephrectomy which resulted in arterial hypertension and albuminuria. Subgroups of animals were treated with the Rac-1 inhibitor EHT1846, the ROCK inhibitor SAR407899 and the ACE inhibitor Ramipril. Only Ramipril reduced hypertension. In contrast, all inhibitors markedly attenuated albumin excretion as well as glomerular and tubulo-interstitial damage. The combination of SAR407899 and Ramipril was more effective in preventing albuminuria than Ramipril alone. To study the involved mechanisms, podocytes were cultured from SV129 mice and exposed to static stretch in the Flexcell device. This activated RhoA and Rac-1 and led via TGFβ to apoptosis and a switch of the cells into a more mesenchymal phenotype, as evident from loss of WT-1 and nephrin and induction of α-SMA and fibronectin expression. Rac-1 and ROCK inhibition as well as blockade of TGFβ dramatically attenuated all these responses. This suggests that Rac-1 and RhoA are mediators of podocyte dysfunction in CKD. Inhibition of Rho-GTPases may be a novel approach for the treatment of CKD.

UR - http://www.scopus.com/inward/record.url?scp=84892375133&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0080328

DO - 10.1371/journal.pone.0080328

M3 - Article

C2 - 24244677

AN - SCOPUS:84892375133

VL - 8

JO - PLOS ONE

JF - PLOS ONE

SN - 1932-6203

IS - 11

M1 - e80328

ER -