Next, we treated cultured podocytes injured by ADR with Notch2 agonistic antibody and assessed the effect of the antibody on apoptosis and examined the pathways involved

in cell survival. We assessed correlation between the number of podocytes expressing activated Notch2 and the number of residual podocytes in nephrotic kidneys. Results: Administration of Notch2 agonistic mAb ameliorates proteinuria and glomerulosclerosis in mouse with ADR-induced nephropathy. Trametinib in vitro In vitro, the specific knockdown of Notch2 leads to increased apoptosis in damaged podocytes. Notch2 agonistic mAb enhances activation of Akt and protects damaged podocytes from apoptosis. Treatment with γ-secretase inhibitor or Akt inhibitor abolishes the protective effect of Notch2 agonistic

mAb. In mice with lipopolysaccaride (LPS)-induced nephropathy, a mouse model of minimal change nephrotic syndrome (MCNS) which does not show podocyte loss, most of the podocytes showed activated Notch2. In vitro, treatment of cultured podocytes with LPS increased cleaved Notch2 and activated Akt. Positive linear correlation between the number of podocytes expressing activated Notch2 and the number of residual podocytes was found in human nephrotic kidneys. Podocytes in MCNS showed more cleaved Notch2 MAPK Inhibitor Library than that in FSGS. Conclusions: Activation of Notch2 rescues injured podocytes from apoptosis. It may represent a novel clinical strategy for the amelioration of nephrosis and glomerulosclerosis. HAMATANI HIROKO1, HIROMURA KEIJU1, SAKAIRI TORU1, Avelestat (AZD9668) TAKAHASHI SATOSHI1, WATANABE MITSUHARU1, MAESHIMA AKITO1, OHSE TAKAMOTO2, PIPPIN JEFFERY W.3, SHANKLAND STUART J.3, NOJIMA YOSHIHISA1 1Department of Medicine and Clinical Science, Gunma University Graduate School of Medicine, Maebashi, Japan; 2Division

of Nephrology and Endocrinology, University of Tokyo School of Medicine, Tokyo, Japan; 3Division of Nephrology, University of Washington, Seattle, Washington Introduction: Sestrin 2, initially identified as a p53 target protein, accumulates in cells exposed to stress and inhibits mammalian target of rapamycin (mTOR) signaling. In this study, we found that sestrin 2 was selectively expressed in rat glomerular parietal cells (PECs) and examined the expression of sestrin 2 and mTOR signaling in the PECs of normal and diseased kidneys. Methods: Adriamycin (ADR), puromycin aminonucleoside (PAN) and anti-glomerular basement membrane (GBM) antibody were used to induce glomerulonephritis in rats and the expression of sestrin 2 was examined immunohistochemically. Activation of mTOR signaling was determined by antibodies against phosphorylated S6RP, 4E-BP1 and p70S6K, which are the downstream targets of mTOR. Results: In the normal rat kidneys, sestrin 2 was selectively expressed in the PECs, similar to PGP9.5, a well-known marker of PECs.