1. Roderick Tan Lab of Youhua Liu Departments of Medicine and Pathology October 24, 2013 EXTRACELLULAR SUPEROXIDE DISMUTASE IN KIDNEY DISEASE

2. OUTLINE Oxidative Stress in Renal Fibrosis Introduction to EC-SOD Role of EC-SOD in Adriamycin Injury Association with Wnt/beta-catenin signaling

3. DEVELOPMENT OF RENAL FIBROSIS Liu, Nat Rev Neph 2011

4. OXIDATIVE STRESS (OS) ROS Antioxidants

5. OXIDATIVE STRESS (OS) ROS Antioxidants Inflammation Angiotensin II NADPH oxidases SOD, Catalase, Glutathione peroxidase

6. EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD) 1 of 3 isoforms of SOD Antioxidant enzyme High expression in normal kidney Folz et al., AJRCMB 1997

7. O 2 - + O 2 - O 2 + H 2 O 2 2H + EC-SOD K ~ 1 x 10 9 Molecular Oxygen (O 2 ) Superoxide (O 2 - ) O ׃׃׃ ׃׃ ●●● O = O ׃ ׃ ׃ ׃ -

8. Inflammatory Cells Activated Fibroblasts, Epithelium, Endothelium O2●-O2●- EC-SOD prevents superoxide from forming potent reactive oxygen species + NO ● ONOO - HO ● NO 2 Fe 3+ Fe 2+ H202H202 HO ● EC-SOD Cell and tissue damage, signaling cascades, aging, etc.

9. Distribution of EC-SOD in kidney Zelko and Folz, Endocrinology 2005

10. ADRIAMYCIN NEPHROPATHY Murine model for FSGS Single IV (tail vein) injection Glomerular injury  proteinuria Tubular damage / fibrosis Superoxide-mediated

11. ADR INJURY IN WILD TYPE MICE serum creatinine (mg/dL) mg Ualb / mg UCr **

12. ADR DEPLETES RENAL EC-SOD * *

13. EC-SOD KNOCKOUT MICE Global knockouts (KO) No overt phenotype under basal conditions

14. EC-SOD KO MICE ARE SENSTIVIE TO ADR INJURY WT ADR KO ADR DHE *

15. NADPH OXIDASE *

16. EC-SOD KO MICE ARE SENSTIVIE TO ADR INJURY * * * * * *

17. EC-SOD KO MICE ARE SENSITIVE TO ADR INJURY

18. * * *

19. WNT/BETA-CATENIN SIGNALING Developmental pathway Quiescent in normal adult kidney tissue Upregulated in glomerular and interstitial injuries Blockade ameliorates of renal injury He, JASN (2009, 2011); Hao, JASN (2011); Dai, JASN (2009)

20. EC-SOD KO MICE ARE SENSTIVIE TO ADR INJURY WT KO β-catenin *

21. SUMMARY EC-SOD is depleted from kidneys after ADR EC-SOD KO mice are sensitized to oxidative stress, proteinuria and fibrosis after ADR EC-SOD KO have increased  -catenin activation after ADR

22. MODEL OF INJURY ADRIAMYCIN PODOCYTE / GLOMERULAR INJURY PROTEINURIA FIBROSISDECREASED RENAL FUNCTION EC-SOD

23. ACKNOWLEDGMENTS Liu Lab Youhua Liu Dong Zhou Liangxiang Xiao Lin Lin Tom Kleyman Tim Oury Donna Beer-Stolz Bruce Freeman Ken Hallows Pittsburgh Center for Kidney Research Center for Biologic Imaging NIDDK R01 DK091239 NIDDK T32 DK061296 AHA FTF 16990086

24. Tokarska-Schlattner, J of Mol and Cell Card (2006) MECHANISM OF ACTION