Feinberg School of Medicine Chicago IL Division of Nephrology and Hypertension
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1 Feinberg School of Medicine Chicago IL Division of Nephrology and Hypertension Daniel Batlle MD Earle, del Greco Levin Professor of Medicine Division of Nephrology and Hypertension Northwestern University Feinberg School of Medicine Chicago IL
2 The great refugee: How Laszlo Kubala became a Barcelona legend 2
3 Blockade of the Renin Angiotensin System: ARB Feedback Loop ARBs impact the feedback loop, resulting in increased PRA, Ang I and Ang II Renin Angiotensinogen AT 1 Receptor ARBs ACE
4
5
6 ACE2
7 7
8 Structural Conservation: ACE-ACE2 Monika Rella, Richard Jackson, Tony Turner
9 Eriksson et al. Current Biology, Vol. 12, R745 R752, 2002
10 Objectives ACE2 localization within the kidney Alteration in ACE2 expression in mouse models of diabetic kidney disease. Impact of ACE2 inhibition and genetic ablation on diabetic kidney disease. Effect of recombinant ACE2 on blood pressure and Angiotensin II metabolism. Potential therapeutic effect of ACE2 amplification.
11 ACE ACE2 Merge
12 ACE ACE2
13
14 Glomerular Cell Markers ACE2 ACE Podocyte Nephrin Podocin ++ - Synaptopodin ++ - Endothelial Cell Pecam-1 +/- +++ Mesangial Cell α-smooth muscle actin ++ +
15 ACE ACE2 Merge Ye et al. JASN 2006
16 ACE Nephrin Merge ACE2 Nephrin Merge
17 ACE2 ACE Ye et al JASN 2006
18 Non-ACE Batlle D, Soler MJ, Wysocki J, Curr Opinion Nephrol and Hypertension2008
19 Objectives ACE2 localization within the kidney Alteration in ACE2 expression in mouse models of diabetic kidney disease. Impact of ACE2 inhibition and genetic ablation on diabetic kidney disease. Effect of recombinant ACE2 on blood pressure and Angiotensin II metabolism. Potential therapeutic effect of ACE2 amplification.
20 db/m db/db
21 Control Diabetic db/db Ye et al. JASN 2006
22 Control Diabetic nephropathy Mizuiri S Am J Kidney disease 2008
23 HYPOTHESIS Glomerular Expression High ACE Low ACE2 = ANG II = Albuminuria
24 Objectives ACE2 localization within the kidney Alteration in ACE2 expression in mouse models of diabetic kidney disease. Impact of ACE2 inhibition pharmacologically and genetic ablation on diabetic kidney disease. Effect of recombinant ACE2 on blood pressure and Angiotensin II metabolism. Potential therapeutic effect of ACE2 amplification.
25 Ye et al ,000 Start of the ACE2 inhibitor (MLN-4760) 0,900 Albumin/creatinine ratio (mg/mg) 0,800 0,700 0,600 0,500 0,400 0,300 0,200 db/m (-) db/m ACE2-I db/db (-) db/db ACE2-I 0,100 0,000 8wk 13wk 17wk 20wk 22wk 24wk Age
26 Rescue of effect of ACE2 inhibition (MLN 4760) on albuminuria by Angiotensin II blockade(telmisartan) Albumin/Creatinine ratio [ug/mg] * * 0 VEHICLE n=13 MLN-4760 n=14 MLN TELMISARTAN n=10 Ye et al 2006
27 Effect of ACE2 inhibition ( MLN4760) on albuminuria in Streptozotocin (STZ) model Control STZ STZ+MLN wk 13wk 17wk 19wk STZ MLN-4760 Age (weeks) Soler et al KI 2008
28 Summary ACE2 is localized in the glomerular epithelial cell (podocyte) ACE2 expression is down-regulated whereas ACE is up-regulated in glomeruli from diabetic mice ACE2 inhibition exacerbates kidney disease in diabetic mice. Studies in ACE2 knockout.
29
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31 Kidney NADPH oxidase activity Males 14 wks 40 wks
32 Heart NADPH oxidase activity in male mice 14 wks 40 wks P<0.05 P<0.01 n=9 n=9 n=4 n=9 Wysocki et al. unpublished
33 Objectives ACE2 localization within the kidney Alteration in ACE2 expression in mouse models of diabetic kidney disease. Impact of ACE2 inhibition and genetic ablation on diabetic kidney disease. Effect of recombinant ACE2 on blood pressure and Angiotensin II metabolism. Potential therapeutic effect of ACE2 amplification.
34 serum ACE2 after 3 and 14 days of treatment ACE2 activity CNTRL AII AII+ACE days
35 Wysocki et al Hypertension 2010
36
37
38 Effect of race2 on blood pressure Exogenous ANG II Exogenous ACE2 ANGIOTENSIN II vasoconstrictor ANGIOTENSIN-(1-7) vasodilator
39
40 Effect of ACE2 on blood pressure Exogenous ANG II Exogenous ACE2 Ang1-7 receptor blocker ANGIOTENSIN II vasoconstrictor ANGIOTENSIN-(1-7) vasodilator
41 Objectives ACE2 localization within the kidney Alteration in ACE2 expression in mouse models of diabetic kidney disease. Impact of ACE2 inhibition and genetic ablation on diabetic kidney disease. Effect of recombinant ACE2 on blood pressure and Angiotensin II metabolism. Potential therapeutic effect of ACE2 amplification.
42 Fig. 1. Angiotensin-converting enzyme (ACE) and ACE2 localization in renal vasculature: immunofluorescence staining of ACE (green; A) and ACE2 (red; B) in a kidney arteriole from mouse kidney Soler, M. J. et al. Am J Physiol Renal Physiol 296: F398-F ; doi: /ajprenal Copyright 2009 American Physiological Society
43 Soler et al AJP 2008
44 Structure-based identification of an ACE2 activator
45 Blockade of the Renin Angiotensin System: ARB Feedback Loop ARBs impact the feedback loop, resulting in increased PRA, Ang I and Ang II Renin Angiotensinogen AT 1 Receptor ARBs ACE
46 Only Direct Renin Inhibition Inhibits the Entire Renin Angiotensin System1-6 Diuretic ACEI ARB Direct Renin Inhibitor (DRI)
47 An Efficient Strategy to Achieve Interruption of the Renin Angiotensin System Is Direct Renin Inhibition1-3
48 ACE Escape With Long-Term ACEI Treatment *
49 Potential ACE2 targets: a large repertoire of disease entities Acute Lung Injury Pulmonary Hypertension Congestive Heart Failure Diabetic Retinopathy Diabetic Kidney Disease Brain injury Stoke Renin Dependent Hypertension Any Angiotensin II excess Syndrome 49
50 Methods of ACE 2 amplification Human Recombinant ACE2 (IP, minipumps, i.v) Murine Recombinant ACE2 (IP, minipumps, i.v ) Lentivirus DNA delivery/adeno-associated Virus delivery ACE2 transgenic models ACE2 Activators ( XNT /DIZE/NCP-2454 ) Minicircle DNA delivery Bioencapsulation of ACE2 Modified race2 with enhanced duration of action 50
51 What is the target of ACE 2 amplification ACE2 deficiency ( relative if inadequate compensation) Excess Angiotensin II (1-8) Deficiency of Ang ( 1-7) All of the above? None of the above? 51
52 Less Ang II or more Ang 1-7? Evidence for the model of Ang II-induced hypertension Jan Wysocki PhD 52
53 Ang 1-7 independent action of ACE2 on blood pressure 53
54 Acute Lungs Injury- Human Recombinant ACE2 Imai et al, Nature,2005
55 Ang II-Induced Cardiac Fibrosis - Studies with Lentivirus ACE2 Huentelman, M. J.,et al Experimental Physiology2005
56 Lung Fibrosis-Asthma model- ACE2 activator (DIZE) Vaibhav Shrirang Dhawale et al, In Toxicology and Applied Pharmacology, 2016
57 Diabetic Retinopathy- Studies with AAV ACE2 Amrisha Verma et al ACE2 and Ang 1-7 diabetic
58 Mini-circles ACE2 DNA delivery by hydrodynamic injection
59 Minicircle ACE2 delivery in the STZ model
60 Minicircle ACE2 delivery in the STZ model
61 Minicircle ACE2 delivery in the STZ model
62 Wysocki et al, KI, 2017
63 Albumin excretion Rates in different model of Kidney Diseases
64 Recovery of ACE2 protein in the urine in the Alport Mice Model
65 Alport Syndrome- Murine Recombinant ACE2 Bae et al, Kidney International, 65
66 66
67 Extending ACE2 half life by Fc-fusion 67
68 Extending ACE2 half life by Fc-fusion 68
69 Extending ACE2 half life by Fc-fusion 69
70 Effect of race2-fc on Ang II-induced cardiac hypertrophy Pan liu et al, unpublished 70
71 Renin Transgenic Mice 71
72 Effect of race2-fc in the Renin transgenic mice (RnTgMK) 72
73 Renin Transgenic Model Untreated and Treated with race2-fc 73
74 Renin Transgenic Model Untreated and Treated with race2-fc 74
75 Ongoing Clinical Trials Hormonal, Metabolic, and Signaling Interactions in PAH Study of GSK on Acute Hypoxia and Exercise Safety and Tolerability Study of APN01 (Recombinant Human Angiotensin Converting Enzyme 2) COMPLETED 75
76 Summary Ace 2 amplification offers spectrum of potential therapeutic applications including: Acute Lung Injury Pulmonary Hypertension Heart Failure and Cardiac Fibrosis Diabetic Retinopathy Certain types of diabetic and non diabetic kidney disease. While available presumed ACE2 activators provide therapeutic benefit the mechanism of action is unclear. Recombinant ACE2 protein with prolonged duration of action provides a therapeutically attractive strategy for ACE 2 amplification and down regulation of hyperactive RAS systemically Effective kidney delivery remains a challenge for early diabetic kidney disease 76
77 L H F P H I Y V R D Angiotensinogen (AOG) Angiotensin (1-10) H F P H I Y V R D Angiotensin (1-9) Angiotensin (1-8) Angiotensin (2-8) F P H I Y V Angiotensin (3-8) Angiotensin (1-5) Angiotensin (1-7) Renin ACE ACE2 ACE2 Chymase PEP PRCP ACE APA ACE Neprilysin PRCP L H F P H I Y V R D F P H I Y V R D P H I Y V R D I Y V R D F P H I Y V R
78 Jan Wysocki MD PHD Tilman Muller Alonso Marquez Peter Daniel Serfozo Jeannette Tang Alejandro Sanchez Sheeba Habeed Jing Jin Pan Liu 78
79 Batlle s Laboratory Feinberg School of Medicine Chicago IL Division of Nephrology and Hypertension Jan Wysocki Minghao Ye Sheeba Habeeb Jeannette Tang Tilman Mueller Alejandro Sanchez Alonso Marquez
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