Targeting intracellular arginine / asymmetric dimethylarginine (ADMA). From bench to practice: Novel anti-atherogenic strategies to improve endothelial function Rainer H. Böger, M.D. Institute of Clinical Pharmacology and Toxicology Center of Experimental Medicine
Declaration of interest: I have been named as inventor on patent appliations relating to analytical methods for analyzing ADMA and related compounds. I receive royalties from such patent licenses.
The endothelial L-arginine / NO Pathway CAT L-arginine NO-Synthase L-citrulline NO Vasodilation Anti-inflammatory effects Anti-proliferatory effects Anti-thrombogenic effects
Lancet 1992; 339: 572-575 Dimethylarginines in human plasma (ADMA+SDMA): Healthy (n = 6) End-stage renal failure (n = 9) 1.15 ± 0.13 µmol/l 8.70 ± 0.70 µmol/l
Chemical Structures of L-Arginine and Methylarginines CH 3 CH 3 CH 3 HN NH 2 HN NH HN N CH 3 H C 3 N NH C C C C NH NH NH NH H 2 N C O H 2 N C O H 2 N C O H 2 N C O OH OH OH OH L-arginine N G -monomethyl- L-arginine (L-NMMA) N G, N G -dimethyl- L-arginine (ADMA) N G, N G`-dimethyl- L-arginine (SDMA) NOS substrate NOS inhibitor NOS inhibitor not a NOS inhibitor
ADMA and SDMA modulate endothelial NO production CAT SDMA ADMA L-arginine NO-Synthase L-citrulline NO Vasodilation Anti-inflammatory effects Anti-proliferatory effects Anti-thrombogenic effects
40 30 Formation of 15 NO 2 [nmol / µg protein] from [guanidino- 15 N 2 ]-L-arginine ADMA inhibits the conversion of [ 15 N 2 ]-L-arginine to NO in vitro 20 10 * * * * * 0 Control -7-6 -5-4 * -3 ADMA [log M] Böger et al.; Circ. Res. 2000; 87: 99-105
Flow-induced vasodilation [%] ADMA plasma concentration inversely correlates with endothelium-dependent vasodilation 16 14 12 10 8 6 4 2 0-2 -4 R = 0.762 p < 0.01-6 -8 0.0 0.5 1.0 1.5 2.0 2.5 3.0 ADMA [µmol/l] Böger et al., Circulation 1998, 98: 1842 1847
ADMA NO is is an a anti-atherogenic pro-atherogenic molecule LDL oxidation vasodilation platelet aggregation ADMA NO O 2 superoxide radical elaboration smooth muscle cell proliferation monocyte adhesion
Effects of systemic ADMA infusion in healthy humans Achan et al., Arterioscler. Thromb. Vasc. Biol. 2003; 23: 1455-1459 Kielstein et al., Circulation 2004, 109: 172-177
ADMA regulates NO production and vascular function Cardounel et al., J. Biol. Chem. 2007; 282: 879-887
ADMA: metabolism and (patho-)physiology methionine L-arginine S-adenosyl-methionine N-methyltransferase(s) -CH 3 S-adenosyl-homocysteine L-arginine CH 3 CH 3 homocysteine proteolysis L-arginine ADMA NO synthase DDAH L-citrulline urine endothelial dysfunction R.H. Böger; Cardiovasc. Res. 2003; 59: 824-833
% change NOS activity % change Effects of DDAH overexpression in mice Increased DDAH activity leads to reduced ADMA levels and reduced systemic vascular resistance 140 120 100 80 60 40 20 * * increased urinary nitrate excretion 0 20 SKM Cardiac Aorta 10 0-10 HR CO MAP SVR Dayoub et al., Circulation 2003; 108: 3042-3047
Disruption of DDAH-1 impairs vascular function in mice Heterozygous knockout of DDAH-1 leads to elevated ADMA plasma level (by some 20%), reduced DDAH activity in kidney and liver, and increased blood pressure with compensatory increase in heart rate. impaired endothelium-dependent relaxation ex vivo Leiper et al.; Nat. Med. 2007; 13: 198-203
Endothelial DDAH-1 account for the major portion of total DDAH activity Homozygous knockout of DDAH-1 leads to Endothelial-specific knockout of DDAH-1 leads to Hu et al.; ATVB 2011; 31: 1540-6. Hu et al.; Circulation 2009; 120: 2222-9.
ADMA: metabolism and (patho-)physiology methionine L-arginine S-adenosyl-methionine N-methyltransferase(s) -CH 3 S-adenosyl-homocysteine L-arginine CH 3 CH 3 homocysteine proteolysis L-arginine ADMA NO synthase DDAH L-citrulline R.H. Böger; Cardiovasc. Res. 2003; 59: 824-833 urine endothelial dysfunction? cardiovascular disease
ADMA is a predictor of total mortality in patients with end-stage renal failure 1.0.9 All cause mortality ADMA percentiles Cumulative survival.8.7 < 50 th (<2.52 µmol/l).6 50 th -75 th (2.52-3.85 µmol/l).5 0 6.7 13.3 20.0 26.7 33.3 40.0 46.7 > 75 th (>3.85 µmol/l) Time (months) Zoccali et al.; Lancet 2001; 358: 2113-2117
1,874 patients with stable coronary artery disease Follow-up 2.6 ± 1.2 years Schnabel et al.; Circ. Res. 2005; 97: e53-59
ADMA: A predictor of mortality in the Framingham Offspring Cohort 3,320 study participants from the general population in Framingham, MA Total mortality HR = 1.21 (1.07-1.37) per SD increment (0.13 µmol/l) P = 0.003 Multivariate model adjusted for established CV risk factors included in the FRS plus BNP, renin, Hcys, CRP, urinary albumin excretion. Böger et al. Circulation 2009
Genome-wide association study of ADMA reveals DDAH-1 as the most important regulatory gene for ADMA in humans Genome-wide association for ADMA (Framingham Heart Study, Gutenberg Health Study, KORA MONICA Study Ddah1
Upregulation of DDAH-1 protects from atherosclerotic vascular injury Control mice ADMA = 0.88 µm DDAH-1 tg mice ADMA = 0.45 µm Jacobi,, Böger. Am J Pathol 2010
Septic shock is correlated with ADMA levels ADMA was elevated more strongly in patients who had greater vasoconstrictor infusion requirements O`Riordan et al.; Crit. Care 2006; 10: R139 RH Böger; Crit. Care 2006; 10: 169
Targeting intracellular arginine / asymmetric dimethylarginine (ADMA) Summary Numerous epidemiological studies have provided evidence for a statistical association between ADMA and cardiovascular disease. Pathophysiological evidence suggests that ADMA may exert its effects by interfering with NO production in the endothelium, resulting in endothelial dysfunction and advanced atherosclerosis. Prospective clinical trials have shown that elevated ADMA predicts major adverse cardiovascular events and mortality throughout the cardiovascular continuum. DDAH-1 appears to be the most prominent regulator of endogenous ADMA. DDAH-1 might thus be a novel target for pharmacotherapeutic intervention in cardiovascular diseases.
Acknowledgments University Medical Center Hamburg-Eppendorf Edzard Schwedhelm Nicole Lüneburg Maike Anderssohn Doro Atzler Anna Steenpaß Mariola Kastner Sandra Maak Daniel Appel Ghainsom Kom Thomas Eschenhagen Karsten Sydow, Kardiologie Thomas Meinertz, Kardiologie Thomas Standl, Anästhesiologie Stanford University John P. Cooke Phil S. Tsao Patrick S. Lin Instituto die Fisiologia, Reggio, IT Carmine Zoccali University of Iowa Steven R. Lentz University of Ulm Wolfgang Koenig University of Erlangen Renke Maas Johannes Jacobi Boston University Vasan S. Ramachandran University of Minnesota Yingjie Chen