key: cord-0697317-h0h6cri8
authors: Soler, María José; Barrios, Clara; Oliva, Raymond; Batlle, Daniel
title: Pharmacologic modulation of ACE2 expression
date: 2008-10-16
journal: Curr Hypertens Rep
DOI: 10.1007/s11906-008-0076-0
sha: b821e513f4b1e9c852516e097ea09a710856450c
doc_id: 697317
cord_uid: h0h6cri8

Angiotensin-converting enzyme 2 (ACE2) is an enzymatically active homologue of angiotensin-converting enzyme that degrades angiotensin I, angiotensin II, and other peptides. Recent studies have shown that under pathologic conditions, ACE2 expression in the kidney is altered. In this review, we briefly summarize recent studies dealing with pharmacologic interventions that modulate ACE2 expression. ACE2 amplification may have a potential therapeutic role for kidney disease and hypertension.

Angiotensin-converting enzyme 2 (ACE2) is a carboxypeptidase that cleaves the peptides angiotensin (Ang) I and II, apelin, and neurotensin [ 1 ] . The ACE2 gene is mapped in the human X chromosome (Xp22) and contains 18 exons, somewhat similar to ACE in exon size and organization [ 2 ] . The human ACE2 protein is a zinc-containing metallopeptidase with 806 amino acids and shares 42% sequence identity and 61% sequence similarity with ACE [ 2 , 3 ]. Within the renin-angiotensin system (RAS), ACE2 promotes the degradation of the vasoconstrictor and proliferative peptide Ang II to the vasodilatory and antiproliferative Ang-(1-7) [ 4 ].

ACE2 expression was initially found in the testis, kidney, and heart [ 2 , 3 ] . Later studies showed widespread ACE2 distribution in the lung, liver, small intestine, brain, and placenta [ 5 , 6• , 7-9 , 10• ]. The distribution of ACE2 in the kidney is specifi cally found in the apical membranes of the proximal tubules and in the glomerular epithelial cells (podocytes) [ 11 , 12 ] . ACE2 alterations have been described in experimental models of hypertension and diabetic kidney disease [ 13•-15• , 16•• ]. ACE2 was found to be decreased in the setting of hypertension. Crackower et al. [ 4 ] showed that ACE2 was reduced in kidneys from three separate hypertensive rat strains: salt-sensitive Sabra hypertensive rats (SBH/y), spontaneous hypertensive rats (SHR), and stroke-prone spontaneously hypertensive rats (SHR-SP). Kidney ACE2 gene and protein expression were decreased in adult SHR as compared with Wistar-Kyoto (WKY) rats [ 17 ] .

Our laboratory showed that in the db/db murine model of experimental diabetes, ACE2 glomerular expression is decreased by immunohistochemistry [ 16•• ]. In agreement with this fi nding, Leehey et al.

[ 18 ] also found decreased glomerular ACE2 protein levels and activity in streptozotocin-induced diabetic rats. The tubular expression of ACE2 is altered, and the glomerular expression is increased in models of diabetic kidney disease [ 3 , 15• ]. It is also known that ACE2 inhibition leads to worsening of albuminuria in diabetic mice [ 11-13 , 15• , 16•• , 18-20 , 21•• ]. ACE2 may be involved in the hemodynamics of pregnancy [ 22 ] . In this report, we discuss conditions in which ACE2 expression is altered, and we review recent studies showing that Ang II receptor blockers, ACE inhibitors, and aldosterone antagonists modulate ACE2 expression.

Studies in kidney and peritoneal cell lines have shown that cytokines can modulate ACE2 expression in vitro [ 23 ] . In Vero E6 cells, which are tubular epithelial cells from monkeys, interferon γ and interleukin 4 decreased ACE2 expression at the protein and gene levels [ 23 ] . In peritoneal macrophages from mice, aldactone (an aldosterone antagonist) decreased ACE2 enzymatic activity and gene expression [ 24•• ] .

The effect of Ang II stimulation on ACE2 expression has been studied using animal experimental models and in vitro cultures. Gallagher et al. [ 25 ] showed that in cultured astrocytes, exposure to Ang II caused a reduction in neural ACE2 mRNA and protein, a response mediated by the Ang II (AT) 1 receptor. In concordance with this study, Ang II was able to up-regulate ACE and down-reg ulate ACE2 in human kidney tubular cells. These effects were blocked by an AT1 receptor antagonist (losartan), but not by an AT2 receptor blocker (PD123319) [ 26• ]. Furthermore, blockade of extracellular signal-regulated kinases 1/2 (ERK1/2) or p38 mitogen-activated protein (MAP) kinases by either specifi c inhibitors or a dominant-negative adenovirus abolished Ang II-induced ACE2 down-regulation in human kidney tubular cells [ 26• ] .

There is evidence that all-trans retinoic acid (at-RA) infl uences gene expression of RAS components. Zhong et al.

[ 17 ] studied this relationship in SHR and WKY rats treated with daily intraperitoneal at-RA injection. They showed that ACE2 expression was markedly decreased in placebo-treated SHR when compared with WKY rats. In at-RA treated SHR, a signifi cant up-regulation of ACE2 expression was observed in the heart and kidney [ 17 ] . Also, at-RA affects regulation of the stem cell marker octomer-4 (Oct-4) and eventually, cellular differentiation. Zulli et al. [ 27 ] showed that cells within atherosclerotic plaques of New Zealand White rabbits co-express ACE2 and the hematopoietic stem cell marker, CD34. Thus, at-RA treatment could affect plaque cellular biology via effects on cellular differentiation and blood pressure via its effect on ACE2 [ 27 ] .

The hormone 17 β -estradiol increased ACE2 protein and gene expression in ovariectomized female rats with the renal wrap model of hypertension. It also prevented glomerular and tubular injury in this experimental hypertensive model [ 28• ] . In an experimental model of acute renal failure after lipopolysaccharide treatment (rat model of endotoxemia), Gupta et al. [ 29 ] showed that activated protein C (APC) injection modulated the RAS by reducing ACE and angiotensinogen. APC was also shown to increase ACE2 mRNA levels in the kidney. This experiment illustrates that the potential protective role of APC in the kidney might be mediated by enhanced renal ACE2 expression, which is consistent with the original proposition that ACE2 may be renoprotective [ 19 ] .

Effect of Renin-Angiotensin-Aldosterone System Blockade on ACE2 Expression Some antihypertensive drugs have been shown to increase ACE2 enzymatic activity and protein and gene expression in different species, tissues, and cells ( Table 1 ) [ 24•• , 30-33 ] .

In the heart, Ang II receptor blockers have been shown to increase ACE2 protein and gene expression in different models of experimental hypertension [ 34 , 35• , 36 ]. In the model of myocardial infarction after left coronary artery ligation and in transgenic Ren-2 rats, the ACE inhibitors enalapril and lisinopril increased heart ACE2 expression [ 31 , 35• ] . In the thoracic aorta of male SHR, ACE2 was increased in association with reversal of vascular hypertrophy in response to olmesartan treatment [ 32 ] . In a study by Whaley-Connell et al. [ 37 ] that examined glomerular fi ltration barrier injury in the Ren-2 transgenic rat, AT1 receptor blockade was associated with increased ACE2 expression. These changes were not observed in atenolol-or hydralazine-treated hypertensive rats [ 32 ] . Taken together, these fi ndings suggest that ACE2 is regulated by AT1 receptors and may be involved in mediating the pressure-independent vascular remodeling effects of Ang II blockers [ 32 ] .

Cardiac Ang II concentration and activity of MAP kinases were markedly increased in response to pressure overload in mice lacking ACE2 (Ace2 -/y mice). Administration of candesartan, an AT1 receptor blocker, attenuated the hypertrophic response and suppressed the activation of MAP kinases in Ace2 -/y mice [ 38• ] . These results suggest that ACE2 plays an important role in regulating the hypertrophic response to pressure overload mediated by Ang II. In a study by Oudit et al. [ 39• ] , Ace2 -/y mutant mice developed a progressive, age-dependent, dilated cardiomyopathy with increased oxidative stress, neutrophilic infi ltration, infl ammatory cytokine and collagenase levels, MAP kinase activation, and pathologic hypertrophy. The AT1 receptor blocker irbesartan prevented the dilated cardiomyopathy in aged Ace2 -/y mutant mice. This confi rms the critical role of Ang II-mediated stimulation of AT1 receptors [ 39• ] .

In the kidney, both lisinopril and losartan increased ACE2 enzymatic activity in the renal cortex of adult Lewis rats [ 33 ] . Our laboratory showed that telmisartan increases ACE2 protein expression in the renal vasculature [ 30 ] . It is unknown if this action is related, in part, to the wellknown peroxisome proliferator-activated receptor (PPAR) effect of telmisartan [ 40 ] . The PPARs are members of the nuclear receptor superfamily of ligand-activated transcription factors. In particular, PPAR-γ plays a critical role in regulating carbohydrate and lipid metabolisms. PPAR-γ ligands have modest antihypertensive effects related to their ability to promote peripheral vasodilation, improve insulin sensitivity, and decrease the risk for atherosclerosis [ 41 ] . The Ang II receptor blocker telmisartan is structurally similar to a PPAR-γ agonist. In fact, telmisartan treatment in vitro augmented PPAR-γ activity. Recently Kobayashi et al. [ 42 ] showed that in Dahl salt-sensitive hypertensive rats, telmisartan stimulates nitric oxide production through PPAR-γ and the Rho-kinase pathway. It also ameliorated cardiac hypertrophy and cardiovascular remodeling. A direct effect of PPARs on ACE2 expression has not been studied, but they could function in synergism with Ang II receptor blockers.

Aldosterone antagonists (eg, spironolactone and eplerenone) have been shown to increase ACE2 enzymatic activity in macrophages from humans and mice [ 24•• ] . Spironolactone treatment increased ACE2 gene expression in human macrophages [ 24•• ] . Eplerenone treatment, on the other hand, increased ACE2 activity and decreased ACE activity in hearts from Balb/C mice as compared with vehicle-treated mice [ 24•• ] . This suggests that aldosterone inhibits ACE2, but the mechanism needs to be elucidated.

Although the foregoing studies suggest that indirect ways exist to infl uence ACE2 activity pharmacologically, namely by using agents that affect the RAS, there is an interest in exploring and developing agents that are primarily inhibitors or activators of ACE2. [ 13• ] found increased albuminuria, glomerular mesangial expansion, and vascular thickness after MLN-4760 treatment. Our fi nding of increased albuminuria in two models of diabetic mice treated with an ACE2 inhibitor suggests a role of this enzyme in regulating Ang II-mediated glomerular permeability. In agreement with this, Tikellis et al. [ 43• ] recently found that in STZ-treated mice, albuminuria was increased after MLN-4760 administration for 4 weeks. ACE2-knockout diabetic mice using STZ experienced a 5.2-fold increase in urinary albumin excretion when compared with untreated ACE2-knockout mice [ 43• ] . Surprisingly, ACE2 inhibition was able to attenuate diabetes-associated changes in osteopontin expression and glomerular fi bronectin accumulation [ 43• ] . The dissociation between the effects on albuminuria and fi brogenesis after ACE2 inhibition is unexpected, and both fi ndings are diffi cult to reconcile. Tikellis et al. [ 43• ] suggested that the decrease in fi bronectin deposition was possibly related to a decrease in renal ACE activity in animals treated with the selective ACE2 inhibitor. The decrease in ACE activity observed by these authors was previously reported in kidney cortex from STZ diabetic mice treated with MLN-4760 [ 13• ]. However, it must be noted that glomeruli from diabetic mice treated with MLN-4760 had an increase in ACE expression [ 13• ]. In contrast, ACE2 expression is decreased (not increased) in kidney cortex, which is mostly composed of proximal tubules [ 15• ]. Another interesting fi nding from Tikellis et al. [ 43• ] is that perindopril reduced renal cortical ACE2 activity in both control and diabetic animals. Although the interaction of ACE and ACE2 appears complex and is not fully understood, ACE2 protein and activity may be infl uenced by the level of ACE protein and activity, and vice versa.

Ongoing studies are intended to develop drugs that enhance ACE2 activity. Recently, Hernandez Prada et al. [ 44•• ] , using structure-base screening, found a compound named xanthenone that enhances ACE2 activity. This compound caused considerable reductions in blood pressure, and a striking reversal of cardiac and renal fi brosis in the SHR model of hypertension [ 44•• ] .

We have used recombinant ACE2 (r-ACE2) as a novel approach to increase Ang II metabolism and reduce Ang II-dependent hypertension [ 45•• ] . We showed that the increase in blood pressure associated with Ang II infusion was abolished in mice infused simultaneously with r-ACE2. Thus, the administration of enzymatically functional ACE2 abrogates Ang II-induced hypertension [ 45•• ] .

ACE2 is an enzymatically active homologue of ACE that plays a signifi cant role in maintaining a balanced status of the RAS. Several studies have shown that ACE2 is altered under pathologic conditions, and its inhibition by pharma-cologic or genetic deletion has been shown to accelerate kidney and heart injury. Drugs based on RAS blockade -ACE inhibitors and AT1 receptor blockers-appear to increase ACE2 expression in the heart and the vasculature. This effect may contribute to their antihypertensive and cardiovascular protective action. New strategies aimed at new drug targets that are more effective in ACE2 amplifi cation may provide a therapeutic approach to protect against cardiovascular disease, kidney disease, and hypertension.

No potential confl icts of interest relevant to this article were reported. Results of this study showed that ACE2 is part of the brain RAS. ACE2 staining is present in the cytoplasm of neuronal cell bodies, but not in glial cells. In the subfornical organ, ACE2 was significantly increased in transgenic mice. ACE2 may have a role in the central regulation of blood pressure and diseases involving the autonomic nervous system (eg, hypertension).

Batlle D, Soler MJ, Wysocki J: New aspects of the reninangiotensin system: angiotensin-converting enzyme 2-a potential target for treatment of hypertension and diabetic nephropathy. 

Mineralocorticoid receptor blocker increases angiotensinconverting enzyme 2 activity in congestive heart failure patients

This paper showed that mineralocorticoid receptor blockade had an effect on ACE and ACE2. ACE activity and mRNA were significantly increased, whereas ACE2 was signifi cantly reduced

Angiotensin II up-regulates angiotensin I-converting enzyme (ACE), but down-regulates ACE2 via the AT1-ERK/p38 MAP kinase pathway

Ang II can act to up-regulate ACE but down-regulate ACE2 via the AT1 receptor-mediated mechanism. Activation of the ERK1/2 and p38 MAP kinase pathway may represent a key mechanism by which Ang II down-regulates ACE2

Co-localization of angiotensin-converting enzyme 2-, octomer-4-and CD34-positive cells in rabbit atherosclerotic plaques

Role of angiotensin-converting enzyme 2 and angiotensin(1-7) in 17beta-oestradiol regulation of renal pathology in renal wrap hypertension

These researchers showed that the hormone 17 β -estradiol increases ACE2 protein and gene expression in ovariectomized female rats with the renal wrap model of hypertension. Furthermore, 17 β -estradiol prevents glomerular and tubular

Activated protein C ameliorates LPS-induced acute kidney injury and downregulates renal INOS and angiotensin 2

ACE2 is preferentially localized in the tunica media layer in renal vasculature and its expression increases after administration of a type 1 receptor antagonist

Enalapril attenuates downregulation of angiotensin-converting enzyme 2 in the late phase of ventricular dysfunction in myocardial infarcted rat

Angiotensin II AT1 receptors regulate ACE2 and angiotensin-(1-7) expression in the aorta of spontaneously hypertensive rats

-7) forming enzymes and receptors

Effects of aldosterone and angiotensin II receptor blockade on cardiac angiotensinogen and angiotensin-converting enzyme 2 expression in Dahl salt-sensitive hypertensive rats

Effect of angiotensin II blockade on a new congenic model of hypertension derived from transgenic Ren-2 rats

This study investigated the effects of type 1 angiotensin II (Ang II) receptor (AT1) blockade and ACE activity inhibition on the Ang-(1

/ACE2 axis of the RAS in the hypertensive model of Ren rats

Olmesartan is an angiotensin II receptor blocker with an inhibitory effect on angiotensin-converting enzyme

Oxidative stress and glomerular fi ltration barrier injury: role of the renin-angiotensin system in the Ren2 transgenic rat

Deletion of angiotensin-converting enzyme 2 accelerates pressure overload-induced cardiac dysfunction by increasing local angiotensin II

Results of this study showed that mice lacking ACE2 (ACE -/y mice) developed reduced cardiac contractility, eventually leading to cardiac dilatation and hypertrophy. Due to chronic overload, the mice developed pulmonary congestion and increased incidence of cardiac death

Angiotensin II-mediated oxidative stress and infl ammation mediate the age-dependent cardiomyopathy in ACE2 null mice

The study showed that age-dependent cardiomyopathy in ACE2-null mice is related to increased Ang II-mediated oxidative stress and neutrophilic infi ltration via AT1 receptors

Identifi cation of telmisartan as a unique angiotensin II receptor antagonist with selective PPARgamma-modulating activity

Telmisartan is a promising cardiometabolic sartan due to its unique PPAR-gammainducing property

Cardioprotective mechanism of telmisartan via PPAR-gamma-eNOS pathway in dahl salt-sensitive hypertensive rats

ACE2 defi ciency modifi es renoprotection afforded by ACE inhibition in experimental diabetes

This study showed that the expression of ACE2 is modifi ed by diabetes and may affect the pathogenesis of kidney disease and responsiveness to RAS blockade

This study provided evidence that a compound named xanthenone, which enhances ACE2 activity, causes considerable reductions in blood pressure and striking reversal of cardiac and renal fi brosis in the SHR model of hypertension

Recombinant ACE2 attenuates angiotensin II induced hypertension

The study showed the effects of r-ACE2 can prevent Ang II-induced hypertension. The data indicated that increasing r-ACE2 activity may provide a novel approach to reduce high blood pressure