Congestive heart failure and expression of myocardial urotensin II

doi:10.1016/S0140-6736(02)08831-1

The Lancet

Volume 359, Issue 9322, 8 June 2002, Pages 1990-1997

https://doi.org/10.1016/S0140-6736(02)08831-1 Get rights and content

Summary

Background

Human urotensin II has several cardiovascular actions, including potent vasoactive, and cardiac inotropic and hypertropic properties. Our aim was to ascertain degree of expression of urotensin II and its receptor GPR14 (now known as UT receptor) in the myocardium of patients with congestive heart failure (CHF).

Methods

We obtained specimens of myocardium from the hearts of 19 patients with end-stage CHF (12 ischaemic heart disease, seven dilated cardiomyopathy), five patients with early-stage CHF, and eight healthy controls. We used immunohistochemistry, in-situ hybridisation, reverse transcriptase-PCR (RT-PCR), and fluorescein isothiocyanate (FITC)-conjugated urotensin II to ascertain degree of myocardial expression of urotensin II and binding urotensin receptor.

Findings

Our results showed strong expression of urotensin II in the cardiomyocytes, and to a lesser extent in the vascular smooth muscle cells, endothelial cells, and inflammatory cells of patients with end-stage CHF. There was significantly less urotensin II expression in the myocardium of patients with early-stage CHF (p<0·0001). Also, there was little to no urotensin II expression in the myocardium of healthy controls. Myocardial expression of urotensin II correlated significantly with left ventricular end-diastolic dimension (p=0·0092), and inversely with ejection fraction (p=0·0002). RT-PCR showed increased concentrations of urotensin II and presence of urotensin receptor mRNA in the myocardium of patients with CHF. The confocal microscopy results showed a significant increase in the binding sites for urotensin in the myocardium of patients with end-stage CHF (p<0·0001).

Interpretation

Our findings suggest a possible role for urotensin II in the cardiac dysfunction and remodelling characteristic of CHF.

Introduction

Congestive heart failure (CHF) is the leading cause of death in the developed world. Several cytokines, vasoactive mediators, and growth factors have been implicated in the disease process. The human form of urotensin II, a cyclic peptide initially isolated from the caudal neurosecretory system of teleost fish,¹ as well as its g-protein-coupled receptors (gprs), now known as UT receptor,² have been isolated.3, 4 Human urotensin II is composed of 11 aminoacid residues with a disulfide linked cyclic octapeptide core region, which is responsible for its biological activity.4, 5 The contractile effects of urotensin II on smooth muscle cells seem to vary dependent on the species and the type of vessel, and are often more potent than that of endothelin 1.3, 6, 7 Results of experiments have shown that human urotensin II causes contraction of all non-human primate arterial vessels,³ aortocoronary vessels of rats7, 8 and dogs,⁷ human trabeculae,⁹ and human coronary, mammary, and radial arteries, saphenous, and umbilical veins.10, 11, 12 Findings of other studies, however, indicate either vasodilatory or no effect of urotensin II in vessels of human beings.13, 14 There therefore seems to be a discrepancy between in-vivo and in-vitro effects of urotensin II on blood vessels of people.

Administration of human urotensin II resulted in deterioration of cardiovascular function in anaesthetised monkeys, causing fatal circulatory collapse and severe myocardial depression (most likely an indirect effect secondary to coronary ischaemia). Indeed, bolus administration of human urotensin II induced a 300% increase in total peripheral resistance in anaesthetised monkeys, without causing systemic hypertension, and an 80% decrease in dP/dt with ST segment changes consistent with concomitant myocardial ischaemia (as would be observed as a result of constriction of coronary arteries).3, 9 The vasoconstrictor effects of human urotensin II are mediated by the binding of the peptide to specific high-affinity receptors, and seem to involve calcium mobilisation and a phospholipase c dependent increase in inositol phosphates.2, 15, 16

Myocardial expression of urotensin II and the UT receptor in the healthy or diseased heart remain to be elucidated. Findings of studies show that human urotensin II causes cardiac dysfunction in mammals, proarrhythmogenic activity in isolated human hearts,⁹ and increased collagen production in neonatal cardiac fibroblasts,¹⁷ suggesting that CHF might be associated with the myocardial urotensin II system. Our aim was to investigate myocardial expression of human urotensin II and its binding to the receptor in the hearts of healthy controls and in patients with CHF. We also correlated cellular expression of urotensin II with clinical characteristics of CHF.

Section snippets

Participants

We obtained tissue from the myocardium of 19 patients with end-stage heart failure (New York Heart Association functional class III-V) at surgery for heart transplantation (through the Quebec Vascular Endothelium Network Tissue Bank). Of the 19 patients, 12 had ischaemic heart disease and seven had dilated cardiomyopathy. Myocardial biopsies (3 mm) from the left ventricle of five patients with mild CHF (New York Heart Association functional class I-II), requiring deep coronary grafts, were

Results

The table shows the clinical characteristics of individuals from whom samples were taken. Patients with CHF were treated with a combination of drugs, including diuretics, digoxin, angiotensin-converting-enzyme inhibitors, nitrates, calcium channel blockers, low-dose aspirin, and antiarrhythmic drugs. Complications included hypertension (n= 10) and diabetes mellitus (n=4).

The myocardium of hearts from healthy controls showed little or no urotensin II mRNA or immunoreactivity to urotensin II

Discussion

Our results indicate that expression of urotensin II is upregulated in the hearts of patients with end-stage CHF when compared with controls. Furthermore, urotensin II expression is localised to the cardiomyocytes, endothelial cells, and vascular smooth muscle cells of the damaged heart, with greatest expression in the subendocardial region and in the surviving cardiomyocytes of infarction and border zones of patients with ischaemic heart disease. Myocardial tissue from patients with

GLOSSARY

g-protein-coupled receptors (gprs): GPRs mediate cellular responses to a variety of molecules, including many drugs. More than 400 GPR have been identified. G stands for Guanosine triphosphate or GTP. When the receptors are activated they bind to G proteins, which through conformational changes and the hydyolysis of GTP then cause intracellular changes.

phospholipase c: Phospholipase C is a an enzyme that releases inositol trisphosphate and diacylglycerol from the plasma membrane to act as

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Cited by (248)

Commentary: Molecular pathogenesis of aortic stenosis: Will the puzzle pieces ever fit together?
2021, Journal of Thoracic and Cardiovascular Surgery
Urotensin II, urotensin-related peptide, and their receptor in aortic valve stenosis
2021, Journal of Thoracic and Cardiovascular Surgery
Aortic valve stenosis (AVS) is the most common cause of surgical valve replacement worldwide. The vasoactive peptide urotensin II (UII) is upregulated in atherosclerosis and several other cardiovascular diseases; however, its role in the pathogenesis of AVS remains to be determined. Here, we investigated the expression of UII, urotensin-related peptide (URP), and the urotensin receptor (UT) and the role this system plays in AVS.
Immunohistochemistry and reverse-transcriptase polymerase chain reaction were used to examine the cellular localization and mRNA expression, of UII, URP, and UT in calcified and noncalcified aortic valves. Human aortic valve interstitial cells were isolated from normal valves and treated with UII or URP, and changes in cell proliferation, cholesterol efflux, calcium deposition, and β-catenin translocation were assessed.
The mRNA expression of UII, URP, and UT was significantly greater in patients with AVS. There was abundant presence of UII, URP, and UT immunostaining in diseased compared with nondiseased valves and correlated significantly with presence of calcification (P < .0001) and fibrosis (P < .0001). Treating human aortic valve interstitial cells with UII or URP significantly increased cell proliferation (P < .0001) and decreased cholesterol efflux (P = .0011 and P = .0002, respectively). UII also significantly reduced ABCA1 protein expression (P = .0457) and increased β-catenin nuclear translocation (P < .0001) and mineral deposition (P < .0001).
Together, these data suggest that the urotensin system plays a role in the pathogenesis of AVS and warrants further investigation.
Identification of novel Urotensin-II receptor antagonists with potent inhibition of U-II induced pressor response in mice
2020, European Journal of Pharmacology
Citation Excerpt :
UT gene is widely expressed in the central nervous system (CNS) and in peripheral tissues including retina, heart, vascular bed, lung, kidney, adrenal medulla, and skeletal muscle (Hunt et al., 2010; Maguire et al., 2008). Various studies have revealed the increased expression of U-II and its receptor UT in the failing hearts and renal diseases (Ng et al., 2002; Douglas et al., 2002; Bousette et al., 2006a,b; Loirand et al., 2008). Genetic deletion of UT in mice renders aortae, refractive to the contractile activity of U-II (You et al., 2012).
Urotensin II (U-II) has been found to be one of the most potent vasoconstrictor (Ames et al., 1999; Bohm et al., 2002) reported till date. U-II exerts its response via activation of a G-protein coupled receptor, Urotensin II receptor(UT). Binding of U-II to UT leads to an instant increase in the inositol phosphate turnover and intracellular Ca²⁺. Such an instant Ca²⁺ release and potent vasoconstriction exerted by U-II is expected to have an important role in the progression of cardiac diseases.
We have previously shown that UT antagonist DS37001789 prevents U-II induced blood pressure elevation in mice (Nishi et al., 2019) in a dose dependent manner, with potent efficacy at 30 and 100 mg/kg. Further to this, we have also shown that DS37001789 ameliorates mortality in pressure-overload mice with heart failure (Nishi et al., 2020). We therefore conducted an extensive structure-activity relationship studies to identify molecules with superior efficacy. In the present manuscript, we report the identification of two potent, non-peptide small molecule antagonists of Urotensin II receptor (UT), RCI-0879 and RCI-0298 which blocked the action of U-II, both in vitro and in vivo. These molecules were found to be very potent in in vitro Ca²⁺ and radioligand binding assays using human and mouse UT over-expressing CHO cells. RCI-0879 and RCI-0298 also exhibited superior efficacy in in vivo mouse pressor response model using C57BL/6 mice, compared to our initial molecules (Nishi et al., 2019) and demonstrated ED₅₀ values of 3.2 mg/kg and 6.8 mg/kg respectively. Our findings reported herewith, further strengthen our concept and belief in UT antagonization as a potential therapeutic approach for the management of chronic heart failure.
Study of the association between Urotensin 2 (p.T21M and p.S89N) variants and breast cancer in Egyptian patients
2020, Gene Reports
In females, breast cancer is the leading cancer regarding both incidence and mortality. Urotensin II (UTS2) is expressed in various tumours, including breast cancer.
To study the association between UTS2 (p.T21M and p.S89N) gene variants and breast cancer.
A total of 106 females with breast cancer and 108 age- and sex-matched healthy volunteers were genotyped for UTS2 (p.T21M and p.S89N) variants by TaqMan real-time PCR and PCR-restriction fragment length polymorphism (RFLP), respectively.
There was no significant difference between cases and controls regarding the distribution of UTS2 p.T21M genotypes. However, frequencies of the heterozygous S/N genotype and the N allele of UTS2 p.S89N were significantly higher in breast cancer patients than in controls (p = 0.018, OR = 5.52, 95% CI = 1.18–25.83; p = 0.020, OR = 5.30, 95% CI = 1.15–24.38, respectively).
The UTS2 p.S89N variant but not the UTS2 p.T21M variant is associated with an increased risk of developing breast cancer in Egyptian patients.
Functional effects of urotensin-II on intracellular pH regulators in human radial artery smooth muscle cells
2020, Peptides
The regulation of intracellular pH (pH_i) plays a vital role in various cellular functions. We previously demonstrated that three different acid extruders, the Na⁺-H⁺ exchanger (NHE), Na⁺−HCO₃⁻ co-transporter (NBC) and H⁺-linked monocarboxylate transporter (MCT), functioned together in cultured human radial artery smooth muscle cells (HRASMCs). However, the functions of acid-loading transporters in HRASMCs remain poorly understood. Urotensin II (U-II), one of the most potent vasoconstrictors, is highly expressed in many cardiovascular diseases. The aim of this present study was to determine the concentration effect of U-II (3 pM∼100 nM) on the functional activity of pH_i regulators in HRASMCs. Cultured HRASMCs were derived from segments of human radial arteries obtained from patients undergoing bypass grafting. Changes in pH_i recovery due to intracellular acidification and alkalization induced by NH₄Cl prepulse and Na-acetate prepulse, respectively, were detected by microspectrofluorimetry with the pH-sensitive fluorescent dye BCECF. Our present study showed that (a) U-II increased the activity of NHE in a concentration-dependent manner but did not change that of NBC or MCT or resting pH_i, (b) the Cl⁻−OH⁻ exchanger (CHE) facilitated base extrusion, and (c) U-II induced a concentration-dependent increase in the activity of CHE. In conclusion, for the first time, our results highlight a concentration-dependent increase in the activity of NHE and CHE, but not NBC and MCT, induced by U-II in HRASMCs.
The urotensin II receptor antagonist DS37001789 ameliorates mortality in pressure-overload mice with heart failure
2020, Heliyon
This study was designed to evaluate the effects of DS37001789, a novel and highly potent urotensin II (U-II) receptor (GPR14) antagonist, against mortality, hypertrophy, and cardiac dysfunction in pressure-overload hypertrophy by transverse aortic constriction (TAC) in mice. In addition, we analyzed the phenotype of GPR14 knockout (KO) mice after TAC induction to confirm the contribution of the U-II/GPR14 system. The oral administration of 0.2% DS37001789 to TAC mice for 12 weeks significantly ameliorated the mortality rate and 0.2% DS37001789 for 4 weeks significantly improved cardiac function by pressure-volume analysis. GPR14 expression was significantly upregulated in the left ventricle in the TAC mice treated with 0.2% DS37001789. Moreover, we confirmed that the significant amelioration of mortality was accomplished by the inhibition of cardiac enlargement and the improvement of cardiac function in GPR14 KO mice after TAC surgery. These results suggest that the U-II/GPR14 system contributes to the progression of heart failure and its blockade ameliorates the mortality via improved cardiac function. The U-II/GPR14 system may thus be an attractive target for treating heart failure with pathological cardiac hypertrophy and DS37001789 may be a novel therapeutic agent for heart failure in patients with pressure-overload conditions such as hypertension and aortic valve stenosis.

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Mechanisms of DiseaseCongestive heart failure and expression of myocardial urotensin II

Summary

Background

Methods

Findings

Interpretation

Introduction

Section snippets

Participants

Results

Discussion

GLOSSARY

Recent Prog Horm Res

Aplysia californica. Peptides

Eur J Pharmacol

Gen Comp Endocrinol

J Am CollCardiol

Urotensin receptors

Human urotensin-II is a potent vasoconstrictor and agonist for the orphan receptor GPR14

Nature

Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord

Proc Natl Acad Sci USA

Molecular modelling and design of analogues of the peptide hormone urotensin II

Biochem Soc Trans

Human urotensin-II is a potent vasoactive peptide: pharmacological characterization in the rat, mouse, dog and primate

J Cardiovasc Pharmacol

Human urotensin-II is an endothelium-dependent vasodilator in rat small arteries

Br J Pharmacol

Cardiostimulant effects of urotensin-II in human heart in vitro

Br J Pharmacol

Orphan-receptor ligand human urotensin II: receptor localization in human tissues and comparison of vasoconstrictor responses with endothelin-1

Br J Pharmacol

Mechanisms of Disease
Congestive heart failure and expression of myocardial urotensin II