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. 2022 Feb 16;16(4):601–608. doi: 10.1007/s12079-022-00672-4

The network map of urotensin-II mediated signaling pathway in physiological and pathological conditions

D A B Rex 1, G P Suchitha 1, Akhina Palollathil 1, Anagha Kanichery 1, T S Keshava Prasad 1,✉, Shobha Dagamajalu 1,✉
PMCID: PMC9733756  PMID: 35174439

Abstract

Urotensin-II is a polypeptide ligand with neurohormone-like activity. It mediates downstream signaling pathways through G-protein-coupled receptor 14 (GPR14) also known as urotensin receptor (UTR). Urotensin-II is the most potent endogenous vasoconstrictor in mammals, promoting cardiovascular remodelling, cardiac fibrosis, and cardiomyocyte hypertrophy. It is also involved in other physiological and pathological activities, including neurosecretory effects, insulin resistance, atherosclerosis, kidney disease, and carcinogenic effects. Moreover, it is a notable player in the process of inflammatory injury, which leads to the development of inflammatory diseases. Urotensin-II/UTR expression stimulates the accumulation of monocytes and macrophages, which promote the adhesion molecules expression, chemokines activation and release of inflammatory cytokines at inflammatory injury sites. Therefore, urotensin-II turns out to be an important therapeutic target for the treatment options and management of associated diseases. The main downstream signaling pathways mediated through this urotensin-II /UTR system are RhoA/ROCK, MAPKs and PI3K/AKT. Due to the importance of urotensin-II systems in biomedicine, we consolidated a network map of urotensin-II /UTR signaling. The described signaling map comprises 33 activation/inhibition events, 31 catalysis events, 15 molecular associations, 40 gene regulation events, 60 types of protein expression, and 11 protein translocation events. The urotensin-II signaling pathway map is made freely accessible through the WikiPathways Database (https://www.wikipathways.org/index.php/Pathway:WP5158). The availability of comprehensive urotensin-II signaling in the public resource will help understand the regulation and function of this pathway in normal and pathological conditions. We believe this resource will provide a platform to the scientific community in facilitating the identification of novel therapeutic drug targets for diseases associated with urotensin-II signaling.

Supplementary Information

The online version contains supplementary material available at 10.1007/s12079-022-00672-4.

Keywords: Intercellular signaling, Ligand-receptor interactions, Omics integration, Pathways, Signaling network

Introduction

Urotensin-II is a neurohormone-like polypeptide molecule (Sun and Liu 2019). It was first isolated in 1980 by D Pearson et al. from the urophysis of the goby fish (Gillichthys mirabilis) (Pearson et al. 1980). Urotensin-II is the most potent vasoconstrictor cyclic peptide, with similar domain sequences to somatostatin (Ames et al. 1999). The urotensin-II system is involved in multiple biological systems, including cardiovascular, nervous, endocrine, and renal (Douglas et al. 2004; Vaudry et al. 2010, 2015). Urotensin-II is encoded by a gene UTS2, which is located on chromosome 1p36 (Tostivint et al. 2006). In humans, the alternative splicing of the UTS2 gene yielding 124 (isoform b, NP_006777) and 139 (isoform a, NP_068835.1) amino acid variants of prepro-urotensin-II. The proteolytic cleavage of preproprotein urotensin-II by a specific urotensin converting enzyme (UCE) results in a mature 11- amino acid sequence peptide urotensin-II. Urotensin-II selectively binds to G-protein -coupled orphan receptor 14 (GPR14), known as the urotensin-II receptor encoded by the UTS2R gene, located on human chromosome 17q25.3 (Muravenko et al. 2000).

In physiological conditions, the vascular effect of urotensin-II is a balance between vasoconstriction and vasodilation, resulting in the release of nitric oxide, prostaglandins, and endothelium-derived hyperpolarizing factors from endothelial cells. In addition, urotensin-II plays a role in the immune response, which leads to activation of cytokine expression and the promotion of immune cell infiltration (Castel et al. 2017). The altered expression of urotensin-II and urotensin receptor (UTR) in plasma, urine and cerebrospinal fluid can be seen in vascular inflammatory diseases, including essential hypertension, chronic heart failure, atherosclerosis, renal failure, renal dysfunction, portal hypertension/cirrhosis, and diabetes mellitus (Tsoukas et al. 2011). It suggests that the  urotensin-II/UTR system may not contribute to the regulation of organ function in healthy states but also plays a significant role in inflammatory diseases (Ong et al. 2005; Tsoukas et al. 2011).

Urotensin-II mediated activation of UTR increases intracellular Ca2+ that causes vasoconstriction (Gibson et al. 1988; Ames et al. 1999; Filipeanu et al. 2002; Song et al. 2006). In addition, the influx of extracellular calcium through L-type calcium channels is also responsible for vasoconstriction (Zhang et al. 2015). Urotensin-II is a well-known potent vasoconstrictor, which turns out to be an important molecule for treatment options and management of the diseases (Douglas and Ohlstein 2000; Zhu et al. 2006). Many peptidic and nonpeptidic urotensin-II antagonists have been developed to reveal the role of urotensinergic system. Urantide is a UTR competitive antagonist that fastidiously blocks the urotensin-II induced effects in the rat aorta and human monocyte-derived macrophages (Patacchini et al. 2003; Watanabe et al. 2005). Another UTR competitive antagonist, BIM-23127 inhibited calcium mobilization in human embryonic kidney cells expressing UTR and inhibited urotensin -II induced hypertrophy in cultured H9c2 cardiomyocytes (Herold et al. 2003; Johns et al. 2004). The treatment of a non-peptide UTR antagonist, SB-611812, improved cardiac function and reduced cardiac remodelling after congenital heart failure (Watanabe et al. 2005; Bousette et al. 2006). Another non-peptide UTR antagonist, Palosuran inhibited urotensin-II-induced inflammation through MAPK phosphorylation in CHO cell transfected with hUT, suggesting an anti-inflammatory role (Clozel et al. 2004). Palosuran has the ability to improve renal dysfunction and injury-induced ischemia, revealing its role in treating diseases in humans (Clozel et al. 2004). A non-peptide UTR antagonist, Piperazino-isoindolinone strongly and specifically bind to human UTR and suggested that this may provide interesting research opportunity to treat diseases caused by the altered expression of urotensin-II (Lawson et al. 2009).

Although urotensin-II mediated UTR activation plays a role in many physiological and pathophysiological conditions, the information pertaining to urotensin-II mediated UTR signaling network is dispersed throughout the literature. In this study, we compiled the research articles containing molecular information about urotensin-II mediated signaling mechanisms and built a detailed pathway map of urotensin-II mediated UTR signaling. We developed a resource of signaling events mediated by UTR similar to the previously published pathways related to cardiac vascular diseases, including endothelin, apelin and elabela (Dagamajalu et al. 2021, 2022a, b). The urotensin-II mediated UTR signaling pathway described in this study comprises 129 proteins undergoing six different molecular reactions stimulated by urotensin-II. These reactions were gathered together through a manual annotation of the scientific literature and depicted as a single pathway map. The urotensin-II mediated UTR signaling pathway map is available through the WikiPathways Database (https://www.wikipathways.org/index.php/Pathway:WP5158).

Methodology

We carried out an extensive literature search in PubMed for the downstream effects of urotensin-II/UTR. The scientific articles were fetched from PubMed using the following search terms (“UTS2” OR “PRO1068” OR “U-II” OR “UCN2” OR “UII” OR “urotensin 2” OR “UTS2R” OR “GPR14” OR “UR-2-R” OR “UTR2” OR “urotensin 2 receptor” OR “urotensin-2 receptor” OR “UR-II-R” OR “UT receptor”) AND (“pathway” OR “signalling” OR “signalling”). Except reviews and short communications, only original research studies were considered for the annotation. The abstract of each of the research articles was manually screened for urotensin-II/UTR mediated signaling reactions that categorized into the following five different groups: (1) molecular association (protein–protein interactions), (2) post-translational modification (PTM), (3) translocation/transport of proteins between subcellular compartments, (4) activation/inhibition, and finally, (5) gene regulation at the mRNA and protein level (both up-and down-regulation). We manually curated the signaling reactions based on the previously published NetPath annotation criteria (Kandasamy et al. 2010). Each signaling event described in urotensin-II mediated UTR pathway was hyperlinked to the PubMed entry of the corresponding articles.

An instructive pathway map was generated using PathVisio, a pathway drawing tool (van Iersel et al. 2008). The map provides a pictorial summary of all the reactions annotated, including downstream effectors at both the transcriptional and translational levels. Annotated pathway reactions were exported to the WiKiPathways with the ID: WP5158.

Results and discussion

The PubMed search retrieved 68 articles, which were relevant to urotensin-II mediated UTR signaling molecular events were used for annotation. The manual annotation yielded a total of 15 molecular associations, 31 post-translational modifications (PTMs), 33 activation/inhibition reactions, 11 translocation events, 60 protein regulation and 40 gene regulation events were induced by rotensin-II mediated UTR signaling. Of the 40 gene regulation events, 38 genes were found to be up-regulated and 2 genes were downregulated. While in the case of 60 induced protein expression events, 8 proteins were downregulated and 52 were up-regulated. These reactions were incorporated into a corresponding map of the urotensin-II mediated UTR signaling pathway (Fig. 1). The pathway datasheet explains the molecular events involved in urotensin-II ligand-mediated UTR signaling (Supplementary data S1). The consolidated pathway map of the UTR-mediated signaling pathway was submitted to the WikiPathways.

Fig. 1.

Fig. 1

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Schematic representation of urotensin-II-mediated signaling pathway. Schematic representation of urotensin-II mediated signaling reactions. The signaling pathway map represents molecules involved in ligand-receptor interactions and UTR activated downstream molecular events including molecular association, enzyme catalysis, translocation, and gene regulation events. In addition, information regarding the post-translational modification site and the residue is also shown in the pathway

Summary of the urotensin pathway map

Urotensin-II binds to a G protein-coupled receptor called GPR14 or UTR signals through intracellular biochemical cascades, inducing many physiological and pathophysiological events (Carlson and Childress 1975; Saetrum Opgaard et al. 2000; Brule et al. 2014; Zhang et al. 2015). The expression of urotensin-II has been elevated in cardiovascular diseases associated with cardiovascular remodeling, including atherosclerosis, fibrosis, systemic, pulmonary hypertension, and congestive heart failure (Djordjevic and Gorlach 2007). Urotensin-II induces ROS production via NADPH oxidases, leading to JNK pathways and insulin resistance in HepG2 cells (Li et al. 2016). The NOX4-mediated ROS production by urotensin-II phosphorylates JNK and 14-3-3 and activates FoxO3a. This pathway leads to the up-regulation of MMP2 and plays an important role in vascular remodeling by enhanced proliferation of smooth muscle cells (SMCs) in human pulmonary artery SMCs (Diebold et al. 2011). Chen et al., 2014, have reported that the protective role of urotensin-II against doxorubicin-induced apoptosis in human umbilical vein endothelial cells (HUVEC) via the upregulation of ATF3, downregulation of p53, via AKT1 (Ser 473) and MAPK1/3 Tyr 204/Thr 202) phosphorylation (Chen et al. 2014; Staff 2015). Li et al. reported that urotensin-II induces collagen synthesis and mediates the proliferation of pulmonary artery smooth muscle cells (PASMCs) by activating MAPK1/3, MAP2K7 and EGR-1 expression. This represents the role of urotensin-II in the pathophysiology of pulmonary hypertension (Li et al. 2015).

Cardiac hypertrophy

Urotensin-II is a small peptide extensively expressed in the cardiovascular system, which induces cardiomyocyte hypertrophy. In cultured rat cardiac myocytes, urotensin-II strongly up-regulated NPPA and NPPB through activation of MAPK1/3 signaling mechanism (Zou et al. 2001). The urotensin-II stimulates UTR and induces the activation of MAPK1/3 and MAPK14 via the tyrosine phosphorylation (Y1068) of epidermal growth factor receptors (EGFRs), which lead to cardiomyocyte hypertrophy in rat cardiomyocytes (Liu et al. 2009). Another study in rat cardiomyocytes reported that urotensin-II-induced cardiac hypertrophy by ROS generation via NADPH oxidase, which inhibits SHP-2 and facilitates the phosphorylation of EGFR (Tyr 1068) and MAPKs (Liu et al. 2009; Shi et al. 2016). Park et al. reported that the urotensin-II induces the activation and phosphorylation of NF-kB and HDAC5 (Ser 493) via MAPK1/3 and GSK3a and GSK3β independent pathways, leading to hypertrophy in rat H9c2 cells (Park et al. 2016). CaMKII/PLN/SERCA2a, signaling pathway contributed in urotensin-II induced cardiomyocyte hypertrophy reported in rat cardiac myocytes. In neonatal rat cardiomyocytes, urotensin-II induces cardiomyocyte hypertrophy via Akt/GSK-3β pathway by inhibiting PTEN and ERK pathways (Chao et al. 2014). The urotensin-II-induced adult cardiomyocyte hypertrophy reported in adult cardiomyocytes involves the Akt/GSK-3β signaling pathways and is accompanied by the stabilization of the beta-catenin (Gruson et al. 2010). Tzanidis et al. reported that urotensin-II stimulation in neonatal cardiac fibroblasts increased mRNA level transcripts for FN1, COL1A1, and COL3A1 through Galpha(q)- and Ras-dependent pathways are implicated in myocardial fibrosis and pathological cardiac hypertrophy (Tzanidis et al. 2003).

Inflammation

Urotensin-II expression is closely related to tissue damage caused by immune inflammation by promoting the production and release of proinflammatory cytokines. A study in primary Kupffer cells (KCs) shows that urotensin-II/UTR system induces the upregulation of TNF and IL1β and RELA via the MAPK pathway, which promotes inflammatory liver injury (Liu et al. 2015a). Another study reported that urotensin-II/UTR system plays a key role in the pathogenesis of acute liver failure (ALF) by promoting the release of inflammatory cytokines including TNF-a, IL-1β and IFN-γ through NF-κB mediated signaling pathway (Liang et al. 2014). In the study in ALF mice, urotensin-II/UTR system induces the secretion of IFN-β via IRF3 signaling and IL-6 and IFN-γ via MAPK/NF-κB signaling, which mediates inflammatory liver injury in ALF (Liu et al. 2016). Furthermore, urotensin-II induces Rho-A activation and cellular adhesion molecules such as ICAM1, VCAM1 and tissue factors expression via NF-κB signaling pathway, which response to athero-thrombosis in human coronary artery endothelial cells (Cirillo et al. 2008). Urotensin-II/UTR system also induces neuropathic pain, a devastating disease related to excessive inflammation in the spinal dorsal horn by protein upregulation of IL1β, IL-6, TNF-alpha via JNK/NF-kB signaling pathway (Liao et al. 2017).

Atherosclerosis and fibrosis

Zhao et al (2020) reported that foam cell formation in the arterial wall plays a key role in the development of atherosclerosis. The increased levels of urotensin-II expression are associated with atherosclerotic myocardial injury in rats by activating MAPKs signaling pathways such as MAPK1/3, MAPK14 and MAPK8/9 (Zhao et al. 2020). Urotensin-II activates MAPK1/3 signaling through NF-kB mediated pathway and suppresses the ATP-binding cassette transporter A1 (ABCA1) protein and gene expression in THP-1 macrophages. This study suggests that urotensin-II may reduce cholesterol efflux to promote macrophage foam cell formation to promote the progression of atherosclerosis (Wang et al. 2014). In THP-1 monocytes, MAPK1/3 induces activation of the IL-1ß pathway, which in turn downregulates expression of ABCA1, which speeds up foam cell formation leads to atherosclerotic lesions (Kim et al. 2017a). Kim et al. (2017a, b) reported that UTR activation by urotensin-II induces ROS generation and MAPK1/3 activation in vascular smooth muscle cells, which are associated with atherosclerosis and restenosis (Kim et al. 2017b). urotensin-II stimulates UTR and induces the overexpression of 5-lipoxygenase (ALOX5) protein and release of leukotriene B4 (LTB4) via ROS/AKT pathways, which initiates macrophage activation and promotes atherosclerosis in mouse RAW264.7 macrophages (Lu et al. 2019). The urotensin-II /UTR system phosphorylates JAK2 (Tyr 1007)/STAT3 (Tyr 705) molecules and activation of MAPKs signaling pathways, p-MAPK1/3 translocated into the nucleus, which induces the atherosclerosis-related kidney injury by overexpression of Agtr1α, Nox4, Cyba and Ncf1 genes in an atherosclerotic rat model (Wang et al. 2020).

Urotensin-II is mainly involved in the occurrence and progression of cardiac fibrosis by stimulating collagen synthesis (Tran et al. 2010; Liu et al. 2015b). Similarly, urotensin-II promotes collagen synthesis via MAPK1/3-dependent and independent TGF-β1 pathway in neonatal cardiac fibroblasts (Zou et al. 2001). Urotensin-II induces enhanced expression of Collagen I (COL1A1) and Collagen III (COL3A1) via the cAMP-PKA signaling pathway, which promotes myocardial fibrosis in the pressure-overload rat model (Liu et al. 2015b). In vascular smooth muscle cells (VSMCs), urotensin-II/UTR stimulates TGF-β1/Smad 2/3 pathway activation, which induces the upregulation of COL1A1 production and vascular fibrosis in VSMCs (Zhao et al. 2013).

Cancer

Recent studies have demonstrated that urotensin-II/UTR system is involved in the pathogenesis of many malignant tumors (Federico et al. 2017). The study by Yu et al. has reported that urotensin-II/UTR system enhanced the proliferation of human hepatoma cells via PKC, MAPK1/3 and MAPK14 pathways (Yu et al. 2014). The study by Yu et al. (2015) has reported that the urotensin-II/UTR system increases ROS production through NADPH oxidase, translocation of NCF1 and NCF2 from the cytoplasm to membrane and upregulated the expression of CYBB, NCF4, CCNE1 and CDK2 via PI3K/AKT and MAPK1/3 pathways. This accelerates the G1/S transition, leading to cell proliferation in human hepatocellular carcinoma (Yu et al. 2015). In another study in hepatocellular carcinoma, urotensin-II/UTR induced ROS generation through NADPH oxidase, activated MAPK8/9 pathways, and increased expression of MMP2, CYBA, NCF4, NCF1, NCF2, CYBB, PCNA, FSCN1, which are involved in migration and invasion (Li et al. 2017). Urotensin-II induced proliferation and migration via MAPK and FAK pathways in TSC2-deficient human angiomyolipoma cells excised from a patient with Lymphangioleiomyomatosis (LAM) (Goldberg et al. 2016). In lung adenocarcinoma tumor-bearing nude mice model, urotensin-II forms an inflammatory microenvironment by increased expression of IL-6, TNF-α and MMP-9 via NF-kB pathway leading to enhanced proliferation (Zhou et al. 2012).

Kidney diseases

The increased levels of urotensin-II and UTR are already reported in kidney specimens of diabetic nephropathy (Langham et al. 2004). In diabetic conditions, UTR activation by urotensin-II induces the elevation of intracellular calcium via the activation of TRPC4 channels and successive activation of CaMKII/CREB signaling pathways by phosphorylation, which contribute to proliferation and extracellular matrix protein accumulation in glomerular mesangial cells (GMCs), which cause diabetic nephropathy (Soni and Adebiyi 2017). Chen et al. (2017) has reported that urotensin-II increased the TGF-β1 expression and secretion during aristolochic acid-induced nephropathy (Chen et al. 2017). In rat proximal tubular epithelial cells, the advanced glycation end product (AGE) stimulates the increased expression of urotensin-II involved in the upregulation of TGFB1, FN1 and COL4A1, which promotes tubulointerstitial nephropathy in diabetes (Tian et al. 2016). In the diabetic nephropathy model, urotensin-II induces the upregulation of AIFM2, HSPA5, DDIT3, ESPL1, ACTA2, which induces ER stress and EMT in human renal proximal tubular epithelial cell line HK-2 cells (Pang et al. 2016).

Conclusions

Urotensin-II mediated signaling is involved in several pathophysiological roles in pro-fibrotic activities, neurosecretory effects, atherosclerosis, kidney disease, cardiomyocyte hypertrophy, carcinogenic effects and inflammatory diseases. Therefore, the availability of comprehensive urotensin-II signaling in the public resource will help understand the regulation and function of this pathway in normal and pathological conditions. Furthermore, we anticipate that this resource about urotensin-II and its signaling system will help and provide more insights to understand the role of urotensin-II signaling in disease progression and proteins with undefined functionality in the pathway, which provides a new hypothesis that will assist them to design better near future therapeutics.

Supplementary Information

Below is the link to the electronic supplementary material.

12079_2022_672_MOESM1_ESM.xlsm (81.4KB, xlsm)

Supplementary file1 (XLSM 81 kb) Supplementary Data S1: A compendium of urotensin-II mediated signaling events

Acknowledgements

We thank Karnataka Biotechnology and Information Technology Services (KBITS), Government of Karnataka, for the support to the Center for Systems Biology and Molecular Medicine at Yenepoya (Deemed to be University) under the Biotechnology Skill Enhancement Programme in Multiomics Technology (BiSEP GO ITD 02 MDA 2017). RDAB is a recipient of the Senior Research Fellowship from the Indian Council of Medical Research (ICMR), Government of India. AP is a recipient of the Junior Research Fellowship from the University Grants Commission (UGC), Government of India.

Declarations

Conflict of interest

The authors report no conflicts of interest.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Contributor Information

D. A. B. Rex, Email: rexprem@yenepoya.edu.in

G. P. Suchitha, Email: suchithagp1@gmail.com

Akhina Palollathil, Email: akhinap@yenepoya.edu.in.

Anagha Kanichery, Email: anaghak0092@gmail.com.

T. S. Keshava Prasad, Email: keshav@yenepoya.edu.in.

Shobha Dagamajalu, Email: shobha_d@yenepoya.edu.in.

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Supplementary file1 (XLSM 81 kb) Supplementary Data S1: A compendium of urotensin-II mediated signaling events

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