DOCSLIB.ORG

Aberrant RSPO3-LGR4 Signaling in Keap1-Deficient Lung

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Aberrant RSPO3-LGR4 Signaling in Keap1-Deficient Lung Oncogene (2015) 34, 4692–4701 © 2015 Macmillan Publishers Limited All rights reserved 0950-9232/15 www.nature.com/onc ORIGINAL ARTICLE Aberrant RSPO3-LGR4 signaling in Keap1-deficient lung adenocarcinomas promotes tumor aggressiveness X Gong1,2,JYi1,2, KS Carmon1,2, CA Crumbley1,2, W Xiong1,2, A Thomas1,2,XFan1,2, S Guo3,ZAn1,2, JT Chang1,4,5,6 and QJ Liu1,2 The four R-spondins (RSPO1–4) and their three related receptors LGR4, 5 and 6 (LGR4–6) have emerged as a major ligand-receptor system with critical roles in development and stem cell survival through modulation of Wnt signaling. Recurrent, gain-of-expression gene fusions of RSPO2 (to EIF3E) and RSPO3 (to PTPRK) occur in a subset of human colorectal cancer. However, the exact roles and mechanisms of the RSPO-LGR system in oncogenesis remain largely unknown. We found that RSPO3 is aberrantly expressed at high levels in approximately half of Keap1-mutated lung adenocarcinomas (ADs). This high RSPO3 expression is driven by a combination of demethylation of its own promoter region and deficiency in Keap1 instead of gene fusion as in colon cancer. Patients with RSPO3-high tumors (~9%, 36/412) displayed much poorer survival than the rest of the cohort (median survival of 28 vs 163 months, log-rank test Po0.0001). Knockdown (KD) of RSPO3, LGR4 or their signaling mediator IQGAP1 in lung cancer cell lines with Keap1 deficiency and high RSPO3-LGR4 expression led to reduction in cell proliferation and migration in vitro, and KD of LGR4 or IQGAP1 resulted in decrease in tumor growth and metastasis in vivo. These findings suggest that aberrant RSPO3-LGR4 signaling potentially acts as a driving mechanism in the aggressiveness of Keap1-deficient lung ADs. Oncogene (2015) 34, 4692–4701; doi:10.1038/onc.2014.417; published online 22 December 2014 INTRODUCTION the formation of a supercomplex between RSPO-LGR4 and Wnt R-spondins are a group of four highly related secreted proteins receptors. In this configuration, IQGAP1 brings in MEK1/2 to (RSPO1–4) with critical roles in embryonic development phosphorylate LRP5/6 for the β-catenin-dependent pathway and and organogenesis as well as in the self-renewal and survival of N-WASP/mDia1 to coordinate actin dynamics for the β-catenin- adult stem cells.1 In particular, loss of RSPO2 led to hypoplasia independent pathway.11 and reduced branching of the lung during mouse Dysregulation of Wnt signaling occurs in nearly every major development.2,3 Work from us and others demonstrated that type of solid tumors. Gain-of-expression gene fusions of RSPO2 (to RSPOs activate three related receptors LGR4–6 (leucine-rich EIF3E) and RSPO3 (to PTPRK) were identified in 10% (7/68) of repeat-containing, G protein-coupled receptor 4, 5 and 6) to human colon cancer.14 The fusions were inferred to have a driving 4–6 potentiate Wnt signaling. LGR4–6 contain a large extracellular role in the carcinogenesis of the affected tumors because of their domain with 17 leucine-rich repeats and a seven transmembrane recurrent occurrence and exclusivity with Apc/β-catenin mutat- domain homologous to members of the rhodopsin family of G 14 7–9 ions. In MMTV-induced mouse models of breast and colon cancer, protein-coupled receptors. LGR4-bound RSPOs directly interact RSPO2 and RSPO3 were two of the most frequent viral integration with two membrane-bound E3 ligases (RNF43 and ZNRF3), which 10 sites, and ectopic expression of RSPO2/3 in mouse mammary otherwise ubiquitinate Fzd receptors for degradation. Formation epithelial cells increased tumor formation and metastasis.15–17 of the LGR4-RSPO-RNF43/ZNRF3 ternary complex induces Furthermore, knockout of LGR4 in mice led to profound hypoplasia the clearance of the E3 ligases, leading to reduced ubiquitination and impaired tubulogenesis in multiple organs during develop- and eventually elevated levels of Wnt receptors on the cell 18–20 surface and increased Wnt signaling.10 Just recently, we ment, suggesting a critical role of LGR4 in the regulation of cell identified IQGAP1 as an LGR4-binding protein and showed that proliferation and migration. Intriguingly, LGR4 was found to be it plays an essential role in RSPO-LGR4-induced potentiation highly upregulated in both adenocarcinomas (AD) and squamous of Wnt signaling.11 IQGAP1 is an intracellular scaffold protein cell carcinomas of non-small-cell lung cancer (NSCLC) despite low 21 that binds to and modulates the activities of a plethora of expression in normal adult lung. We found that RSPO3 was highly signaling molecules to regulate cell adhesion and migration.12,13 expressed in a subset of ADs. Here we show that the aberrant RSPO3 We found that RSPO-LGR4 not only induces the clearance of expression in lung ADs was not driven by PTPRK fusion as in colon RNF43/ZNRF3 but also increases the affinity of IQGAP1 for cancer, and that RSPO3-LGR4 signaling plays a major role in the dishelvelled (DVL) bound to the Wnt signalosome. This leads to aggressiveness of RSPO3-high tumors. 1Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USA; 2Texas Therapeutics Institute, University of Texas Health Science Center at Houston, Houston, TX, USA; 3Divison of Oncology, University of Texas Health Science Center at Houston, Houston, TX, USA; 4Center for Clinical and Translational Sciences, University of Texas Health Science Center at Houston, Houston, TX, USA; 5Department of Integrative Biology and Pharmacology, University of Texas Health Science Center at Houston, Houston, TX, USA and 6School of Biomedical Informatics, University of Texas Health Science Center at Houston, Houston, TX, USA. Correspondence: Dr QJ Liu, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, 1825 Pressler Street, Suite 530A, Houston, TX 77030, USA. E-mail: [email protected] Received 29 April 2014; revised 31 October 2014; accepted 3 November 2014; published online 22 December 2014 RSPO3-LGR4 signaling in lung cancer X Gong et al 4693 RESULTS (CCLE) project24 (Supplementary Figure 1b). Using our recently 25 RSPO3 is aberrantly expressed in a subset of lung ADs and its high characterized LGR4 monoclonal antibody 7E7, we analyzed the expression is associated with poor survival expression of LGR4 in normal and AD samples of the lung on We mined the RNA-Seq data of LGR4–6, RSPO1–4, and other tissue microarrays. LGR4 staining was detected in all AD samples genes encoding Wnt ligands, receptors and modulators in TCGA’s with two representatives shown (Figures 1a and b). No expression of LGR4 was observed in respiratory epithelial or alveolar cells in provisional Lung AD cohort (LUAD, 230 samples) as provided by 22 normal adult lung (Figure 1c), consistent with previous report of the cBio Portal website. Based on the RSEM values (RNA-Seq by 21 23 LGR4 upregulation in lung ADs. Expectation-Maximization) of the transcripts, LGR4 was Of the potential RSPO ligands, RSPO3 was expressed at expressed at moderate to high levels in ~ 90% of the cases, significant levels with a highly skewed pattern, that is, high levels whereas LGR5 and LGR6 were expressed at much lower levels in in ~ 10% (25/230) of the tumors with no to little expression in the nearly all samples (Supplementary Figure 1a). A similar pattern of rest of the cases (Figure 1d). The other three RSPOs were expression for LGR4–6 was also found in the 186 lung cancer cell expressed at much lower, insignificant levels with two exceptions line collection characterized by the Cancer Cell Line Encyclopedia in RSPO2 (Supplementary Figure 1c). Comparison of the RSPO3 Tumor-1 Tumor-2 Normal lung 15 15 RSPO3-CCLE 100 RSPO3-high (One-way ANOVA p = 0.2) RSPO3-TCGA RSPO3-norm 12 LGR4-TCGA 80 (Logrank test p = 0.003) 10 60 9 40 % Surviving 5 Exp (log2[signal]) 6 20 N = 173 RSPO3 Exp (log2[RSEM]) N = 22 3 0 0 0 50 100 150 200 250 0 50 100 150 200 250 Sample # Months of Survival Stage-I Stge-II Stage-III Stage-IV 120 120 RSPO3-high RSPO3-high 100 RSPO3-high 100 100 RSPO3-norm RSPO3-norm RSPO3-norm 80 (Logrank test p < 0.0001) 80 (Logrank test p = 0.01) 80 (Logrank test p = 0.01) 60 60 60 40 % Surviving % Surviving 40 % Surviving 40 N = 376 N = 44 20 20 N = 127 20 N = 36 N = 14 N = 5 0 0 0 0 50 100 150 200 250 0 50 100 150 200 250 0 50 100 150 200 250 Months of Survival Months of Survival Months of Survival Figure 1. High level of RSPO3 expression is correlated with poor survival in lung AD. (a-c) Immunohistochemistry staining of LGR4 in two lung AD samples (a, b) and normal adult lung (c). (d) Expression distribution of RSPO3 and LGR4 in 230 lung AD samples of TCGA’s LUAD cohort and RSPO3 in CCLE’s 186 lung cancer cell lines. Values in log2 of expression signal (RSEM for TCGA samples and RMA for CCLE samples) were sorted from low to high for each gene and plotted. (e) Kaplan–Meier survival plot of patients with tumors of high RSPO3 expression vs the rest of the cohort in the TCGA LUAD set. RSPO3-high was defined as samples with log2[RSEM] values at 1 s.d. above the population mean (Z 4= +1) while the remaining samples were defined as RSPO-norm. (f) Scatter plot of RSPO3 expression among different tumor stages in TCGA’s LUAD cohort. (g) Kaplan–Meier survival plot of patients with RSPO3-high vs RSPO3-norm tumors in stages I-II of TCGA’s LUAD cohort.
Load more

Recommended publications
  • Edinburgh Research Explorer
    Edinburgh Research Explorer International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein-coupled receptor list Citation for published version: Davenport, AP, Alexander, SPH, Sharman, JL, Pawson, AJ, Benson, HE, Monaghan, AE, Liew, WC, Mpamhanga, CP, Bonner, TI, Neubig, RR, Pin, JP, Spedding, M & Harmar, AJ 2013, 'International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein-coupled receptor list: recommendations for new pairings with cognate ligands', Pharmacological reviews, vol. 65, no. 3, pp. 967-86. https://doi.org/10.1124/pr.112.007179 Digital Object Identifier (DOI): 10.1124/pr.112.007179 Link: Link to publication record in Edinburgh Research Explorer Document Version: Publisher's PDF, also known as Version of record Published In: Pharmacological reviews Publisher Rights Statement: U.S. Government work not protected by U.S. copyright General rights Copyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. Take down policy The University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorer content complies with UK legislation. If you believe that the public display of this file breaches copyright please contact [email protected] providing details, and we will remove access to the work immediately and investigate your claim. Download date: 02. Oct. 2021 1521-0081/65/3/967–986$25.00 http://dx.doi.org/10.1124/pr.112.007179 PHARMACOLOGICAL REVIEWS Pharmacol Rev 65:967–986, July 2013 U.S.
    [Show full text]
  • And LPA 3-Mediated Recruitment of Leukocytes In
    Supplemental Material can be found at: http://www.jlr.org/content/suppl/2011/04/26/jlr.M008045.DC1 .html ␣ TNF- promotes LPA1 - and LPA3 -mediated recruitment of leukocytes in vivo through CXCR2 ligand chemokines Chenqi Zhao , 1, * Anne Sardella , 1, * Jerold Chun , † Patrice E. Poubelle , * Maria J. Fernandes , * and Sylvain G. Bourgoin 2, * Rheumatology and Immunology Research Center,* CHUQ-CHUL Research Center and Faculty of † Medicine, Laval University , Québec City, Québec, Canada ; and Department of Molecular Biology, The Scripps Research Institute , La Jolla, CA Downloaded from Abstract Lysophosphatidic acid (LPA) is a bioactive lyso- Lysophosphatidic acid (LPA) is a simple lipid with a sin- phospholipid present in low concentrations in serum and gle fatty acyl chain, a glycerol backbone, and a free phos- biological fl uids but in high concentrations at sites of infl am- phate group ( 1 ). Despite the simplicity of its structure, www.jlr.org mation. LPA evokes a variety of cellular responses via binding LPA has been implicated in various physiological (cell to and activation of its specifi c G protein-coupled receptors growth, differentiation, migration, and survival) and path- (GPCR), namely LPA1-6 . Even though LPA is a chemoattrac- tant for infl ammatory cells in vitro, such a role for LPA in ological (angiogenesis, cancer, and autoimmunity) situa- at Kresge Library, The Scripps Research Institute, on November 14, 2012 vivo remains largely unexplored. In the present study, we tions. LPA is present in micromolar concentrations in used the murine air pouch model to study LPA-mediated serum and biological fl uids and in higher concentrations leukocyte recruitment in vivo using selective LPA receptor at sites of infl ammation and tumor ( 2 ).
    [Show full text]
  • G Protein-Coupled Receptors
    S.P.H. Alexander et al. The Concise Guide to PHARMACOLOGY 2015/16: G protein-coupled receptors. British Journal of Pharmacology (2015) 172, 5744–5869 THE CONCISE GUIDE TO PHARMACOLOGY 2015/16: G protein-coupled receptors Stephen PH Alexander1, Anthony P Davenport2, Eamonn Kelly3, Neil Marrion3, John A Peters4, Helen E Benson5, Elena Faccenda5, Adam J Pawson5, Joanna L Sharman5, Christopher Southan5, Jamie A Davies5 and CGTP Collaborators 1School of Biomedical Sciences, University of Nottingham Medical School, Nottingham, NG7 2UH, UK, 2Clinical Pharmacology Unit, University of Cambridge, Cambridge, CB2 0QQ, UK, 3School of Physiology and Pharmacology, University of Bristol, Bristol, BS8 1TD, UK, 4Neuroscience Division, Medical Education Institute, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY, UK, 5Centre for Integrative Physiology, University of Edinburgh, Edinburgh, EH8 9XD, UK Abstract The Concise Guide to PHARMACOLOGY 2015/16 provides concise overviews of the key properties of over 1750 human drug targets with their pharmacology, plus links to an open access knowledgebase of drug targets and their ligands (www.guidetopharmacology.org), which provides more detailed views of target and ligand properties. The full contents can be found at http://onlinelibrary.wiley.com/doi/ 10.1111/bph.13348/full. G protein-coupled receptors are one of the eight major pharmacological targets into which the Guide is divided, with the others being: ligand-gated ion channels, voltage-gated ion channels, other ion channels, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading.
    [Show full text]
  • G Protein-Coupled Receptors in Cancer
    International Journal of Molecular Sciences Review G Protein-Coupled Receptors in Cancer Rachel Bar-Shavit 1,*, Myriam Maoz 1, Arun Kancharla 1, Jeetendra Kumar Nag 1, Daniel Agranovich 1, Sorina Grisaru-Granovsky 2 and Beatrice Uziely 1 1 Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Jerusalem 91120, Israel; [email protected] (M.M.); [email protected] (A.K.); [email protected] (J.K.N.); [email protected] (D.A.); [email protected] (B.U.) 2 Department of Obstetrics and Gynecology, Shaare-Zedek Medical Center, Jerusalem 91031, Israel; [email protected] * Correspondence: [email protected]; Tel.: +972-2-677-7563; Fax: +972-2-642-7485 Academic Editors: Kathleen Van Craenenbroeck and Gregor Drummen Received: 6 June 2016; Accepted: 8 August 2016; Published: 12 August 2016 Abstract: Despite the fact that G protein-coupled receptors (GPCRs) are the largest signal-conveying receptor family and mediate many physiological processes, their role in tumor biology is underappreciated. Numerous lines of evidence now associate GPCRs and their downstream signaling targets in cancer growth and development. Indeed, GPCRs control many features of tumorigenesis, including immune cell-mediated functions, proliferation, invasion and survival at the secondary site. Technological advances have further substantiated GPCR modifications in human tumors. Among these are point mutations, gene overexpression, GPCR silencing by promoter methylation and the number of gene copies. At this point, it is imperative to elucidate specific signaling pathways of “cancer driver” GPCRs. Emerging data on GPCR biology point to functional selectivity and “biased agonism”; hence, there is a diminishing enthusiasm for the concept of “one drug per GPCR target” and increasing interest in the identification of several drug options.
    [Show full text]
  • Multi-Functionality of Proteins Involved in GPCR and G Protein Signaling: Making Sense of Structure–Function Continuum with In
    Cellular and Molecular Life Sciences (2019) 76:4461–4492 https://doi.org/10.1007/s00018-019-03276-1 Cellular andMolecular Life Sciences REVIEW Multi‑functionality of proteins involved in GPCR and G protein signaling: making sense of structure–function continuum with intrinsic disorder‑based proteoforms Alexander V. Fonin1 · April L. Darling2 · Irina M. Kuznetsova1 · Konstantin K. Turoverov1,3 · Vladimir N. Uversky2,4 Received: 5 August 2019 / Revised: 5 August 2019 / Accepted: 12 August 2019 / Published online: 19 August 2019 © Springer Nature Switzerland AG 2019 Abstract GPCR–G protein signaling system recognizes a multitude of extracellular ligands and triggers a variety of intracellular signal- ing cascades in response. In humans, this system includes more than 800 various GPCRs and a large set of heterotrimeric G proteins. Complexity of this system goes far beyond a multitude of pair-wise ligand–GPCR and GPCR–G protein interactions. In fact, one GPCR can recognize more than one extracellular signal and interact with more than one G protein. Furthermore, one ligand can activate more than one GPCR, and multiple GPCRs can couple to the same G protein. This defnes an intricate multifunctionality of this important signaling system. Here, we show that the multifunctionality of GPCR–G protein system represents an illustrative example of the protein structure–function continuum, where structures of the involved proteins represent a complex mosaic of diferently folded regions (foldons, non-foldons, unfoldons, semi-foldons, and inducible foldons). The functionality of resulting highly dynamic conformational ensembles is fne-tuned by various post-translational modifcations and alternative splicing, and such ensembles can undergo dramatic changes at interaction with their specifc partners.
    [Show full text]
  • RSPO–LGR4 Functions Via IQGAP1 to Potentiate Wnt Signaling
    RSPO–LGR4 functions via IQGAP1 to potentiate PNAS PLUS Wnt signaling Kendra S. Carmon, Xing Gong, Jing Yi, Anthony Thomas, and Qingyun Liu1 Brown Foundation Institute of Molecular Medicine and Texas Therapeutics Institute, University of Texas Health Science Center at Houston, Houston, TX 77030 Edited by Roeland Nusse, Stanford University School of Medicine, Stanford, CA, and approved February 20, 2014 (received for review December 11, 2013) R-spondins (RSPOs) and their receptor leucine-rich repeat-contain- typical of the rhodopsin family of G-protein–coupled receptors (26). ing G-protein coupled receptor 4 (LGR4) play pleiotropic roles in Stimulation of LGRs with RSPO1–4 greatly potentiates the activity normal and cancer development as well as the survival of adult of Wnt ligands in both the canonical (β-catenin–dependent) and stem cells through potentiation of Wnt signaling. Current evidence noncanonical (β-catenin–independent; planar cell polarity) path- indicates that RSPO–LGR4 functions to elevate levels of Wnt recep- ways (3–5). This function of the LGRs is independent of hetero- tors through direct inhibition of two membrane-bound E3 ligases trimeric G proteins and β-arrestin (3, 4). Instead, it was shown that (RNF43 and ZNRF3), which otherwise ubiquitinate Wnt receptors the RSPOs function as an extracellular bridge to bring LGR4 and for degradation. Whether RSPO–LGR4 is coupled to intracellular E3 ligases RNF43/ZNRF3 together to form a ternary complex signaling proteins to regulate Wnt pathways remains unknown. (LGR4–RSPO–RNF43/ZNRF3), which induces clearance of the We identified the intracellular scaffold protein IQ motif containing E3 ligases (27).
    [Show full text]
  • 2-Carba Cyclic Phosphatidic Acid Suppresses Inflammation Via
    www.nature.com/scientificreports OPEN 2-carba cyclic phosphatidic acid suppresses infammation via regulation of microglial polarisation Received: 13 March 2018 Accepted: 14 June 2018 in the stab-wounded mouse Published: xx xx xxxx cerebral cortex Kei Hashimoto1,2,4, Mari Nakashima1,2, Ayana Hamano1,2, Mari Gotoh2,3, Hiroko Ikeshima- Kataoka5,6, Kimiko Murakami-Murofushi3 & Yasunori Miyamoto 1,2 Traumatic brain injury (TBI) is caused by physical damage to the brain and it induces blood-brain barrier (BBB) breakdown and infammation. To diminish the sequelae of TBI, it is important to decrease haemorrhage and alleviate infammation. In this study, we aimed to determine the efects of 2-carba- cyclic phosphatidic acid (2ccPA) on the repair mechanisms after a stab wound injury as a murine TBI model. The administration of 2ccPA suppressed serum immunoglobulin extravasation after the injury. To elucidate the efects of 2ccPA on infammation resulting from TBI, we analysed the mRNA expression of infammatory cytokines. We found that 2ccPA prevents a TBI-induced increase in the mRNA expression of Il-1β, Il-6, Tnf-α and Tgf-β1. In addition, 2ccPA reduces the elevation of Iba1 levels. These data suggest that 2ccPA attenuates the infammation after a stab wound injury via the modulation of pro-infammatory cytokines release from microglial cells. Therefore, we focused on the function of 2ccPA in microglial polarisation towards M1 or M2 phenotypes. The administration of 2ccPA decreased the number of M1 and increased the number of M2 type microglial cells, indicating that 2ccPA modulates the microglial polarisation and shifts them towards M2 phenotype.
    [Show full text]
  • Lysophosphatidic Acid Signaling in Cancer Cells: What Makes LPA So Special?
    cells Review Lysophosphatidic Acid Signaling in Cancer Cells: What Makes LPA So Special? Pravita Balijepalli, Ciera C. Sitton and Kathryn E. Meier * Department of Pharmaceutical Sciences, College of Pharmacy and Pharmaceutical Sciences, Washington State University, Spokane, WA 98686, USA; [email protected] (P.B.); [email protected] (C.C.S.) * Correspondence: [email protected]; Tel.: +1-509-358-7631 Abstract: Lysophosphatidic acid (LPA) refers to a family of simple phospholipids that act as ligands for G protein-coupled receptors. While LPA exerts effects throughout the body in normal physiolog- ical circumstances, its pathological role in cancer is of great interest from a therapeutic viewpoint. The numerous LPA receptors (LPARs) are coupled to a variety of G proteins, and more than one LPAR is typically expressed on any given cell. While the individual receptors signal through conventional GPCR pathways, LPA is particularly efficacious in stimulating cancer cell proliferation and migration. This review addresses the mechanistic aspects underlying these pro-tumorigenic effects. We provide examples of LPA signaling responses in various types of cancers, with an emphasis on those where roles have been identified for specific LPARs. While providing an overview of LPAR signaling, these examples also reveal gaps in our knowledge regarding the mechanisms of LPA action at the receptor level. The current understanding of the LPAR structure and the roles of LPAR interactions with other receptors are discussed. Overall, LPARs provide insight into the potential molecular mechanisms that underlie the ability of individual GPCRs (or combinations of GPCRs) to elicit a unique spectrum of responses from their agonist ligands.
    [Show full text]
  • Repertoires of G Protein-Coupled Receptors for Ciona-Specific Neuropeptides
    Repertoires of G protein-coupled receptors for Ciona-specific neuropeptides Akira Shiraishia, Toshimi Okudaa, Natsuko Miyasakaa, Tomohiro Osugia, Yasushi Okunob, Jun Inouec, and Honoo Satakea,1 aBioorganic Research Institute, Suntory Foundation for Life Sciences, 619-0284 Kyoto, Japan; bDepartment of Biomedical Intelligence, Graduate School of Medicine, Kyoto University, 606-8507 Kyoto, Japan; and cMarine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, 904-0495 Okinawa, Japan Edited by Thomas P. Sakmar, The Rockefeller University, New York, NY, and accepted by Editorial Board Member Jeremy Nathans March 11, 2019 (received for review September 26, 2018) Neuropeptides play pivotal roles in various biological events in the conservesagreaternumberofneuropeptide homologs than proto- nervous, neuroendocrine, and endocrine systems, and are corre- stomes (e.g., Caenorhabditis elegans and Drosophila melanogaster) lated with both physiological functions and unique behavioral and other invertebrate deuterostomes (7–13), confirming the evo- traits of animals. Elucidation of functional interaction between lutionary and phylogenetic relatedness of ascidians to vertebrates. neuropeptides and receptors is a crucial step for the verification of The second group includes Ciona-specific novel neuropeptides, their biological roles and evolutionary processes. However, most namely Ci-NTLPs, Ci-LFs, and Ci-YFV/Ls (SI Appendix,Fig. receptors for novel peptides remain to be identified. Here, we S1 and Table S1), which share neither consensus motifs nor se- show the identification of multiple G protein-coupled receptors quence similarity with any other peptides (8, 9). The presence of (GPCRs) for species-specific neuropeptides of the vertebrate sister both homologous and species-specific neuropeptides highlights this group, Ciona intestinalis Type A, by combining machine learning phylogenetic relative of vertebrates as a prominent model organism and experimental validation.
    [Show full text]
  • Overexpression of G Protein-Coupled Receptor GPR87 Promotes
    Wang et al. Molecular Cancer (2017) 16:61 DOI 10.1186/s12943-017-0627-6 RESEARCH Open Access Overexpression of G protein-coupled receptor GPR87 promotes pancreatic cancer aggressiveness and activates NF-κB signaling pathway Li Wang1†, Wei Zhou2†, Yunfeng Zhong2, Yongbao Huo2, Ping Fan2, Sudong Zhan2, Jun Xiao2, Xin Jin2, Shanmiao Gou2, Tao Yin2, Heshui Wu2 and Tao Liu2,3* Abstract Background: Pancreatic cancer is a highly lethal disease and has the worst prognosis of any major malignancy. G protein-coupled receptor GPR87 is reported to be overexpressed in multiple cancers. The clinical significance and biological role of GPR87 in pancreatic cancer, however, remain to be established. Methods: GPR87 expression in pancreatic cancer cell lines, paired patient tissues were determined using western blotting and Real-time PCR. Ninety-six human pancreatic cancer tissue samples were analyzed by immunochemistry (IHC) to investigate the association between GPR87 expression and the clinicopathological characteristics of pancreatic cancer. Functional assays, such as anchorage-independent growth, chicken chorioallantoic membrane (CAM) assay, transwell matrix penetration assay, and Annexin V-FITCandPIstainingandaxenografttumormodelwereused to determine the oncogenic role of GPR87 in human pancreatic cancer progression. The effect of GPR87 on NF-κB signaling pathway was further investigated using the luciferase reporter assays, and by detection of the NF-κB signaling downstream genes. Results: Herein, we reported that GPR87 was markedly overexpressed in pancreatic cancer cells and clinical tissues. Immunohistochemical analysis showed that the expression of GPR87 significantly correlated with patients’ clinicopathologic features, including clinical stage and tumor-nodule-metastasis (TNM) classification. Pancreatic cancer patients with higher levels of GPR87 expression had shorter overall survival compared to patients with lower GPR87 levels.
    [Show full text]
  • G Protein-Coupled Receptor 87 (GPR87) Promotes Cell Proliferation in Human Bladder Cancer Cells
    Int. J. Mol. Sci. 2015, 16, 24319-24331; doi:10.3390/ijms161024319 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Article G Protein-Coupled Receptor 87 (GPR87) Promotes Cell Proliferation in Human Bladder Cancer Cells Xia Zhang 1,*, Dage Liu 2, Yushi Hayashida 1, Homare Okazoe 1, Takeshi Hashimoto 3, Nobufumi Ueda 1, Mikio Sugimoto 1 and Yoshiyuki Kakehi 1,* 1 Department of Urology, Kagawa University Faculty of Medicine, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0793, Japan; E-Mails: [email protected] (Y.H.); [email protected] (H.O.); [email protected] (N.U.); [email protected] (M.S.) 2 Department of General Thoracic Surgery, Kagawa University Faculty of Medicine, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0793, Japan; E-Mail: [email protected] 3 Department of Cardiovascular Physiology, Kagawa University Faculty of Medicine, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0793, Japan; E-Mail: [email protected] * Authors to whom correspondence should be addressed; E-Mails: [email protected] (X.Z.); [email protected] (Y.K.); Tel.: +81-87-891-2202 (X.Z. & Y.K.); Fax: +81-87-891-2203 (X.Z. & Y.K.). Academic Editor: Kathleen Van Craenenbroeck Received: 3 July 2015 / Accepted: 24 September 2015 / Published: 14 October 2015 Abstract: G protein-coupled receptor 87 (GPR87) is a newly deorphanized member of the cell surface molecule G protein-coupled receptor family. GPR signaling was shown to play a role in promotion of cell growth and survival, metastasis, and drug resistance.
    [Show full text]
  • Introduction
    GPCR Introduction G protein-coupled receptors (GPCRs) are the GPCRs Super-Family largest family of transmembrane receptors and are 375 GPCR Drug Targets, 225 with Known Ligands, 150 Orphan Targets responsible for the transduction of a diverse range of extracellular signals. The range of physiological processes mediated by GPCRs makes them one GRM7 GRM8 GLUTAMATE (15) SECRETIN GRM2 FZD7 TAS1R3 FZD2 of the most important classes of proteins for drug (15) GRM4 GRM3 TAS1R1FZD1 GLP2R GIPR GRM6 GRPC6A FRIZZED/TAS2 discovery. ADHESION GLP1R GCGR GRM5 FZD3 PTHR2 GRM1 (24) LEC1 VIPR2 PTHR1 (24) TAS1R2 FZD6 LEC2 TAS2R13 CELSR2PACAP FZD8 The study of GPCRs may involve ligand binding, G CRHR2 FZD5 TAS2R16 TAS2R14 CALCRL LEC3 VIPR1 CRHR1 CASR GABBR2 FZD10 TAS2R1 TAS2R10 EMR3EMR2 CELSR3 BAI2 CALCR FZD4 TAS2R5 TAS2R3 SCTR FZD9 protein activation, internalization, and downstream ETL BAI3 GPR60 TAS2R9 CELSR1 GHRHR GABBR1 GPR59 TAS2R8 TAS2R4 TAS2R7 EMR1 BAI1 SMOH CXCR3 second messenger events. Each step of the CXCR5 CCR11 CXCR2 CD97 SSTR1 SSTR3 CCR10 CCR6 SSTR5 CXCR1 GPR111 CXCR6 pathway offers an opportunity to exploit the full SSTR2 CCR9 GPR115 SSTR4 CCR7 GPR116 GPR112 GPR8 CCRL2 GPR113 GPR7 CXC3R1CCR8 therapeutic potential of targeting GPCRs. GPR110 CCR4 HE6 NTSR2 CCR1 TM7XN1 GPR114 NMU1R GPR54 GALR1 CCBP2 GHSR GALR2 RDC1 CCR3 GPR97 NPY1R XCR1 PPYR1 NMU2R MTLR MCHR1 GALR3 ADMR NPY2R AGTR1 TACR3 UR2R MCHR2 AGTRL1 AGTR2 CCR5 TAC3RL PrRP γ GPR26 BDKRB2 CCR2 GRM7 GRM8 GLUTAMATE (15) TACR1 TACR2 GRP72 OR1A1 SALPR OLFACTORY GPR15 NPFF1 NPY5R OR1D2 (388) GPTH2
    [Show full text]