Wnt Inhibitor Dickkopf-1 As a Target for Passive Cancer Immunotherapy

Total Page:16

File Type:pdf, Size:1020Kb

Wnt Inhibitor Dickkopf-1 As a Target for Passive Cancer Immunotherapy Published OnlineFirst June 15, 2010; DOI: 10.1158/0008-5472.CAN-09-3879 Published OnlineFirst on June 15, 2010 as 10.1158/0008-5472.CAN-09-3879 Molecular and Cellular Pathobiology Cancer Research Wnt Inhibitor Dickkopf-1 as a Target for Passive Cancer Immunotherapy Nagato Sato1,2, Takumi Yamabuki1, Atsushi Takano1,6, Junkichi Koinuma1, Masato Aragaki1, Ken Masuda1, Nobuhisa Ishikawa1, Nobuoki Kohno3, Hiroyuki Ito4, Masaki Miyamoto2, Haruhiko Nakayama4, Yohei Miyagi5, Eiju Tsuchiya5, Satoshi Kondo2, Yusuke Nakamura1, and Yataro Daigo1,6 Abstract Dickkopf-1 (DKK1) is an inhibitor of Wnt/β-catenin signaling that is overexpressed in most lung and esoph- ageal cancers. Here, we show its utility as a serum biomarker for a wide range of human cancers, and we offer evidence favoring the potential application of anti-DKK1 antibodies for cancer treatment. Using an original ELISA system, high levels of DKK1 protein were found in serologic samples from 906 patients with cancers of the pancreas, stomach, liver, bile duct, breast, and cervix, which also showed elevated expression levels of DKK1. Additionally, anti-DKK1 antibody inhibited the invasive activity and the growth of cancer cells in vitro and suppressed the growth of engrafted tumors in vivo. Tumor tissues treated with anti-DKK1 displayed sig- nificant fibrotic changes and a decrease in viable cancer cells without apparent toxicity in mice. Our findings suggest DKK1 as a serum biomarker for screening against a variety of cancers, and anti-DKK1 antibodies as potential theranostic tools for diagnosis and treatment of cancer. Cancer Res; 70(13); OF1–11. ©2010 AACR. Introduction growth factor (VEGF) used for the treatment of metastatic colorectal and lung cancers in combination with chemother- The concept of specific molecular targeting therapy has been apy (4). The results were promising when compared with applied to the development of innovative cancer-treatment responses of advanced cancers to conventional cytotoxic strategies. At present, two main approaches are available in agents, but the proportion of patients showing good response clinical practice: therapeutic monoclonal antibodies (mAb) is still limited, and in some cases, the medication caused se- and small-molecule agents (1). There is an increasing interest rious adverse effects including life-threatening ones. Hence, in the use of antibody-based immunotherapy for the treat- the development of new therapeutic antibodies targeting ment of malignant diseases, and some dramatic clinical transmembrane and/or secreted proteins, which show a responses have enhanced the activity in this field (1). For cancer-specific expression pattern and have an oncogenic example, rituximab (Rituxan) is the chimeric anti-CD20 anti- function, is eagerly awaited. body that revolutionized lymphoma treatment (2). Trastuzu- To develop new diagnostics and therapeutics improving mab (Herceptin) is the humanized Ab against the human cancer treatment, we have been screening the candidate tar- epidermal growth factor receptor (HER)/ERBB2 for the treat- get molecules by the following strategy: (a) identifying genes ment of patients with metastatic breast cancer in which HER/ upregulated in lung and esophageal cancers by genome-wide ERBB2 gene was highly overexpressed (3). Bevacizumab cDNA microarray system (5–11), (b) verifying the candidate (Avastin) is the humanized Ab against vascular endothelial genes for the absence of or very low level of expression in nor- mal tissues (5), (c) validating biological significance of their overexpression by means of tissue microarray containing Authors' Affiliations: 1Laboratory of Molecular Medicine, Human hundreds of archived lung cancer samples and functional as- Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo, Japan; 2Department of Surgical Oncology, Hokkaido University says for examining their activity in cell growth and/or invasive Graduate School of Medicine, Sapporo, Japan; 3Department of activity (12–33), (d) evaluating them for usefulness as a serum Molecular and Internal Medicine, Graduate School of Biomedical diagnostic/prognostic biomarker for lung cancer by ELISA Sciences, Hiroshima University, Hiroshima, Japan; Divisions of – 4Thoracic Surgery and 5Molecular Pathology and Genetics, Kanagawa (29 33), if they are tumor-specific transmembrane or secretory Cancer Center, Yokohama, Japan; and 6Department of Medical proteins, and (e) screening the epitopes recognized by human Oncology, Shiga University of Medical Science, Otsu, Japan histocompatibility leukocyte HLA-*A0201/*A2402-restricted Note: Supplementary data for this article are available at Cancer Research CTL, if they are categorized into cancer testis antigens and Online (http://cancerres.aacrjournals.org/). are indispensable for cancer cell growth/survival (34–36). Corresponding Author: Yataro Daigo, Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Using this approach, we had identified Dickkopf-1 (DKK1) Japan. Phone: 81-3-5449-5457; Fax: 81-3-5449-5406; E-mail: ydaigo@ as a novel serologic and histochemical biomarker as well as ims.u-tokyo.ac.jp. a possible therapeutic target for lung and esophageal cancers doi: 10.1158/0008-5472.CAN-09-3879 (31). Our data indicated that DKK1 expression was associated ©2010 American Association for Cancer Research. with poor prognosis for patients with non–small cell lung www.aacrjournals.org OF1 Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 2010 American Association for Cancer Research. Published OnlineFirst June 15, 2010; DOI: 10.1158/0008-5472.CAN-09-3879 Sato et al. cancer (NSCLC) or esophageal squamous cell carcinoma tained at 37°C in humidified air with 5% CO2. Surgically (ESCC), and that serum DKK1 levels were significantly higher resected primary cancers from six pancreatic cancer pa- in lung and esophageal cancer patients than in healthy con- tients, five gastric cancer patients, seven HCC patients, trols; the proportions of the serum DKK1–positive cases was six bile duct cancers, six breast cancer patients, and five 74.1% for lung adenocarcinoma (83 of 112), 63.2% for lung cervical cancer patients were obtained with informed con- squamous cell carcinoma (43 of 68), 69.4% for small cell lung sent at Kanagawa Cancer Center (Yokohama, Japan). Pri- cancer (59 of 85), and 63.0% for ESCC (51 of 81; ref. 31). In mary bile duct cancers were obtained with informed addition, we identified that the exogenous expression of consent from six patients who had undergone surgery at DKK1 increased the invasive activity of mammalian cells. Hokkaido University Hospital (Sapporo, Japan). Primary The evidence further prompted us to focus on DKK1 as a lung cancers in the same set of 18 lung cancer cases whose potential target for therapeutic antibodies as well as a serum serum had been collected before surgery (nine patients biomarker for a wide range of human cancers. with DKK1-positive tumors and nine with DKK1-negative DKK1 encodes a secreted protein that plays a crucial role tumors) were obtained at Hiroshima University Hospital in head formation in vertebrate development (37), and is (Hiroshima, Japan) and Kanagawa Cancer Center. This known as a negative regulator of the Wnt-signaling path- study and the use of all clinical materials used were way in colon cancer cell lines (38). During specific period approved by individual institutional Ethical Committees. of embryogenesis, DKK1 is secreted and binds to the LRP5/6 coreceptor (39), resulting in the blocking of the Serum samples interaction with secreted Wnt protein, inducing degrada- Serum samples were obtained with informed consent from tion of β-catenin, and then reducing the expression of 207 healthy control individuals (168 males and 39 females; genes regulated by T-cell factor. This mechanism of DKK1 median age of 50.3 y with a range of 31–61 y) and from action is important in limb and head development (37). 179 pancreatic cancer patients (114 males and 65 females; Inhibition of the Wnt pathway by secreted DKK1 is also median age of 66 y with a range of 30–83 y), 101 gastric can- known to initiate cardiogenesis in vertebrate embryos (40). cer patients (71 males and 30 females; median age of 62 y On the other hand, the overexpression of DKK1 was de- with a range of 29–82 y), 168 HCC patients (146 males and scribed in multiple myeloma, hepatoblastoma, Wilms' 22 females; median age of 70 y with a range of 32–84 y), 107 tumor, prostate cancer, kidney cancer, and breast cancer bile duct cancer patients (75 males and 32 females; median as well as lung and esophageal cancers (31, 41–44). Serum age of 67 y with a range of 45–75 y), 169 breast cancer concentrations of DKK1 protein were reported to be in- patients (169 females; median age of 51 y with a range of creased in patients with multiple myeloma or osteosarcoma 27–72 y), and 182 cervical cancer patients (182 females; me- (41, 45). In spite of recent accumulating reports that indi- dian age of 48 y with a range of 31–77 y) who were registered cate the oncogenic potential of DKK1 activation and its in the Japanese Project for Personalized Medicine (BioBank usefulness as a cancer biomarker, there was no report indi- Japan). These serum samples from a total of 906 cancer pa- cating the direct antitumor effect of anti-DKK1 antibodies. tients were selected for the study on the basis of the follow- We here show evidence suggesting DKK1 as a diagnostic ing criteria: (a) patients were newly diagnosed and (b) their biomarker for a wide variety of cancers and results implicat- tumors were pathologically diagnosed as cancers (stages I–IV). ing the therapeutic potential of anti-DKK1 antibody to neu- Serum was obtained at the time of diagnosis and stored at tralize the activity of DKK1 function for cancer cell invasion −150°C. The sample size of each cancer group (pancreas, and growth. stomach, liver, bile duct, breast, and cervix) and a healthy control group provided 90% statistical power (α =0.05; Materials and Methods β = 0.1) to detect group differences in mean levels of serum DKK1.
Recommended publications
  • Harnessing Low-Density Lipoprotein Receptor Protein 6 (LRP6) Genetic Variation and Wnt Signaling for Innovative Diagnostics in Complex Diseases
    OPEN The Pharmacogenomics Journal (2018) 18, 351–358 www.nature.com/tpj REVIEW Harnessing low-density lipoprotein receptor protein 6 (LRP6) genetic variation and Wnt signaling for innovative diagnostics in complex diseases Z-M Wang1,2, J-Q Luo1,2, L-Y Xu3, H-H Zhou1,2 and W Zhang1,2 Wnt signaling regulates a broad variety of processes in both embryonic development and various diseases. Recent studies indicated that some genetic variants in Wnt signaling pathway may serve as predictors of diseases. Low-density lipoprotein receptor protein 6 (LRP6) is a Wnt co-receptor with essential functions in the Wnt/β-catenin pathway, and mutations in LRP6 gene are linked to many complex human diseases, including metabolic syndrome, cancer, Alzheimer’s disease and osteoporosis. Therefore, we focus on the role of LRP6 genetic polymorphisms and Wnt signaling in complex diseases, and the mechanisms from mouse models and cell lines. It is also highly anticipated that LRP6 variants will be applied clinically in the future. The brief review provided here could be a useful resource for future research and may contribute to a more accurate diagnosis in complex diseases. The Pharmacogenomics Journal (2018) 18, 351–358; doi:10.1038/tpj.2017.28; published online 11 July 2017 INTRODUCTION signaling pathways and expressed in various target organs.1 LDLR- The Wnt1 gene was identified in 1982. Ensuing studies in related proteins 5/6 (LRP5/6) belong to this large family and Drosophila and Xenopus unveiled a highly conserved Wnt/ function as co-receptors of the Wnt/β-catenin pathway. These β-catenin pathway, namely, canonical Wnt signaling.
    [Show full text]
  • Lgr5 Homologues Associate with Wnt Receptors and Mediate R-Spondin Signalling
    ARTICLE doi:10.1038/nature10337 Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling Wim de Lau1*, Nick Barker1{*, Teck Y. Low2, Bon-Kyoung Koo1, Vivian S. W. Li1, Hans Teunissen1, Pekka Kujala3, Andrea Haegebarth1{, Peter J. Peters3, Marc van de Wetering1, Daniel E. Stange1, Johan E. van Es1, Daniele Guardavaccaro1, Richard B. M. Schasfoort4, Yasuaki Mohri5, Katsuhiko Nishimori5, Shabaz Mohammed2, Albert J. R. Heck2 & Hans Clevers1 The adult stem cell marker Lgr5 and its relative Lgr4 are often co-expressed in Wnt-driven proliferative compartments. We find that conditional deletion of both genes in the mouse gut impairs Wnt target gene expression and results in the rapid demise of intestinal crypts, thus phenocopying Wnt pathway inhibition. Mass spectrometry demonstrates that Lgr4 and Lgr5 associate with the Frizzled/Lrp Wnt receptor complex. Each of the four R-spondins, secreted Wnt pathway agonists, can bind to Lgr4, -5 and -6. In HEK293 cells, RSPO1 enhances canonical WNT signals initiated by WNT3A. Removal of LGR4 does not affect WNT3A signalling, but abrogates the RSPO1-mediated signal enhancement, a phenomenon rescued by re-expression of LGR4, -5 or -6. Genetic deletion of Lgr4/5 in mouse intestinal crypt cultures phenocopies withdrawal of Rspo1 and can be rescued by Wnt pathway activation. Lgr5 homologues are facultative Wnt receptor components that mediate Wnt signal enhancement by soluble R-spondin proteins. These results will guide future studies towards the application of R-spondins for regenerative purposes of tissues expressing Lgr5 homologues. The genes Lgr4, Lgr5 and Lgr6 encode orphan 7-transmembrane 4–5 post-induction onwards.
    [Show full text]
  • Recombinant Human Dkk-1 Catalog Number: 5439-DK
    Recombinant Human Dkk-1 Catalog Number: 5439-DK DESCRIPTION Source Spodoptera frugiperda, Sf 21 (baculovirus)­derived human Dkk­1 protein Thr32­His266 Accession # O94907 N­terminal Sequence Thr32 Analysis Predicted Molecular 25.8 kDa Mass SPECIFICATIONS SDS­PAGE 33­38 kDa, reducing conditions Activity Measured by its ability to inhibit Wnt induced TCF reporter activity in HEK293 human embryonic kidney cells. Recombinant Human Dkk­1 (Catalog # 5439­DK) inhibits a constant dose of 500 ng/mL of Recombinant Human Wnt­3a (Catalog # 5036­WN). The ED50 for this effect is 10­60 ng/mL. Endotoxin Level <1.0 EU per 1 μg of the protein by the LAL method. Purity >95%, by SDS­PAGE with silver staining. Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. See Certificate of Analysis for details. PREPARATION AND STORAGE Reconstitution Reconstitute at 100 μg/mL in PBS containing at least 0.1% human or bovine serum albumin. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. Stability & Storage Use a manual defrost freezer and avoid repeated freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 3 months, ­20 to ­70 °C under sterile conditions after reconstitution. DATA Bioactivity SDS­PAGE Recombinant Human Wnt­3a 1 μg/lane of Recombinant Human Dkk­1 was (Catalog # 5036­WN) induces a resolved with SDS­PAGE under reducing (R) dose responsive increase in Wnt conditions and visualized by silver staining, reporter activity in HEK293 cells showing major bands at 33­38 kDa.
    [Show full text]
  • Dickkopf-1 (DKK1) Reveals That Fibronectin Is a Major Target of Wnt Signaling in Branching Morphogenesis of the Mouse Embryonic Lung
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector Developmental Biology 277 (2005) 316–331 www.elsevier.com/locate/ydbio Dickkopf-1 (DKK1) reveals that fibronectin is a major target of Wnt signaling in branching morphogenesis of the mouse embryonic lung Stijn P. De Langhea, Fre´de´ric G. Salaa, Pierre-Marie Del Morala, Timothy J. Fairbanksa, Kenneth M. Yamadab, David Warburtona, Robert C. Burnsa, Saverio Belluscia,* aDevelopmental Biology Program, Department of Surgery, USC Keck School of Medicine and the Saban Research Institute of Children’s Hospital Los Angeles, Los Angeles, CA 90027, USA bCraniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA Received for publication 6 April 2004, revised 16 September 2004, accepted 20 September 2004 Available online 22 October 2004 Abstract Members of the Dickkopf (Dkk) family of secreted proteins are potent inhibitors of Wnt/h-catenin signaling. In this study we show that Dkk1, -2, and -3 are expressed distally in the epithelium, while Kremen1, the needed co-receptor, is expressed throughout the epithelium of the developing lung. Using TOPGAL mice [DasGupta, R., Fuchs, E., 1999. Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation. Development 126, 4557–4568] to monitor the Wnt pathway, we show that canonical Wnt signaling is dynamic in the developing lung and is active throughout the epithelium and in the proximal smooth muscle cells (SMC) until E12.5. However, from E13.5 onwards, TOPGAL activity is absent in the SMC and is markedly reduced in the distal epithelium coinciding with the onset of Dkk-1 expression in the distal epithelium.
    [Show full text]
  • Structural Characterization and Association of Ovine Dickkopf-1 Gene with Wool Production and Quality Traits in Chinese Merino
    G C A T T A C G G C A T genes Article Structural Characterization and Association of Ovine Dickkopf-1 Gene with Wool Production and Quality Traits in Chinese Merino Fang Mu 1,†, Enguang Rong 1,†, Yang Jing 1, Hua Yang 2, Guangwei Ma 1, Xiaohong Yan 1, Zhipeng Wang 1, Yumao Li 1, Hui Li 1 and Ning Wang 1,* 1 Key Laboratory of Chicken Genetics and Breeding at Ministry of Agriculture, Key Laboratory of Animal Genetics, Breeding and Reproduction at Education Department of Heilongjiang Province, Key Laboratory of Animal Cellular and Genetic Engineering of Heilongjiang Province, Harbin 150030, China; [email protected] (F.M.); [email protected] (E.R.); [email protected] (Y.J.); [email protected] (G.M.); [email protected] (X.Y.); [email protected] (Z.W.); [email protected] (Y.L.); [email protected] (H.L.) 2 Institute of Animal Husbandry and Veterinary, Xinjiang Academy of Agriculture and Reclamation Science, Shihezi 832000, China; [email protected] * Correspondence: [email protected]; Tel.: +86-0451-5519-1770 † These authors contributed equally to this work. Received: 20 October 2017; Accepted: 15 December 2017; Published: 20 December 2017 Abstract: Dickkopf-1 (DKK1) is an inhibitor of canonical Wnt signaling pathway and regulates hair follicle morphogenesis and cycling. To investigate the potential involvement of DKK1 in wool production and quality traits, we characterized the genomic structure of ovine DKK1, performed polymorphism detection and association analysis of ovine DKK1 with wool production and quality traits in Chinese Merino. Our results showed that ovine DKK1 consists of four exons and three introns, which encodes a protein of 262 amino acids.
    [Show full text]
  • CNN1 Regulates the DKK1/Wnt/Β‑Catenin/C‑Myc Signaling Pathway by Activating TIMP2 to Inhibit the Invasion, Migration and EMT of Lung Squamous Cell Carcinoma Cells
    EXPERIMENTAL AND THERAPEUTIC MEDICINE 22: 855, 2021 CNN1 regulates the DKK1/Wnt/β‑catenin/c‑myc signaling pathway by activating TIMP2 to inhibit the invasion, migration and EMT of lung squamous cell carcinoma cells WUSHENG LIU1, XIAOGANG FU2 and RUMEI LI3 1Department of Respiratory Medicine, The Affiliated Xinyu Hospital of Nanchang University; Departments of 2Respiratory Medicine and 3Endocrinology, Xinyu People's Hospital, Xinyu, Jiangxi 338000, P.R. China Received October 23, 2020; Accepted February 12, 2021 DOI: 10.3892/etm.2021.10287 Abstract. The present study aimed to investigate the effect tumors has increased over the last decades and this type of of calponin 1 (CNN1) on the invasion and migration of lung cancer is responsible for >1.3 million deaths worldwide annu‑ squamous cell carcinoma (LUSC) cells and the associations ally (1). Lung tumors are one of the most frequent malignant between CNN1, tissue inhibitor of metalloproteinases 2 tumors types in China (2,3). Lung cancer is mainly divided (TIMP2), Dickkopf‑1 (DKK1) and the Wnt/β‑catenin/c‑myc into non‑small cell lung cancer (NSCLC) and small cell lung signaling pathway. The expression levels of CNN1 and cancer, with NSCLC accounting for ~80% of all cases (4). TIMP2 in LUSC cells and the association between CNN1 The two main subtypes of NSCLC are lung adenocarcinoma and TIMP2 were predicted using the GEPIA database. The and lung squamous cell carcinoma (LUSC), and LUSC is cells were transiently transfected to overexpress CNN1, which insidious and develops rapidly (5). A subset of patients with resulted in inhibition of DKK1 and TIMP2 expression levels.
    [Show full text]
  • Targeting Wnt-Driven Cancer Through the Inhibition of Porcupine by LGK974
    Targeting Wnt-driven cancer through the inhibition of Porcupine by LGK974 Jun Liua,1, Shifeng Pana, Mindy H. Hsieha, Nicholas Nga, Fangxian Suna, Tao Wangb, Shailaja Kasibhatlaa, Alwin G. Schullerc, Allen G. Lia, Dai Chenga, Jie Lia, Celin Tompkinsa, AnneMarie Pferdekampera, Auzon Steffya, Jane Chengc, Colleen Kowalc, Van Phunga, Guirong Guoa, Yan Wanga, Martin P. Grahamd, Shannon Flynnd, J. Chad Brennerd, Chun Lia, M. Cristina Villarroele, Peter G. Schultza,2,XuWua,3, Peter McNamaraa, William R. Sellersc, Lilli Petruzzellie, Anthony L. Borale, H. Martin Seidela, Margaret E. McLaughline, Jianwei Chea, Thomas E. Careyd, Gary Vanassee, and Jennifer L. Harrisa,1 aGenomics Institute of Novartis Research Foundation, San Diego, CA 92121; bPreclinical Safety, Novartis Institutes for Biomedical Research, Emeryville, CA 94608; cOncology Research and eOncology Translational Medicine, Novartis Institutes for Biomedical Research, Cambridge, MA 02139; and dDepartment of Otolaryngology – Head and Neck Surgery, University of Michigan, Ann Arbor, MI 48109 Edited by Marc de la Roche, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom, and accepted by the Editorial Board October 31, 2013 (received for review July 31, 2013) Wnt signaling is one of the key oncogenic pathways in multiple Cytoplasmic and nuclear β-catenin have also been correlated with cancers, and targeting this pathway is an attractive therapeutic triple-negative and basal-like breast cancer subtypes (9, 10), and approach. However, therapeutic success has been limited because Wnt signaling has also been implicated in cancer-initiating cells in of the lack of therapeutic agents for targets in the Wnt pathway multiple cancer types (11–14). Wnt pathway signaling activity is and the lack of a defined patient population that would be dependent on Wnt ligand.
    [Show full text]
  • R-Spondin1 Regulates Wnt Signaling by Inhibiting Internalization of LRP6
    R-Spondin1 regulates Wnt signaling by inhibiting internalization of LRP6 Minke E. Binnerts, Kyung-Ah Kim, Jessica M. Bright, Sejal M. Patel, Karolyn Tran, Mei Zhou, John M. Leung, Yi Liu, Woodrow E. Lomas III, Melissa Dixon, Sophie A. Hazell, Marie Wagle, Wen-Sheng Nie, Nenad Tomasevic, Jason Williams, Xiaoming Zhan, Michael D. Levy, Walter D. Funk, and Arie Abo* Nuvelo, Inc., 201 Industrial Road, Suite 310, San Carlos, CA 94070-6211 Communicated by Lewis T. Williams, Five Prime Therapeutics, Inc., Emeryville, CA, March 13, 2007 (received for review December 1, 2006) The R-Spondin (RSpo) family of secreted proteins act as potent Wnt signaling by modulating levels of LRP6 on the cell surface, activators of the Wnt/␤-catenin signaling pathway. We have previ- through inhibition of DKK1-dependent internalization of LRP6. ously shown that RSpo proteins can induce proliferative effects on the gastrointestinal epithelium in mice. Here we provide a mechanism Results and Discussion whereby RSpo1 regulates cellular responsiveness to Wnt ligands by To further investigate the role of RSpo1 in the activation of the Wnt modulating the cell-surface levels of the coreceptor LRP6. We show pathway, we generated a HEK-293 stable cell line expressing a TCF that RSpo1 activity critically depends on the presence of canonical luciferase reporter plasmid. In agreement with reported results Wnt ligands and LRP6. Although RSpo1 does not directly activate (16), both RSpo1 and Wnt3A proteins induced a comparable LRP6, it interferes with DKK1/Kremen-mediated internalization of TCF-mediated reporter activity [supporting information (SI) Fig. LRP6 through an interaction with Kremen, resulting in increased LRP6 7].
    [Show full text]
  • Catenin Pathway (DKK1) in a STAT 3-Dependent Manner This Information Is Current As of September 24, 2021
    IFN-γ Mediates Enhancement of HIV Replication in Astrocytes by Inducing an Antagonist of the β-Catenin Pathway (DKK1) in a STAT 3-Dependent Manner This information is current as of September 24, 2021. Wei Li, Lisa J. Henderson, Eugene O. Major and Lena Al-Harthi J Immunol 2011; 186:6771-6778; Prepublished online 11 May 2011; doi: 10.4049/jimmunol.1100099 Downloaded from http://www.jimmunol.org/content/186/12/6771 References This article cites 41 articles, 10 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/186/12/6771.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 24, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology IFN-g Mediates Enhancement of HIV Replication in Astrocytes by Inducing an Antagonist of the b-Catenin Pathway (DKK1) in a STAT 3-Dependent Manner Wei Li,*,1 Lisa J. Henderson,* Eugene O.
    [Show full text]
  • Expression of Cancer Stem Cell-Associated DKK1 Mrna
    ANTICANCER RESEARCH 37 : 4881-4888 (2017) doi:10.21873/anticanres.11897 Expression of Cancer Stem Cell-associated DKK1 mRNA Serves as Prognostic Marker for Hepatocellular Carcinoma TOMOHIKO SAKABE 1,2 , JUNYA AZUMI 1, YOSHIHISA UMEKITA 2, KAN TORIGUCHI 3, ETSURO HATANO 3, YASUAKI HIROOKA 4 and GOSHI SHIOTA 1 1Division of Molecular and Genetic Medicine, Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medicine, Tottori University, Yonago, Japan; 2Division of Organ Pathology, Department of Pathology, Faculty of Medicine, Tottori University, Yonago, Japan; 3Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan; 4Department of Pathobiological Science and Technology, School of Health Science, Faculty of Medicine, Tottori University, Yonago, Japan Abstract. Background/Aim: Cancer stem cells (CSCs) are Hepatocellular carcinoma (HCC) is the sixth most common associated with prognosis of hepatocellular carcinoma (HCC). cancer, and the third most frequent cause of death worldwide In our previous study, we created cDNA microarray databases (1). Since biomarkers are useful for early diagnosis and on the CSC population of human HuH7 cells. In the present prediction of prognosis (2), they may provide effective study, we identified genes that might serve as prognostic treatment options. Although several biomarkers, including markers of HCC by employing existing databases. Materials alpha-fetoprotein (AFP), protein induced by vitamin K and Methods: Expressions of glutathione S-transferase pi 1 deficiency or antagonist-II (PIVKAII), and glypican-3, were (GSTP1), lysozyme (LYZ), C-X-C motif chemokine ligand 5 reported as being useful (2, 3), identification of novel (CXCL5), interleukin-8 (IL8) and dickkopf WNT signaling biomarkers for HCC is expected to improve prognosis of pathway inhibitor 1 (DKK1), the five most highly expressed patients with HCC.
    [Show full text]
  • The Activation of the WNT Signaling Pathway Is a Hallmark in Neurofibromatosis Type 1 Tumorigenesis
    Published OnlineFirst November 11, 2013; DOI: 10.1158/1078-0432.CCR-13-0780 Clinical Cancer Human Cancer Biology Research The Activation of the WNT Signaling Pathway Is a Hallmark in Neurofibromatosis Type 1 Tumorigenesis Armelle Luscan1,4, Ghjuvan'Ghjacumu Shackleford5, Julien Masliah-Planchon1, Ingrid Laurendeau1, Nicolas Ortonne10, Jennifer Varin1, Francois¸ Lallemand14, Karen Leroy11, Valerie Dumaine6, Mikael Hivelin2,12, Didier Borderie7, Thomas De Raedt15, Laurence Valeyrie-Allanore13,Fred erique Larousserie8, Beno^t Terris3,9, Laurent Lantieri2, Michel Vidaud1,4, Dominique Vidaud1,4, Pierre Wolkenstein13,Beatrice Parfait1,4, Ivan Bieche 1,14, Charbel Massaad5, and Eric Pasmant1,4 Abstract Purpose: The hallmark of neurofibromatosis type 1 (NF1) is the onset of dermal or plexiform neurofibromas, mainly composed of Schwann cells. Plexiform neurofibromas can transform into malignant peripheral nerve sheath tumors (MPNST) that are resistant to therapies. Experimental Design: The aim of this study was to identify an additional pathway in the NF1 tumorigenesis. We focused our work on Wnt signaling that is highly implicated in cancer, mainly in regulating the proliferation of cancer stem cells. We quantified mRNAs of 89 Wnt pathway genes in 57 NF1- associated tumors including dermal and plexiform neurofibromas and MPNSTs. Expression of two major stem cell marker genes and five major epithelial–mesenchymal transition marker genes was also assessed. The expression of significantly deregulated Wnt genes was then studied in normal human Schwann cells, fibroblasts, endothelial cells, and mast cells and in seven MPNST cell lines. Results: The expression of nine Wnt genes was significantly deregulated in plexiform neurofibromas in comparison with dermal neurofibromas. Twenty Wnt genes showed altered expression in MPNST biopsies and cell lines.
    [Show full text]
  • Gene Associations in Human Cancers by Vitamin D and Sulforaphane
    J Cancer Sci Clin Ther 2020; 4 (3): 237-244 DOI: 10.26502/jcsct.5079068 Research Article DICKKOPF-1 (DKK-1) Gene Associations in Human Cancers by Vitamin D and Sulforaphane Sharmin Hossain1*, Zhenhua Liu2, Richard J. Wood2 1National Institute on Aging (NIA/NIH), Baltimore, Maryland, USA 2Department of Nutrition, University of Massachusetts, Amherst, Massachusetts, USA *Corresponding Author: Dr. Sharmin Hossain, National Institute on Aging (NIA/NIH), Baltimore, Maryland, USA, Tel: 410-558-8545; E-mail: [email protected] Received: 06 July 2020; Accepted: 28 July 2020; Published: 03 August 2020 Citation: Sharmin Hossain, Zhenhua Liu, Richard J. DICKKOPF-1 (DKK-1) Gene Associations in Human Cancers by Vitamin D and Sulforaphane. Journal of Cancer Science and Clinical Therapeutics 4 (2020): 237-244. Abstract while in MCF-7 cells saw no effect of the treatments. Co- Dickkopf Wnt Signaling Pathway Inhibitor-1 gene (DKK- treating the cells with both compounds seemed to attenuate 1) encodes a member of the Dickkopf family of proteins the D effect, implying competing needs along the signaling and is involved primarily in the embryonic development pathway and require further investigation. and bone formation in adults. Based on its known relationship to Wnt signaling pathway, we set out to Keywords: Sulforaphane; Trichostatin A; Vitamin-D determine how this gene is affected by dietary agents- receptor; DICKKOPF-1 (DKK-1) gene; Colon cancer; vitamin D and sulforaphane (SFN). Cells were exposed for Breast cancer 24 hours to vitamin D3 [1,25(OH)2D3] (100nM) either alone or in combination with L-sulforaphane and TSA Abbreviations: 1,25(OH)2D3- 1,25 di-hydroxyvitamin (20μM and 1μM respectively) at 70% confluency.
    [Show full text]