DOCSLIB.ORG

Cancer Stem Cells Extensively and Form New Tumors

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cancer Stem Cells Extensively and Form New Tumors Targeting mammary stem cells for breast cancer prevention Max Wicha M.D. Cancer Stem Cell Hypothesis Cancers Arise from Cells with Dysregulated Self-Renewal Cancers Are “Driven” By Cells With Stem Cell Properties Stem cells as targets for cancer prevention HYPOTHESIES If cancers arise in self-renewing cells then: • Stem/Progenitor cell number may be a cancer risk factor • Targeting tissue stem/progenitor cell self- renewal pathways may reduce cancer risk • Tissue stem cell and self-renewal pathway components may be useful biomarkers for cancer prevention studies characteristics of stem cells • self renewal • differentiation into multiple cell types • types of stem cells embryonic adult • cancer Stem cells and breast carcinogenesis Stem cell Cancer stem cell Quiescent pool of stem cell Early Progenitor Late Progenitor Cancer stem cell DIFFERENTIATION Luminal cells Myoepithelial cells Alveolar cells Reduction mammoplasties “Mammosphere” Suspension culture (Serum-free, EGF, …) Culture Systems Mammospheres Dissociated to single cells Acinar Ductal- acinar Regeneration of the mammary N generation of spheres Mixed Myoepithelial Epithelial structure structure gland in the cleared fat- pad B. Differentiation in C. Differentiation in A. Self- renewal D. Differentiation and in vitro vitro in 2- D culture vitro in 3- D culture self- renewal in vivo (Suspension) (Collagen) (Matrigel ) (NOD/SCID mice) Dontu & Wicha, 2003 Dontu & Wicha, 2003 Weaver & Bissell, 1999 Kuperwasser & Weinberg, 2004 Dontu & Wicha, 2003 Liu & Wicha, 2006 ALDEFLUOR + cells in Normal breast epithelium BAAA ALDH DEAB With DEAB Without DEAB 6% of ALDEFLUOR positive cells in Normal breast epithelium 6% Ginestier et al Cell Stem Cell 2007 ALDEFLUOR+ cells have stem cell properties Culture in suspension With DEAB Without DEAB Only ALDEFLUOR + cells generate mammospheres in suspension ALDEFLUOR+ cells have stem cell properties Without DEAB CK18 SMA ALDEFLUOR+ Luminal cells CK18+ Myoepithelial cells SMA+ ALDEFLUOR- ALDEFLUOR- ALDEFLUOR+ Ginestier, 2007 Models of Tumor Heterogeneity Stochastic model Cancer stem cell model CSC CSC CSC CSC Cancer cells are heterogeneous, Cancer cells are heterogeneous, but most cells can proliferate and only rare cancer stem cells extensively and form new tumors. have the ability to proliferate extensively and form new tumors. Reya et al 2001 Tumor Formation by Human Breast Cancer “Stem Cells” 20,000 200 CD44- CD44+ CD24+ CD24- Cells Cells Al Hajj et al PNAS 2003 Stem Cell Markers • CD44 • CD24 • Aldehyde Dehydrogenase ( Christophe Ginestier) • Oct-4 • Bmi-1 • Nuclear B-Catenin ALDEFLUOR+ population and tumorigenicity With DEAB Without DEAB ALDEFLUOR- ALDEFLUOR+ R1 R2 R1 R2 5% to 10% of ALDEFLUOR + population in Breast tumors ALDEFLUOR- ALDEFLUOR+ 50,000 cells 50,000 cells ALDEFLUOR – ALDEFLUOR + 5000 cells 5000 cells 50,000 cells 500 cells 500 cells 500 cells ALDEFLUOR+ population and tumorigenicity ALDEFLUOR - ALDEFLUOR + ALDEFLUOR+ population regenerates heterogeneity of the initial tumor Overlap between CD44+CD24- and ALDEFLUOR+ Populations Ginestier et al Cell Stem Cell 2007 Breast Cancer Development Luminal Myoepithelial Normal duct Hyperplasia In situ carcinoma Invasive carcinoma Aldefluor + cells are invasive and metastatic U (-) (+) HCC1954HCC1954 (100k) 500 100,000 ALDELFUOR-positive 450 100,00 ALDEFLUOR-negative 400 350 100,000 Unseparated 300 Positive E 250 Negative 200 Unsorted 150 100 50 Normalized Flux Photon Normalized Photon Flux Photon Normalized 0 1 2 3 4 5 6 7 8 F WeeksWeeks after after injection injection B 600 MDAMDAMB453-MB-453 (100k) K 500 500 450 100,000 ALDELFUOR-positive 400 400 100,00 ALDEFLUOR-negative 350 300 300 100,000 Unseparated 250 Positive G 200 200 Negative 150 Unsorted 100 100 Relative invasion rate Normalized Flux Photon 50 Normalized Photon Flux Photon Normalized 0 0 L 1 2 3 4 1 2 3 4 5 ALDEFLUOR + + + + + + - - - - - - Weeks after injection BrCa-MZ-01 MDA-MB-453 SUM159 Weeks after injection C H IL8 - - - + + + - - - + + + SUM159SUM159 (100k) HCC1954 MDA-MB-453 SUM159 30 A 100,000 ALDELFUOR-positive 25 100,00 ALDEFLUOR-negative 20 100,000 Unseparated Positive 15 Negative Unsorted I M 10 5 Normalized Flux Photon Normalized Photon Flux Photon Normalized 0 1 2 3 4 5 Weeks after after injection injection D J Cancer Stem Cells: Implications For Metastasis TDC only No Metastases CSC=Cancer Stem Cell TDC=Terminally Differentiated Cell NO CSC 1o Tumor CSC with FULL malignant potential Metastases in months to few years TDC TDC Subsequently to other sites TDC CSC TDC TDC TDC CSC with PARTIAL malignant potential Dormancy followed by Metastases after many years: Secondary Oncogenic “Hits” and/or Changes in Microenvironment Carcinogenesis Stem cell regulation Dysregulation Cancer TUMOR Microenvironment Stem Cell Generation Tumor Microenviornment and the “CSC Niche” • The Microenvironment (“Stem cell niche”) contains fibroblasts, endothelial cells, inflammatory cells and mesenchymal stem cells recruited from bone marrow. • The Microenvironment plays an important role in normal mammary development as well as tumor growth and metastasis. > Bone marrow derived mesenchymal stem cells are recruited to murine breast carcinomas (Karnoub 2007). • Cytokines regulate breast cancer stem cells. > IL6 increases breast cancer stem cells (Sanson 2008). > IL8/CXCR1 axis regulates breast cancer stem cells (Charafe-Jaffret Cancer Research 2009) Effects of Cytokines on breast cancer stem/progenitor cell population A 14 12 10 positive - 8 6 4 SUM159 cells (%) ALDEFLOUR-positive cells (%) ALDEFLOUR 2 0 Co- Control 2 ug/ml 1 ug/ml 10 ng/ml 10 ng/ml 100 ng/ml 100 ng/ml Co- 100 ng/ml culture N.T. CXCL12 ug/ml CXCL51 ug/ml CXCL610 ng/ml CXCL710 ng/ml IL6 100IL8 ng/ml culture CXCL1 CXCL5 CXCL6 CXCL7 IL6 IL8 SUM159 B 16 12 positive - 8 4 SUM159 cells (%) ALDEFLOUR ALDEFLOUR-positive cells (% cells ALDEFLOUR-positive 0 10 ug/ml SUM159SUM159 ControlN.T. 100100 ug/ml ug/ml 5050 ug/ml ug/ml 2020 ug/ml ug/ml 100100 ug/ml ug/ml 10 ug/ml anti-IL8 alonealone anti-CXCL5anti-CXCL5 anti-CXCL6anti-CXCL6 anti-CXCL7anti-CXCL7 anti-IL6anti-IL6 anti-IL8 Co-culture Model of cytokine networks between MSC and breast cancer cells CXCL7 Bulk Tumor Mesenchymal Cells Cells ALDH- CXCR2 ALDH+ ALDH- IL-6R ALDH- gp130 IL-6 CXCL1 CXCL5 CXCL6 IL-8 IL-6 CXCR1 IL-6R gp130 Stem cell self Cancer renewal Stem Cell S. Liu Canc. Res. 2011 Regulation of CSC’s by cytokine loops in the tumor microenviornment Korkaya & Wicha JCI, 121(10):3804-09, 2011 Diet ,exercise and breast cancer risk Neilson H K et al. Cancer Epidemiol Biomarkers Prev 2009;18:11-27 Fas-Ligand IL-8 IL-8 Fas-Ligand IL-8 IL-8 Chemotherapy Fas CXCR1 Fas FADD PI3-K FADD FAK PTEN AKT Apoptosis Self- FOXO3a IL-8 IL-8 β-catenin IL-8 Renewal GSK3β β-catenin Wnt pathway IL-8 TCF targets, Fas-Ligand Self-renewal Fas-Ligand A Cancer stem cell Bystander effect Bulk tumor cells Repertaxin Fas-Ligand Fas-Ligand Chemotherapy Fas Fas CXCR1 FADD FADD FAK FOXO3a Apoptosis Apoptosis IL-8 IL-8 AKT Death IL-8 FOXO3a IL-8 Fas-Ligand Fas-Ligand Fas-Ligand Cancer stem cell Bystander effect Bulk tumor cells B Stem Cell Equilibria miR-93 EMT MET Differentiation TGF-b Il6 Quiescent Cycling Vimentin + Vimentin - Epcam (ESA) - Epcam (ESA) + p53 p53 p21 p21 CD44+/CD24- Aldefluor + E Cadherin - E Cadherin + 44+/24- Highly Tumorigenic Aldefluor+ Recapitulation of Tumors Via CSC Metastasis Breast Tumor Micrometastasis Macrometastasis Vasculature Aldefluor + Invasive Edge of Tumor Time CD44+/CD24- Distal Healthy Organ Necrotic Distal Colonized Organ Zone EMT MET Strategic Approach to Preventive Therapeutics Genetic Estimated Toxicity Background Lifetime Risk I N C R E A S E S A N C R E I NUTRITION E S A N C R E I BOTANICAL Sporadic <10% BASED Haplotype-SNP 10%-40% I1307K PHARMACEUTICAL Familial Syndrome >40% BASED (BRCA) Kakalara Self Renewal and Differientiation Pathways In Breast Stem Cells ER- stem cells SC SC SC Her2 Self renewal SC Notch Bmi-1 BRCA-1 Hedgehog PTEN ER+ Progenitor cells Differentiation Myoepithelial cells Alveolar epithelial cells Ductal epithelial cells A Conceptual Link Between Hereditary & Sporatic Breast Cancers Wnt Wnt IL-8 IL-8 IL-8 IL-8 LRP CXCR1 Repertaxin HH PI3-K HH PTCH Frizzled Perifosine HH FAK DSH GSK3β AKT FU SMO PTEN Cyclopamine β-catenin RTKs Src HER2 SUFU β-catenin GLI TCF Trastuzumab GLI Bmi-1 STAT3 NOTCH HES DSL HER2 DSL NICD STAT3 Anti-DLL4 γ-secretase JAK3 gp130 IL-6R GSI CXCR2 IL6 IL6 IL6 Cancer stem cell Breast Tumor Monoclonal Abs IL-8 CXCL7 IL-6 IL-6R gp130 Targeting Self-Renewal Pathways in CSC’S Mesenchymal Cells Broccoli could stop breast cancer spreading by targeting stem cells Broccoli contains sulforaphane, which seemed to kill off cancer stem cells in tests Figure 2. A Primary Mammospheres B Mammosphere Size 120.0 MCF7 0 µM 0.5 µM 1 µM 5 µM 100.0 SUM1… SF SF SF SF 80.0 MCF7 ** 60.0 ** ** ** 40.0 **** Sphere formation Sphere formation 20.0 SUM159 normalized to control (%) 0.0 0 0.5 1 5 Concentration of SF (µM) C 2nd Passage 3rd Passage 120.0 140.0 MCF7 MCF7 100.0 SUM159 120.0 SUM159 100.0 80.0 ** 80.0 * 60.0 ** 60.0 * 40.0 ** ** 40.0 Sphere formation Sphere formation 20.0 ** Sphere formation 20.0 ** ** ** normalized to control (%) 0.0 normalized to control (%) 0.0 0 0.5 1 5 0 0.5 1 5 Li CCR 2010 Concentration of SF (µM) Concentration of SF (µM) Figure 3. A P=0.003 4 P=0.008 3.5 3 2.5 2 positive cells positive - 1.5 1 0.5 % ALDH % 0 0 1 5 B Concentration of SF (µm Control 1 µM SF 5 µM SF 3.01% 1.47% 0.49% B A 25 ) 500 1st Generation 1st Generation 3 400 Control 20 50 mg/kg SF 300 P=0.018
Load more

Recommended publications
  • The CXCL7/CXCR1/2 Axis Is a Key Driver in the Growth of Clear Cell Renal Cell Carcinoma
    Published OnlineFirst December 12, 2013; DOI: 10.1158/0008-5472.CAN-13-1267 Cancer Therapeutics, Targets, and Chemical Biology Research The CXCL7/CXCR1/2 Axis Is a Key Driver in the Growth of Clear Cell Renal Cell Carcinoma Renaud Grepin 5,Melanie Guyot1, Sandy Giuliano1, Marina Boncompagni1, Damien Ambrosetti1,2, Emmanuel Chamorey3, Jean-Yves Scoazec4, Sylvie Negrier4,Hel ene Simonnet4, and Gilles Pages 1 Abstract Mutations in the von Hippel–Lindau gene upregulate expression of the central angiogenic factor VEGF, which drives abnormal angiogenesis in clear cell renal cell carcinomas (ccRCC). However, the overexpression of VEGF in these tumors was not found to correlate with overall survival. Here, we show that the proangiogenic, proin- flammatory cytokine CXCL7 is an independent prognostic factor for overall survival in this setting. CXCL7 antibodies strongly reduced the growth of ccRCC tumors in nude mice. Conversely, conditional overexpression of CXCL7 accelerated ccRCC development. CXCL7 promoted cell proliferation in vivo and in vitro, in which expression of CXCL7 was induced by the central proinflammatory cytokine interleukin (IL)-1b. ccRCC cells normally secrete low amounts of CXCL7; it was more highly expressed in tumors due to high levels of IL-1b there. We found that a pharmacological inhibitor of the CXCL7 receptors CXCR1 and CXCR2 (SB225002) was sufficient to inhibit endothelial cell proliferation and ccRCC growth. Because CXCR1 and CXCR2 are present on both endothelial and ccRCC cells, their inhibition affected both the tumor vasculature and the proliferation of tumor cells. Our results highlight the CXCL7/CXCR1/CXCR2 axis as a pertinent target for the treatment of ccRCC.
    [Show full text]
  • The Effect of Hypoxia on the Expression of CXC Chemokines and CXC Chemokine Receptors—A Review of Literature
    International Journal of Molecular Sciences Review The Effect of Hypoxia on the Expression of CXC Chemokines and CXC Chemokine Receptors—A Review of Literature Jan Korbecki 1 , Klaudyna Kojder 2, Patrycja Kapczuk 1, Patrycja Kupnicka 1 , Barbara Gawro ´nska-Szklarz 3 , Izabela Gutowska 4 , Dariusz Chlubek 1 and Irena Baranowska-Bosiacka 1,* 1 Department of Biochemistry and Medical Chemistry, Pomeranian Medical University in Szczecin, Powsta´nców Wielkopolskich 72 Av., 70-111 Szczecin, Poland; [email protected] (J.K.); [email protected] (P.K.); [email protected] (P.K.); [email protected] (D.C.) 2 Department of Anaesthesiology and Intensive Care, Pomeranian Medical University in Szczecin, Unii Lubelskiej 1, 71-281 Szczecin, Poland; [email protected] 3 Department of Pharmacokinetics and Therapeutic Drug Monitoring, Pomeranian Medical University in Szczecin, Powsta´nców Wielkopolskich 72 Av., 70-111 Szczecin, Poland; [email protected] 4 Department of Medical Chemistry, Pomeranian Medical University in Szczecin, Powsta´nców Wlkp. 72 Av., 70-111 Szczecin, Poland; [email protected] * Correspondence: [email protected]; Tel.: +48-914661515 Abstract: Hypoxia is an integral component of the tumor microenvironment. Either as chronic or cycling hypoxia, it exerts a similar effect on cancer processes by activating hypoxia-inducible factor-1 (HIF-1) and nuclear factor (NF-κB), with cycling hypoxia showing a stronger proinflammatory influ- ence. One of the systems affected by hypoxia is the CXC chemokine system. This paper reviews all available information on hypoxia-induced changes in the expression of all CXC chemokines (CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8 (IL-8), CXCL9, CXCL10, CXCL11, CXCL12 Citation: Korbecki, J.; Kojder, K.; Kapczuk, P.; Kupnicka, P.; (SDF-1), CXCL13, CXCL14, CXCL15, CXCL16, CXCL17) as well as CXC chemokine receptors— Gawro´nska-Szklarz,B.; Gutowska, I.; CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7 and CXCR8.
    [Show full text]
  • Exploration of Prognostic Biomarkers and Therapeutic Targets in the Microenvironment of Bladder Cancer Based on CXC Chemokines
    Exploration of Prognostic Biomarkers and Therapeutic Targets in The Microenvironment of Bladder Cancer Based on CXC Chemokines Xiaoqi Sun Department of Urology, Kaiping Central Hospital, Kaiping, 529300, China Qunxi Chen Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, 510060, China Lihong Zhang Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, 510060, China Jiewei Chen Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, 510060, China Xinke Zhang ( [email protected] ) Sun Yat-sen University Cancer Center Research Keywords: Bladder cancer, Biomarkers, CXC Chemokines, Microenvironment Posted Date: February 24th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-223127/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/29 Abstract Background: Bladder cancer (BLCA) has a high rate of morbidity and mortality, and is considered as one of the most malignant tumors of the urinary system. Tumor cells interact with surrounding interstitial cells, playing a key role in carcinogenesis and progression, which is partly mediated by chemokines. CXC chemokines exert anti‐tumor biological roles in the tumor microenvironment and affect patient prognosis. Nevertheless, their expression and prognostic values patients with BLCA remain unclear. Methods: We used online tools, including Oncomine, UALCAN, GEPIA, GEO databases, cBioPortal, GeneMANIA, DAVID 6.8, Metascape, TRUST (version 2.0), LinkedOmics, TCGA, and TIMER2.0 to perform the relevant analysis. Results: The mRNA levels of C-X-C motif chemokine ligand (CXCL)1, CXCL5, CXCL6, CXCL7, CXCL9, CXCL10, CXCL11, CXCL13, CXCL16, and CXCL17 were increased signicantly increased, and those of CXCL2, CXCL3, and CXCL12 were decreased signicantly in BLCA tissues as assessed using the Oncomine, TCGA, and GEO databases.
    [Show full text]
  • Ncomms1239.Pdf
    ARTICLE Received 10 Nov 2010 | Accepted 15 Feb 2011 | Published 15 Mar 2011 DOI: 10.1038/ncomms1239 Inflammation driven by tumour-specific Th1 cells protects against B-cell cancer Ole Audun Werner Haabeth1, Kristina Berg Lorvik1, Clara Hammarström2, Ian M. Donaldson3,4, Guttorm Haraldsen2, Bjarne Bogen1 & Alexandre Corthay1 The immune system can both promote and suppress cancer. Chronic inflammation and proinflammatory cytokines such as interleukin (IL)-1 and IL-6 are considered to be tumour promoting. In contrast, the exact nature of protective antitumour immunity remains obscure. Here, we quantify locally secreted cytokines during primary immune responses against myeloma and B-cell lymphoma in mice. Strikingly, successful cancer immunosurveillance mediated by tumour-specific CD4 + T cells is consistently associated with elevated local levels of both proinflammatory (IL-1α, IL-1β and IL-6) and T helper 1 (Th1)-associated cytokines (interferon-γ (IFN-γ), IL-2 and IL-12). Cancer eradication is achieved by a collaboration between tumour- specific Th1 cells and tumour-infiltrating, antigen-presenting macrophages. Th1 cells induce secretion of IL-1β and IL-6 by macrophages. Th1-derived IFN-γ is shown to render macrophages directly cytotoxic to cancer cells, and to induce macrophages to secrete the angiostatic chemokines CXCL9/MIG and CXCL10/IP-10. Thus, inflammation, when driven by tumour- specific Th1 cells, may prevent rather than promote cancer. 1 Centre for Immune Regulation, Institute of Immunology, University of Oslo and Oslo University Hospital Rikshospitalet, PO Box 4950 Nydalen, 0424 Oslo, Norway. 2 Department of Pathology, Institute of Pathology, Oslo University Hospital Rikshospitalet and University of Oslo, PO Box 4950 Nydalen, 0424 Oslo, Norway.
    [Show full text]
  • Cxcl9l and Cxcr3.2 Regulate Recruitment of Osteoclast Progenitors to Bone Matrix in a Medaka Osteoporosis Model
    Cxcl9l and Cxcr3.2 regulate recruitment of osteoclast progenitors to bone matrix in a medaka osteoporosis model Quang Tien Phana,b,1, Wen Hui Tana,b,1, Ranran Liua,b, Sudha Sundarama,b, Anita Buettnera,b, Susanne Kneitzc, Benedict Cheonga,b, Himanshu Vyasa,b, Sinnakaruppan Mathavand,e, Manfred Schartlc,f, and Christoph Winklera,b,2 aDepartment of Biological Sciences, National University of Singapore, Singapore 117543, Singapore; bCentre for Bioimaging Sciences, National University of Singapore, Singapore 117543, Singapore; cDepartment of Developmental Biochemistry, Biocenter, University of Würzburg, 97080 Würzburg, Germany; dGenome Institute of Singapore, Singapore 138672, Singapore; eLee Kong Chian School of Medicine, Nanyang Technological University, Singapore 639798, Singapore; and fThe Xiphophorus Genetic Stock Center, Department of Chemistry and Biochemistry, Texas State University, San Marcos, TX 78666 Edited by Clifford J. Tabin, Harvard Medical School, Boston, MA, and approved July 4, 2020 (received for review April 1, 2020) Bone homeostasis requires continuous remodeling of bone matrix demonstrating RANKL’s important role as a coupling factor to maintain structural integrity. This involves extensive communi- (6–8). However, more coupling factors remain to be identified as cation between bone-forming osteoblasts and bone-resorbing os- osteoclasts also form in a RANKL-independent manner (9). teoclasts to orchestrate balanced progenitor cell recruitment and Zebrafish and medaka have become popular models for hu- activation. Only a few mediators controlling progenitor activation man skeletal disorders (10). Both species are amenable to ad- are known to date and have been targeted for intervention of vanced forward and reversed genetics and genome modification bone disorders such as osteoporosis. To identify druggable path- and uniquely suited for live bioimaging, which makes them ideal ways, we generated a medaka (Oryzias latipes) osteoporosis for bone research.
    [Show full text]
  • Platelet-Derived Chemokine CXCL7 Dimer Preferentially Exists in the Glycosaminoglycan-Bound Form: Implications for Neutrophil–Platelet Crosstalk
    ORIGINAL RESEARCH published: 02 October 2017 doi: 10.3389/fimmu.2017.01248 Platelet-Derived Chemokine CXCL7 Dimer Preferentially Exists in the Glycosaminoglycan-Bound Form: Implications for Neutrophil–Platelet Crosstalk Aaron J. Brown1,2, Krishna Mohan Sepuru1,2, Kirti V. Sawant1 and Krishna Rajarathnam1,2,3* 1 Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, United States, 2 Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX, United States, 3 Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, United States Platelet-derived chemokine CXCL7 (also known as NAP-2) plays a crucial role in orches- trating neutrophil recruitment in response to vascular injury. CXCL7 exerts its function by activating the CXC chemokine receptor 2 (CXCR2) receptor and binding sulfated glycosaminoglycans (GAGs) that regulate receptor activity. CXCL7 exists as monomers, dimers, and tetramers, and previous studies have shown that the monomer dominates Edited by: Deirdre R. Coombe, at lower and the tetramer at higher concentrations. These observations then raise the Curtin University, Australia question: what, if any, is the role of the dimer? In this study, we make a compelling Reviewed by: observation that the dimer is actually the favored form in the GAG-bound state. Further, James R. Whiteford, we successfully characterized the structural basis of dimer binding to GAG heparin using Queen Mary University of London, United Kingdom solution nuclear magnetic resonance (NMR) spectroscopy. The chemical shift assign- Philipp Von Hundelshausen, ments were obtained by exploiting heparin binding-induced NMR spectral changes Ludwig-Maximilians-Universität München, Germany in the WT monomer and dimer and also using a disulfide-linked obligate dimer.
    [Show full text]
  • Human CXCL4/PF4 Immunoassay Quantikine
    Quantikine® ELISA Human CXCL4/PF4 Immunoassay Catalog Number DPF40 For the quantitative determination of human Platelet Factor 4 (PF4) concentrations in cell culture supernates, serum, and platelet-poor plasma. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures. TABLE OF CONTENTS SECTION PAGE INTRODUCTION ....................................................................................................................................................................1 PRINCIPLE OF THE ASSAY ..................................................................................................................................................2 LIMITATIONS OF THE PROCEDURE ................................................................................................................................2 TECHNICAL HINTS ................................................................................................................................................................2 MATERIALS PROVIDED & STORAGE CONDITIONS ..................................................................................................3 OTHER SUPPLIES REQUIRED ............................................................................................................................................3 PRECAUTIONS ........................................................................................................................................................................4 SAMPLE
    [Show full text]
  • And Nuclear Factor-Κb–Regulated CXC Chemokine Gene Expression in Lung Carcinogenesis
    Cancer Prevention Research Cyclic AMP-Responsive Element Binding Protein– and Nuclear Factor-κB–Regulated CXC Chemokine Gene Expression in Lung Carcinogenesis Hongxia Sun, Wen-Cheng Chung, Seung-Hee Ryu, Zhenlin Ju, Hai T. Tran, Edward Kim, Jonathan M. Kurie and Ja Seok Koo Abstract The recognition of the importance of angiogenesis in tumor progression has led to the development of antiangiogenesis as a new strategy for cancer treatment and prevention. By modulating tumor microenvironment and inducing angiogenesis, the proinflammatory cytokine interleukine (IL)-1β has been reported to promote tumor development. However, the factors mediating IL-1β–induced angiogenesis in non–small cell lung cancer (NSCLC) and the regulation of these angiogenicfactorsby IL-1 β are less clear. Here, we report that IL-1β up-regulated an array of proangiogenic CXC chemokine genes in the NSCLC cell line A549 and in normal human tracheobronchial epithelium cells, as determined by microarray analysis. Further analysis revealed that IL-1β induced much higher protein levels of CXC chemokines in NSCLC cells than in normal human tracheobronchial epithelium cells. Con- ditioned medium from IL-1β–treated A549 cells markedly increased endothelial cell migra- tion, which was suppressed by neutralizing antibodies against CXCL5 and CXCR2. We also found that IL-1β–induced CXC chemokine gene overexpression in NSCLC cells was abro- gated with the knockdown of cyclic AMP-responsive element binding protein (CREB) or nuclear factor κB (NF-κB). Moreover, the expression of the CXC chemokine genes as well as CREB and NF-κB activities was greatly increased in the tumorigenic NSCLC cell line compared with normal, premalignant immortalized or nontumorigenic cell lines.
    [Show full text]
  • COMPREHENSIVE INVITED REVIEW Chemokines and Their Receptors
    COMPREHENSIVE INVITED REVIEW Chemokines and Their Receptors Are Key Players in the Orchestra That Regulates Wound Healing Manuela Martins-Green,* Melissa Petreaca, and Lei Wang Department of Cell Biology and Neuroscience, University of California, Riverside, California. Significance: Normal wound healing progresses through a series of over- lapping phases, all of which are coordinated and regulated by a variety of molecules, including chemokines. Because these regulatory molecules play roles during the various stages of healing, alterations in their presence or function can lead to dysregulation of the wound-healing process, potentially leading to the development of chronic, nonhealing wounds. Recent Advances: A discovery that chemokines participate in a variety of disease conditions has propelled the study of these proteins to a level that potentially could lead to new avenues to treat disease. Their small size, ex- posed termini, and the fact that their only modifications are two disulfide Manuela Martins-Green, PhD bonds make them excellent targets for manipulation. In addition, because they bind to G-protein-coupled receptors (GPCRs), they are highly amenable to Submitted for publication January 9, 2013. *Correspondence: Department of Cell Biology pharmacological modulation. and Neuroscience, University of California, Riv- Critical Issues: Chemokines are multifunctional, and in many situations, their erside, Biological Sciences Building, 900 Uni- functions are highly dependent on the microenvironment. Moreover, each versity Ave., Riverside, CA 92521 (email: [email protected]). specific chemokine can bind to several GPCRs to stimulate the function, and both can function as monomers, homodimers, heterodimers, and even oligo- mers. Activation of one receptor by any single chemokine can lead to desen- Abbreviations sitization of other chemokine receptors, or even other GPCRs in the same cell, and Acronyms with implications for how these proteins or their receptors could be used to Ang-2 = angiopoietin-2 manipulate function.
    [Show full text]
  • CXC Chemokines and Their Receptors in Early Metanephric Development
    BASIC RESEARCH www.jasn.org ELR؉-CXC Chemokines and Their Receptors in Early Metanephric Development Zoia B. Levashova,* Nirmala Sharma,* Olga A. Timofeeva,* Jeffrey S. Dome,† and Alan O. Perantoni* *Laboratory of Comparative Carcinogenesis, National Cancer Institute, National Institutes of Health, Frederick, Maryland; and †Division of Oncology, Children’s National Medical Center, Washington, DC ABSTRACT Although originally identified as mediators of inflammation, it is now apparent that chemokines play a ϩ fundamental role in tissue development. In this study, ELR -CXC chemokine family members CXCL2 and CXCL7, along with their preferred receptor CXCR2, were expressed at the earliest stages of metaneph- ric development in the rat, and signaling through this receptor was required for the survival and maintenance of the undifferentiated metanephric mesenchyme (MM). A specific antagonist of the CXCR2 receptor SB225002 induced apoptosis in this population but did not affect more mature structures or cells in the ureteric bud. CXCL7 treatment of isolated MM elicited an angiogenic response by upregulation of matrix metalloprotease 9 and endothelial and mesangial markers (platelet-endothelial cell adhesion molecule, Megsin, Thy-1, PDGF receptor ␣, and vascular ␣-actin) and induced SB225002- sensitive cell invasion through a matrix. Because Wilms’ tumor cells may similarly depend on CXCR2 signaling for survival, primary tumor samples were analyzed, and 15 of 16 Wilms’ tumors were found to be CXCR2 positive, whereas grossly normal kidney tissues from tumor patients or renal cell carcinomas were CXCR2 negative. Furthermore, cell lines derived from Wilms’ tumors but not those from renal cell carcinomas were sensitive to SB225002-induced apoptosis. These data provide evidence for a prosur- ϩ vival and proangiogenic role of ELR -CXC chemokines and their receptor CXCR2 during metanephric development and suggest a novel mechanism for chemotherapeutic intervention in Wilms’ tumor.
    [Show full text]
  • Defining a Transcriptional Fingerprint of Murine Splenic B-Cell Development
    Genes and Immunity (2008) 9, 706–720 & 2008 Macmillan Publishers Limited All rights reserved 1466-4879/08 $32.00 www.nature.com/gene ORIGINAL ARTICLE Defining a transcriptional fingerprint of murine splenic B-cell development I Debnath1, KM Roundy1, DM Dunn2, RB Weiss2, JJ Weis1 and JH Weis1 1Division of Cell Biology and Immunology, Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA and 2Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA B-cell development occurs in a stepwise fashion that can be followed by the expression of B cell-specific surface markers. In this study, we wished to identify proteins that could contribute to the changes in expression of such markers. By using RNA from freshly isolated B220 þ cells, we hoped to reduce the effect of artifacts that occur during the isolation and amplification steps necessary to use flow cytometry analysis-sorted subsets in microarray experiments. Analyses comparing expression patterns from B220 þ 2-week bone marrow (pro-B, pre-B, immature B cells), 2-week spleen (predominantly transitional cells) and 8-week spleen (mainly mature B cells) yielded hundreds of genes. We also examined the B cell-activating factor (BAFF)- dependent effects on immature splenic B cells by comparing expression patterns in the spleen between 2-week A/J vs 2-week A/WySnJ mice, which lack functional BAFF receptor signaling. Genes that showed the expression differences between spleen and bone marrow samples were then analyzed through quantitative PCR on B-cell subsets isolated using two different sorting protocols.
    [Show full text]
  • Products for Chemokine Research
    RnDSy-lu-2945 Products for Chemokine Research The Chemokine Superfamily Chemokines are small cell surface-localized or secreted chemotactic cytokines that bind to and activate specific G protein-coupled chemokine receptors. Most chemokines have at least four conserved N-terminal cysteine residues that form two intramolecular disulfide bonds. Four chemokine subfamilies (CXC, CC, C and CX3C) have been defined based upon the placement of the first two cysteine residues. The CXC chemokine subfamily is characterized by two cysteine residues separated by one amino acid. Within this subfamily, two CXC classes are further defined by the presence or absence of an ELR motif sequence. ELR– CXC chemokines act as chemoattractants for lymphocytes, while ELR+ CXC chemokines are chemoattractants for neutrophils. Additionally, CXC chemokines can mediate angiogenesis.1 The CC chemokine subfamily is defined by two adjacent cysteine CCR1 H: CCL3-5, 7, 8, 13-16, 23, CCL3L1, CCL3L3, CCL4L1, CCL4L2 M: CCL3-7, 9/10 residues. CC chemokines induce inflammatory H: XCL1, 2 XCR1 responses via regulation of monocyte, M: XCL1 macrophage, mast cell, and T cell migration.2 H: CCL2, 7, 8, 13, 16 CCR2 (A or B) M: CCL2, 7, 12 C chemokines are characterized by a single H: CCL26 CX3CR1 cysteine residue and are constitutively expressed H: CX3CL1 in the thymus where they regulate T cell H: CCL5, 7, 8, 11, 13, 14, 15, 24, 26, 28, CCL3L1, CCL3L3 M: CX3CL1 CCR3 M: CCL5, 7, 9/10, 11, 24 differentiation.3 The CX3C chemokine subfamily is defined by two cysteine residues separated by H: CXCL6-8 CXCR1 three amino acids.
    [Show full text]