Abstract
In order to explore the mechanism(s) underlying the pro-tumorigenic capacity of heparanase, we established an inducible Tet-on system. Heparanase expression was markedly increased following addition of doxycycline (Dox) to the culture medium of CAG human myeloma cells infected with the inducible heparanase gene construct, resulting in increased colony number and size in soft agar. Moreover, tumor xenografts produced by CAG-heparanase cells were markedly increased in mice supplemented with Dox in their drinking water compared with control mice maintained without Dox. Consistently, we found that heparanase induction is associated with decreased levels of CXCL10, suggesting that this chemokine exerts tumor-suppressor properties in myeloma. Indeed, recombinant CXCL10 attenuated the proliferation of CAG, U266 and RPMI-8266 myeloma cells. Similarly, CXCL10 attenuated the proliferation of human umbilical vein endothelial cells, implying that CXCL10 exhibits anti-angiogenic capacity. Strikingly, development of tumor xenografts produced by CAG-heparanase cells overexpressing CXCL10 was markedly reduced compared with control cells. Moreover, tumor growth was significantly attenuated in mice inoculated with human or mouse myeloma cells and treated with CXCL10–Ig fusion protein, indicating that CXCL10 functions as a potent anti-myeloma cytokine.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Arvatz G, Shafat I, Levy-Adam F, Ilan N, Vlodavsky I . The heparanase system and tumor metastasis: is heparanase the seed and soil? Cancer Metastasis Rev 2011; 30: 253–268.
Barash U, Cohen-Kaplan V, Dowek I, Sanderson RD, Ilan N, Vlodavsky I . Proteoglycans in health and disease: new concepts for heparanase function in tumor progression and metastasis. FEBS J 2010; 277: 3890–3903.
Ilan N, Elkin M, Vlodavsky I . Regulation, function and clinical significance of heparanase in cancer metastasis and angiogenesis. Intl J Biochem Cell Biol 2006; 38: 2018–2039.
Vlodavsky I, Ilan N, Naggi A, Casu B . Heparanase: structure, biological functions, and inhibition by heparin-derived mimetics of heparan sulfate. Curr Pharm Des 2007; 13: 2057–2073.
Vreys V, David G . Mammalian heparanase: what is the message? J Cell Mol Med 2007; 11: 427–452.
Cohen I, Pappo O, Elkin M, San T, Bar-Shavit R, Hazan R et al. Heparanase promotes growth, angiogenesis and survival of primary breast tumors. Intl J Cancer 2006; 118: 1609–1617.
Fux L, Ilan N, Sanderson RD, Vlodavsky I . Heparanase: busy at the cell surface. Trends Biochem Sci 2009; 34: 511–519.
Lerner I, Baraz L, Pikarsky E, Meirovitz A, Edovitsky E, Peretz T et al. Function of heparanase in prostate tumorigenesis: potential for therapy. Clin Cancer Res 2008; 14: 668–676.
Cassinelli G, Lanzi C, Tortoreto M, Cominetti D, Petrangolini G, Favini E et al. Antitumor efficacy of the heparanase inhibitor SST0001 alone and in combination with antiangiogenic agents in the treatment of human pediatric sarcoma models. Biochemical Pharmacol 2013; 85: 1424–1432.
Dredge K, Hammond E, Handley P, Gonda TJ, Smith MT, Vincent C et al. PG545, a dual heparanase and angiogenesis inhibitor, induces potent anti-tumour and anti-metastatic efficacy in preclinical models. Br J Cancer 2011; 104: 635–642.
Ritchie JP, Ramani VC, Ren Y, Naggi A, Torri G, Casu B et al. SST0001, a chemically modified heparin, inhibits myeloma growth and angiogenesis via disruption of the heparanase/syndecan-1 axis. Clin Cancer Res 2011; 17: 1382–1393.
Shafat I, Ben-Arush MW, Issakov J, Meller I, Naroditsky I, Tortoreto M et al. Pre-clinical and clinical significance of heparanase in Ewing’s sarcoma. J Cell Mol Med 2011; 15: 1857–1864.
Elkin M, Ilan N, Ishai-Michaeli R, Friedmann Y, Papo O, Pecker I et al. Heparanase as mediator of angiogenesis: mode of action. FASEB J 2001; 15: 1661–1663.
Folkman J, Klagsbrun M, Sasse J, Wadzinski M, Ingber D, Vlodavsky I . A heparin-binding angiogenic protein—basic fibroblast growth factor—is stored within basement membrane. Am J Pathol 1988; 130: 393–400.
Barash U, Cohen-Kaplan V, Arvatz G, Gingis-Velitski S, Levy-Adam F, Nativ O et al. A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics. FASEB J 2010; 24: 1239–1248.
Fux L, Feibish N, Cohen-Kaplan V, Gingis-Velitski S, Feld S, Geffen C et al. Structure-function approach identifies a COOH-terminal domain that mediates heparanase signaling. Cancer Res 2009; 69: 1758–1767.
Cohen-Kaplan V, Doweck I, Naroditsky I, Vlodavsky I, Ilan N . Heparanase augments epidermal growth factor receptor phosphorylation: correlation with head and neck tumor progression. Cancer Res 2008; 68: 10077–10085.
Cohen-Kaplan V, Jrbashyan J, Yanir Y, Naroditsky I, Ben-Izhak O, Ilan N et al. Heparanase induces signal transducer and activator of transcription (STAT) protein phosphorylation: preclinical and clinical significance in head and neck cancer. J Biol Chem 2012; 287: 6668–6678.
Riaz A, Ilan N, Vlodavsky I, Li JP, Johansson S . Characterization of heparanase-induced phosphatidylinositol 3-kinase-AKT activation and its integrin dependence. J Biol Chem 2013; 288: 12366–12375.
Cohen-Kaplan V, Naroditsky I, Zetser A, Ilan N, Vlodavsky I, Doweck I . Heparanase induces VEGF C and facilitates tumor lymphangiogenesis. Intl J Cancer 2008; 123: 2566–2573.
Nadir Y, Brenner B, Zetser A, Ilan N, Shafat I, Zcharia E et al. Heparanase induces tissue factor expression in vascular endothelial and cancer cells. J Thromb Haemost 2006; 4: 2443–2451.
Yang Y, Ren Y, Ramani VC, Nan L, Suva LJ, Sanderson RD . Heparanase enhances local and systemic osteolysis in multiple myeloma by upregulating the expression and secretion of RANKL. Cancer Res 2010; 70: 8329–8338.
Okawa T, Naomoto Y, Nobuhisa T, Takaoka M, Motoki T, Shirakawa Y et al. Heparanase is involved in angiogenesis in esophageal cancer through induction of cyclooxygenase-2. Clin Cancer Res 2005; 11: 7995–8005.
Ramani VC, Yang Y, Ren Y, Nan L, Sanderson RD . Heparanase plays a dual role in driving hepatocyte growth factor (HGF) signaling by enhancing HGF expression and activity. J Biol Chem 2011; 286: 6490–6499.
Nadav L, Katz BZ, Baron S, Cohen N, Naparstek E, Geiger B . The generation and regulation of functional diversity of malignant plasma cells. Cancer Res 2006; 66: 8608–8616.
Purushothaman A, Uyama T, Kobayashi F, Yamada S, Sugahara K, Rapraeger AC et al. Heparanase-enhanced shedding of syndecan-1 by myeloma cells promotes endothelial invasion and angiogenesis. Blood 2010; 115: 2449–2457.
Gingis-Velitski S, Zetser A, Flugelman MY, Vlodavsky I, Ilan N . Heparanase induces endothelial cell migration via protein kinase B/Akt activation. J Biol Chem 2004; 279: 23536–23541.
Beider K, Begin M, Abraham M, Wald H, Weiss ID, Wald O et al. CXCR4 antagonist 4F-benzoyl-TN14003 inhibits leukemia and multiple myeloma tumor growth. Exp Hematol 2011; 39: 282–292.
Zetser A, Bashenko Y, Edovitsky E, Levy-Adam F, Vlodavsky I, Ilan N . Heparanase induces vascular endothelial growth factor expression: correlation with p38 phosphorylation levels and Src activation. Cancer Res 2006; 66: 1455–1463.
Meiron M, Zohar Y, Anunu R, Wildbaum G, Karin N . CXCL12 (SDF-1alpha) suppresses ongoing experimental autoimmune encephalomyelitis by selecting antigen-specific regulatory T cells. J Exp Med 2008; 205: 2643–2655.
Vlodavsky I . Preparation of extracellular matrices produced by cultured corneal endothelial and PF-HR9 endodermal cells. In: Bonifacino MD JS, Hartford JB, Lippincott-Schwartz J, Yamada KM, (eds) Protocols in Cell Biology vol. 1. John Wiley & Sons: New York, pp 10.14.11–10.14.14 1999.
Barash U, Arvatz G, Farfara R, Naroditsky I, Doweck I, Feld S et al. Clinical significance of heparanase splice variant (t5) in renal cell carcinoma: evaluation by a novel t5-specific monoclonal antibody. PLoS One 2012; 7: e51494.
Sanderson RD, Yang Y, Kelly T, Macleod V, Dai Y, Theus A . Enzymatic remodeling of heparan sulfate proteoglycans within the tumor microenvironment: growth regulation and the prospect of new cancer therapies. J Cell Biochem 2005; 96: 897–905.
Arvatz G, Barash U, Nativ O, Ilan N, Vlodavsky I . Post-transcriptional regulation of heparanase gene expression by a 3′ AU-rich element. FASEB J 2011; 24: 4969–4976.
Miao HQ, Liu H, Navarro E, Kussie P, Zhu Z . Development of heparanase inhibitors for anti-cancer therapy. Curr Med Chem 2006; 13: 2101–2111.
Purushothaman A, Babitz SK, Sanderson RD . Heparanase enhances the insulin receptor signaling pathway to activate extracellular signal-regulated kinase in multiple myeloma. J Biol Chem 2012; 287: 41288–41296.
Purushothaman A, Chen L, Yang Y, Sanderson RD . Heparanase stimulation of protease expression implicates it as a master regulator of the aggressive tumor phenotype in myeloma. J Biol Chem 2008; 283: 32628–32636.
Ramani VC, Purushothaman A, Stewart MD, Thompson CA, Vlodavsky I, Au JL et al. The heparanase/syndecan-1 axis in cancer: mechanisms and therapies. FEBS J 2013; 280: 2294–2306.
Sanderson RD, Iozzo RV . Targeting heparanase for cancer therapy at the tumor-matrix interface. Matrix Biol 2012; 31: 283–284.
Sanderson RD, Yang Y . Syndecan-1: a dynamic regulator of the myeloma microenvironment. Clin Exp Metastasis 2008; 25: 149–159.
He YQ, Sutcliffe EL, Bunting KL, Li J, Goodall KJ, Poon IK et al. The endoglycosidase heparanase enters the nucleus of T lymphocytes and modulates H3 methylation at actively transcribed genes via the interplay with key chromatin modifying enzymes. Transcription 2012; 3: 130–145.
Li RW, Freeman C, Yu D, Hindmarsh EJ, Tymms KE, Parish CR et al. Dramatic regulation of heparanase activity and angiogenesis gene expression in synovium from patients with rheumatoid arthritis. Arthritis Rheum 2008; 58: 1590–1600.
Giuliani N, Bonomini S, Romagnani P, Lazzaretti M, Morandi F, Colla S et al. CXCR3 and its binding chemokines in myeloma cells: expression of isoforms and potential relationships with myeloma cell proliferation and survival. Haematologica 2006; 91: 1489–1497.
Liu M, Guo S, Stiles JK . The emerging role of CXCL10 in cancer (Review). Oncol Lett 2011; 2: 583–589.
Rosenkilde MM, Schwartz TW . The chemokine system - a major regulator of angiogenesis in health and disease. APMIS 2004; 112: 481–495.
Bodnar RJ, Yates CC, Rodgers ME, Du X, Wells A . IP-10 induces dissociation of newly formed blood vessels. J Cell Sci 2009; 122: 2064–2077.
Luster AD, Greenberg SM, Leder P . The IP-10 chemokine binds to a specific cell surface heparan sulfate site shared with platelet factor 4 and inhibits endothelial cell proliferation. J Exp Med 1995; 182: 219–231.
Feldman AL, Friedl J, Lans TE, Libutti SK, Lorang D, Miller MS et al. Retroviral gene transfer of interferon-inducible protein 10 inhibits growth of human melanoma xenografts. Intl J Cancer 2002; 99: 149–153.
Man K, Ng KT, Xu A, Cheng Q, Lo CM, Xiao JW et al. Suppression of liver tumor growth and metastasis by adiponectin in nude mice through inhibition of tumor angiogenesis and downregulation of Rho kinase/IFN-inducible protein 10/matrix metalloproteinase 9 signaling. Clin Cancer Res 2010; 16: 967–977.
Sun Y, Finger C, Alvarez-Vallina L, Cichutek K, Buchholz CJ . Chronic gene delivery of interferon-inducible protein 10 through replication-competent retrovirus vectors suppresses tumor growth. Cancer Gene Ther 2005; 12: 900–912.
Tannenbaum CS, Tubbs R, Armstrong D, Finke JH, Bukowski RM, Hamilton TA . The CXC chemokines IP-10 and Mig are necessary for IL-12-mediated regression of the mouse RENCA tumor. J Immunol 1998; 161: 927–932.
Sgadari C, Angiolillo AL, Cherney BW, Pike SE, Farber JM, Koniaris LG et al. Interferon-inducible protein-10 identified as a mediator of tumor necrosis in vivo. Proc Natl Acad Sci USA 1996; 93: 13791–13796.
Arenberg DA, White ES, Burdick MD, Strom SR, Strieter RM . Improved survival in tumor-bearing SCID mice treated with interferon-gamma-inducible protein 10 (IP-10/CXCL10). Cancer Immunol Immunother 2001; 50: 533–538.
Lamy L, Ngo VN, Emre NC, Shaffer AL 3rd, Yang Y, Tian E et al. Control of autophagic cell death by caspase-10 in multiple myeloma. Cancer Cell 2013; 23: 435–449.
Veitonmaki N, Hansson M, Zhan F, Sundberg A, Lofstedt T, Ljungars A et al. A human ICAM-1 antibody isolated by a function-first approach has potent macrophage-dependent antimyeloma activity in vivo. Cancer Cell 2013; 23: 502–515.
Acknowledgements
We acknowledge the devoted help of Dr Liat Linde and Dr Boaz Kigel (Rappaport Faculty of Medicine) in performing the gene array methodology and purification of the CXCL10–Ig fusion protein, respectively. This study was supported (in part) by research funding from the Israel Science Foundation (grant 593/10); National Cancer Institute, NIH (grant CA106456); the Israel Cancer Research Fund (ICRF); and the Rappaport Family Institute Fund to I Vlodavsky. I Vlodavsky is a research professor of the ICRF.
Author contributions
UB, GW and KB designed and performed experiments, analyzed and interpreted data; YZ, NK and AN provided valuable reagents, designed, analyzed and interpreted data; NI co-directed the study, designed, analyzed and interpreted data and wrote the manuscript; IV directed the study, designed, analyzed and interpreted data and co-wrote the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Leukemia website
Rights and permissions
About this article
Cite this article
Barash, U., Zohar, Y., Wildbaum, G. et al. Heparanase enhances myeloma progression via CXCL10 downregulation. Leukemia 28, 2178–2187 (2014). https://doi.org/10.1038/leu.2014.121
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/leu.2014.121