Top

Published in:

01-10-2011 | Original article

Ectonucleotidases CD39 and CD73 on OvCA cells are potent adenosine-generating enzymes responsible for adenosine receptor 2A-dependent suppression of T cell function and NK cell cytotoxicity

Authors: Sebastian F. M. Häusler, Itsaso Montalbán del Barrio, Jenny Strohschein, P. Anoop Chandran, Jörg B. Engel, Arnd Hönig, Monika Ossadnik, Evi Horn, Birgitt Fischer, Mathias Krockenberger, Stefan Heuer, Ahmed Adel Seida, Markus Junker, Hermann Kneitz, Doris Kloor, Karl-Norbert Klotz, Johannes Dietl, Jörg Wischhusen

Published in: Cancer Immunology, Immunotherapy | Issue 10/2011

Abstract

The ectonucleotidases CD39 and CD73 degrade immune stimulatory ATP to adenosine that inhibits T and NK cell responses via the A_2A adenosine receptor (ADORA2A). This mechanism is used by regulatory T cells (T_reg) that are associated with increased mortality in OvCA. Immunohistochemical staining of human OvCA tissue specimens revealed further aberrant expression of CD39 in 29/36 OvCA samples, whereas only 1/9 benign ovaries showed weak stromal CD39 expression. CD73 could be detected on 31/34 OvCA samples. While 8/9 benign ovaries also showed CD73 immunoreactivity, expression levels were lower than in tumour specimens. Infiltration by CD4⁺ and CD8⁺ T cells was enhanced in tumour specimens and significantly correlated with CD39 and CD73 levels on stromal, but not on tumour cells. In vitro, human OvCA cell lines SK-OV-3 and OaW42 as well as 11/15 ascites-derived primary OvCA cell cultures expressed both functional CD39 and CD73 leading to more efficient depletion of extracellular ATP and enhanced generation of adenosine as compared to activated T_reg. Functional assays using siRNAs against CD39 and CD73 or pharmacological inhibitors of CD39, CD73 and ADORA2A revealed that tumour-derived adenosine inhibits the proliferation of allogeneic human CD4⁺ T cells in co-culture with OvCA cells as well as cytotoxic T cell priming and NK cell cytotoxicity against SK-OV3 or OAW42 cells. Thus, both the ectonucleotidases CD39 and CD73 and ADORA2A appear as possible targets for novel treatments in OvCA, which may not only affect the function of T_reg but also relieve intrinsic immunosuppressive properties of tumour and stromal cells.

Pectasides D, Pectasides E (2006) Maintenance or consolidation therapy in advanced ovarian cancer. Oncology 70(5):315–324. doi:10.1159/000097943 PubMedCrossRef

Sugiyama T, Konishi I (2008) Emerging drugs for ovarian cancer. Expert Opin Emerg Drugs 13(3):523–536. doi:10.1517/14728214.2010.502888 PubMedCrossRef

Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, Evdemon-Hogan M, Conejo-Garcia JR, Zhang L, Burow M, Zhu Y, Wei S, Kryczek I, Daniel B, Gordon A, Myers L, Lackner A, Disis ML, Knutson KL, Chen L, Zou W (2004) Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med 10(9):942–949. doi:10.1038/nm1093 PubMedCrossRef

Blay J, White TD, Hoskin DW (1997) The extracellular fluid of solid carcinomas contains immunosuppressive concentrations of adenosine. Cancer Res 57(13):2602–2605PubMed

Ohta A, Gorelik E, Prasad SJ, Ronchese F, Lukashev D, Wong MK, Huang X, Caldwell S, Liu K, Smith P, Chen JF, Jackson EK, Apasov S, Abrams S, Sitkovsky M (2006) A2A adenosine receptor protects tumors from antitumor T cells. Proc Natl Acad Sci USA 103(35):13132–13137. doi:10.1073/pnas.0605251103 PubMedCrossRef

Hasko G, Cronstein BN (2004) Adenosine: an endogenous regulator of innate immunity. Trends Immunol 25(1):33–39. doi:10.1016/j.it.2003.11.003 PubMedCrossRef

Borsellino G, Kleinewietfeld M, Di Mitri D, Sternjak A, Diamantini A, Giometto R, Hopner S, Centonze D, Bernardi G, Dell’Acqua ML, Rossini PM, Battistini L, Rötzschke O, Falk K (2007) Expression of ectonucleotidase CD39 by Foxp3⁺ Treg cells: hydrolysis of extracellular ATP and immune suppression. Blood 110(4):1225–1232. doi:10.1182/blood-2006-12-064527 PubMedCrossRef

Deaglio S, Dwyer KM, Gao W, Friedman D, Usheva A, Erat A, Chen JF, Enjyoji K, Linden J, Oukka M, Kuchroo VK, Strom TB, Robson SC (2007) Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory T cells mediates immune suppression. J Exp Med 204(6):1257–1265. doi:10.1084/jem.20062512 PubMedCrossRef

Resta R, Yamashita Y, Thompson LF (1998) Ecto-enzyme and signaling functions of lymphocyte CD73. Immunol Rev 161:95–109. doi:10.1111/j.1600-065X.1998.tb01574.x PubMedCrossRef

10.

Csoka B, Himer L, Selmeczy Z, Vizi ES, Pacher P, Ledent C, Deitch EA, Spolarics Z, Nemeth ZH, Hasko G (2008) Adenosine A2A receptor activation inhibits T helper 1 and T helper 2 cell development and effector function. FASEB J 22(10):3491–3499PubMedCrossRef

11.

Elliott MR, Chekeni FB, Trampont PC, Lazarowski ER, Kadl A, Walk SF, Park D, Woodson RI, Ostankovich M, Sharma P, Lysiak JJ, Harden TK, Leitinger N, Ravichandran KS (2009) Nucleotides released by apoptotic cells act as a find-me signal to promote phagocytic clearance. Nature 461(7261):282–286PubMedCrossRef

12.

Aymeric L, Apetoh L, Ghiringhelli F, Tesniere A, Martins I, Kroemer G, Smyth MJ, Zitvogel L (2010) Tumor cell death and ATP release prime dendritic cells and efficient anticancer immunity. Cancer Res 70(3):855–858. doi:10.1158/0008-5472.CAN-09-3566 PubMedCrossRef

13.

Künzli BM, Berberat PO, Giese T, Csizmadia E, Kaczmarek E, Baker C, Halaceli I, Büchler MW, Friess H, Robson SC (2007) Upregulation of CD39/NTPDases and P2 receptors in human pancreatic disease. Am J Physiol Gastrointest Liver Physiol 292(1):G223–G230. doi:10.1152/ajpgi.00259.2006 PubMedCrossRef

14.

Dzhandzhugazyan KN, Kirkin AF, Thor Straten P, Zeuthen J (1998) Ecto-ATP diphosphohydrolase/CD39 is overexpressed in differentiated human melanomas. FEBS Lett 430(3):227–230PubMedCrossRef

15.

Shi XJ, Knowles AF (1994) Prevalence of the mercurial-sensitive EctoATPase in human small cell lung carcinoma: characterization and partial purification. Arch Biochem Biophys 315(1):177–184PubMedCrossRef

16.

Morrone FB, Oliveira DL, Gamermann P, Stella J, Wofchuk S, Wink MR, Meurer L, Edelweiss MI, Lenz G, Battastini AM (2006) In vivo glioblastoma growth is reduced by apyrase activity in a rat glioma model. BMC Cancer 6:226PubMedCrossRef

17.

Braganhol E, Morrone FB, Bernardi A, Huppes D, Meurer L, Edelweiss MI, Lenz G, Wink MR, Robson SC, Battastini AM (2009) Selective NTPDase2 expression modulates in vivo rat glioma growth. Cancer Sci 100(8):1434–1442PubMedCrossRef

18.

Zhou X, Zhi X, Zhou P, Chen S, Zhao F, Shao Z, Ou Z, Yin L (2007) Effects of ecto-5′-nucleotidase on human breast cancer cell growth in vitro and in vivo. Oncol Rep 17(6):1341–1346PubMed

19.

Stagg J, Divisekera U, McLaughlin N, Sharkey J, Pommey S, Denoyer D, Dwyer KM, Smyth MJ (1547) Anti-CD73 antibody therapy inhibits breast tumor growth and metastasis. Proc Natl Acad Sci USA 107(4):1547–1552. doi:10.1073/pnas.0908801107 CrossRef

20.

Wang L, Zhou X, Zhou T, Ma D, Chen S, Zhi X, Yin L, Shao Z, Ou Z, Zhou P (2008) Ecto-5′-nucleotidase promotes invasion, migration and adhesion of human breast cancer cells. J Cancer Res Clin Oncol 134(3):365–372. doi:10.1007/s00432-007-0292-z PubMedCrossRef

21.

Jin D, Fan J, Wang L, Thompson LF, Liu A, Daniel BJ, Shin T, Curiel TJ, Zhang B (2010) CD73 on tumor cells impairs antitumor T-Cell responses: a novel mechanism of tumor-induced immune suppression. Cancer Res 70(6):2245–2255

22.

Panjehpour M, Castro M, Klotz KN (2005) Human breast cancer cell line MDA-MB-231 expresses endogenous A2B adenosine receptors mediating a Ca2+ signal. Br J Pharmacol 145(2):211–218PubMedCrossRef

23.

Krockenberger M, Dombrowski Y, Weidler C, Ossadnik M, Honig A, Häusler S, Voigt H, Becker JC, Leng L, Steinle A, Weller M, Bucala R, Dietl J, Wischhusen J (2008) Macrophage migration inhibitory factor contributes to the immune escape of ovarian cancer by down-regulating NKG2D. J Immunol 180(11):7338–7348PubMed

24.

Aversa GG, Suranyi MG, Waugh JA, Bishop AG, Hall BM (1988) Detection of a late lymphocyte activation marker by A1, a new monoclonal antibody. Transplant Proc 20(1):49–52PubMed

25.

Dörken B, Möller P, Pezzuto A, Schwartz-Albiez R, Moldenhauer G (1989) Part I: B-cell antigens. In: Knapp W, Dörken B, Gilks WR et al (eds) Leukocyte typing IV: white cell differentiation antigens. Oxford University Press, New York

26.

Buira SP, Albasanz JL, Dentesano G, Moreno J, Martin M, Ferrer I, Barrachina M (2010) DNA methylation regulates adenosine A(2A) receptor cell surface expression levels. J Neurochem 112(5):1273–1285PubMedCrossRef

27.

Neufeld HA, Towner RD, Pace J (1975) A rapid method for determining ATP by the firefly luciferin-luciferase system. Experientia 31(3):391–392PubMedCrossRef

28.

Häusler SF, Ossadnik M, Horn E, Heuer S, Dietl J, Wischhusen J (2010) A cell-based luciferase-dependent assay for the quantitative determination of free extracellular adenosine with paracrine signaling activity. J Immunol Methods 361(1–2):51–56PubMedCrossRef

29.

Dyer BW, Ferrer FA, Klinedinst DK, Rodriguez R (2000) A noncommercial dual luciferase enzyme assay system for reporter gene analysis. Anal Biochem 282(1):158–161. doi:10.1006/abio.2000.4605 PubMedCrossRef

30.

Crack BE, Pollard CE, Beukers MW, Roberts SM, Hunt SF, Ingall AH, McKechnie KC, Ijzerman AP, Leff P (1995) Pharmacological and biochemical analysis of FPL 67156, a novel, selective inhibitor of ecto-ATPase. Br J Pharmacol 114(2):475–481PubMed

31.

Krug F, Parikh I, Illiano G, Cuatrecasas P (1973) α,β-methylene-adenosine 5′-triphosphate. A competitive inhibitor of adenylate cyclase in fat and liver cell membranes. J Biol Chem 248(4):1203–1206PubMed

32.

Ongini E, Dionisotti S, Gessi S, Irenius E, Fredholm BB (1999) Comparison of CGS 15943, ZM 241385 and SCH 58261 as antagonists at human adenosine receptors. Naunyn Schmiedebergs Arch Pharmacol 359(1):7–10PubMedCrossRef

33.

Brown CE, Wright CL, Naranjo A, Vishwanath RP, Chang WC, Olivares S, Wagner JR, Bruins L, Raubitschek A, Cooper LJ, Jensen MC (2005) Biophotonic cytotoxicity assay for high-throughput screening of cytolytic killing. J Immunol Methods 297(1–2):39–52. doi:10.1016/j.jim.2004.11.021 PubMedCrossRef

34.

Hoskin DW, Mader JS, Furlong SJ, Conrad DM, Blay J (2008) Inhibition of T cell and natural killer cell function by adenosine and its contribution to immune evasion by tumor cells (review). Int J Oncol 32(3):527–535PubMed

35.

Shi J, Wan Y, Di W (2008) Effect of hypoxia and re-oxygenation on cell invasion and adhesion in human ovarian carcinoma cells. Oncol Rep 20(4):803–807. doi:10.3892/or_00000077 PubMed

36.

Zhang L, Conejo-Garcia JR, Katsaros D, Gimotty PA, Massobrio M, Regnani G, Makrigiannakis A, Gray H, Schlienger K, Liebman MN, Rubin SC, Coukos G (2003) Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 348(3):203–213. doi:10.1056/NEJMoa020177 PubMedCrossRef

37.

Havre PA, Abe M, Urasaki Y, Ohnuma K, Morimoto C, Dang NH (2008) The role of CD26/dipeptidyl peptidase IV in cancer. Front Biosci 13:1634–1645. doi:10.2741/2787 PubMedCrossRef

38.

Eltzschig HK, Abdulla P, Hoffman E, Hamilton KE, Daniels D, Schönfeld C, Löffler M, Reyes G, Duszenko M, Karhausen J, Robinson A, Westerman KA, Coe IR, Colgan SP (2005) HIF-1-dependent repression of equilibrative nucleoside transporter (ENT) in hypoxia. J Exp Med 202(11):1493–1505PubMedCrossRef

39.

Mandapathil M, Hilldorfer B, Szczepanski MJ, Czystowska M, Szajnik M, Ren J, Lang S, Jackson EK, Gorelik E, Whiteside TL (2010) Generation and accumulation of immunosuppressive adenosine by human CD4⁺CD25^highFOXP3⁺ regulatory T cells. J Biol Chem 285(10):7176–7186PubMedCrossRef

40.

Mandapathil M, Szczepanski MJ, Szajnik M, Ren J, Lenzner DE, Jackson EK, Gorelik E, Lang S, Johnson JT, Whiteside TL (2009) Increased ectonucleotidase expression and activity in regulatory T cells of patients with head and neck cancer. Clin Cancer Res 15(20):6348–6357. doi:1078-0432.CCR-09-1143 PubMedCrossRef

41.

Eltzschig HK, Ibla JC, Furuta GT, Leonard MO, Jacobson KA, Enjyoji K, Robson SC, Colgan SP (2003) Coordinated adenine nucleotide phosphohydrolysis and nucleoside signaling in posthypoxic endothelium: role of ectonucleotidases and adenosine A2B receptors. J Exp Med 198(5):783–796. doi:10.1084/jem.20030891 PubMedCrossRef

42.

Enjyoji K, Kotani K, Thukral C, Blumel B, Sun X, Wu Y, Imai M, Friedman D, Csizmadia E, Bleibel W, Kahn BB, Robson SC (2008) Deletion of CD39/ENTPD1 results in hepatic insulin resistance. Diabetes 57(9):2311–2320PubMedCrossRef

43.

Synnestvedt K, Furuta GT, Comerford KM, Louis N, Karhausen J, Eltzschig HK, Hansen KR, Thompson LF, Colgan SP (2002) Ecto-5′-nucleotidase (CD73) regulation by hypoxia-inducible factor-1 mediates permeability changes in intestinal epithelia. J Clin Invest 110(7):993–1002. doi:10.1172/JCI15337 PubMed

44.

Tanganelli S, Sandager Nielsen K, Ferraro L, Antonelli T, Kehr J, Franco R, Ferre S, Agnati LF, Fuxe K, Scheel-Krüger J (2004) Striatal plasticity at the network level. Focus on adenosine A2A and D2 interactions in models of Parkinson’s disease. Parkinsonism Relat Disord 10(5):273–280. doi:10.1016/j.parkreldis.2004.02.015 PubMedCrossRef

45.

Sheehy ME, McDermott AB, Furlan SN, Klenerman P, Nixon DF (2001) A novel technique for the fluorometric assessment of T lymphocyte antigen specific lysis. J Immunol Methods 249(1–2):99–110. doi:10.1016/S0022-1759(00)00329-X PubMedCrossRef

Title: Ectonucleotidases CD39 and CD73 on OvCA cells are potent adenosine-generating enzymes responsible for adenosine receptor 2A-dependent suppression of T cell function and NK cell cytotoxicity
Authors: Sebastian F. M. Häusler
Itsaso Montalbán del Barrio
Jenny Strohschein
P. Anoop Chandran
Jörg B. Engel
Arnd Hönig
Monika Ossadnik
Evi Horn
Birgitt Fischer
Mathias Krockenberger
Stefan Heuer
Ahmed Adel Seida
Markus Junker
Hermann Kneitz
Doris Kloor
Karl-Norbert Klotz
Johannes Dietl
Jörg Wischhusen
Publication date: 01-10-2011
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 10/2011
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-011-1040-4

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 10/2011

Elevated myeloid-derived suppressor cells in pancreatic, esophageal and gastric cancer are an independent prognostic factor and are associated with significant elevation of the Th2 cytokine interleukin-13

Dendritic cells combining with cytokine-induced killer cells synergize chemotherapy in patients with late-stage non-small cell lung cancer

Serum levels of cytoplasmic melanoma-associated antigen at diagnosis may predict clinical relapse in neuroblastoma patients

An ADP ribosylation factor-GTPase activating protein negatively regulates the production of proinflammatory mediators in response to lipopolysaccharide

The n3-polyunsaturated fatty acid docosahexaenoic acid induces immunogenic cell death in human cancer cell lines via pre-apoptotic calreticulin exposure

Th17 and Th17-stimulated CD8+ T cells play a distinct role in Th17-induced preventive and therapeutic antitumor immunity

Keynote webinar | Spotlight on antibody–drug conjugates in cancer