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Published in: BMC Cancer 1/2015

Open Access 01-12-2015 | Research article

Liposome encapsulated zoledronate favours M1-like behaviour in murine macrophages cultured with soluble factors from breast cancer cells

Authors: Sofia Sousa, Seppo Auriola, Jukka Mönkkönen, Jorma Määttä

Published in: BMC Cancer | Issue 1/2015

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Abstract

Background

Tumour stromal macrophages differentiate to tumour-associated macrophages (TAMs) with characteristics of immunosuppressive M2-type macrophages, having a central role in promoting tumour vascularisation, cancer cell dissemination and in suppressing anti-cancer immune responses. Bisphosphonates (BPs) are a group of drugs commonly used as anti-resorptive agents. Further, nitrogen containing BPs like Zoledronate (ZOL), are known to cause unspecific inflammatory reactions hence the hypothesis that its use could modulate TAMs polarization toward a more inflammatory phenotype.

Methods

We studied the in vitro polarization of J774 murine macrophages upon culture in 4T1 breast cancer cell-conditioned medium (4T1CM) and stimulation with LPS and free and liposome-encapsulated bisphosphonates.

Results

In this system, breast cancer soluble factors reduced the pro-inflammatory activation of macrophages but increased the secretion of matrix metalloproteinases (MMPs). In the presence of 4T1CM, a non-cytotoxic dose of liposome-encapsulated ZOL (ZOL-LIP) enhanced the expression of iNOS and TNF-α, markers of M1 activation, but did not diminish the expression of M2-type markers. In contrast, clodronate treatment either as a free drug (CLO) or liposome-encapsulated (CLO-LIP) decreased the expression of the M1-type markers and was highly cytotoxic to the macrophages.

Conclusions

Breast cancer cells soluble factors modulate macrophages toward M2 activation state. Bisphosphonates may be applied to counteract this modulation. We propose that ZOL-LIP may be suitable for favouring cytotoxic immune responses by TAMs in breast cancer, whereas CLO-LIP may be appropriate for TAM depletion.
Literature
1.
Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M. The chemokine system in diverse forms of macrophage activation and polarization. Trends Immunol. 2004;25(12):677–86.PubMedCrossRef
2.
Mantovani A, Sica A. Macrophages, innate immunity and cancer: balance, tolerance, and diversity. Curr Opin Immunol. 2010;22(2):231–7.PubMedCrossRef
3.
Wang T, Ge Y, Xiao M, Lopez-Coral A, Azuma R, Somasundaram R, et al. Melanoma-derived conditioned media efficiently induce the differentiation of monocytes to macrophages that display a highly invasive gene signature. Pigm Cell Melanoma Res. 2012;25(4):493–505.CrossRef
4.
Klug F, Prakash H, Huber P, Seibel T, Bender N, Halama N, et al. Low-dose irradiation programs macrophage differentiation to an iNOS+/M1 phenotype that orchestrates effective T cell immunotherapy. Cancer Cell. 2013;24(5):589–602.PubMedCrossRef
5.
Guiducci C, Vicari AP, Sangaletti S, Trinchieri G, Colombo MP. Redirecting in vivo elicited tumor infiltrating macrophages and dendritic cells towards tumor rejection. Cancer Res. 2005;65(8):3437–46.PubMed
6.
Klimp AH, de Vries EGE, Scherphof GL, Daemen T. A potential role of macrophage activation in the treatment of cancer. Crit Rev Oncol. 2002;44(2):143–61.CrossRef
7.
Mantovani A, Schioppa T, Porta C, Allavena P, Sica A. Role of tumor-associated macrophages in tumor progression and invasion. Cancer Metastasis Rev. 2006;25(3):315–22.PubMedCrossRef
8.
Mantovani A, Sozzani S, Locati M, Allavena P, Sica A. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol. 2002;23(11):549–55.PubMedCrossRef
9.
Su S, Liu Q, Chen J, Chen J, Chen F, He C, et al. A Positive feedback loop between mesenchymal-like cancer cells and macrophages is essential to breast cancer metastasis. Cancer Cell. 2014;25(5):605–20.PubMedCrossRef
10.
Pollard JW. Tumour-educated macrophages promote tumour progression and metastasis. Nat Rev Cancer. 2004;4(1):71–8.PubMedCrossRef
11.
Rogers TL, Holen I. Tumour macrophages as potential targets of bisphosphonates. J Transl Med. 2011;9:177-5876-9-177.CrossRef
12.
Thompson K, Rogers MJ, Coxon FP, Crockett JC. Cytosolic entry of bisphosphonate drugs requires acidification of vesicles after fluid-phase endocytosis. Mol Pharmacol. 2006;69(5):1624–32.PubMedCrossRef
13.
Rogers MJ, Crockett JC, Coxon FP, Mönkkönen J. Biochemical and molecular mechanisms of action of bisphosphonates. Bone. 2011;49(1):34–41.PubMedCrossRef
14.
Auriola S, Frith J, Rogers MJ, Koivuniemi A, Mönkkönen J. Identification of adenine nucleotide-containing metabolites of bisphosphonate drugs using ion-pair liquid chromatography-electrospray mass spectrometry. J Chromatogr B Biomed Appl. 1997;704(1–2):187–95.CrossRef
15.
Mitrofan LM, Pelkonen J, Mönkkönen J. The level of ATP analog and isopentenyl pyrophosphate correlates with zoledronic acid-induced apoptosis in cancer cells in vitro. Bone. 2009;45(6):1153–60.PubMedCrossRef
16.
Roelofs AJ, Jauhiainen M, Mönkkönen H, Rogers MJ, Mönkkönen J, Thompson K. Peripheral blood monocytes are responsible for γδ T cell activation induced by zoledronic acid through accumulation of IPP/DMAPP. Br J Haematol. 2009;144(2):245–50.PubMedPubMedCentralCrossRef
17.
Mönkkönen H, Auriola S, Lehenkari P, Kellinsalmi M, Hassinen IE, Vepsäläinen J, et al. A new endogenous ATP analog (ApppI) inhibits the mitochondrial adenine nucleotide translocase (ANT) and is responsible for the apoptosis induced by nitrogen-containing bisphosphonates. Br J Pharmacol. 2006;147(4):437–45.PubMedPubMedCentralCrossRef
18.
Melani C, Sangaletti S, Barazzetta FM, Werb Z, Colombo MP. Amino-biphosphonate-mediated MMP-9 inhibition breaks the tumor-bone marrow axis responsible for myeloid-derived suppressor cell expansion and macrophage infiltration in tumor stroma. Cancer Res. 2007;67(23):11438–46.PubMedPubMedCentralCrossRef
19.
Coscia M, Quaglino E, Iezzi M, Curcio C, Pantaleoni F, Riganti C, et al. Zoledronic acid repolarizes tumour-associated macrophages and inhibits mammary carcinogenesis by targeting the mevalonate pathway. J Cell Mol Med. 2010;14(12):2803–15.PubMedCrossRef
20.
Mönkkönen J, Valjakka R, Hakasalo M, Urtti A. Optimization of liposome formulation for the delivery of clodronate to cells in vitro. Proc Control Release Soc. 1994;21:224–5.
21.
Bartlett GR. Colorimetric assay methods for free and phosphorylated glyceric acids. J Biol Chem. 1959;234(3):469–71.PubMed
22.
Nargi FE, Yang TJ. Optimization of the L-M cell bioassay for quantitating tumor necrosis factor alpha in serum and plasma. J Immunol Methods. 1993;159(1–2):81–91.PubMedCrossRef
23.
24.
Mönkkönen H, Moilanen P, Mönkkönen J, Frith JC, Rogers MJ, Auriola S. Analysis of an adenine nucleotide-containing metabolite of clodronate using ion pair high-performance liquid chromatography-electrospray ionisation mass spectrometry. J Chromatogr B Biomed Sci Appl. 2000;738(2):395–403.PubMedCrossRef
25.
Räikkönen J, Mönkkönen H, Auriola S, Mönkkönen J. Mevalonate pathway intermediates downregulate zoledronic acid-induced isopentenyl pyrophosphate and ATP analog formation in human breast cancer cells. Biochem Pharmacol. 2010;79(5):777–83.PubMedCrossRef
26.
Anne-Valérie R, Christelle D, Yannick F, Norbert P, Marc P, Dominique H. Human growth hormone stimulates proteinase activities of rabbit bone cells via IGF-I. Biochem Biophys Res Commun. 2000;268(3):875–81.PubMedCrossRef
27.
Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000;132:365–86.PubMed
28.
Muller PY, Janovjak H, Miserez AR, Dobbie Z. Processing of gene expression data generated by quantitative real-time RT-PCR. Biotechniques. 2002;32(6):1372. -4, 1376, 1378-9.PubMed
29.
Frith JC, Mönkkönen J, Blackburn GM, Russell RGG, Rogers MJ. Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5′-(β, γ-dichloromethylene) triphosphate, by mammalian cells in vitro. J Bone Miner Res. 1997;12(9):1358–67.PubMedCrossRef
30.
MacMicking J, Xie Q, Nathan C. Nitric oxide and macrophage function. Annu Rev Immunol. 1997;15:323–50.PubMedCrossRef
31.
Ellies LG, Fishman M, Hardison J, Kleeman J, Maglione JE, Manner CK, et al. Mammary tumor latency is increased in mice lacking the inducible nitric oxide synthase. Int J Cancer. 2003;106(1):1–7.PubMedCrossRef
32.
Lin C, Shen S, Ko C, Lin H, Chen Y. Reciprocal activation of macrophages and breast carcinoma cells by nitric oxide and colony-stimulating factor-1. Carcinogenesis. 2010;31(12):2039–48.PubMedCrossRef
33.
Gallet M, Mentaverri R, Sévenet N, Brazier M, Kamel S. Ability of breast cancer cell lines to stimulate bone resorbing activity of mature osteoclasts correlates with an anti-apoptotic effect mediated by macrophage colony stimulating factor. Apoptosis. 2006;11(11):1909–21.PubMedCrossRef
34.
Mancino AT, Klimberg VS, Yamamoto M, Manolagas SC, Abe E. Breast cancer increases osteoclastogenesis by secreting M-CSF and upregulating RANKL in stromal cells. J Surg Res. 2001;100(1):18–24.PubMedCrossRef
35.
Goswami S, Sahai E, Wyckoff JB, Cammer M, Cox D, Pixley FJ, et al. Macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop. Cancer Res. 2005;65(12):5278–83.PubMedCrossRef
36.
Neumark E, Sagi-Assif O, Shalmon B, Ben-Baruch A, Witz IP. Progression of mouse mammary tumors: MCP-1-TNFα cross-regulatory pathway and clonal expression of promalignancy and antimalignancy factors. Int J Cancer. 2003;106(6):879–86.PubMedCrossRef
37.
Hagemann T, Robinson SC, Schulz M, Trümper L, Balkwill FR, Binder C. Enhanced invasiveness of breast cancer cell lines upon co-cultivation with macrophages is due to TNF-α dependent up-regulation of matrix metalloproteases. Carcinogenesis. 2004;25(8):1543–9.PubMedCrossRef
38.
Barbera-Guillem E, Nyhus JK, Wolford CC, Friece CR, Sampsel JW. Vascular endothelial growth factor secretion by tumor-infiltrating macrophages essentially supports tumor angiogenesis, and IgG immune complexes potentiate the process. Cancer Res. 2002;62(23):7042–9.PubMed
39.
Randow F, Syrbe U, Meisel C, Krausch D, Zuckermann H, Platzer C, et al. Mechanism of endotoxin desensitization: involvement of interleukin 10 and transforming growth factor beta. J Exp Med. 1995;181(5):1887–92.PubMedCrossRef
40.
Pennanen N, Lapinjoki S, Urtti A, Mönkkönen J. Effect of liposomal and free bisphosphonates on the IL-1β, IL-6 and TNFα secretion from RAW 264 cells in vitro. Pharm Res. 1995;12(6):916–22.PubMedCrossRef
41.
Mönkkönen J, Similä J, Rogers MJ. Effects of tiludronate and ibandronate on the secretion of proinflammatory cytokines and nitric oxide from macrophages in vitro. Life Sci. 1998;62(8):L95-PL102.CrossRef
42.
Makkonen N, Hirvonen M, Teräväinen T, Savolainen K, Mönkkönen J. Different effects of three bisphosphonates on nitric oxide production by RAW 264 macrophage-like cells in vitro. J Pharmacol Exp Ther. 1996;277(2):1097–102.PubMed
43.
Thiebaud D, Sauty A, Burckhardt P, Leuenberger P, Sitzler L, Green JR, et al. An in vitro and in vivo study of cytokines in the acute-phase response associated with bisphosphonates. Calcif Tissue Int. 1997;61(5):386–92.PubMedCrossRef
44.
Töyräs A, Ollikainen J, Taskinen M, Mönkkönen J. Inhibition of mevalonate pathway is involved in alendronate-induced cell growth inhibition, but not in cytokine secretion from macrophages in vitro. Eur J Pharm Sci. 2003;19(4):223–30.PubMedCrossRef
45.
Takagi K, Takagi M, Kanangat S, Warrington KJ, Shigemitsu H, Postlethwaite AE. Modulation of TNF-α gene expression by IFN-γ and pamidronate in murine macrophages: regulation by STAT1-dependent pathways. J Immunol. 2005;174(4):1801–10.PubMedCrossRef
46.
Carbone LD, Warrington KJ, Barrow KD, Pugazhenthi M, Watsky MA, Somes G, et al. Pamidronate infusion in patients with systemic sclerosis results in changes in blood mononuclear cell cytokine profiles. Clin Exp Immunol. 2006;146(3):371–80.PubMedPubMedCentralCrossRef
47.
Makkonen N, Salminen A, Rogers MJ, Frith JC, Urtti A, Azhayeva E, et al. Contrasting effects of alendronate and clodronate on RAW 264 macrophages: the role of a bisphosphonate metabolite. Eur J Pharm Sci. 1999;8(2):109–18.PubMedCrossRef
48.
Senzer N, Mani S, Rosemurgy A, Nemunaitis J, Cunningham C, Guha C, et al. TNFerade biologic, an Adenovector with a radiation-inducible promoter, carrying the human tumor necrosis factor alpha gene: a phase I study in patients with solid tumors. J Clin Oncol. 2004;22(4):592–601.PubMedCrossRef
49.
Valleala H, Hanemaaijer R, Mandelin J, Salminen A, Teronen O, Mönkkönen J, et al. Regulation of MMP-9 (gelatinase B) in activated human monocyte/macrophages by two different types of bisphosphonates. Life Sci. 2003;73(19):2413–20.PubMedCrossRef
50.
Teronen O, Heikkilä P, Konttinen YT, Laitinen M, Salo T, Hanemaaijer R, et al. MMP inhibition and downregulation by bisphosphonates. Ann N Y Acad Sci. 1999;878(1):453–65.PubMedCrossRef
51.
Leifler KS, Svensson S, Abrahamsson A, Bendrik C, Robertson J, Gauldie J, et al. Inflammation induced by mmp-9 enhances tumor regression of experimental breast cancer. J Immunol. 2013;190(8):4420–30.PubMedPubMedCentralCrossRef
52.
Wyckoff JB, Wang Y, Lin EY, Li J, Goswami S, Stanley ER, et al. Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res. 2007;67(6):2649–56.PubMedCrossRef
Metadata
Title
Liposome encapsulated zoledronate favours M1-like behaviour in murine macrophages cultured with soluble factors from breast cancer cells
Authors
Sofia Sousa
Seppo Auriola
Jukka Mönkkönen
Jorma Määttä
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2015
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-015-1005-7

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