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Breast Cancer Research
Published in: Breast Cancer Research 1/2018

Open Access 01-12-2018 | Research article

Expression of ID4 protein in breast cancer cells induces reprogramming of tumour-associated macrophages

Authors: Sara Donzelli, Elisa Milano, Magdalena Pruszko, Andrea Sacconi, Silvia Masciarelli, Ilaria Iosue, Elisa Melucci, Enzo Gallo, Irene Terrenato, Marcella Mottolese, Maciej Zylicz, Alicja Zylicz, Francesco Fazi, Giovanni Blandino, Giulia Fontemaggi

Published in: Breast Cancer Research | Issue 1/2018

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Abstract

Background

As crucial regulators of the immune response against pathogens, macrophages have been extensively shown also to be important players in several diseases, including cancer. Specifically, breast cancer macrophages tightly control the angiogenic switch and progression to malignancy. ID4, a member of the ID (inhibitors of differentiation) family of proteins, is associated with a stem-like phenotype and poor prognosis in basal-like breast cancer. Moreover, ID4 favours angiogenesis by enhancing the expression of pro-angiogenic cytokines interleukin-8, CXCL1 and vascular endothelial growth factor. In the present study, we investigated whether ID4 protein exerts its pro-angiogenic function while also modulating the activity of tumour-associated macrophages in breast cancer.

Methods

We performed IHC analysis of ID4 protein and macrophage marker CD68 in a triple-negative breast cancer series. Next, we used cell migration assays to evaluate the effect of ID4 expression modulation in breast cancer cells on the motility of co-cultured macrophages. The analysis of breast cancer gene expression data repositories allowed us to evaluate the ability of ID4 to predict survival in subsets of tumours showing high or low macrophage infiltration. By culturing macrophages in conditioned media obtained from breast cancer cells in which ID4 expression was modulated by overexpression or depletion, we identified changes in the expression of ID4-dependent angiogenesis-related transcripts and microRNAs (miRNAs, miRs) in macrophages by RT-qPCR.

Results

We determined that ID4 and macrophage marker CD68 protein expression were significantly associated in a series of triple-negative breast tumours. Interestingly, ID4 messenger RNA (mRNA) levels robustly predicted survival, specifically in the subset of tumours showing high macrophage infiltration. In vitro and in vivo migration assays demonstrated that expression of ID4 in breast cancer cells stimulates macrophage motility. At the molecular level, ID4 protein expression in breast cancer cells controls, through paracrine signalling, the activation of an angiogenic programme in macrophages. This programme includes both the increase of angiogenesis-related mRNAs and the decrease of members of the anti-angiogenic miR-15b/107 group. Intriguingly, these miRNAs control the expression of the cytokine granulin, whose enhanced expression in macrophages confers increased angiogenic potential.

Conclusions

These results uncover a key role for ID4 in dictating the behaviour of tumour-associated macrophages in breast cancer.
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Literature
1.
Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136(5):E359–86.CrossRefPubMed
2.
Sorlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, et al. Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci U S A. 2001;98(19):10869–74.CrossRefPubMedPubMedCentral
3.
McAllister SS, Weinberg RA. The tumour-induced systemic environment as a critical regulator of cancer progression and metastasis. Nat Cell Biol. 2014;16:717–27.CrossRefPubMed
4.
5.
Lin EY, Li JF, Gnatovskiy L, Deng Y, Zhu L, Grzesik DA, et al. Macrophages regulate the angiogenic switch in a mouse model of breast cancer. Cancer Res. 2006;66(23):11238–46.CrossRefPubMed
6.
7.
Franklin RA, Liao W, Sarkar A, Kim MV, Bivona MR, Liu K, et al. The cellular and molecular origin of tumor-associated macrophages. Science. 2014;344(6186):921–5.CrossRefPubMedPubMedCentral
8.
Campbell MJ, Wolf D, Mukhtar RA, Tandon V, Yau C, Au A, et al. The prognostic implications of macrophages expressing proliferating cell nuclear antigen in breast cancer depend on immune context. PLoS One. 2013;8(10):e79114.CrossRefPubMedPubMedCentral
9.
Mukhtar RA, Moore AP, Tandon VJ, Nseyo O, Twomey P, Adisa CA, et al. Elevated levels of proliferating and recently migrated tumor-associated macrophages confer increased aggressiveness and worse outcomes in breast cancer. Ann Surg Oncol. 2012;19(12):3979–86.CrossRefPubMed
10.
11.
Joyce JA, Pollard JW. Microenvironmental regulation of metastasis. Nat Rev Cancer. 2009;9(4):239–52.CrossRefPubMed
12.
13.
Pollard JW. Tumour-educated macrophages promote tumour progression and metastasis. Nat Rev Cancer. 2004;4:71–8.CrossRefPubMed
14.
Condeelis J, Pollard JW. Macrophages: obligate partners for tumor cell migration, invasion, and metastasis. Cell. 2006;124(2):263–6.CrossRefPubMed
15.
Bingle L, Brown NJ, Lewis CE. The role of tumour-associated macrophages in tumour progression: implications for new anticancer therapies. J Pathol. 2002;196:254–65.CrossRefPubMed
16.
Perk J, Iavarone A, Benezra R. Id family of helix-loop-helix proteins in cancer. Nat Rev Cancer. 2005;5(8):603–14.CrossRefPubMed
17.
Badve S, Dabbs DJ, Schnitt SJ, Baehner FL, Decker T, Eusebi V, et al. Basal-like and triple-negative breast cancers: a critical review with an emphasis on the implications for pathologists and oncologists. Mod Pathol. 2011;24(2):157–67.CrossRefPubMed
18.
Wen YH, Ho A, Patil S, Akram M, Catalano J, Eaton A, et al. Id4 protein is highly expressed in triple-negative breast carcinomas: possible implications for BRCA1 downregulation. Breast Cancer Res Treat. 2012;135(1):93–102.CrossRefPubMed
19.
Hu Z, Fan C, Oh DS, Marron JS, He X, Qaqish BF, et al. The molecular portraits of breast tumors are conserved across microarray platforms. BMC Genomics. 2006;7:96.CrossRefPubMedPubMedCentral
20.
Thike AA, Tan PH, Ikeda M, Iqbal J. Increased ID4 expression, accompanied by mutant p53 accumulation and loss of BRCA1/2 proteins in triple-negative breast cancer, adversely affects survival. Histopathology. 2016;68(5):702–12.CrossRefPubMed
21.
Junankar S, Baker LA, Roden DL, Nair R, Elsworth B, Gallego-Ortega D, et al. ID4 controls mammary stem cells and marks breast cancers with a stem cell-like phenotype. Nat Commun. 2015;6:6548.CrossRefPubMed
22.
Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature. 2012;490(7418):61–70.CrossRef
23.
Baker LA, Holliday H, Swarbrick A. ID4 controls luminal lineage commitment in normal mammary epithelium and inhibits BRCA1 function in basal-like breast cancer. Endocr Relat Cancer. 2016;23(9):R381–92.CrossRefPubMed
24.
Beger C, Pierce LN, Kruger M, Marcusson EG, Robbins JM, Welcsh P, et al. Identification of Id4 as a regulator of BRCA1 expression by using a ribozyme-library-based inverse genomics approach. Proc Natl Acad Sci U S A. 2001;98(1):130–5.CrossRefPubMedPubMedCentral
25.
Roldán G, Delgado L, Musé IM. Tumoral expression of BRCA1, estrogen receptor α and ID4 protein in patients with sporadic breast cancer. Cancer Biol Ther. 2006;5(5):505–10.CrossRefPubMed
26.
Crippa E, Lusa L, De Cecco L, Marchesi E, Calin GA, Radice P, et al. miR-342 regulates BRCA1 expression through modulation of ID4 in breast cancer. PLoS One. 2014;9(1):e87039.CrossRefPubMedPubMedCentral
27.
Prat A, Cruz C, Hoadley KA, Díez O, Perou CM, Balmaña J. Molecular features of the basal-like breast cancer subtype based on BRCA1 mutation status. Breast Cancer Res Treat. 2014;147:185–91.CrossRefPubMedPubMedCentral
28.
Fontemaggi G, Dell’Orso S, Trisciuoglio D, Shay T, Melucci E, Fazi F, et al. The execution of the transcriptional axis mutant p53, E2F1 and ID4 promotes tumor neo-angiogenesis. Nat Struct Mol Biol. 2009;16(10):1086–93.CrossRefPubMed
29.
Dell’Orso S, Ganci F, Strano S, Blandino G, Fontemaggi G. ID4: a new player in the cancer arena. Oncotarget. 2010;1(1):48–58.PubMedPubMedCentral
30.
Pruszko M, Milano E, Forcato M, Donzelli S, Ganci F, Di Agostino S, et al. The mutant p53-ID4 complex controls VEGFA isoforms by recruiting lncRNA MALAT1. EMBO Rep. 2017;18(8):1331–51.CrossRefPubMedPubMedCentral
31.
Doedens AL, Stockmann C, Rubinstein MP, Liao D, Zhang N, DeNardo DG, et al. Macrophage expression of hypoxia-inducible factor-1 α suppresses T-cell function and promotes tumor progression. Cancer Res. 2010;70(19):7465–75.CrossRefPubMedPubMedCentral
32.
Jian J, Li G, Hettinghouse A, Progranulin LC. A key player in autoimmune diseases. Cytokine. 2016;101:48–55.CrossRefPubMed
33.
Fontemaggi G, Bellissimo T, Donzelli S, Iosue I, Benassi B, Bellotti G, et al. Identification of post-transcriptional regulatory networks during myeloblast-to-monocyte differentiation transition. RNA Biol. 2015;12(7):690–700.CrossRefPubMedPubMedCentral
34.
Novelli F, Milella M, Melucci E, Di Benedetto A, Sperduti I, Perrone-Donnorso R, et al. A divergent role for estrogen receptor-β in node-positive and node-negative breast cancer classified according to molecular subtypes: an observational prospective study. Breast Cancer Res. 2008;10(5):R74.CrossRefPubMedPubMedCentral
35.
Zhao X, Qu J, Sun Y, Wang J, Liu X, Wang F, et al. Prognostic significance of tumor-associated macrophages in breast cancer: a meta-analysis of the literature. Oncotarget. 2017;8(18):30576–86.PubMedPubMedCentral
36.
Gyorffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q, et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1809 patients. Breast Cancer Res Treat. 2010;123(3):725–31.CrossRefPubMed
37.
38.
Montrucchio G, Lupia E, Battaglia E, Passerini G, Bussolino F, Emanuelli G, et al. Tumor necrosis factor α-induced angiogenesis depends on in situ platelet-activating factor biosynthesis. J Exp Med. 1994;180(1):377–82.CrossRefPubMed
39.
Anghelina M, Krishnan P, Moldovan L, Moldovan NI. Monocytes/macrophages cooperate with progenitor cells during neovascularization and tissue repair: conversion of cell columns into fibrovascular bundles. Am J Pathol. 2006;168(2):529–41.CrossRefPubMedPubMedCentral
40.
Murao S, Gemmell MA, Callaham MF, Anderson NL, Huberman E. Control of macrophage cell differentiation in human promyelocytic HL-60 leukemia cells by 1,25-dihydroxyvitamin D3 and phorbol-12-myristate-13-acetate. Cancer Res. 1983;43(10):4989–96.PubMed
41.
Mangelsdorf DJ, Koeffler HP, Donaldson CA, Pike JW, Haussler MR. 1,25-Dihydroxyvitamin D3-induced differentiation in a human promyelocytic leukemia cell line (HL-60): receptor-mediated maturation to macrophage-like cells. J Cell Biol. 1984;98(2):391–8.CrossRefPubMed
42.
Wang WX, Kyprianou N, Wang X, Nelson PT. Dysregulation of the mitogen granulin in human cancer through the miR-15/107 microRNA gene group. Cancer Res. 2010;70(22):9137–42.CrossRefPubMedPubMedCentral
43.
Finnerty JR, Wang WX, Hébert SS, Wilfred BR, Mao G, Nelson PT. The miR-15/107 group of microRNA genes: evolutionary biology, cellular functions, and roles in human diseases. J Mol Biol. 2010;402(3):491–509.CrossRefPubMedPubMedCentral
44.
Yamakuchi M, Lotterman CD, Bao C, Hruban RH, Karim B, Mendell JT, et al. P53-induced microRNA-107 inhibits HIF-1 and tumor angiogenesis. Proc Natl Acad Sci U S A. 2010;107(14):6334–9.CrossRefPubMedPubMedCentral
45.
Chen L, Li ZY, Xu SY, Zhang XJ, Zhang Y, Luo K, et al. Upregulation of miR-107 inhibits glioma angiogenesis and VEGF expression. Cell Mol Neurobiol. 2016;36(1):113–20.CrossRefPubMed
46.
Wang R, Zhao N, Li S, Fang JH, Chen MX, Yang J, et al. MicroRNA-195 suppresses angiogenesis and metastasis of hepatocellular carcinoma by inhibiting the expression of VEGF, VAV2, and CDC42. Hepatology. 2013;58(2):642–53.CrossRefPubMed
47.
Wang Y, Zhang X, Zou C, Kung HF, Lin MC, Dress A, et al. miR-195 inhibits tumor growth and angiogenesis through modulating IRS1 in breast cancer. Biomed Pharmacother. 2016;80:95–101.CrossRefPubMed
48.
Shi Y, Zhao Y, Shao N, Ye R, Lin Y, Zhang N, et al. Overexpression of microRNA-96-5p inhibits autophagy and apoptosis and enhances the proliferation, migration and invasiveness of human breast cancer cells. Oncol Lett. 2017;13(6):4402–12.CrossRefPubMedPubMedCentral
49.
Wang WX, Wilfred BR, Madathil SK, Tang G, Hu Y, Dimayuga J, et al. miR-107 regulates granulin/progranulin with implications for traumatic brain injury and neurodegenerative disease. Am J Pathol. 2010;177(1):334–45.CrossRefPubMedPubMedCentral
50.
Elkabets M, Gifford AM, Scheel C, Nilsson B, Reinhardt F, Bray MA, et al. Human tumors instigate granulin-expressing hematopoietic cells that promote malignancy by activating stromal fibroblasts in mice. J Clin Invest. 2011;121(2):784–99.CrossRefPubMedPubMedCentral
51.
Eguchi R, Nakano T, Wakabayashi I. Progranulin and granulin-like protein as novel VEGF-independent angiogenic factors derived from human mesothelioma cells. Oncogene. 2017;36(5):714–22.CrossRefPubMed
52.
Mundra JJ, Jian J, Bhagat P, Liu CJ. Progranulin inhibits expression and release of chemokines CXCL9 and CXCL10 in a TNFR1 dependent manner. Sci Rep. 2016;6:21115.CrossRefPubMedPubMedCentral
53.
Sawamiphak S, Seidel S, Essmann CL, Wilkinson GA, Pitulescu ME, Acker T, et al. Ephrin-B2 regulates VEGFR2 function in developmental and tumour angiogenesis. Nature. 2010;465(7297):487–91.CrossRefPubMed
54.
Neufeld G, Kessler O, Herzog Y. The interaction of neuropilin-1 and neuropilin-2 with tyrosine-kinase receptors for VEGF. Adv Exp Med Biol. 2002;515:81–90.CrossRefPubMed
55.
Sulpice E, Plouët J, Bergé M, Allanic D, Tobelem G, Merkulova-Rainon T. Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity. Blood. 2008;111:2036–45.CrossRefPubMed
56.
Gao B, Hao S, Tian W, Jiang Y, Zhang M, Guo L, et al. MicroRNA-107 is downregulated and having tumor suppressive effect in breast cancer by negatively regulating BDNF. J Gene Med. 2016;19(12):e2932.CrossRef
57.
Li XY, Luo QF, Wei CK, Li DF, Li J, Fang L. MiRNA-107 inhibits proliferation and migration by targeting CDK8 in breast cancer. Int J Clin Exp Med. 2014;7(1):32–40.PubMedPubMedCentral
58.
Polytarchou C, Iliopoulos D, Struhl K. An integrated transcriptional regulatory circuit that reinforces the breast cancer stem cell state. Proc Natl Acad Sci U S A. 2012;109(36):14470–5.CrossRefPubMedPubMedCentral
59.
Thakur S, Grover RK, Gupta S, Yadav AK, Das BC. Identification of specific miRNA signature in paired sera and tissue samples of Indian women with triple negative breast cancer. PLoS One. 2016;11(7):e0158946.CrossRefPubMedPubMedCentral
60.
Ven't Veer LJ, Dai H, van de Vijver MJ, He YD, Hart AA, Mao M, Peterse HL, van der Kooy K, Marton MJ, Witteveen AT: Gene expression profiling predicts clinical outcome of breast cancer. Nature. 2002, 415 (6871): 530-536. https://​doi.​org/​10.​1038/​415530a.
Metadata
Title
Expression of ID4 protein in breast cancer cells induces reprogramming of tumour-associated macrophages
Authors
Sara Donzelli
Elisa Milano
Magdalena Pruszko
Andrea Sacconi
Silvia Masciarelli
Ilaria Iosue
Elisa Melucci
Enzo Gallo
Irene Terrenato
Marcella Mottolese
Maciej Zylicz
Alicja Zylicz
Francesco Fazi
Giovanni Blandino
Giulia Fontemaggi
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2018
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/s13058-018-0990-2

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