Disseminated BRAFV600E melanoma responds to BRAF inhibitors (BRAFi) but easily develops resistance with poor prognosis. Secretome plays a pivotal role during tumour progression causing profound effects on therapeutic efficacy. Secreted M-CSF is involved in both cytotoxicity suppression and tumour progression in melanoma. We aimed to analyse the M-CSF contribution in resistant metastatic melanoma to BRAF-targeted therapies.
Conditioned media from melanoma cells were analysed by citoarray. Viability and migration/invasion assays were performed with paired melanoma cells and tumour growth in xenografted SCID mice. We evaluated the impact of M-CSF plasma levels with clinical prognosis from 35 metastatic BRAFV600E-mutant melanoma patients.
BRAFi-resistant melanoma cells secretome is rich in pro-tumour cytokines. M-CSF secretion is essential to induce a Vemurafenib-resistant phenotype in melanoma cells. Further, we demonstrated that M-CSF mAb in combination with Vemurafenib and autophagy blockers synergistically induce apoptosis, impair migration and reduce tumour growth in BRAFi-resistant melanoma cells. Interestingly, lower M-CSF plasma levels are associated with better prognosis in metastatic melanoma patients.
Secreted M-CSF induces a BRAFi-resistant phenotype and means worse prognosis in BRAFV600E metastatic melanoma patients. These results identify secreted M-CSF as a promising therapeutic target toward BRAFi-resistant melanomas.
This is a preview of subscription content
Subscribe to Journal
Get full journal access for 1 year
only $4.96 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Kosmopoulou M, Giannopoulou AF, Iliou A, Benaki D, Panagiotakis A, Velentzas AD, et al. Human melanoma‐cell metabolic profiling: Identification of novel biomarkers indicating metastasis. Int J Mol Sci. 2020;21:2436. https://doi.org/10.3390/ijms21072436.
Mishra H, Mishra PK, Ekielski A, Jaggi M, Iqbal Z, Talegaonkar S. Melanoma treatment: from conventional to nanotechnology. J Cancer Res Clin Oncol. 2018;144:2283–302.
Trubini S, Ubiali A, Paties C, Cavanna L. Novel BRAF mutation in melanoma: a case report. Mol Clin Oncol. 2018;8:460–2. https://doi.org/10.3892/mco.2018.1555. Epub 2018 Jan 12.
Kakadia S, Yarlagadda N, Awad R, Kundranda M, Niu J, Naraev B, et al. Mechanisms of resistance to BRAF and MEK inhibitors and clinical update of us food and drug administration-approved targeted therapy in advanced melanoma. Onco Targets Ther. 2018;11:7095–107.
Das A, Pushparaj C, Herreros J, Nager M, Vilella R, Portero M, et al. T-type calcium channel blockers inhibit autophagy and promote apoptosis of malignant melanoma cells. Pigment Cell Melanoma Res. 2013;26:874–85.
Maiques O, Barceló C, Panosa A, Pijuan J, Orgaz JL, Rodriguez-Hernandez I, et al. T-type calcium channels drive migration/invasion in BRAFV600E melanoma cells through Snail1. Pigment Cell Melanoma Res. 2018;31:484–95.
Yang X, Yu DD, Yan F, Jing YY, Han ZP, Sun K, et al. The role of autophagy induced by tumor microenvironment in different cells and stages of cancer. Cell Biosci. 2015;5:1–11.
Ndoye A, Weeraratna AT. Autophagy—an emerging target for melanoma therapy [version 1; referees: 2 approved]. F1000Research. 2016;5:1–9.
Barceló C, Sisó P, Maiques O, García-Mulero S, Sanz-Pamplona R, Navaridas R, et al. T-type calcium channels as potential therapeutic targets in vemurafenib-resistant BRAFV600E melanoma. J Invest Dermatol. 2020;140:1253–65.
Barceló C, Sisó P, Maiques O, de la Rosa I, Martí RM, Macià A. T-type calcium channels: a potential novel target in melanoma. Cancers (Basel). 2020;12:1–13.
Di Leo L, Bodemeyer V, De, Zio D. The complex role of autophagy in melanoma evolution: new perspectives from mouse models. Front Oncol. 2020;9:1–9.
Wang M, Zhao J, Zhang L, Wei F, Lian Y, Wu Y, et al. Role of tumor microenvironment in tumorigenesis. J Cancer. 2017;8:761–73.
Jin MZ, Jin WL. The updated landscape of tumor microenvironment and drug repurposing. Signal Transduct Target Ther. 2020;5. https://doi.org/10.1038/s41392-020-00280-x.
Di Blasio S, van Wigcheren GF, Becker A, van Duffelen A, Gorris M, Verrijp K, et al. The tumour microenvironment shapes dendritic cell plasticity in a human organotypic melanoma culture. Nat Commun. 2020;11:1–17. https://doi.org/10.1038/s41467-020-16583-0.
Peinado H, Zhang H, Matei IR, Costa-Silva B, Hoshino A, Rodrigues G, et al. Pre-metastatic niches: organ-specific homes for metastases. Nat Rev Cancer. 2017;17:302–17. https://doi.org/10.1038/nrc.2017.6.
Herraiz C, Jiménez-Cervantes C, Sánchez-Laorden B, García-Borrón JC. Functional interplay between secreted ligands and receptors in melanoma. Semin Cell Dev Biol. 2018;78:73–84. https://doi.org/10.1016/j.semcdb.2017.06.021.
Klemm F, Joyce JA. Microenvironmental regulation of therapeutic response in cancer. Trends Cell Biol. 2015;25:198–213.
Obenauf AC, Zou Y, Ji AL, Vanharanta S, Shu W, Shi H, et al. Therapy-induced tumour secretomes promote resistance and tumour progression. Nature. 2015;520:368–72.
da Cunha BR, Domingos C, Buzzo Stefanini AC, Henrique T, Polachini GM, Castelo-Branco P, et al. Cellular interactions in the tumor microenvironment: the role of secretome. J Cancer. 2019;10:4574–87.
Madden EC, Gorman AM, Logue SE, Samali A. Tumour cell secretome in chemoresistance and tumour recurrence. Trends Cancer. 2020;6:489–505. https://doi.org/10.1016/j.trecan.2020.02.020.
Wu T, Dai Y. Tumor microenvironment and therapeutic response. Cancer Lett. 2017;387:61–8. https://doi.org/10.1016/j.canlet.2016.01.043.
Priceman SJ, Sung JL, Shaposhnik Z, Burton JB, Torres-Collado AX, Moughon DL, et al. Targeting distinct tumor-infiltrating myeloid cells by inhibiting CSF-1 receptor: combating tumor evasion of antiangiogenic therapy. Blood. 2010;115:1461–71.
Chockalingam S, Ghosh SS. Macrophage colony-stimulating factor and cancer: a review. Tumor Biol. 2014;35:10635–44.
Laoui D, van Overmeire E, de Baetselier P, van Ginderachter JA, Raes G. Functional relationship between tumor-associated macrophages and macrophage colony-stimulating factor as contributors to cancer progression. Front Immunol. 2014;5:489. https://doi.org/10.3389/fimmu.2014.00489.
Neubert NJ, Schmittnaegel M, Bordry N, Nassiri S, Martignier C, Tillé L, et al. T cell–induced CSF1 promotes melanoma resistance to PD1 blockade Natalie. Sci Transl Med. 2018;10:1–30.
Dwyer AR, Greenland EL, Pixley FJ. Promotion of tumor invasion by tumor-associated macrophages: The role of CSF-1-activated phosphatidylinositol 3 kinase and Src family kinase motility signaling. Cancers (Basel). 2017;9:1–15.
Baghdadi M, Endo H, Takano A, Ishikawa K, Kameda Y, Wada H, et al. High co-expression of IL-34 and M-CSF correlates with tumor progression and poor survival in lung cancers. Sci Rep. 2018;8:1–10. https://doi.org/10.1038/s41598-017-18796-8.
Giricz O, Mo Y, Dahlman KB, Cotto-Rios XM, Vardabasso C, Nguyen H, et al. The RUNX1/IL-34/CSF-1R axis is an autocrinally regulated modulator of resistance to BRAF-V600E inhibition in melanoma. JCI Insight. 2018;3:e120422. https://doi.org/10.1172/jci.insight.120422.
Mok S, Tsoi J, Koya RC, Hu-Lieskovan S, West BL, Bollag G, et al. Inhibition of colony stimulating factor-1 receptor improves antitumor efficacy of BRAF inhibition. BMC Cancer. 2015;15:1–10.
Liberato T, Pessotti DS, Fukushima I, Kitano ES, Serrano SMT, Zelanis A. Signatures of protein expression revealed by secretome analyses of cancer associated fibroblasts and melanoma cell lines. J Proteom. 2018;174:1–8. https://doi.org/10.1016/j.jprot.2017.12.013.
Roma-Rodrigues C, Mendes R, Baptista PV, Fernandes AR. Targeting tumor microenvironment for cancer therapy. Int J Mol Sci. 2019;20:840. https://doi.org/10.3390/ijms20040840.
Pijuan J, Barceló C, Moreno DF, Maiques O, Sisó P, Marti RM, et al. In vitro cell migration, invasion, and adhesion assays: From cell imaging to data analysis. Front Cell Dev Biol. 2019;7:107. https://doi.org/10.3389/fcell.2019.00107.
Eritja N, Chen BJ, Rodríguez-Barrueco R, Santacana M, Gatius S, Vidal A, et al. Autophagy orchestrates adaptive responses to targeted therapy in endometrial cancer. Autophagy. 2017;13:608–24. https://doi.org/10.1080/15548627.2016.1271512.
Kimura S, Noda T, Yoshimori T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy. 2007;3:452–60.
Maiques O, Macià A, Moreno S, Barceló C, Santacana M, Vea A, et al. Immunohistochemical analysis of T-type calcium channels in acquired melanocytic naevi and melanoma. Br J Dermatol. 2017;176:1247–58.
Bankhead P, Loughrey MB, Fernández JA, Dombrowski Y, McArt DG, Dunne PD, et al. QuPath: open source software for digital pathology image analysis. Sci Rep. 2017;7:1–7.
Bindea G, Mlecnik B, Hackl H, Charoentong P, Tosolini M, Kirilovsky A, et al. ClueGO: a cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics. 2009;25:1091–3. https://pubmed.ncbi.nlm.nih.gov/19237447/.
Sanz-Moreno V, Gaggioli C, Yeo M, Albrengues J, Wallberg F, Viros A, et al. ROCK and JAK1 signaling cooperate to control actomyosin contractility in tumor cells and stroma. Cancer Cell. 2011;20(Aug):229–45. https://pubmed.ncbi.nlm.nih.gov/21840487/.
Neubert NJ, Schmittnaegel M, Bordry N, Nassiri S, Wald N, Martignier C, et al. T cell-induced CSF1 promotes melanoma resistance to PD1 blockade. Sci Transl Med. 2018;10:eaan3311. https://doi.org/10.1126/scitranslmed.aan3311.
Dvořánková B, Szabo P, Kodet O, Strnad H, Kolář M, Lacina L, et al. Intercellular crosstalk in human malignant melanoma. Vol. 254, Protoplasma. 2017;254:1143–50. https://pubmed.ncbi.nlm.nih.gov/27807664/.
Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646–74. https://doi.org/10.1016/j.cell.2011.02.013.
Kodet O, Dvořánková B, Bendlová B, Sýkorová V, Krajsová I, Štork J, et al. Microenvironment-driven resistance to B-Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts. Int J Mol Med. 2018;41:2687–703.
Qu Y, Dou B, Tan H, Feng Y, Wang N, Wang D. Tumor microenvironment-driven non-cell-autonomous resistance to antineoplastic treatment. Mol Cancer. 2019;18:1–16.
Chinnasamy D, Yu Z, Theoret MR, Zhao Y, Shrimali RRK, Morgan RA, et al. Gene therapy using genetically modified lymphocytes targeting VEGFR-2 inhibits the growth of vascularized syngenic tumors in mice. J Clin Invest. 2010;120:3953–68.
Landskron G, De La Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res. 2014;2014:149185. https://doi.org/10.1155/2014/149185. Epub 2014 May 13.
Bielenberg DR, Zetter BR. The contribution of angiogenesis to the process of metastasis. Cancer J (U S). 2015;21:267–73.
Kubota Y, Takubo K, Shimizu T, Ohno H, Kishi K, Shibuya M, et al. M-CSF inhibition selectively targets pathological angiogenesis and lymphangiogenesis. J Exp Med. 2009;206(May):1089–102. https://pubmed.ncbi.nlm.nih.gov/19398755/.
Fend L, Accart N, Kintz J, Cochin S, Reymann C, Le Pogam F, et al. Therapeutic effects of anti-CD115 monoclonal antibody in mouse cancer models through dual inhibition of tumor-associated macrophages and osteoclasts. PLoS ONE. 2013. https://pubmed.ncbi.nlm.nih.gov/24019914/.
Cannarile MA, Weisser M, Jacob W, Jegg AM, Ries CH, Rüttinger D. Colony-stimulating factor 1 receptor (CSF1R) inhibitors in cancer therapy. J Immunother Cancer. 2017;5:1–13.
Chapman PB, Hauschild A, Robert C, Haanen JB, Ascierto P, Larkin J, et al. Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N. Engl J Med. 2011;364:2507–16.
Sun C, Wang L, Huang S, Heynen GJJE, Prahallad A, Robert C. et al. Reversible and adaptive resistance to BRAF(V600E) inhibition in melanoma. Nature. 2014;508:118–22. https://doi.org/10.1038/nature13121.
Ngiow SF, Meeth KM, Stannard K, Barkauskas DS, Bollag G, Bosenberg M, et al. Co-inhibition of colony stimulating factor-1 receptor and BRAF oncogene in mouse models of BRAFV600Emelanoma. Oncoimmunology. 2016;5:1–11. https://doi.org/10.1080/2162402X.2015.1089381.
Solinas G, Germano G, Mantovani A, Allavena P. Tumor-associated macrophages (TAM) as major players of the cancer-related inflammation. J Leukoc Biol. 2009;86:1065–73.
Qian B-Z, Pollard JW. Macrophage diversity enhances tumor progression and metastasis. Cell. 2010;141:1477–90.
This work was supported by grants from ISCIII/FEDER “Una manera de hacer Europa” (PI1500711 to R.M.M. & PI18/00573 to RMM & AM and PI20/00502 to NE) and CIBERONC-CB16/12/00231 to XMG, Fundació la Marató de TV3 (FMTV 201331-31) to RM and Generalitat de Catalunya (2014/SGR138) to XMG. CB and PS hold a predoctoral fellowship from UdL-IRBLleida. IR holds a predoctoral fellowship from AECC. AM holds a postdoctoral fellowship from AECC. The cell culture experiments were performed in the Cell Culture Scientific & Technical Service from Universitat de Lleida (UdL), Lleida, Spain. Work supported by the Xarxa de Bancs de Tumours de Catalunya sponsored by Pla Director d’Oncología de Catalunya (XBTC)”, IRBLleida Biobank (B.0000682) and PLATAFORMA BIOBANCOS (PT17/0015/0027; PT20/00021) and HCB-IDIBAPS Biobank (R120904-090) integrated in the Spanish National Biobank Network (ISCIII Code C 0.000.334). The research at the Melanoma Unit from Hospital Clinic of Barcelona was partially funded by Insituto de Salud Carlos III (ISCIII), Spain, through projects PI18/00419 and PI18/01077, and co-funded by the European Union; by the grant AC16/00081, integrated in the Plan Estatal I + D + I, IMMUSPHINX-Transcan-2; by the CIBER de Enfermedades Raras of ISCIII, Spain, cofinanced by European Development Regional Fund “A way to achieve Europe” ERDF; and by the Generalitat de Catalunya (AGAUR 2017/SGR1134 and CERCA Program). We are grateful to our patients and relatives, to physicians and nurses from the Melanoma Unit of Hospital Clínic of Barcelona for collecting patients samples and data, and to Judit Mateu from the “Melanoma: image, genetics and immunology” group at IDIBAPS for her technical assistance. NCL holds a predoctoral fellowship from Ministerio de Educación, Cultura y Deportes, Spain (FPU17/05453).
The authors declare no competing interests.
Ethics approval and consent to participate
The animal experimental committee approved animal work procedures (CEEA from University of Lleida). The Clinical Research Ethics Committee of the HCB (study registry 2013/8305) approved this study and each study participant signed written informed consent in accordance with the Declaration of Helsinki.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Barceló, C., Sisó, P., de la Rosa, I. et al. M-CSF as a therapeutic target in BRAFV600E melanoma resistant to BRAF inhibitors. Br J Cancer (2022). https://doi.org/10.1038/s41416-022-01886-4