{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T17:50:37Z","timestamp":1777053037622,"version":"3.51.4"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T00:00:00Z","timestamp":1776988800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["T32OD01043"],"award-info":[{"award-number":["T32OD01043"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Colorado School of Medicine, Anschutz Medical Campus Cancer Center Genomics Shared Resource Core Facility","award":["P30CA046934"],"award-info":[{"award-number":["P30CA046934"]}]},{"DOI":"10.13039\/100013989","name":"Anschutz Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100013989","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Tumor-associated macrophages (TAMs) drive cancer progression and metastasis. However, the mechanisms by which tumor cells shape TAM phenotypes in canine cancers remain poorly understood. We investigated correlations between cancer cell gene expression and macrophage polarization to identify potential biomarkers and therapeutic targets. Tumor-conditioned media from 25 canine cancer cell lines were applied to monocyte-derived macrophages from three canine donors for 24 hours. Following washout, supernatants were analyzed for immunomodulatory cytokines and chemokines. Each cell line\u2019s polarization capacity was ranked using modified z-scores, then correlated with RNA-sequencing data through Spearman\u2019s correlation and differential expression analysis. Cancer cell lines showed marked heterogeneity in macrophage polarization capacity, largely independent of histologic type.\n                    <jats:italic>MVB12A,<\/jats:italic>\n                    a gene involved in exosome biogenesis, strongly correlated with vascular endothelial growth factor (VEGF) stimulation, suggesting exosome-mediated polarization mechanisms. Exosome fractionation experiments confirmed that purified exosomes induced significantly more macrophage VEGF secretion than other conditions, and high-\n                    <jats:italic>MVB12A<\/jats:italic>\n                    cell lines showed greater VEGF enrichment in exosomes.\n                    <jats:italic>C-C motif chemokine ligand<\/jats:italic>\n                    <jats:italic>3<\/jats:italic>\n                    \u00a0(\n                    <jats:italic>CCL3)<\/jats:italic>\n                    was strongly correlated with tumor necrosis factor-alpha (TNF-\u03b1) secretion exclusively in histiocytic sarcoma cells, and recombinant CCL3 induced dose-dependent TNF-\u03b1 secretion from macrophages. High-polarizing cell lines exhibited upregulation of macrophage activation, epithelial-to-mesenchymal transition (EMT), and metabolic reprogramming genes, and downregulation of immune surveillance and cell adhesion genes. Gene set enrichment analysis confirmed pathways for immune suppression, EMT, and extracellular matrix remodeling. These findings identify exosome-associated VEGF stimulation as a previously uncharacterized mechanism in canine tumors and highlight CCL3 as a potential histiocytic sarcoma-specific driver of macrophage TNF-\u03b1 secretion. Further validation in canine clinical cohorts will determine whether these pathways can serve as biomarkers or therapeutic targets in veterinary oncology.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0346239","type":"journal-article","created":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T17:32:03Z","timestamp":1777051923000},"page":"e0346239","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Distinct tumor genomic signatures underlie canine macrophage polarization"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5688-3872","authenticated-orcid":true,"given":"Rachel V.","family":"Brady","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8941-1468","authenticated-orcid":true,"given":"Sunetra","family":"Das","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5310-0078","authenticated-orcid":true,"given":"Dawn L.","family":"Duval","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristen B.","family":"Farrell","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric P.","family":"Palmer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Douglas H.","family":"Thamm","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,4,24]]},"reference":[{"issue":"3","key":"pone.0346239.ref001","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.ccell.2023.02.016","article-title":"The evolving tumor microenvironment: From cancer initiation to metastatic outgrowth","volume":"41","author":"KE de Visser","year":"2023","journal-title":"Cancer Cell"},{"key":"pone.0346239.ref002","doi-asserted-by":"crossref","first-page":"107753","DOI":"10.1016\/j.pharmthera.2020.107753","article-title":"Tumor microenvironment as a therapeutic target in cancer","volume":"221","author":"Y Xiao","year":"2021","journal-title":"Pharmacol Ther"},{"issue":"10","key":"pone.0346239.ref003","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1101\/gr.273300.120","article-title":"A single-cell tumor immune atlas for precision oncology","volume":"31","author":"P Nieto","year":"2021","journal-title":"Genome Res"},{"issue":"3","key":"pone.0346239.ref004","doi-asserted-by":"crossref","DOI":"10.1016\/j.cell.2021.01.010","article-title":"A pan-cancer single-cell transcriptional atlas of tumor infiltrating myeloid cells","volume":"184","author":"S Cheng","year":"2021","journal-title":"Cell"},{"key":"pone.0346239.ref005","doi-asserted-by":"crossref","first-page":"1176807","DOI":"10.3389\/fimmu.2023.1176807","article-title":"Tumor-associated macrophages: prognostic and therapeutic targets for cancer in humans and dogs","volume":"14","author":"RV Brady","year":"2023","journal-title":"Front Immunol"},{"key":"pone.0346239.ref006","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.3389\/fonc.2019.01291","article-title":"Emerging translational opportunities in comparative oncology with companion canine cancers: radiation oncology","volume":"9","author":"MW Nolan","year":"2019","journal-title":"Front Oncol"},{"issue":"12","key":"pone.0346239.ref007","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.2460\/javma.24.08.0532","article-title":"The promise of immunotherapeutic strategies to advance cancer treatment in pet dogs","volume":"262","author":"JM Hampel","year":"2024","journal-title":"J Am Vet Med Assoc"},{"key":"pone.0346239.ref008","doi-asserted-by":"crossref","first-page":"1130215","DOI":"10.3389\/fonc.2023.1130215","article-title":"Naturally occurring canine sarcomas: bridging the gap from mouse models to human patients through cross-disciplinary research partnerships","volume":"13","author":"M Klosowski","year":"2023","journal-title":"Front Oncol"},{"issue":"9","key":"pone.0346239.ref009","doi-asserted-by":"crossref","first-page":"2133","DOI":"10.1158\/1078-0432.CCR-15-2347","article-title":"Defining the value of a comparative approach to cancer drug development","volume":"22","author":"AK LeBlanc","year":"2016","journal-title":"Clin Cancer Res"},{"key":"pone.0346239.ref010","doi-asserted-by":"crossref","first-page":"1593333","DOI":"10.3389\/fvets.2025.1593333","article-title":"Human immuno-therapeutics for cancer treatment of dogs?","volume":"12","author":"H Klingemann","year":"2025","journal-title":"Front Vet Sci"},{"issue":"11","key":"pone.0346239.ref011","doi-asserted-by":"crossref","first-page":"2003","DOI":"10.3390\/cancers16112003","article-title":"Checkpoint inhibitors in dogs: are we there yet?","volume":"16","author":"A Giuliano","year":"2024","journal-title":"Cancers (Basel)"},{"issue":"3","key":"pone.0346239.ref012","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1208\/s12248-019-0322-1","article-title":"CAR T cell immunotherapy in human and veterinary oncology: changing the odds against hematological malignancies","volume":"21","author":"JP Mochel","year":"2019","journal-title":"AAPS J"},{"issue":"12","key":"pone.0346239.ref013","doi-asserted-by":"crossref","first-page":"1376","DOI":"10.1038\/s41423-024-01232-z","article-title":"Targeting of TAMs: can we be more clever than cancer cells?","volume":"21","author":"J Kzhyshkowska","year":"2024","journal-title":"Cell Mol Immunol"},{"issue":"4","key":"pone.0346239.ref014","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1177\/1040638711408064","article-title":"Polymerase chain reaction-based species verification and microsatellite analysis for canine cell line validation","volume":"23","author":"LE O\u2019Donoghue","year":"2011","journal-title":"J Vet Diagn Invest"},{"issue":"2","key":"pone.0346239.ref015","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1111\/vco.12192","article-title":"The Flint Animal Cancer Center (FACC) canine tumour cell line panel: a resource for veterinary drug discovery, comparative oncology and translational medicine","volume":"15","author":"JS Fowles","year":"2017","journal-title":"Vet Comp Oncol"},{"key":"pone.0346239.ref016","volume-title":"R: A language and environment for statistical computing","author":"R Core Team","year":"2025"},{"key":"pone.0346239.ref017","volume-title":"RStudio: integrated development environment for R","author":"Posit Team","year":"2025"},{"key":"pone.0346239.ref018","volume-title":"Volume 16: How to detect and handle outliers","author":"B Iglewicz","year":"1993"},{"issue":"8","key":"pone.0346239.ref019","doi-asserted-by":"crossref","first-page":"1460","DOI":"10.1158\/1535-7163.MCT-18-1346","article-title":"Identifying candidate druggable targets in canine cancer cell lines using whole-exome sequencing","volume":"18","author":"S Das","year":"2019","journal-title":"Mol Cancer Ther"},{"issue":"5","key":"pone.0346239.ref020","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0303470","article-title":"VDX-111 targets proliferative pathways in canine cancer cell lines","volume":"19","author":"KB Farrell","year":"2024","journal-title":"PLoS One"},{"issue":"12","key":"pone.0346239.ref021","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1093\/bioinformatics\/btr260","article-title":"Molecular signatures database (MSigDB) 3.0","volume":"27","author":"A Liberzon","year":"2011","journal-title":"Bioinformatics"},{"key":"pone.0346239.ref022","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/978-1-0716-2217-9_5","article-title":"Measurement of VEGF content in exosomes and subsequent tumor tubulogenesis and in vivo angiogenesis functional assays","volume":"2475","author":"Y Zeng","year":"2022","journal-title":"Methods Mol Biol"},{"issue":"2","key":"pone.0346239.ref023","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.bbrc.2010.07.060","article-title":"Distinct functions of human MVB12A and MVB12B in the ESCRT-I dependent on their posttranslational modifications","volume":"399","author":"T Tsunematsu","year":"2010","journal-title":"Biochem Biophys Res Commun"},{"issue":"19","key":"pone.0346239.ref024","doi-asserted-by":"crossref","DOI":"10.15252\/embj.2019101704","article-title":"The TRAPP complex mediates secretion arrest induced by stress granule assembly","volume":"38","author":"F Zappa","year":"2019","journal-title":"EMBO J"},{"key":"pone.0346239.ref025","article-title":"Genenames.org: the HGNC resources in 2023","volume":"51","author":"RL Seal","year":"2023","journal-title":"Nucleic Acids Res"},{"issue":"8","key":"pone.0346239.ref026","doi-asserted-by":"crossref","first-page":"7262","DOI":"10.3390\/ijms24087262","article-title":"Involvement in tumorigenesis and clinical significance of CXCL1 in reproductive cancers: breast cancer, cervical cancer, endometrial cancer, ovarian cancer and prostate cancer","volume":"24","author":"J Korbecki","year":"2023","journal-title":"Int J Mol Sci"},{"key":"pone.0346239.ref027","doi-asserted-by":"crossref","first-page":"692142","DOI":"10.3389\/fonc.2021.692142","article-title":"Tumor-induced inflammatory cytokines and the emerging diagnostic devices for cancer detection and prognosis","volume":"11","author":"AER Kartikasari","year":"2021","journal-title":"Front Oncol"},{"issue":"16","key":"pone.0346239.ref028","doi-asserted-by":"crossref","first-page":"8530","DOI":"10.3390\/ijms22168530","article-title":"Roles of CCL2-CCR2 axis in the tumor microenvironment","volume":"22","author":"S Kadomoto","year":"2021","journal-title":"Int J Mol Sci"},{"key":"pone.0346239.ref029","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.semcancer.2023.10.004","article-title":"TGF-\u03b2, EMT, and resistance to anti-cancer treatment","volume":"97","author":"X Wang","year":"2023","journal-title":"Semin Cancer Biol"},{"issue":"1","key":"pone.0346239.ref030","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1158\/1078-0432.CCR-22-1366","article-title":"Molecular mechanisms and future implications of VEGF\/VEGFR in cancer therapy","volume":"29","author":"SA Patel","year":"2023","journal-title":"Clin Cancer Res"},{"key":"pone.0346239.ref031","doi-asserted-by":"crossref","first-page":"990568","DOI":"10.3389\/fonc.2022.990568","article-title":"IL-8 and its role as a potential biomarker of resistance to anti-angiogenic agents and immune checkpoint inhibitors in metastatic renal cell carcinoma","volume":"12","author":"M Rizzo","year":"2022","journal-title":"Front Oncol"},{"key":"pone.0346239.ref032","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.cca.2009.06.029","article-title":"TNF-alpha\/IL-10 ratio and C-reactive protein as markers of the inflammatory response in CAD-prone North Indian patients with acute myocardial infarction","volume":"408","author":"B Goswami","year":"2009","journal-title":"Clin Chim Acta"},{"key":"pone.0346239.ref033","doi-asserted-by":"crossref","first-page":"118886","DOI":"10.1016\/j.biopha.2025.118886","article-title":"Exosomal proteins as drivers of metastasis to distant organs","volume":"193","author":"I Mart\u00ednez-Espinosa","year":"2025","journal-title":"Biomed Pharmacother"},{"key":"pone.0346239.ref034","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.omtm.2021.05.016","article-title":"Exosomes and organ-specific metastasis","volume":"22","author":"L Zhao","year":"2021","journal-title":"Mol Ther Methods Clin Dev"},{"issue":"3","key":"pone.0346239.ref035","first-page":"209","article-title":"Effects of inhibitors of vascular endothelial growth factor receptor 2 and downstream pathways of receptor tyrosine kinases involving phosphatidylinositol 3-kinase\/Akt\/mammalian target of rapamycin or mitogen-activated protein kinase in canine hemangiosarcoma cell lines","volume":"80","author":"M Adachi","year":"2016","journal-title":"Can J Vet Res"},{"issue":"4","key":"pone.0346239.ref036","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1002\/1878-0261.13744","article-title":"Genomic landscape and preclinical models of angiosarcoma","volume":"19","author":"A Benton","year":"2025","journal-title":"Mol Oncol"},{"issue":"3","key":"pone.0346239.ref037","article-title":"Enriching CCL3 in the tumor microenvironment facilitates T cell responses and improves the efficacy of anti-PD-1 therapy","volume":"21","author":"TG Kang","year":"2021","journal-title":"Immune Netw"},{"issue":"5","key":"pone.0346239.ref038","doi-asserted-by":"crossref","first-page":"650","DOI":"10.5858\/arpa.2018-0349-RS","article-title":"Histiocytic sarcoma","volume":"144","author":"YP Hung","year":"2020","journal-title":"Arch Pathol Lab Med"},{"issue":"1","key":"pone.0346239.ref039","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.cvsm.2022.07.010","article-title":"Histiocytic diseases","volume":"53","author":"PF Moore","year":"2023","journal-title":"Vet Clin North Am Small Anim Pract"},{"issue":"5","key":"pone.0346239.ref040","doi-asserted-by":"crossref","DOI":"10.1152\/ajpregu.00019.2019","article-title":"CCR5 is a required signaling receptor for macrophage expression of inflammatory genes in response to viral double-stranded RNA","volume":"316","author":"ZR Shaheen","year":"2019","journal-title":"Am J Physiol Regul Integr Comp Physiol"},{"key":"pone.0346239.ref041","first-page":"1079851","article-title":"TNF-\u03b1 autocrine feedback loops in human monocytes: the pro- and anti-inflammatory roles of the TNF-\u03b1 receptors support the concept of selective TNFR1 blockade in vivo","volume":"2016","author":"JM Gane","year":"2016","journal-title":"J Immunol Res"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0346239","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T17:32:11Z","timestamp":1777051931000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0346239"}},"subtitle":[],"editor":[{"given":"Daotai","family":"Nie","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,4,24]]},"references-count":41,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4,24]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0346239","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,24]]}}}