FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2033820276> ?p ?o ?g. }

• W2033820276 endingPage "829" @default.
• W2033820276 startingPage "815" @default.
• W2033820276 abstract "•Monocytes and macrophages show a protumor phenotype in a human cancer setting•The protumor phenotype is regulated by an IL-1 receptor-dependent mechanism•Targeting this mechanism abrogated macrophage protumor phenotype and tumor in vivo Monocytes and macrophages are major components of the tumor microenvironment, but their contributions to human cancer are poorly understood. We used molecular profiling combined with functional assays to investigate the role of these cells in human renal cell carcinoma (RCC). Blood monocytes from RCC patients displayed a tumor-promoting transcriptional profile that supported functions like angiogenesis and invasion. Induction of this protumor phenotype required an interleukin-1 receptor (IL-1R)-dependent mechanism. Indeed, targeting of IL-1-IL-1R axis in a human RCC xenograft model abrogated the protumor phenotype of tumor-associated macrophages (TAMs) and reduced tumor growth in vivo. Supporting this, meta-analysis of gene expression from human RCC tumors showed IL1B expression to correlate with myelomonocytic markers, protumor genes, and tumor staging. Analyzing RCC patient tumors confirmed the protumor phenotype of TAMs. These data provide direct evidence for a tumor-promoting role of monocytes and macrophages in human cancer and indicate IL-1-IL-1R as a possible therapeutic target. Monocytes and macrophages are major components of the tumor microenvironment, but their contributions to human cancer are poorly understood. We used molecular profiling combined with functional assays to investigate the role of these cells in human renal cell carcinoma (RCC). Blood monocytes from RCC patients displayed a tumor-promoting transcriptional profile that supported functions like angiogenesis and invasion. Induction of this protumor phenotype required an interleukin-1 receptor (IL-1R)-dependent mechanism. Indeed, targeting of IL-1-IL-1R axis in a human RCC xenograft model abrogated the protumor phenotype of tumor-associated macrophages (TAMs) and reduced tumor growth in vivo. Supporting this, meta-analysis of gene expression from human RCC tumors showed IL1B expression to correlate with myelomonocytic markers, protumor genes, and tumor staging. Analyzing RCC patient tumors confirmed the protumor phenotype of TAMs. These data provide direct evidence for a tumor-promoting role of monocytes and macrophages in human cancer and indicate IL-1-IL-1R as a possible therapeutic target. The causal link between inflammation and cancer is now well established (Coussens et al., 2013Coussens L.M. Zitvogel L. Palucka A.K. Neutralizing tumor-promoting chronic inflammation: a magic bullet?.Science. 2013; 339: 286-291Crossref PubMed Scopus (795) Google Scholar, Grivennikov et al., 2010Grivennikov S.I. Greten F.R. Karin M. Immunity, inflammation, and cancer.Cell. 2010; 140: 883-899Abstract Full Text Full Text PDF PubMed Scopus (7427) Google Scholar, Hanahan and Weinberg, 2011Hanahan D. Weinberg R.A. Hallmarks of cancer: the next generation.Cell. 2011; 144: 646-674Abstract Full Text Full Text PDF PubMed Scopus (42748) Google Scholar, Mantovani et al., 2008Mantovani A. Allavena P. Sica A. Balkwill F. Cancer-related inflammation.Nature. 2008; 454: 436-444Crossref PubMed Scopus (7861) Google Scholar). Monocytes and macrophages represent the major inflammatory infiltrate associated within most solid tumors (Mantovani et al., 2008Mantovani A. Allavena P. Sica A. Balkwill F. Cancer-related inflammation.Nature. 2008; 454: 436-444Crossref PubMed Scopus (7861) Google Scholar, Noy and Pollard, 2014Noy R. Pollard J.W. Tumor-associated macrophages: from mechanisms to therapy.Immunity. 2014; 41: 49-61Abstract Full Text Full Text PDF PubMed Scopus (2392) Google Scholar), and their recruitment and activation at these sites is largely regulated by tumor-derived signals including chemokines, cytokines, and endogenous signals (Mantovani et al., 2008Mantovani A. Allavena P. Sica A. Balkwill F. Cancer-related inflammation.Nature. 2008; 454: 436-444Crossref PubMed Scopus (7861) Google Scholar). The role of these cells in promoting tumor progression has been revealed primarily by studies involving their depletion or accumulation in spontaneous and transplanted murine tumor models (Biswas and Mantovani, 2010Biswas S.K. Mantovani A. Macrophage plasticity and interaction with lymphocyte subsets: cancer as a paradigm.Nat. Immunol. 2010; 11: 889-896Crossref PubMed Scopus (2578) Google Scholar, Biswas et al., 2008Biswas S.K. Sica A. Lewis C.E. Plasticity of macrophage function during tumor progression: regulation by distinct molecular mechanisms.J. Immunol. 2008; 180: 2011-2017Crossref PubMed Scopus (332) Google Scholar, Coussens et al., 2000Coussens L.M. Tinkle C.L. Hanahan D. Werb Z. MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis.Cell. 2000; 103: 481-490Abstract Full Text Full Text PDF PubMed Scopus (1138) Google Scholar, Lin et al., 2006Lin E.Y. Li J.F. Gnatovskiy L. Deng Y. Zhu L. Grzesik D.A. Qian H. Xue X.N. Pollard J.W. Macrophages regulate the angiogenic switch in a mouse model of breast cancer.Cancer Res. 2006; 66: 11238-11246Crossref PubMed Scopus (811) Google Scholar, Noy and Pollard, 2014Noy R. Pollard J.W. Tumor-associated macrophages: from mechanisms to therapy.Immunity. 2014; 41: 49-61Abstract Full Text Full Text PDF PubMed Scopus (2392) Google Scholar). In contrast, our knowledge of the role played by monocytes and macrophages in human cancers remains limited. Most of the available human data come from epidemiological studies that demonstrate a correlation between increased macrophage density and poor prognosis in various cancers (including thyroid, breast, cervix, lung, and liver) (Bingle et al., 2002Bingle L. Brown N.J. Lewis C.E. The role of tumour-associated macrophages in tumour progression: implications for new anticancer therapies.J. Pathol. 2002; 196: 254-265Crossref PubMed Scopus (1576) Google Scholar, Zhang et al., 2012Zhang Q.W. Liu L. Gong C.Y. Shi H.S. Zeng Y.H. Wang X.Z. Zhao Y.W. Wei Y.Q. Prognostic significance of tumor-associated macrophages in solid tumor: a meta-analysis of the literature.PLoS ONE. 2012; 7: e50946Crossref PubMed Scopus (652) Google Scholar) and with poor responses to chemotherapy (DeNardo et al., 2011DeNardo D.G. Brennan D.J. Rexhepaj E. Ruffell B. Shiao S.L. Madden S.F. Gallagher W.M. Wadhwani N. Keil S.D. Junaid S.A. et al.Leukocyte complexity predicts breast cancer survival and functionally regulates response to chemotherapy.Cancer Discov. 2011; 1: 54-67Crossref PubMed Scopus (531) Google Scholar). High circulating monocyte counts have also been associated with poor survival in patients with other cancers such as melanoma, head and neck cancer, and malignant pleural mesothelioma (Burt et al., 2011Burt B.M. Rodig S.J. Tilleman T.R. Elbardissi A.W. Bueno R. Sugarbaker D.J. Circulating and tumor-infiltrating myeloid cells predict survival in human pleural mesothelioma.Cancer. 2011; 117: 5234-5244Crossref PubMed Scopus (82) Google Scholar). However, an in-depth characterization of monocytes and macrophages in human cancer together with the mechanism(s) responsible for “educating” these cells to a tumor-promoting phenotype in vivo is still lacking. We investigated the role played by monocytes and macrophages in human renal cell carcinoma (RCC), which is the most common type of kidney cancer in humans and the third most common urological cancer after prostate and bladder cancer. The incidence of RCC has steadily increased over the last 20 years, leading to estimates that approximately 58,240 patients will have been diagnosed by the start of this decade, with 13,040 cases presenting in the United States alone (Chow et al., 2010Chow W.H. Dong L.M. Devesa S.S. Epidemiology and risk factors for kidney cancer.Nat. Rev. Urol. 2010; 7: 245-257Crossref PubMed Scopus (983) Google Scholar, Howlader et al., 2000Howlader N. Noone A.M. Krapcho M. Neyman N. Aminou R. Waldron W. Altekruse S.F. Kosary C.L. Ruhl J. Tatalovich Z. et al.SEER Cancer Statistics Review, 1975-2008. National Cancer Institute, Bethesda, MD2000Google Scholar). RCC is characterized by a lack of early warning symptoms, a variety of clinical manifestations, resistance to chemo- and radiation therapy, and most importantly, a high rate of metastasis (Koul et al., 2011Koul H. Huh J.S. Rove K.O. Crompton L. Koul S. Meacham R.B. Kim F.J. Molecular aspects of renal cell carcinoma: a review.Am. J. Cancer Res. 2011; 1: 240-254PubMed Google Scholar). Indeed, around 50% of RCC patients with localized disease subsequently develop metastatic disease, and the 5-year survival for metastatic disease is only 9%. Although targeted antiangiogenic agents in the form of mammalian target of rapamycin (mTOR) inhibitors, tyrosine kinase inhibitors, and monoclonal antibodies are increasingly being used (Motzer et al., 2007aMotzer R.J. Hudes G.R. Curti B.D. McDermott D.F. Escudier B.J. Negrier S. Duclos B. Moore L. O’Toole T. Boni J.P. Dutcher J.P. Phase I/II trial of temsirolimus combined with interferon alfa for advanced renal cell carcinoma.J. Clin. Oncol. 2007; 25: 3958-3964Crossref PubMed Scopus (113) Google Scholar, Motzer et al., 2007bMotzer R.J. Michaelson M.D. Rosenberg J. Bukowski R.M. Curti B.D. George D.J. Hudes G.R. Redman B.G. Margolin K.A. Wilding G. Sunitinib efficacy against advanced renal cell carcinoma.J. Urol. 2007; 178: 1883-1887Abstract Full Text Full Text PDF PubMed Scopus (180) Google Scholar) and have delivered improvements in progression-free survival, RCC still remains a challenging disease to treat. A better understanding of the cellular and molecular interactions that contribute to RCC progression is therefore necessary to support the future development of more effective therapeutic strategies. The present study reports a tumor-promoting role for monocytes and macrophages in human RCC and identifies a molecular mechanism responsible for polarizing these cells toward a protumor phenotype. We further demonstrate that in vitro and in vivo targeting of this mechanism not only prevents monocytes and macrophages from adopting a protumor phenotype but also favors the acquisition of an antitumor phenotype, resulting in decreased tumor growth in a xenograft model of human RCC. These findings define a critical component of tumor progression in human RCC and identify a potential target pathway for future therapeutic interventions. In order to clarify the role played by monocytes in human RCC progression, we first performed transcriptomal profiling of these cells in RCC patients. Blood monocytes from RCC patients (RCC monocytes) and healthy donors (monocytes) were isolated and compared by microarray analysis, using the 48K genome-wide human Illumina HT-12v4 microarray (as described in the Supplemental Experimental Procedures section and shown in Figure S1A available online). Principal component analysis (PCA) and hierarchical clustering clearly segregated the monocyte trancriptome profile of healthy controls from that of RCC patients, suggesting the populations to be transcriptomally distinct (Figures 1A and 1B ). Limma differentially expressed gene (DEG) analysis of the transcriptome revealed differential modulation of 2,384 genes (1,054 upregulated; 1,330 downregulated; FDR < 0.05) in RCC monocytes compared with control monocytes (Figure S1B). These DEGs were then grouped using gene ontology (GO) biological processes, which identified immune-related genes as the most significant upregulated gene function group in RCC monocytes (Figure 1C). We initially focused on the expression of immune-related genes that encode cytokines, chemokines, and growth factors because these are known to shape the tumor microenvironment (Biswas et al., 2008Biswas S.K. Sica A. Lewis C.E. Plasticity of macrophage function during tumor progression: regulation by distinct molecular mechanisms.J. Immunol. 2008; 180: 2011-2017Crossref PubMed Scopus (332) Google Scholar, Lewis and Pollard, 2006Lewis C.E. Pollard J.W. Distinct role of macrophages in different tumor microenvironments.Cancer Res. 2006; 66: 605-612Crossref PubMed Scopus (1708) Google Scholar, Mantovani et al., 2008Mantovani A. Allavena P. Sica A. Balkwill F. Cancer-related inflammation.Nature. 2008; 454: 436-444Crossref PubMed Scopus (7861) Google Scholar). As indicated in Table 1, RCC monocytes consistently displayed upregulation of a large number of proinflammatory cytokine and chemokine genes (e.g., TNF, IL1A, IL1B, IL24, CCL3, CCL3L1, CCL5, CCL7, CCL20) relative to monocytes obtained from healthy controls. Importantly, RCC monocytes also exhibited upregulation of several “protumor” genes including PTGS2 (encoding COX2), IL8, VEGFA, MMP19, MMP10, CXCR4, and HIF1A, which are known to mediate key processes in tumor development (Table 1; Mantovani et al., 2008Mantovani A. Allavena P. Sica A. Balkwill F. Cancer-related inflammation.Nature. 2008; 454: 436-444Crossref PubMed Scopus (7861) Google Scholar, Murdoch et al., 2008Murdoch C. Muthana M. Coffelt S.B. Lewis C.E. The role of myeloid cells in the promotion of tumour angiogenesis.Nat. Rev. Cancer. 2008; 8: 618-631Crossref PubMed Scopus (1236) Google Scholar). Figure 1D shows a heatmap representation of selected proinflammatory and protumor genes that were differentially expressed in RCC monocytes compared with healthy control monocytes.Table 1Upregulation of Selected Inflammatory and Protumor Genes in RCC MonocytesaAll genes shown here are DEGs derived from the transcriptome analysis of RCC monocytes versus monocytes, as described in the Supplemental Experimental Procedures and Results sections. Upregulated DEGs are defined by an adjusted p value ≤ 0.05 and an absolute log2 fold change > = 1 (i.e., an absolute fold change > = 2) See also Table S1.Gene SymbolFull NameFold change (Log 2)Inflammatory cytokine/chemokine genesCCL3Homo sapiens chemokine (C-C motif) ligand 3 (CCL3), mRNA.2.43CCL3L1Homo sapiens chemokine (C-C motif) ligand 3-like 1 (CCL3L1), mRNA.2.39CCL4L2Homo sapiens chemokine (C-C motif) ligand 4-like 2 (CCL4L2), mRNA.3.16CCL5Homo sapiens chemokine (C-C motif) ligand 5 (CCL5), mRNA.2.14CCL7Homo sapiens chemokine (C-C motif) ligand 7 (CCL7), mRNA.3.16CCL20Homo sapiens chemokine (C-C motif) ligand 20 (CCL20), mRNA.4.05CXCL2Homo sapiens chemokine (C-X-C motif) ligand 2 (CXCL2), mRNA.3.11TNFHomo sapiens tumor necrosis factor (TNF superfamily, member 2) (TNF), mRNA.2.83IL1AHomo sapiens interleukin 1, alpha (IL-1A), mRNA.2.65IL1BHomo sapiens interleukin 1, beta (IL-1B), mRNA.3.48IL24Homo sapiens interleukin 24 (IL-24), transcript variant 2, mRNA.1.55Protumoral genesIL8Homo sapiens interleukin 8 (IL-8), mRNA.1.58VEGFAHomo sapiens vascular endothelial growth factor A (VEGFA), transcript variant 2, mRNA.1.28PTGS2Homo sapiens prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) (PTGS2), mRNA.3.44MMP10Homo sapiens matrix metallopeptidase 10 (stromelysin 2) (MMP10), mRNA.1.86MMP19Homo sapiens matrix metallopeptidase 19 (MMP19), transcript variant 1, mRNA.1.70CXCR4Homo sapiens chemokine (C-X-C motif) receptor 4 (CXCR4), transcript variant 1, mRNA.2.05HIF1AHomo sapiens hypoxia-inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor) (HIF1A), transcript variant 2, mRNA.1.18a All genes shown here are DEGs derived from the transcriptome analysis of RCC monocytes versus monocytes, as described in the Supplemental Experimental Procedures and Results sections. Upregulated DEGs are defined by an adjusted p value ≤ 0.05 and an absolute log2 fold change > = 1 (i.e., an absolute fold change > = 2) See also Table S1. Open table in a new tab Because the tumor microenvironment can polarize myelomonocytic cells (Biswas and Mantovani, 2010Biswas S.K. Mantovani A. Macrophage plasticity and interaction with lymphocyte subsets: cancer as a paradigm.Nat. Immunol. 2010; 11: 889-896Crossref PubMed Scopus (2578) Google Scholar), we next aimed to determine whether the gene-expression profile of RCC monocytes was indicative of their polarization status. RCC monocytes were screened for the differential expression of a panel of M1- or M2-polarization related genes (Martinez et al., 2006Martinez F.O. Gordon S. Locati M. Mantovani A. Transcriptional profiling of the human monocyte-to-macrophage differentiation and polarization: new molecules and patterns of gene expression.J. Immunol. 2006; 177: 7303-7311Crossref PubMed Scopus (1732) Google Scholar, Martinez et al., 2013Martinez F.O. Helming L. Milde R. Varin A. Melgert B.N. Draijer C. Thomas B. Fabbri M. Crawshaw A. Ho L.P. et al.Genetic programs expressed in resting and IL-4 alternatively activated mouse and human macrophages: similarities and differences.Blood. 2013; 121: e57-e69Crossref PubMed Scopus (347) Google Scholar, Murray et al., 2014Murray P.J. Allen J.E. Biswas S.K. Fisher E.A. Gilroy D.W. Goerdt S. Gordon S. Hamilton J.A. Ivashkiv L.B. Lawrence T. et al.Macrophage activation and polarization: nomenclature and experimental guidelines.Immunity. 2014; 41: 14-20Abstract Full Text Full Text PDF PubMed Scopus (3534) Google Scholar). The profiling data for these genes indicated that RCC monocytes express a mixture of both M1 and M2 genes rather than exhibiting a distinct M1 or M2 phenotype (Figure 1E). Taken together, the transcriptome data indicated that RCC monocytes possess a distinct gene expression profile suggestive of altered function in human cancer. We therefore sought to further validate these data and to functionally characterize RCC monocytes in subsequent experiments. A complete list of the DEGs can be found in Table S1 available online. To further validate the gene-expression profile of RCC monocytes, a panel of differentially modulated genes was chosen from Table 1 and assessed by quantitative PCR (qPCR). qPCR analysis confirmed significant increase in the expression of proinflammatory cytokine and chemokine genes TNF, IL1A, IL1B, CCL3, CCL5, CCL20, and IL6 in RCC monocytes compared with monocytes from healthy controls (Figure 2A). Accordingly, we were able to confirm the upregulated expression of TNF-α, IL-1β, IL-6, and CCL3 proteins in RCC monocyte culture supernatants (Figure 2B). Further confirmation of the microarray data was achieved by qPCR validation of the upregulated expression of protumor genes IL8, VEGFA, PTGS2, CXCR4, and MMP10 in RCC monocytes compared with control monocytes (Figure 2C). Consistent with these data, elevated protein expression of proangiogenic factors VEGFA and IL-8 were also detected in RCC monocyte culture supernatants as compared with control monocytes (Figure 2D). These data indicated that RCC monocytes upregulate key protumor genes and proteins that have been reported to support angiogenesis and metastasis (Murdoch et al., 2008Murdoch C. Muthana M. Coffelt S.B. Lewis C.E. The role of myeloid cells in the promotion of tumour angiogenesis.Nat. Rev. Cancer. 2008; 8: 618-631Crossref PubMed Scopus (1236) Google Scholar). To test whether RCC monocytes could actually promote such tumor-promoting functions, in vitro functional assays for angiogenesis and invasion were performed. RCC monocyte supernatant markedly enhanced tube formation by HUVEC cells (a measure of angiogenesis) as compared to control monocyte supernatant, indicating their proangiogenic property (Figure 2E, left). This effect was inhibited by adding an α-VEGFR2 antibody indicating that the proangiogenic ability of RCC monocytes was dependent on VEGF (Figure 2E, right), consistent with the upregulation of VEGFA by RCC monocytes (Figures 2C and 2D). Additionally, data from an in vitro invasion assay using matrix-gel-coated transwells revealed a significant increase in invasion by RCC cells in the presence of RCC monocyte supernatant (compared with control monocyte supernatant), suggesting that RCC monocytes are also capable of enhancing tumor cell invasion (Figure 2F, left). This effect was abrogated by adding a matrix metalloproteinases (MMP) inhibitor, indicating that the ability of RCC monocytes to promote tumor cell invasion was dependent on MMP (Figure 2F, right), which is consistent with the upregulation of MMP10 by RCC monocytes (Figure 2C). Collectively, these gene-expression, protein, and in vitro functional data strongly suggested that RCC monocytes exhibit an inflammatory, protumor phenotype with potential to promote cancer progression. We next sought to determine the molecular mechanism(s) that support monocytes acquisition of a protumor phenotype. Because NF-κB is a master regulator of many genes known to modulate tumor development, e.g., TNFA, IL6, VEGFA, PTGS2, and MMP (hereafter termed “protumor genes”) (Biswas and Lewis, 2010Biswas S.K. Lewis C.E. NF-κB as a central regulator of macrophage function in tumors.J. Leukoc. Biol. 2010; 88: 877-884Crossref PubMed Scopus (105) Google Scholar, Karin and Greten, 2005Karin M. Greten F.R. NF-kappaB: linking inflammation and immunity to cancer development and progression.Nat. Rev. Immunol. 2005; 5: 749-759Crossref PubMed Scopus (2517) Google Scholar), we began by investigating whether NF-κB activation was responsible for inducing a protumor phenotype in RCC monocytes. In order to achieve this, we first generated “tumor-conditioned” monocytes by coculturing monocytes obtained from healthy donors with a RCC tumor cell line (RCC4) in a transwell plate. After 48 hr coculture, RCC-conditioned monocytes exhibited marked increase in the expression of protumor genes, angiogenic activity, and invasive function compared with unconditioned monocytes (Figures S2A–S2C). Similar results were obtained for monocytes cocultured with another RCC cell line (Caki-2) (Figure S2D). These data indicate that RCC cells can “condition” normal monocytes to adopt a protumor phenotype similar to that observed in RCC patients and could therefore be used to probe the mechanisms that support monocyte acquisition of tumor-promoting functions. RCC-conditioned monocytes displayed evidence of NF-κB activation as indicated by enhanced I-κBα phosphorylation and nuclear translocation of p65 NF-κB (Figures 3A and 3B ). The role of NF-κB activation in RCC-conditioned monocytes was further investigated using a specific inhibitory peptide for the NF-κB regulator, IKKγ. RCC-conditioned monocytes treated with the inhibitory peptide displayed a marked decrease in protumor genes TNF, IL6, VEGFA, and PTGS2 expression compared with conditioned monocytes that received a control peptide (Figure 3C). Functionally, these inhibitor-treated cells also showed marked downregulation of angiogenic activity and invasive behavior (Figures 3D and 3E). Together, these results suggest that the protumor phenotype of RCC-conditioned monocytes is regulated by NF-κB activation. We further analyzed whether the protumor phenotype of RCC-conditioned monocytes relied on the MyD88 signaling pathway, upstream of NF-κB. RCC-conditioned monocytes exposed to a MyD88 inhibitory peptide displayed a marked decrease in the expression of TNF, PTGS2, IL6, and VEGFA compared with monocytes exposed to the control peptide (Figure 3F). Functionally, RCC-conditioned monocytes were impaired in their ability to stimulate angiogenesis and tumor cell invasion in vitro after treatment with the MyD88 inhibitory peptide (Figures 3G and 3H). Collectively, these observations support the involvement of the MyD88 signaling pathway in driving the protumor phenotype of RCC-conditioned monocytes. Elevated IL-1β expression was detected in the plasma of RCC patients (Figure S3A). Because IL-1 signals through the MyD88-NF-κB pathway (Biswas and Lewis, 2010Biswas S.K. Lewis C.E. NF-κB as a central regulator of macrophage function in tumors.J. Leukoc. Biol. 2010; 88: 877-884Crossref PubMed Scopus (105) Google Scholar) and could induce protumor genes (Figure S3B), we focused subsequent studies on investigating the involvement of IL-1-IL-1R in mediating the protumor phenotype of monocytes. In order to assess the role of IL-1-IL-1R in inducing protumor gene expression by RCC-conditioned monocytes, we supplemented Monocyte:RCC cocultures with human recombinant IL-1 receptor antagonist (IL-1RA), to inhibit IL-1-IL-1R signaling. IL-1RA treatment significantly inhibited the expression of protumor genes in RCC-conditioned monocytes (Figure 4A). Because TNF is another primary cytokine that is implicated in RCC (Chuang et al., 2008Chuang M.J. Sun K.H. Tang S.J. Deng M.W. Wu Y.H. Sung J.S. Cha T.L. Sun G.H. Tumor-derived tumor necrosis factor-alpha promotes progression and epithelial-mesenchymal transition in renal cell carcinoma cells.Cancer Sci. 2008; 99: 905-913Crossref PubMed Scopus (126) Google Scholar), we checked in parallel the effect of adding TNF neutralizing antibody in addition to IL-1RA in the above experiments. However, addition of anti-TNF antibody (α-TNF+IL-1RA) did not have any further inhibitory effect (Figure 4A, compare IL-1RA versus α-TNF+IL-1RA). Similarly, adding anti-TNF antibody alone failed to show a significant inhibition in the expression of most protumor genes (Figure S3C). We also examined whether IL-1RA treatment altered the protumor functions of RCC-conditioned monocytes. Figures 4B and 4C show that supernatants from RCC-conditioned monocytes that had received IL-1RA treatment displayed a significant reduction of angiogenesis and tumor cell invasion. Similar results were reproduced in monocytes cocultured with another metastatic RCC cell line, A498 (Figures 4D–4F). Together, these results clearly indicate a role for the IL-1-IL-1R pathway in promoting a protumor phenotype in RCC monocytes. In order to validate our in vitro data in an in vivo system, we established a human RCC4 xenograft model in SCID mice (detailed in Supplemental Experimental Procedures). The xenograft tumors showed substantial infiltration of F4/80+ TAMs (constituting >10% of the total live tumor cell population) (Figures S4A and S4B, left panel). To trace the origin of these TAMs, we used a previously reported fate-mapping approach for labeling blood monocytes in vivo with fluorescent latex beads that could then be tracked in the tumors (Movahedi et al., 2010Movahedi K. Laoui D. Gysemans C. Baeten M. Stangé G. Van den Bossche J. Mack M. Pipeleers D. In’t Veld P. De Baetselier P. Van Ginderachter J.A. Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes.Cancer Res. 2010; 70: 5728-5739Crossref PubMed Scopus (835) Google Scholar). Figure S4C shows the infiltration of labeled inflammatory monocytes (CD11b+Ly6ChiF4/80lo) and TAMs (CD11b+Ly6Chi/loF4/80hi) in the RCC tumors with monocyte percentage decreasing, while TAM percentage increases from day 8 to 14 of tumor growth. These results indicate inflammatory blood monocytes to infiltrate the RCC tumors and differentiate into TAMs. Moreover, TAMs showed protumor gene expression, as revealed by upregulation of TNF, IL6, IL1B, PTGS2, VEGFA, and MMP10, compared to peritoneal macrophages (PECs) from tumor-free animals (Figure S4D). Further evidence for their protumor role came from a monocyte-macrophage depletion experiment, using liposome-clodronate injection, which significantly reduced tumor growth in our RCC4 xenograft model (Figure S4E). We next investigated the involvement of IL-1-IL-1R signaling in driving the protumor phenotype of TAMs in vivo using the above xenograft tumor model. Initial immunohistochemistry revealed the presence of IL-1β in the RCC4 tumor tissues (Figure S4B, right). To directly assess the role of IL-1R signaling in driving TAMs to adopt a protumor phenotype and support disease progression in vivo, we injected tumor-bearing animals with recombinant IL-1RA or PBS intratumorally on days 7, 9, and 11 after tumor implantation (as described in Supplemental Experimental Procedures). Tumor take was monitored throughout and TAMs were analyzed after sacrifice on day 20-21. IL-1RA treatment resulted in a decrease in tumor growth (Figure 5A). TAMs from IL-1RA-treated mice showed a marked downregulation of protumor genes as compared to TAMs from PBS-treated mice (Figure 5B). In line with this, culture supernatants of TAMs from IL-1RA-treated tumor showed lesser angiogenic activity and tumor cell invasion, indicating diminished protumor functions (Figures 5C and 5D). The ability of α-VEGFR2 antibody and MMP inhibitor to block angiogenesis and tumor cell invasion induced by TAM supernatants mechanistically links these functions to VEGFA and MMP10 expression by TAMs. Additionally, in vivo imaging confirmed reduced angiogenesis and MMP activity (= invasion) in situ in the tumors of IL-1RA-treated mice (Figure 5E). Finally, our observations were also reproduced in another RCC xenograft model using the aggressive A498 line (Figure S5A). Decreased tumor growth (Figure S5B) and abrogation of TAM protumor gene expression and function were noted in the IL-1RA-treated animals (Figures S5C–S5E). However, TAM infiltration did not show any significant change between PBS- and IL-1RA-treated tumors in both our xenograft models (Figure S5F). As TAMs have been considered as polarized macrophages, we wondered whether IL-1RA could change the polarization status of these cells. TAMs from PBS-treated, tumor-bearing mice displayed a IL12BloIL10hi/NOS2loARG1hi gene-expression profile, which is characteristic to murine M2 macrophages. In contrast, TAMs from IL-1RA-treated tumor-bearing mice showed increased IL12B and NOS2 expression accompanied by decreased IL10 and ARG1 expression, suggestive of a skewing toward the so-called M1-like phenotype with respect to these markers (Figure 5F). Taken together, the multiple evidences present above demonstrate that in vivo targeting of the IL-1-IL-1R pathway by IL-1RA could indeed abrogate the protumor gene expression and functions of TAMs resulting in decreased tumor growth. Co" @default.
• W2033820276 created "2016-06-24" @default.
• W2033820276 creator A5002364273 @default.
• W2033820276 creator A5006185510 @default.
• W2033820276 creator A5007295641 @default.
• W2033820276 creator A5008473414 @default.
• W2033820276 creator A5009692472 @default.
• W2033820276 creator A5024654014 @default.
• W2033820276 creator A5028518285 @default.
• W2033820276 creator A5040482537 @default.
• W2033820276 creator A5042429669 @default.
• W2033820276 creator A5050267398 @default.
• W2033820276 creator A5054557853 @default.
• W2033820276 creator A5058268636 @default.
• W2033820276 creator A5071896688 @default.
• W2033820276 creator A5076434867 @default.
• W2033820276 creator A5077446473 @default.
• W2033820276 creator A5080409424 @default.
• W2033820276 creator A5088406118 @default.
• W2033820276 creator A5090123939 @default.
• W2033820276 date "2014-11-01" @default.
• W2033820276 modified "2023-10-10" @default.
• W2033820276 title "Molecular Profiling Reveals a Tumor-Promoting Phenotype of Monocytes and Macrophages in Human Cancer Progression" @default.
• W2033820276 cites W1502240334 @default.
• W2033820276 cites W1649938880 @default.
• W2033820276 cites W1923917571 @default.
• W2033820276 cites W1963904011 @default.
• W2033820276 cites W1968594028 @default.
• W2033820276 cites W1969057487 @default.
• W2033820276 cites W1971620413 @default.
• W2033820276 cites W1976043651 @default.
• W2033820276 cites W1977918192 @default.
• W2033820276 cites W1982593115 @default.
• W2033820276 cites W1988924112 @default.
• W2033820276 cites W1991863108 @default.
• W2033820276 cites W2004150448 @default.
• W2033820276 cites W2009771778 @default.
• W2033820276 cites W2017742161 @default.
• W2033820276 cites W2021857124 @default.
• W2033820276 cites W2023164440 @default.
• W2033820276 cites W2034400622 @default.
• W2033820276 cites W2056666823 @default.
• W2033820276 cites W2061123122 @default.
• W2033820276 cites W2061294496 @default.
• W2033820276 cites W2062736692 @default.
• W2033820276 cites W2071507111 @default.
• W2033820276 cites W2094659815 @default.
• W2033820276 cites W2101327682 @default.
• W2033820276 cites W2104104624 @default.
• W2033820276 cites W2105724560 @default.
• W2033820276 cites W2111161262 @default.
• W2033820276 cites W2112057120 @default.
• W2033820276 cites W2117692326 @default.
• W2033820276 cites W2118278569 @default.
• W2033820276 cites W2123105952 @default.
• W2033820276 cites W2130085747 @default.
• W2033820276 cites W2130587596 @default.
• W2033820276 cites W2131464730 @default.
• W2033820276 cites W2132125330 @default.
• W2033820276 cites W2136188258 @default.
• W2033820276 cites W2138670677 @default.
• W2033820276 cites W2145040145 @default.
• W2033820276 cites W2149142217 @default.
• W2033820276 cites W2159560463 @default.
• W2033820276 cites W2159803625 @default.
• W2033820276 cites W2159999569 @default.
• W2033820276 cites W2160042784 @default.
• W2033820276 cites W2163035374 @default.
• W2033820276 cites W2169281418 @default.
• W2033820276 cites W2318706214 @default.
• W2033820276 cites W2387025069 @default.
• W2033820276 doi "https://doi.org/10.1016/j.immuni.2014.09.014" @default.
• W2033820276 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/25453823" @default.
• W2033820276 hasPublicationYear "2014" @default.
• W2033820276 type Work @default.
• W2033820276 sameAs 2033820276 @default.
• W2033820276 citedByCount "211" @default.
• W2033820276 countsByYear W20338202762015 @default.
• W2033820276 countsByYear W20338202762016 @default.
• W2033820276 countsByYear W20338202762017 @default.
• W2033820276 countsByYear W20338202762018 @default.
• W2033820276 countsByYear W20338202762019 @default.
• W2033820276 countsByYear W20338202762020 @default.
• W2033820276 countsByYear W20338202762021 @default.
• W2033820276 countsByYear W20338202762022 @default.
• W2033820276 countsByYear W20338202762023 @default.
• W2033820276 crossrefType "journal-article" @default.
• W2033820276 hasAuthorship W2033820276A5002364273 @default.
• W2033820276 hasAuthorship W2033820276A5006185510 @default.
• W2033820276 hasAuthorship W2033820276A5007295641 @default.
• W2033820276 hasAuthorship W2033820276A5008473414 @default.
• W2033820276 hasAuthorship W2033820276A5009692472 @default.
• W2033820276 hasAuthorship W2033820276A5024654014 @default.
• W2033820276 hasAuthorship W2033820276A5028518285 @default.
• W2033820276 hasAuthorship W2033820276A5040482537 @default.
• W2033820276 hasAuthorship W2033820276A5042429669 @default.
• W2033820276 hasAuthorship W2033820276A5050267398 @default.
• W2033820276 hasAuthorship W2033820276A5054557853 @default.

• next