Dissection of cancer cell and stromal cell-derived signals in melanoma xenografts following treatment with the stromal-targeting agent DMXAA
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PROVIDER: PRJNA136749 | ENA |
REPOSITORIES: ENA
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Project description:5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent, is shown here to have substantial activity as a single agent against human A375 melanoma xenografts in nude mice (94 % hemorrhagic necrosis after 24 h, and 26 days growth delay following single dose at 25 mg/kg). CD45+ cells in tumor tissue increased 5-fold over the first 3 days after treatment, which was due largely to an influx of CD11b+ Ly6G+ neutrophils. Using murine and human multiplex cytokine assays to dissect the cytokines produced by host stromal cells or by the melanoma cells, it was shown that both the stromal cells and the A375 melanoma cells produced cytokines capable of attracting neutrophils into the tumor. The same xenografts were also analyzed using human and mouse Affymetrix microarrays to separately identify tumor cell-specific (human) and stromal cell-specific (mouse) gene expression changes. DMXAA induced numerous stromal cytokine mRNAs, including IP-10, IL-6, MIP-1α/β, MIP-2, KC, RANTES, MIG, MCP-1 and IL-1β, many of which were also elevated at the protein level. Numerous human cytokine mRNAs were also induced including MCP-1, IL-8, GRO, VEGF, GM-CSF and IL-6, which again was in line with our protein data. Pathway analysis indicated that significant numbers of the stromal mRNAs induced by DMXAA are regulated downstream of TNF-α, interferon-β and NFκB. Our results suggest that DMXAA may have utility in combination therapy for human melanoma through the activation of pro-inflammatory signalling pathways and cytokine expression from both stromal and tumor cells, leading to haemorrhagic necrosis, neutrophil influx and growth inhibition. Microarrays were used to identify separately the abundance of tumour cell and stromal cell RNA in A375 melanoma xenografts, with and without treatment by the stromal targetting drug DMXAA.
2012-03-15 | E-GEOD-26308 | biostudies-arrayexpress
Project description:5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent, is shown here to have substantial activity as a single agent against human A375 melanoma xenografts in nude mice (94 % hemorrhagic necrosis after 24 h, and 26 days growth delay following single dose at 25 mg/kg). CD45+ cells in tumor tissue increased 5-fold over the first 3 days after treatment, which was due largely to an influx of CD11b+ Ly6G+ neutrophils. Using murine and human multiplex cytokine assays to dissect the cytokines produced by host stromal cells or by the melanoma cells, it was shown that both the stromal cells and the A375 melanoma cells produced cytokines capable of attracting neutrophils into the tumor. The same xenografts were also analyzed using human and mouse Affymetrix microarrays to separately identify tumor cell-specific (human) and stromal cell-specific (mouse) gene expression changes. DMXAA induced numerous stromal cytokine mRNAs, including IP-10, IL-6, MIP-1α/β, MIP-2, KC, RANTES, MIG, MCP-1 and IL-1β, many of which were also elevated at the protein level. Numerous human cytokine mRNAs were also induced including MCP-1, IL-8, GRO, VEGF, GM-CSF and IL-6, which again was in line with our protein data. Pathway analysis indicated that significant numbers of the stromal mRNAs induced by DMXAA are regulated downstream of TNF-α, interferon-β and NFκB. Our results suggest that DMXAA may have utility in combination therapy for human melanoma through the activation of pro-inflammatory signalling pathways and cytokine expression from both stromal and tumor cells, leading to haemorrhagic necrosis, neutrophil influx and growth inhibition.
2012-03-15 | GSE26308 | GEO
Project description:5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a tumor vascular disrupting agent, is shown here to have substantial activity as a single agent against human A375 melanoma xenografts in nude mice (94 % hemorrhagic necrosis after 24 h, and 26 days growth delay following single dose at 25 mg/kg). CD45+ cells in tumor tissue increased 5-fold over the first 3 days after treatment, which was due largely to an influx of CD11b+ Ly6G+ neutrophils. Using murine and human multiplex cytokine assays to dissect the cytokines produced by host stromal cells or by the melanoma cells, it was shown that both the stromal cells and the A375 melanoma cells produced cytokines capable of attracting neutrophils into the tumor. The same xenografts were also analyzed using human and mouse Affymetrix microarrays to separately identify tumor cell-specific (human) and stromal cell-specific (mouse) gene expression changes. DMXAA induced numerous stromal cytokine mRNAs, including IP-10, IL-6, MIP-1α/β, MIP-2, KC, RANTES, MIG, MCP-1 and IL-1β, many of which were also elevated at the protein level. Numerous human cytokine mRNAs were also induced including MCP-1, IL-8, GRO, VEGF, GM-CSF and IL-6, which again was in line with our protein data. Pathway analysis indicated that significant numbers of the stromal mRNAs induced by DMXAA are regulated downstream of TNF-α, interferon-β and NFκB. Our results suggest that DMXAA may have utility in combination therapy for human melanoma through the activation of pro-inflammatory signalling pathways and cytokine expression from both stromal and tumor cells, leading to haemorrhagic necrosis, neutrophil influx and growth inhibition. Microarrays were used to identify separately the abundance of tumour cell and stromal cell RNA in A375 melanoma xenografts, with and without treatment by the stromal targetting drug DMXAA.
| S-ECPF-GEOD-26308 | biostudies-other
Project description:BackgroundThe non-malignant cells of the tumour stroma have a critical role in tumour biology. Studies dissecting the interplay between cancer cells and stromal cells are required to further our understanding of tumour progression and methods of intervention. For proof-of-principle of a multi-modal approach to dissect the differential effects of treatment on cancer cells and stromal cells, we analysed the effects of the stromal-targeting agent 5,6-dimethylxanthenone-4-acetic acid on melanoma xenografts.MethodsFlow cytometry and multi-colour immunofluorescence staining was used to analyse leukocyte numbers in xenografts. Murine-specific and human-specific multiplex cytokine panels were used to quantitate cytokines produced by stromal and melanoma cells, respectively. Human and mouse Affymetrix microarrays were used to separately identify melanoma cell-specific and stromal cell-specific gene expression.Results5,6-Dimethylxanthenone-4-acetic acid activated pro-inflammatory signalling pathways and cytokine expression from both stromal and cancer cells, leading to neutrophil accumulation and haemorrhagic necrosis and a delay in tumour re-growth of 26 days in A375 melanoma xenografts.Conclusion5,6-Dimethylxanthenone-4-acetic acid and related analogues may potentially have utility in the treatment of melanoma. The experimental platform used allowed distinction between cancer cells and stromal cells and can be applied to investigate other tumour models and anti-cancer agents.
| S-EPMC3304430 | biostudies-literature
Project description:Flavonoid derivative DMXAA, also known as vadimezan or ASA404, is considered one of the most promising vascular disrupting agents that displays remarkable antitumor properties. To investigate the effect of DMXAA on gene expression in renal tissues, C57BL/6J mice were intraperitoneal injected with DMXAA or vehicle (10 mg/kg daily) for three days, then renal tissues were harvested and performed RNA-seq.
2023-11-01 | GSE216295 | GEO
Project description:Glioblastoma is the most common and most devastating adult primary brain tumor. Despite aggressive multimodal therapy, the median survival is approximately one year after diagnosis. In recent years bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have shown promise as cell-based delivery vehicles for anti-glioma therapeutics. This is largely due to their innate ability to migrate towards gliomas. Several studies have successfully demonstrated their use as delivery vehicles for anti-glioma agents. However, it is now evident that BM-hMSCs demonstrate variable tropism towards gliomas based on a clinically relevant glioma stem cell (GSC) model of GBM. In this study, we compared the proteomic profile of cancer and stromal cell populations in GSC xenografts that attract BM-hMSCs (‘attractors’) with those to do not (‘non-attractors’) in order to identify cell-signaling pathways that may modulate BM-hMSCs homing followed by targeted transcriptomic analysis of human gene sets related to glioma biology. We identified lower protein expression of fatty acid metabolism and glucose-dependent metabolic pathways in attractors. While transcriptomic analysis suggested that N-linked glycosylation was increased. Conversely, glucose metabolic pathways, including oxidative phosphorylation, were increased in the stromal cells present in attractors. The results presented here provide the first evidence for glucose metabolism, reactive oxidative species and lipid-mediated tumor inflammatory response, and N-linked glycosylation in the homing of BM-hMSC to GSC xenografts. Reciprocal expression of these pathways in the stromal cell population may suggest microenvironment cross-talk. Our studies provide new insights on the signaling correlates underlying the differential homing capacity of BM-hMSCs to GSC xenografts.
2016-02-17 | PXD001778 | Pride
Project description:This study is designed to uncover the mechanism by which STING agonist (DMXAA) changes the tumor microenivorment of mammary tumor to support Th/Tc17 CAR-T cells treatment.
2020-11-25 | GSE162073 | GEO
Project description:Vascular disrupting agents (VDA) represent a novel approach to the treatment of cancer, resulting in collapse of tumor vasculature and tumor death. 5,6-Dimethylxanthenone-4-acetic acid (DMXAA) is a VDA currently in advanced Phase II clinical trials, yet its precise mechanism of action is unknown despite extensive preclinical and clinical investigations. The data presented herein demonstrate that DMXAA is a novel and specific activator of the TBK1-IRF-3 signaling pathway. DMXAA treatment of primary murine macrophages resulted in robust IRF-3 activation, a ~750-fold increase in IFN-beta mRNA and, in contrast to the potent Toll-like receptor 4 (TLR4) agonist, lipopolysaccharide (LPS), signaling was independent of mitogen-activated protein kinase (MAPK) activation and elicited minimal NF-kappaB-dependent gene expression. DMXAA-induced signaling was critically dependent on the IRF-3 kinase, TBK1, and IRF-3, but MyD88-, TRIF-, IPS-1/MAVS-, and IKKbeta-independent, thus excluding all known TLRs and cytosolic helicase receptors. DMXAA pretreatment of murine macrophages induced a state of tolerance to LPS and vice versa. In contrast to LPS stimulation, DMXAA-induced IRF-3 dimerization and IFN-beta expression were inhibited by salicylic acid (SA). These findings detail a novel pathway for TBK-1-mediated IRF-3 activation and provide new insights into the mechanism of this new class of chemotherapeutic drugs. Keywords: DMXAA, IRF-3, TBK1, IFN-beta, Salicylic acid, TLR
2007-03-08 | GSE7194 | GEO
Project description:Bulk RNAseq of pancreatic stellate cells (PSC) treated with DMXAA, interferon gamma or DMSO control. PSC were cultured alone as mono-cultures or in co-culture with mouse tumour organoids, The PSC were recovered by FACS using fluorecent protein mKate-2.
2025-03-19 | E-MTAB-14943 | biostudies-arrayexpress
Project description:The mechanisms by which tumors develop resistance to angiogenesis inhibitors, and the relative contributions of tumor cells and stroma to resistance, are not completely understood. We developed three human lung adenocarcinoma murine models of resistance to the VEGF inhibitor bevacizumab and, using species-specific profiling, separately investigated tumor cell and stromal molecules associated with resistance. Gene expression changes associated with acquired resistance occurred predominantly in stromal (mouse) and not tumor (human) cells. Components of the EGFR and FGFR2 pathways were significantly upregulated in stroma, but not in tumor cells. Increased activated EGFR was detected on pericytes of xenografts that acquired resistance and on endothelium of tumors with relative primary resistance. Acquired resistance was associated with a pattern of pericyte-covered, normalized revascularization, whereas tortuous, uncovered vessels were observed in relative primary resistance. Dual targeting of VEGF and EGFR pathways with bevacizumab and erlotinib, or the VEGFR/EGFR inhibitor vandetanib, reduced pericyte coverage and increased progression-free survival. These findings demonstrate that alterations in tumor stromal pathways, including EGFR and FGFR2, are associated with, and may contribute to VEGF inhibitor resistance and that targeting these pathways may improve therapeutic efficacy. Understanding stromal signaling may be critical for developing biomarkers for angiogenesis inhibitors and improving combination regimens. To identify changes in stromal and tumor gene expression associated with acquired resistance to anti-VEGF therapy, we performed RNA microarray analyses comparing H1975 control (vehicle-treated) and BV-resistant (bevacizumab-treated until progression) xenografts (n=3 biologic replicates per group of treatment) using Illumina mouse (WG-6 v2) and human (WG-6 v3)-specific expression arrays. Probes in these arrays have been designed to minimize cross-species reactivity and consistent with this essentially no cross reactivity was observed in experiments mixing human and mouse cell lines.Total RNA was extracted from snap-frozen tissues using the mirVanaTM miRNA Isolation Kit (Ambion, Austin, TX) according to manufacturer’s protocol. Biotin labeled cRNA samples for hybridization were prepared by using Illumina Total Prep RNA Amplification Kit (Ambion Inc., Austin, TX). One microgram of total RNA was used for the synthesis of cDNA and followed by an amplification and biotin labeling. Each of 1.5 μg of biotinylated cRNAs was hybridized to both mouseWG-6 v2 and human WG-6v3 Expression BeadChips (Illumina®, San Diego, CA) at the same time, for analysis of murine and human transcriptomes. Signals were developed by Amersham fluorolink streptavidin-Cy3 (GE Health care Bio-Sciences, Little Chalfont, UK). Gene expression data were collected by using Illumina bead Array Reader confocal scanner (BeadStation 500GXDW; Illumina Inc.). Data were analyzed using the BRB-ArrayTools Version 3.7.0 Beta platform developed by Dr. Richard Simon and the BRB-Array Tools development team. A log base 2 transformation was applied to the data set prior to data normalization. A median array was selected as the reference array for normalization and statistical significance was set using a p<0.01. Genes differentially expressed between groups were determined applying univariate t-test with estimation of the false discovery rate (FDR). Genes were determined using selection criteria of a p<0.005 and a fold-change ≥1.5.
2011-04-22 | E-GEOD-26644 | biostudies-arrayexpress