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Tumor-cell-derived ECM proteins influence the invasiveness of primary mammary tumors. Primary tumor sections were stained with Masson's trichrome (blue: collagen fibers, red: cells) to evaluate fibrosis (indicated by vertical double-dashed line) and encapsulation (bracket) or invasiveness (white arrowhead) into the skin of the primary tumors. Note that tumors in which LTBP3 or SNED1 are knocked down are less invasive and more encapsulated than in the control tumors. CYR61, EGLN1, or S100A2 knockdown did not affect tumors' invasiveness. DOI: 10.7554/eLife.01308.016 The following figure supplements are available for figure 6: Figure supplement 1. Tumor-cell-derived ECM proteins do not influence vascularization of primary mammary tumors. DOI: 10.7554/eLife.01308.017
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The extracellular matrix (ECM) is a major component of tumors and a significant contributor to cancer progression. In this study, we use proteomics to investigate the ECM of human mammary carcinoma xenografts and show that primary tumors of differing metastatic potential differ in ECM composition. Both tumor cells and stromal cells contribute to th...
Contexts in source publication
Context 1
... of any of the five ECM genes selected did not affect significantly primary tumor vascularization ( Figure 6-figure supplement 1). Control tumors markedly invaded the skin ( Figure 6) and the surrounding muscles (not shown), and were significantly fibrotic, as indicated by staining tumor sections with Masson's Trichrome ( Figure 6). Cells in which CYR61, EGLN1, or S100A2 were knocked down formed highly invasive (white arrow) and desmoplastic tumors resembling the control tumors, suggesting that these proteins did not influence tumor invasion or fibrosis ( Figure 6). ...
Context 2
... of any of the five ECM genes selected did not affect significantly primary tumor vascularization ( Figure 6-figure supplement 1). Control tumors markedly invaded the skin ( Figure 6) and the surrounding muscles (not shown), and were significantly fibrotic, as indicated by staining tumor sections with Masson's Trichrome ( Figure 6). Cells in which CYR61, EGLN1, or S100A2 were knocked down formed highly invasive (white arrow) and desmoplastic tumors resembling the control tumors, suggesting that these proteins did not influence tumor invasion or fibrosis ( Figure 6). ...
Context 3
... tumors markedly invaded the skin ( Figure 6) and the surrounding muscles (not shown), and were significantly fibrotic, as indicated by staining tumor sections with Masson's Trichrome ( Figure 6). Cells in which CYR61, EGLN1, or S100A2 were knocked down formed highly invasive (white arrow) and desmoplastic tumors resembling the control tumors, suggesting that these proteins did not influence tumor invasion or fibrosis ( Figure 6). Interestingly, LTBP3 and SNED1 knockdown tumors were charac- terized by deposition of collagen at the periphery of the tumor forming a thick capsule. ...
Context 4
... of EGLN1 or S100A2 strongly inhibited the formation of pulmonary metastases ( Figure 7A-7C). In contrast, knockdown of LTBP3 or SNED1 in LM2 cells did not affect lung coloniza- tion ( Figure 7A,B), consistent with our finding that these proteins influence tumor cell invasion at the primary site ( Figure 6). Although LTBP3 knockdown gave rise to the same number of metastases, the metastases were significantly smaller (based on the size of the ZsGreen-positive foci) and consistently, the Alu PCR signal was significantly lower than in the control lungs ( Figure 7C). ...
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The tumor microenvironment (TME) is a complex ecosystem, including blood vessels, immune cells, fibroblasts, extracellular matrix, cytokines, hormones, and so on. The TME differs from the normal tissue environment (NTE) in many aspects, such as tissue architecture, chronic inflammation, level of oxygen and pH, nutritional state of the cells, as wel...
Currently, metastasis remains the primary cause of death of patients with breast cancer despite the important advances in the treatment of this disease. In the complex tumour microenvironment network, several malignant and non-malignant cell types as well as components of extracellular matrix cooperate in promoting the metastatic spread of breast c...
Citations
... Our work challenges this notion, indicating that the tumour cells within the metastatic niche appear to play a critical role in the secretion and deposition of fibronectin into the ECM. Consistent with our results, at least one previous study found fibronectin in both tumour cells 39,47 and the stroma of the primary tumour niche 47 , implicating fibronectin in metastatic spread. ...
Identifying the adaptive mechanisms of metastatic cancer cells remains an elusive question in the treatment of metastatic disease, particularly in pancreatic cancer (pancreatic adenocarcinoma, PDA). A loss-of-function shRNA targeted screen in metastatic-derived cells identified Gstt1, a member of the glutathione S-transferase superfamily, as uniquely required for dissemination and metastasis, but dispensable for primary tumour growth. Gstt1 is expressed in latent disseminated tumour cells (DTCs), is retained within a subpopulation of slow-cycling cells within existing metastases, and its inhibition leads to complete regression of macrometastatic tumours. This distinct Gstt1high population is highly metastatic and retains slow-cycling phenotypes, epithelial–mesenchymal transition features and DTC characteristics compared to the Gstt1low population. Mechanistic studies indicate that in this subset of cancer cells, Gstt1 maintains metastases by binding and glutathione-modifying intracellular fibronectin, in turn promoting its secretion and deposition into the metastatic microenvironment. We identified Gstt1 as a mediator of metastasis, highlighting the importance of heterogeneity and its influence on the metastatic tumour microenvironment.
... For example, H3K27ac, H3K4me1, and H3K4me3 differential signals in the differential cRAMs were highly enriched in the ECM-receptor interaction pathway. ECM pathway and its remodeling by cancer have been substantiated to be highly associated with cancer cells growth, survival, and metastasis, and thus it would be a prognostic signature of non-small lung cancer [42][43][44][45] . In addition, we observed pathways such as "TGFBR1KD mutants in cancer", "loss of function of TGFBR1 in cancer", "loss of function of smad2/3 in cancer", and "signaling by TGF-beta receptor complex in cancer", which are critical in cancer development and progress and considered therapeutic targets in clinical studies 46,47 . ...
Epigenomic mechanisms are critically involved in mediation of genetic and environmental factors that underlie cancer development. Histone modifications represent highly informative epigenomic marks that reveal activation and repression of gene activities and dysregulation of transcriptional control due to tumorigenesis. Here, we present a comprehensive epigenomic and transcriptomic mapping of 18 tumor and 20 non-neoplastic tissues from non-small cell lung adenocarcinoma patients. Our profiling covers 5 histone marks including activating (H3K4me3, H3K4me1, and H3K27ac) and repressive (H3K27me3 and H3K9me3) marks and the transcriptome using only 20 mg of tissue per sample, enabled by low-input omic technologies. Using advanced integrative bioinformatic analysis, we uncovered cancer-driving signaling cascade networks, changes in 3D genome modularity, and differential expression and functionalities of transcription factors and noncoding RNAs. Many of these identified genes and regulatory molecules showed no significant change in their expression or a single epigenomic modality, emphasizing the power of integrative multimodal and multiomic analysis using patient samples.
... The intersection of DEGs also revealed five correlated genes (ADRA2A, SNED1, RASL12, LIMS2, and JAM2). These screened genes have been reported as intermediate elements in tumour biological processes and are involved in tumour metastasis and drug resistance (37)(38)(39). Potential regulatory mechanisms exist among these five correlated genes in the interaction networks, in which ADCY2 might also affect tumour malignancies. ...
Background
The adenylyl cyclase (ADCY) gene family encodes enzymes responsible for the synthesis of cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP), which comprises nine transmembrane isoforms (ADCYs 1–9). Although ADCYs correlate with intracellular signalling and tumorigenesis in different malignancies, their roles in bladder cancer remain unclear.
Methods
Utilizing the bladder urothelial carcinoma (BLCA) dataset from The Cancer Genome Atlas (TCGA), we employed the R package ‘limma’ to identify differential genes. Subsequent correlation analysis with corresponding clinical data was conducted. Prognostic significance of ADCY family genes was assessed through survival analysis. Univariate and multivariate Cox regression determined ADCY2 as a potential independent risk factor for BLCA. Validation was performed using immunohistochemistry results from independent cohorts. Additionally, we delved into the mechanism of genetic variations, methylation modifications, and signalling pathways of ADCY family genes. Evaluation of their role in the immune microenvironment was achieved through R packages single-sample gene set enrichment analysis (ssGSEA), CIBERPORT, and ESTIMATE.
Results
Cases of bladder cancer were retrieved from TCGA, and the transcriptionally differentially expressed members of ADCY were identified (members 2, 4, and 5). Genomic alteration, epigenomic modification, clinicopathological characteristics and clinical survival were systematically investigated. A co-expression network was established based on the intersection of correlated genes, which was centred around ADCY2, ADCY4, and ADCY5. Enrichment analysis revealed that correlated genes were involved in epithelial-mesenchymal transition (EMT). The ADCY2 was selected as the most representative biomarker for prognosis in bladder cancer. Bladder tumour with higher ADCY2 expression had higher prognostic risk and worse survival outcomes. Moreover, ADCY2 was correlated with classic immune checkpoints, and a better responsiveness to immunotherapy was exhibited in high-expression subsets. To ameliorate universality of the conclusion, our study also included several real-world cohorts into the preliminary validation, using datasets from the Gene Expression Omnibus (GEO; GSE13507), tissue microarray (TMA) with 80 bladder cancer inclusion and clinical trial IMvigor210, which were associated with immunotherapy sensitivity, prognosis, and common biomarker presentation.
Conclusions
Our study reveals that ADCY family has prognostic value in patients with bladder cancer; the ADCY2 is a prominent prognostic biomarker. The bioinformatics analyses and validation provide direction for further functional and mechanistic studies on the screened members of ADCY family.
... However, there is emerging evidence showing support of cancer cell-derived collagen in the TME. For example, one group found that Hep3 cancer cells were able to produce type 3 collagen, and another group found matrix signatures highlighted in tumor cells through proteomic analyses of tumors (85,86). The proteomic analysis also revealed upregulation of several types of collagens in both tumor cells and stromal cells (86). ...
... For example, one group found that Hep3 cancer cells were able to produce type 3 collagen, and another group found matrix signatures highlighted in tumor cells through proteomic analyses of tumors (85,86). The proteomic analysis also revealed upregulation of several types of collagens in both tumor cells and stromal cells (86). Building upon these previous findings, we identified a mechanistic link by which SPON1 + TIMs signal through LRP8 + cancer cells to activate TGF-β1, resulting in robust cancer cell production of fibrillar collagens. ...
Lung cancer is the leading cause of cancer-related deaths in the world, and non-small cell lung cancer (NSCLC) is the most common subset. We previously found that infiltration of tumor inflammatory monocytes (TIMs) into lung squamous carcinoma (LUSC) tumors is associated with increased metastases and poor survival. To further understand how TIMs promote metastases, we compared RNA-Seq profiles of TIMs from several LUSC metastatic models with inflammatory monocytes (IMs) of non-tumor-bearing controls. We identified Spon1 as upregulated in TIMs and found that Spon1 expression in LUSC tumors corresponded with poor survival and enrichment of collagen extracellular matrix signatures. We observed SPON1+ TIMs mediate their effects directly through LRP8 on NSCLC cells, which resulted in TGF-β1 activation and robust production of fibrillar collagens. Using several orthogonal approaches, we demonstrated that SPON1+ TIMs were sufficient to promote NSCLC metastases. Additionally, we found that Spon1 loss in the host, or Lrp8 loss in cancer cells, resulted in a significant decrease of both high-density collagen matrices and metastases. Finally, we confirmed the relevance of the SPON1/LRP8/TGF-β1 axis with collagen production and survival in patients with NSCLC. Taken together, our study describes how SPON1+ TIMs promote collagen remodeling and NSCLC metastases through an LRP8/TGF-β1 signaling axis.
... For instance, dense ECM that forms part of the scar tissue in wound healing is similar to collagen deposits known as desmoplasia in cancer, which correlate with poor prognosis [5,6]. During tumor development, desmoplasia is induced by cancer-associated fibroblasts (CAF), macrophages and tumor cells and the produced collagen is resistant to enzymatic degradation [7][8][9][10][11]. CAFs are the main players in collagen remodeling in cancer. ...
... The ECM also influences the migration of immune cells that promote or prevent tumor growth, depending on tumor type and disease stage [7,9,18,19]. T-cell migration is reduced in dense, stiff matrices compared to less dense, more flexible matrices. ...
Tumors are highly complex and heterogenous ecosystems where malignant cells interact with healthy cells and the surrounding extracellular matrix (ECM). Solid tumors contain large ECM deposits that can constitute up to 60% of the tumor mass. This supports the survival and growth of cancerous cells and plays a critical role in the response to immune therapy. There is untapped potential in targeting the ECM and cell-ECM interactions to improve existing immune therapy and explore novel therapeutic strategies. The most abundant proteins in the ECM are the collagen family. There are 28 different collagen subtypes that can undergo several post-translational modifications (PTMs), which alter both their structure and functionality. Here, we review current knowledge on tumor collagen composition and the consequences of collagen PTMs affecting receptor binding, cell migration and tumor stiffness. Furthermore, we discuss how these alterations impact tumor immune responses and how collagen could be targeted to treat cancer.
... 2D cell adhesion assays are commonly used to investigate cell phenotypic behavior because they are relatively inexpensive and have low equipment requirements. Cells were left to adhere on either Collagen I, the most abundant ECM protein in breast tissue [35,36], or tissue culture plastic for 4 or 24 h, and their morphology was analyzed: size (cell area and perimeter), irregularity (solidity and form factor), and elongation (eccentricity and compactness) were quantified (Fig. 4A). On Collagen I for 4 and 24 h the 468 cells were the smallest, as measured by cell area and perimeter (Fig. 4B, G, Additional file 1: Fig S4A, D), they were also least irregular indicating fewer protrusions measured by solidity and form factor, but also rounder and less elongated, measured by eccentricity and compactness. ...
... We have shown that different ECM proteins' impact on morphology can be used to predict 3D invasion [22]. In our current study, we used Collagen I as a substrate, as it is the most abundant ECM protein in breast tissues [35,36] In conjunction with existing studies, our data further support the use of cell morphological measurements to dissect the role of individual genes and pathways in metastasis. ...
Background
Metastasis is the leading cause of death in breast cancer patients. For metastasis to occur, tumor cells must invade locally, intravasate, and colonize distant tissues and organs, all steps that require tumor cell migration. The majority of studies on invasion and metastasis rely on human breast cancer cell lines. While it is known that these cells have different properties and abilities for growth and metastasis, the in vitro morphological, proliferative, migratory, and invasive behavior of these cell lines and their correlation to in vivo behavior is poorly understood. Thus, we sought to classify each cell line as poorly or highly metastatic by characterizing tumor growth and metastasis in a murine model of six commonly used human triple-negative breast cancer xenografts, as well as determine which in vitro assays commonly used to study cell motility best predict in vivo metastasis.
Methods
We evaluated the liver and lung metastasis of human TNBC cell lines MDA-MB-231, MDA-MB-468, BT549, Hs578T, BT20, and SUM159 in immunocompromised mice. We characterized each cell line's cell morphology, proliferation, and motility in 2D and 3D to determine the variation in these parameters between cell lines.
Results
We identified MDA-MB-231, MDA-MB-468, and BT549 cells as highly tumorigenic and metastatic, Hs578T as poorly tumorigenic and metastatic, BT20 as intermediate tumorigenic with poor metastasis to the lungs but highly metastatic to the livers, and SUM159 as intermediate tumorigenic but poorly metastatic to the lungs and livers. We showed that metrics that characterize cell morphology are the most predictive of tumor growth and metastatic potential to the lungs and liver. Further, we found that no single in vitro motility assay in 2D or 3D significantly correlated with metastasis in vivo.
Conclusions
Our results provide an important resource for the TNBC research community, identifying the metastatic potential of 6 commonly used cell lines. Our findings also support the use of cell morphological analysis to investigate the metastatic potential and emphasize the need for multiple in vitro motility metrics using multiple cell lines to represent the heterogeneity of metastasis in vivo.
... The ECM, made up of structural proteins such as collagen and proteoglycans, undergoes remodeling by TME cells, affecting both its structure and function (18). It serves not just as a scaffold for tumor cells but also in roles like intercellular adhesion and paracrine signaling, contributing to tumor growth, immune evasion, and metastasis (19)(20)(21)(22)(23). CAFs are crucial for ECM upkeep, fibrosis, angiogenesis, immune suppression, invasion, and chemoresistance. ...
Colorectal cancer (CRC), known for its high metastatic potential, remains a leading cause of cancer-related death. This review emphasizes the critical role of immune responses in CRC metastasis, focusing on the interaction between immune cells and tumor microenvironment. We explore how immune cells, through cytokines, chemokines, and growth factors, contribute to the CRC metastasis cascade, underlining the tumor microenvironment’s role in shaping immune responses. The review addresses CRC’s immune evasion tactics, especially the upregulation of checkpoint inhibitors like PD-1 and CTLA-4, highlighting their potential as therapeutic targets. We also examine advanced immunotherapies, including checkpoint inhibitors and immune cell transplantation, to modify immune responses and enhance treatment outcomes in CRC metastasis. Overall, our analysis offers insights into the interplay between immune molecules and the tumor environment, crucial for developing new treatments to control CRC metastasis and improve patient prognosis, with a specific focus on overcoming immune evasion, a key aspect of this special issue.
... RRAD methylation is correlated with prognosis in lung cancer, esophageal cancer, prostate cancer, nasopharyngeal carcinoma and breast cancers (Suzuki et antigen-like 1) is a newly discovered matricellular protein and is involved in cancer progression (Shan et al., 2021). TINAGL1 is generally dysregulated in highly metastatic tumors (Naba et al., 2014). Umeyama et al. (2014) have found that methylation-related TINAGL1 is a potential therapy target with a suppressive role in NSCLC metastasis. ...
We aimed to identify prognostic methylation genes associated with lymph node metastasis (LNM) in lung squamous cell carcinoma (LUSC). Bioinformatics methods were used to obtain optimal prognostic genes for risk model construction using data from the Cancer Genome Atlas database. ROC curves were adopted to predict the prognostic value of the risk model. Multivariate regression was carried out to identify independent prognostic factors and construct a prognostic nomogram. The differences in overall survival, gene mutation and pathways between high- and low-risk groups were analyzed. Finally, the expression and methylation level of the optimal prognostic genes among different LNM stages were analyzed. FGA, GPR39, RRAD and TINAGL1 were identified as the optimal prognostic genes and were applied to establish a prognostic risk model. Significant differences were found among the different LNM stages. The risk model could predict overall survival, showing a moderate performance with AUC of 0.64–0.68. The model possessed independent prognostic value, and could accurately predict 1-, 3- and 5-year survival. Patients with a high risk score showed poorer survival. Lower gene mutation frequencies and enrichment of leukocyte transendothelial migration and the VEGF signaling pathway in the high-risk group may lead to the poor prognosis. This study identified several specific methylation markers associated with LNM in LUSC and generated a prognostic model to predict overall survival for LUSC patients.
... Growing evidence suggests that extracellular signals originating from the TME can surpass cancer cell intrinsic signaling and influence tumor growth independently of tumor clonal heterogeneity [9,10]. ECM, undergoing modification by cancer cells, stromal elements, and immune cells, has been shown to enhance cancer cell proliferation, survival, and metastasis [11][12][13][14]. The deregulation of ECM components has also been connected to HGSOC development and progression [15][16][17][18][19][20]. ...
Background
High-grade serous ovarian cancer (HGSOC) is a challenging malignancy characterized by complex interactions between tumor cells and the surrounding microenvironment. Understanding the immune landscape of HGSOC, particularly the role of the extracellular matrix (ECM), is crucial for improving prognosis and guiding therapeutic interventions.
Methods and results
Using univariate Cox regression analysis, we identified 71 ECM genes associated with prognosis in seven HGSOC populations. The ECMscore signature, consisting of 14 genes, was validated using Cox proportional hazards regression with a lasso penalty. Cox regression analyses demonstrated that ECMscore is an excellent indicator for prognostic classification in prevalent malignancies, including HGSOC. Moreover, patients with higher ECMscores exhibited more active stromal and carcinogenic activation pathways, including apical surface signaling, Notch signaling, apical junctions, Wnt signaling, epithelial-mesenchymal transition, TGF-beta signaling, and angiogenesis. In contrast, patients with relatively low ECMscores showed more active immune-related pathways, such as interferon alpha response, interferon-gamma response, and inflammatory response. The relationship between the ECMscore and genomic anomalies was further examined. Additionally, the correlation between ECMscore and immune microenvironment components and signals in HGSOC was examined in greater detail. Moreover, the expression of MGP, COL8A2, and PAPPA and its correlation with FAP were validated using qRT-PCR on samples from HGSOC. The utility of ECMscore in predicting the prospective clinical success of immunotherapy and its potential in guiding the selection of chemotherapeutic agents were also explored. Similar results were obtained from pan-cancer research.
Conclusion
The comprehensive evaluation of the ECM may help identify immune activation and assist patients in HGSOC and even pan-cancer in receiving proper therapy.
... The dysregulated expression of BM proteins was recognized as an important driver of tumor progression [37]. Naba, Alexandra et al. reported 43 ECM proteins characterized by the highly metastasic phenotype of breast cancer, such as AGRN, CST3, TIMP1, and TINAGL1 [38]. The upregulation of LTBG3 expression in metastatic melanoma compared with benign nevi was identified; this condition exhibited a significant correlation with lymph node metastasis in melanoma [39]. ...
Background
Hepatocellular carcinoma (HCC), which is characterized by its high malignancy, generally exhibits poor response to immunotherapy. As part of the tumor microenvironment, basement membranes (BMs) are involved in tumor development and immune activities. Presently, there is no integrated analysis linking the basement membrane with immune checkpoints, especially from the perspective of lncRNA.
Methods
Based on transcriptome data from The Cancer Genome Atlas, BMs-related and immune checkpoint-related lncRNAs were identified. By applying univariable Cox regression and Machine learning (LASSO and SVM-RFE algorithm), a 10-lncRNA prognosis signature was constructed. The prognostic significance of this signature was assessed by survival analysis. GSEA, ssGSEA, and drug sensitivity analysis were conducted to investigate potential functional pathways, immune status, and clinical implications of guiding individual treatments in HCC. Finally, the promoting migration effect of LINC01224 was validated via in vitro experiments.
Results
The multiple Cox regression, receiver operating characteristic curves, and stratified survival analysis of clinical subgroups exhibited the robust prognostic ability of the lncRNA signature. Results of the GSEA and drug sensitivity analysis revealed significant differences in potential functional pathways and response to drugs between the two risk groups. In addition, the risk level of HCC patients was distinctly correlated with immune cell infiltration status. More importantly, LINC01224 was independently associated with the OS of HCC patients (P < 0.05), suppressing the expression of LINC01224 inhibited the migration of HCC cells.
Conclusion
This study developed a reliable signature for the prognosis of HCC based on BM and immune checkpoint related lncRNA, revealing that LINC01224 might be a prognostic biomarker for HCC associated with the progression of HCC.
