Risk Of Breast Cancer Metastasis Research Articles

1α,25-Dihydroxyvitamin D Downregulates Adipocyte Impact on Breast Cancer Cell Migration and Adipokine Release

Background/Objectives: Excess adiposity is associated with a higher risk of breast cancer metastasis and mortality. Evidence suggests that dietary vitamin D inhibits breast cancer metastasis. However, the mechanistic link between vitamin D’s regulation of adipocyte metabolism and metastasis has not been previously investigated. Therefore, the purpose of these experiments was to examine the effect of the active form of vitamin D, 1α,25-dihydroxyvitamin D (1,25(OH)2D), on adipocyte release of bioactive compounds and whether the impact on adipocytes leads to inhibition of breast cancer cell migration, an important step of metastasis. Methods: Differentiated 3T3-L1 adipocytes were treated with 1,25(OH)2D for two days, followed by either harvesting the adipocytes or collecting adipocyte-conditioned media without 1,25(OH)2D. A transwell migration assay was conducted with vehicle- or 1,25(OH)2D-conditioned media. In order to explore the mechanism underlying effects on breast cancer metastatic capability, the mRNA expression of leptin, adiponectin, insulin-like growth factor (IGF-1), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) was measured in adipocytes following either vehicle or 1,25(OH)2D treatment. Results: Conditioned media from 1,25(OH)2D-treated adipocytes inhibited the migration of metastatic MDA-MB-231 breast cancer cells compared to conditioned media from vehicle-treated adipocytes. Treatment of adipocytes with 1,25(OH)2D decreased mRNA expression of leptin, adiponectin, IGF-1, IL-6, and MCP-1. Consistent with mRNA expression, concentrations of leptin, adiponectin, IGF-1, and IL-6 in adipocyte-conditioned media were decreased with 1,25(OH)2D treatment, although MCP-1 remained unchanged. Conclusions: In summary, these results suggest that 1,25(OH)2D alters adipocyte secretions to prevent breast cancer metastasis.

CCDC88C, an O-GalNAc glycosylation substrate of GALNT6, drives breast cancer metastasis by promoting c-JUN-mediated CEMIP transcription

Coiled-coil domain containing 88C (CCDC88C) is a component of non-canonical Wnt signaling, and its dysregulation causes colorectal cancer metastasis. Dysregulated expression of CCDC88C was observed in lymph node metastatic tumor tissues of breast cancer. However, the role of CCDC88C in breast cancer metastasis remains unclear. To address this, the stable BT549 and SKBR3 cell lines with CCDC88C overexpression or knockdown were developed. Loss/gain-of-function experiments suggested that CCDC88C drove breast cancer cell motility in vitro and lung and liver metastasis in vivo. We found that CCDC88C led to c-JUN-induced transcription activation. Overlapping genes were identified from the genes modulated by CCDC88C and c-JUN. CEMIP, one of these overlapping genes, has been confirmed to confer breast cancer metastasis. We found that CCDC88C regulated CEMIP mRNA levels via c-JUN and it exerted pro-metastatic capabilities in a CEMIP-dependent manner. Moreover, we identified the CCDC88C as a substrate of polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6). GALNT6 was positively correlated with CCDC88C protein abundance in the normal breast and breast cancer tissues, indicating that GALNT6 might be associated with expression patterns of CCDC88C in breast cancer. Our data demonstrated that GALNT6 maintained CCDC88C stability by promoting its O-linked glycosylation, and the modification was critical for the pro-metastatic potential of CCDC88C. CCDC88C also could mediate the pro-metastatic potential of GALNT6 in breast cancer. Collectively, our findings uncover that CCDC88C may increase the risk of breast cancer metastasis and elucidate the underlying molecular mechanisms.

Abstract 1390: Pulmonary viral infection promotes the awakening of dormant metastatic breast cancer cells in lungs

Abstract Background: Breast cancer is the most common form of cancer and the second cancer-causing death in females. Although remission rates are high if detected early, survival rates drop substantially when breast cancer becomes metastatic. The most common sites of metastatic breast cancer are bone, liver, and lung. Respiratory viral infections inflict illnesses on countless people, as evidenced by annual flu seasons and the latest pandemic caused by SARS-CoV-2. Respiratory viral infections result in increased inflammation with immune cell influx to facilitate viral clearance. Prior studies have shown that inflammation can contribute to dormant cancer cells awakening and outgrowth. However, how respiratory viral infections contribute to breast cancer lung metastasis remains to be unraveled. Methods: MMTV/NEU mouse model of dormant breast cancer was used, with influenza virus as the model respiratory virus. Lungs and bronchoalveolar lavage fluid were harvested for immunohistochemistry, flow cytometry, cytokines, and single-cell RNA-seq analysis. Flatiron Health Database was mined for epidemiological data to assess the risk of breast cancer metastasis to lung with COVID infection. Results: We have shown a dramatic increase in disseminated cancer cell (DCC) awakening in the lungs following influenza and SARS-CoV-2 infections. Influenza infection results in loss of a pro-dormancy mesenchymal phenotype and increased proliferation of DCC in the lungs within days post-infection, with a more than 1000-fold expansion of carcinoma cells over a couple of weeks. Strikingly, by 15 days post-infection, the lesions that expanded from solitary HER2+ DCCs almost homogeneously return to a quiescent state. Our preliminary data indicate that post-infection increases in IL-6 are required for DCC expansion post-influenza. Acute influenza infection also contributed to an accumulation of CD4+ T cells around expanding tumor cells for as long as 28 days after the initial infection. Depletion of CD4+ T cells but not CD8+ cells during infection with influenza virus prevents the expansion of the DCCs in the lung. Co-depletion of CD8 cells with CD4 cells partially restored the DCC burden in the lungs, indicating a role for CD8 cells in eliminating DCCs when released from CD4-mediated suppression. Single-cell RNA-seq analyses of CD8+ T cells with CD4 depletion showed an increased IL2/STAT5, MTORC1, and Ox/Phos activation by 15 dpi. The analyses also showed an immunosuppressive effect by the tumor cells. Finally, analysis of female breast cancer patients who tested positive with SARS-CoV-2 after their initial diagnosis exhibited a hazard ratio of 1.44 (p=0.043) for subsequent diagnosis of metastatic breast cancer in lungs. Conclusions: These studies reveal the potential risks to cancer survivors who experience respiratory viral infections, with mechanistic insight that could lead to novel prevention or intervention strategies. Citation Format: Shi Biao Chia, Bryan Johnson, Meher Boorgula, Varsha Sreekanth, Andrew Goodspeed, Bennett J. Davenport, Junxiao Hu, Dexiang Gao, Michael Papanicolaou, Thomas E. Morrison, Julio A. Aguirre-Ghiso, Mercedes Rincon, James V. DeGregori. Pulmonary viral infection promotes the awakening of dormant metastatic breast cancer cells in lungs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1390.

Construction and Validation of a Novel Nomogram for Predicting the Risk of Metastasis in a Luminal B Type Invasive Ductal Carcinoma Population.

Postoperative distant metastasis is the main cause of death in breast cancer patients. We aimed to construct a nomogram to predict the risk of metastasis of luminal B type invasive ductal carcinoma. We applied the data of 364 luminal B type breast cancer patients between 2008 and 2013. Patients were categorized into modeling group and validation group randomly (1:1). The breast cancer metastasis nomogram was developed from the logistic regression model using clinicopathological variables. The area under the receiver-operating characteristic curve (AUC) was calculated in modeling group and validation group to evaluate the predictive accuracy of the nomogram. The multivariate logistic regression analysis showed that tumor size, No. of the positive level 1 axillary lymph nodes, human epidermal growth factor receptor 2 (HER2) status and Ki67 index were the independent predictors of the breast cancer metastasis. The AUC values of the modeling group and the validation group were 0.855 and 0.818, respectively. The nomogram had a well-fitted calibration curve. The positive and negative predictive values were 49.3% and 92.7% in the modeling group, and 47.9% and 91.0% in the validation group. Patients who had a score of 60 or more were thought to have a high risk of breast cancer metastasis. The nomogram has a great predictive accuracy of predicting the risk of breast cancer metastasis. If patients had a score of 60 or more, necessary measures, like more standard treatment methods and higher treatment adherence of patients, are needed to take to lower the risk of metastasis and improve the prognosis.

Cancer Metastasis Prediction and Genomic Biomarker Identification through Machine Learning and eXplainable Artificial Intelligence in Breast Cancer Research.

Method: This research presents a model combining machine learning (ML) techniques and eXplainable artificial intelligence (XAI) to predict breast cancer (BC) metastasis and reveal important genomic biomarkers in metastasis patients. A total of 98 primary BC samples was analyzed, comprising 34 samples from patients who developed distant metastases within a 5-year follow-up period and 44 samples from patients who remained disease-free for at least 5 years after diagnosis. Genomic data were then subjected to biostatistical analysis, followed by the application of the elastic net feature selection method. This technique identified a restricted number of genomic biomarkers associated with BC metastasis. A light gradient boosting machine (LightGBM), categorical boosting (CatBoost), Extreme Gradient Boosting (XGBoost), Gradient Boosting Trees (GBT), and Ada boosting (AdaBoost) algorithms were utilized for prediction. To assess the models' predictive abilities, the accuracy, F1 score, precision, recall, area under the ROC curve (AUC), and Brier score were calculated as performance evaluation metrics. To promote interpretability and overcome the "black box" problem of ML models, a SHapley Additive exPlanations (SHAP) method was employed. The LightGBM model outperformed other models, yielding remarkable accuracy of 96% and an AUC of 99.3%. In addition to biostatistical evaluation, in XAI-based SHAP results, increased expression levels of TSPYL5, ATP5E, CA9, NUP210, SLC37A1, ARIH1, PSMD7, UBQLN1, PRAME, and UBE2T (p ≤ 0.05) were found to be associated with an increased incidence of BC metastasis. Finally, decreased levels of expression of CACTIN, TGFB3, SCUBE2, ARL4D, OR1F1, ALDH4A1, PHF1, and CROCC (p ≤ 0.05) genes were also determined to increase the risk of metastasis in BC. The findings of this study may prevent disease progression and metastases and potentially improve clinical outcomes by recommending customized treatment approaches for BC patients.

The Potential Effect of General Anesthetics in Cancer Surgery: Meta-Analysis of Postoperative Metastasis and Inflammatory Cytokines.

Metastasis or recurrence following curative surgery is the main indicator of tumor progress and is the main cause of patient death. For more than three decades, the potential for general anesthesia to affect cancer outcomes has been a subject of concern with considerable research interest. Here, we conducted this systematic review and meta-analysis to summarize the effect of inhalational anesthesia (IHNA) vs. propofol-based total intravenous anesthesia (TIVA) on metastasis and recurrence after cancer surgery from clinical and pre-clinical studies. The relative risk for metastasis/recurrence in TIVA is 0.61 (95% confidence interval (95% CI) 0.46 to 0.82, p = 0.0009) compared to IHNA. Inflammatory cytokines have been implicated in cancer metastasis following cancer surgery, thus we analyzed inflammatory cytokines levels after surgery under IHNA or TIVA. Based on pooled analysis, a lower IL-6 level was noticed in TIVA in comparison to IHNA (standardized mean difference (SMD) = 0.77, 95% CI = 0.097 to 1.44, I2 = 92%, p = 0.02) but not TNF-α or IL-10. Preclinical animal model studies show that inhalational anesthetics increase the risk of breast cancer metastasis compared to propofol. In conclusion, the current evidence suggests intravenous anesthetic propofol is associated with less metastasis/recurrence and lower postoperative IL-6 level over inhaled anesthetics in the oncological surgery. We urge more well-designed clinical and preclinical studies in this field.

Abstract NG06: Investigating biological mediators of social and environmental risk factors on cancer health disparities

Abstract NG06: Investigating biological mediators of social and environmental risk factors on cancer health disparities

Abstract 6123: High-fat diets promote lung metastasis of breast cancer by activating lung fibroblasts

Abstract Metastasis is the main cause of patient death among many tumor types including breast cancer and breast cancer patients with obesity have higher risk of distant metastases during 10-year follow-up. Diet is one of the main and manageable factors that cause obesity and regular intake of high-fat diets can induce obesity. In addition, fibroblasts, as the most abundant stromal cell type in tumor microenvironment, are involved in cancer metastasis. However, the role of fibroblasts and the downstream mechanism in metastasis of patients with high-fat diet remains unclear. In this study, we fed MMTV-PyMT transgenic mice with high-fat diet to obtain a high-fat tumorigenesis model, and isolate primary lung fibroblasts and then coculture them with breast cancer cells to explore the interaction. We found that a high-fat diet can significantly promote progression of primary breast cancer and lung metastasis, and that before weight difference shows, there is a different chemotactic influence of lung fibroblasts on breast cancer cell between high-fat diet and normal diet group. And we observed increased chemotactic function of primary lung fibroblasts in high-fat diet group. Moreover, tumor cells treated with primary lung fibroblasts in high-fat diet group show increased ability of adhesion, suggesting that high-fat diet chemoattracts breast cancer cell colonization by activating lung fibroblasts. Collectively, we explored the functional differences of lung fibroblast and the key factors for the lung colonization of recruited breast cancer cells. Our findings suggest that dietary patterns are associated with the risk of breast cancer metastasis and targeting activated fibroblasts in pre-metastatic microenvironment may serve as potential early therapeutic strategy to reduce breast cancer lung metastasis. Citation Format: Huiping Chen, Penghan Huang, Jianing Chen, Erwei Song. High-fat diets promote lung metastasis of breast cancer by activating lung fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6123.

Aspirin suppresses breast cancer metastasis to lung by targeting anoikis resistance.

Clinical evidence recently suggests that the regular use of aspirin is associated with a lower risk of breast cancer metastasis, but mechanisms remain unclear. Resistance to anoikis has been implicated in malignant transformation and metastasis. Here, we investigated whether aspirin might prevent breast cancer metastasis to lung by targeting anoikis resistance. Aspirin sensitized breast cancer cells to anoikis in vitro and lowered the circulating tumor cells as well as distant metastasis in vivo. Mechanistically, thromboxane A2 (TXA2) pathway was identified as the relevant molecular target for aspirin in anoikis sensitization. Upon detachment, both thromboxane A2 receptor (TP) and thromboxane A2 synthase 1 (TBXAS1) were up-regulated in metastatic breast cancer cells, conferred anoikis resistance through persistent activation of Akt, thereby facilitated breast cancer metastasis to lung. Consistently, either knockdown of TP in cancer cells or genetic deletion of TP in mice protected against lung metastasis in vivo. Collectively, TXA2 pathway plays a critical role in anoikis resistance and might serve as potential target for chemoprevention of breast cancer metastasis.

Aspirin Suppresses Breast Cancer Metastasis by Targeting Anoikis Resistance

Background: Aspirin intake lowers the risk of breast cancer metastasis, but mechanisms remain unclear. Anoikis resistance has been recently implicated in metastasis. Here we investigated whether aspirin might prevent breast cancer metastasis by targeting anoikis resistance. MethodsA:noikis was induced by culturing breast cancer cells on 1% agar treated plate for 24 h, and apoptotic cell death was determined by Annexin V and PI staining. Chemoprevention of breast cancer metastasis was evaluated by using a 4T1 mouse mammary tumor model. Findings: Aspirin sensitized breast cancer cells to anoikis in vitro, and lowered the circulating tumor cells as well as distant metastasis in vivo. Thromboxane A2 (TXA2) pathway was identified as the relevant molecular target for aspirin in anoikis sensitization. Mechanistically, thromboxane A2 receptor (TP) was up-regulated in detached breast cancer cells, conferred anoikis resistance through Akt activation, thereby facilitated breast cancer metastasis to lung. Interpretation: TXA2 pathway plays a critical role in anoikis resistance, and might serve as potential target for chemoprevention of breast cancer metastasis. Funding Information: his work was supported by the National Natural Science Foundation of China (81773064, 31972973 and 32021005). Declaration of Interests: No potential conflicts of interest were disclosed. Ethics Approval Statement: All animal study was conducted under protocols approved by the Institutional Animal Ethics Committee of Jiangnan University. (Issue No. 20200430c0501101[020], 20190330c0700615[58]).

The diagnostic value of DNA repair gene in breast cancer metastasis

Breast cancer is the most common malignant tumor in China and even in the world. DNA repair genes can lead to tumor metastasis by affecting cancer cell resistance. Studies have preliminarily shown that DNA repair genes are related to breast cancer metastasis, but it is not clear whether they can be used as a prediction of the risk of breast cancer metastasis. Therefore, this study mainly discusses the predictive value of DNA repair genes in postoperative metastasis of breast cancer. The nested case–control method was used in patients with breast cancer metastasis after surgery (n = 103) and patients without metastasis after surgery (n = 103). The proteins and mRNA of DNA repair genes were detected by immunohistochemistry and Real-time PCR respectively. In protein expression, PARP1 (OR 1.147, 95% CI 1.067 ~ 1.233, P < 0.05), XRCC4 (OR 1.088, 95% CI 1.015 ~ 1.166, P < 0.05), XRCC1 (OR 1.114, 95% CI 1.021 ~ 1.215, P < 0.05), ERCC1 (OR 1.068, 95% CI 1.000 ~ 1.141, P < 0.10) were risk factors for postoperative metastasis of breast cancer. In addition, we used the ROC curve to study the optimal critical values of MSH2, MLH1, PARP1, XRCC1, XRCC4, 53BP1, ERCC1 and XPA combined with the Youden index, and the effects of MSH2, MLH1, PARP1, XRCC1, XRCC4, 53BP1, ERCC1 and XPA on breast cancer metastasis were verified again. Among them, the risk of metastasis in the PARP1 high expression group was 3.286 times that of the low expression group (OR 3.286, 95% CI 2.013 ~ 5.364, P < 0.05). The risk of metastasis in the XRCC4 high expression group was 1.779 times that of the low expression group (OR 1.779, 95% CI 1.071 ~ 2.954, P < 0.05). The risk of metastasis in patients with ERCC1 high expression group was 2.012 times that of the low expression group (OR 2.012, 95% CI 1.056 ~ 3.836, P < 0.05). So we can conclude that protein expression of PARP1 (cut-off value = 6, Se = 76.70%, Sp = 79.61%), XRCC4 (cut-off value = 6, Se = 78.64%0, Se = 79.61%), ERCC1 (cut-off value = 3, Se = 89.32%, Sp = 50.49%), suggesting that when the PARP1 score is higher than 6 or the XRCC4 score is higher than 6 or the ERCC1 score is higher than 3, the risk of metastasis will increases. Due to PARP1, XRCC4 and ERCC1 belong to a part of DNA repair gene system, and the three proteins are positively correlated by correlation analysis (rPARP1-XRCC4 = 0.343; rPAPR1-ERCC1 = 0.335; rXRCC4-ERCC1 = 0.388). The combined diagnosis of the PARR1, XRCC4 and ERCC1 have greater predictive value for the risk of metastasis of breast cancer (Se = 94.17%, Sp = 75.73%; OR 11.739, 95% CI 2.858 ~ 40.220, P < 0.05). The postoperative metastasis of breast cancer could be effectively predicted when the immunohistochemical scores met PARP1 (IHC score) > 6, XRCC4 (IHC score) > 6 and ERCC1 (IHC score) > 3. In addition, the combined diagnosis of PARP1, XRCC4 and ERCC1 has great predictive value for the risk of breast cancer metastasis.

Cellular Mechanisms of Circulating Tumor Cells During Breast Cancer Metastasis.

Circulating tumor cells (CTCs) are cancer cells that detach from the primary site and travel in the blood stream. A higher number of CTCs increases the risk of breast cancer metastasis, and it is inversely associated with the survival rates of patients with breast cancer. Although the numbers of CTCs are generally low and the majority of CTCs die in circulation, the survival of a few CTCs can seed the development of a tumor at a secondary location. An increasing number of studies demonstrate that CTCs undergo modification in response to the dynamic biophysical environment in the blood due in part to fluid shear stress. Fluid shear stress generates reactive oxygen species (ROS), triggers redox-sensitive cell signaling, and alters the function of intracellular organelles. In particular, the mitochondrion is an important target organelle in determining the metastatic phenotype of CTCs. In healthy cells, mitochondria produce adenosine triphosphate (ATP) via oxidative phosphorylation in the electron transport chain, and during oxidative phosphorylation, they produce physiological levels of ROS. Mitochondria also govern death mechanisms such as apoptosis and mitochondrial permeability transition pore opening to, in order eliminate unwanted or damaged cells. However, in cancer cells, mitochondria are dysregulated, causing aberrant energy metabolism, redox homeostasis, and cell death pathways that may favor cancer invasiveness. In this review, we discuss the influence of fluid shear stress on CTCs with an emphasis on breast cancer pathology, then discuss alterations of cellular mechanisms that may increase the metastatic potentials of CTCs.

Association of polymorphisms in metastasis suppressor genes NME1 and KISS1 with breast cancer development and metastasis

BackgroundNME1 and KISS1 genes are two tumor metastasis suppressor genes, mapped to chromosomes 17q21.3 and 1q32 respectively. Here, we analyzed the association of EcoR1 (rs34214448—G/T) polymorphism in NME1 gene and 9 del T (rs5780218—A/-) polymorphism in KISS1 gene with breast cancer development and metastasis.ResultsThe study included 75 women newly diagnosed with breast cancer recruited from Oncology Center at Mansoura University Hospitals and 37 age-matched healthy female volunteers as a control group. DNA was extracted from peripheral blood samples and genotyping of rs34214448 and rs5780218 SNPs was carried out by PCR-RFLP technique. NME1 EcoR1 (rs34214448) polymorphism has a statistically significant association with breast cancer risk (P < 0.001). Most of breast cancer group (55%) had heterozygous (G/T) genotype while most of control group (95%) had homozygous wild (G/G) genotype (P < 0.0005). Also, KISS1 rs5780218 polymorphism has a statistically significant association with breast cancer risk. The wild (A/A) genotype was associated with lower risk of breast cancer (A/- + -/- vs. A/A: OR = 3.1, 95% CI = 1.15–8.36, P = 0.025). EcoR1 (rs34214448) polymorphism revealed a significant association with tumor stage and distant metastasis as patients. Carriers of the wild (G/G) genotype were more likely to present with advanced disease stages and distant metastasis.ConclusionsBoth EcoR1 (rs34214448) polymorphism of NME1 gene and rs5780218 polymorphism of KISS1 gene revealed significant association with increased risk of breast cancer development. The (G/G) genotype of EcoR1 polymorphism was associated with higher risk of breast cancer metastasis.

Single Tumor Cells With Epithelial-Like Morphology Are Associated With Breast Cancer Metastasis.

Introduction: The identification of tumor cells that can be potential metastatic seeds would reach two key aims—prognosis of metastasis risk and appointment of the optimal adjuvant therapy to prevent metastatic disease. Single tumor cells (STCs) located out of multicellular structures can most likely demonstrate features that are needed to initiate metastasis.Methods: One-hundred-and-thirty-five patients with invasive breast carcinoma of no special type have been enrolled. Molecular subtypes of breast cancer were categorized according to St. Gallen recommendations. Hematoxylin and eosin staining was used to identify STCs with epithelial-like morphology (eSTCs) in breast tumors. Immunofluorescence staining was applied to evaluate stemness and epithelial–mesenchymal transition (EMT) in STCs. The correlation between STCs and recurrence and metastasis-free survival (MFS) was performed using the Kaplan–Meier method and the log-rank test.Results: Distant metastasis was more frequent in eSTC-positive than eSTC-negative patients (28.0% vs. 9.4%, p = 0.007). When tumor types were analyzed separately, distant metastasis tended to be more frequent in eSTC-positive than eSTC-negative patients for HER2-positive cancer [75.0% (3/4) vs. 12.5% (1/8), p = 0.066]. In luminal A [22.7% (5/22) vs. 10.0% (3/30), p = 0.259], luminal B [21.1% (4/19) vs. 6.7% (2/30), p = 0.189], and triple-negative [40.0% (2/5) vs. 11.8% (2/17), p = 0.209] cancers, distance metastasis was not associated with eSTCs. Median MFS was not reached in eSTC-positive and eSTC-negative patients. eSTC-positive patients had a higher risk of breast cancer metastasis [hazard ratio (HR) 3.57, 95% confidence interval (CI): 1.46–8.71; p = 0.001]. When tumor types were analyzed separately, a higher risk of breast cancer metastasis occurred only in HER2-positive patients (HR 8.49, 95% CI: 1.29–55.59; p = 0.016). Immunofluorescence analysis revealed mesenchymal-like STCs (mSTCs) and inter- and intra-tumor heterogeneity in STCs. There were breast tumors with either eSTCs or mSTCs and tumors with both types of STCs. Both eSTCs and mSTCs were represented by cells with different stem and/or EMT phenotypes.Conclusions: STCs with epithelial-like morphology contribute to breast cancer metastasis and represent an attractive model for studying mechanisms of metastatic seeding. The assessment of STCs in histological sections of breast tumors can be a simple and effective method for the prediction of metastasis risk.

Associations between persistent organic pollutants and risk of breast cancer metastasis

BackgroundBreast cancer (BC) is a major public health concern with over 2 million new cases diagnosed and over 600,000 deaths in 2018 in women worldwide. When distant metastases are present at diagnosis, the 5-year survival rate is only 26%. Recent studies have suggested that persistent organic pollutants (POPs) that accumulate in adipose tissue (AT) can influence tumor phenotype and stimulate cellular processes important for metastasis such as invasion. We, therefore, tested the hypothesis that POP exposure is associated with BC metastasis. MethodsWe conducted an exploratory case-control study in which the concentrations of 49 POPs were measured in both AT and serum samples from BC patients, with or without lymph node metastasis, who underwent partial or total mastectomies, lymph node biopsies and sampling of the adipocytic tumor microenvironment. Adjusted, unconditional logistic models were used to study the associations between the POP concentrations and the risk of metastasis and other hallmarks of cancer aggressiveness. Results2.3.7.8-TCDD concentrations in AT are positively associated with the risk of metastasis in 43 patients who have BMIs equal or higher than 25 kg/m2 (odds ratio: 4.48 (1.32–20.71)). Furthermore, the concentrations of 2.3.7.8-TCDD and two coplanar PCBs (77&169) in AT also were positively associated with the risk of lymph node metastasis and the tumor size. ConclusionOur study suggests that 2.3.7.8-TCDD and some PCBs contribute to the development of tumor metastasis and other hallmarks of cancer aggressiveness. While these results should be considered with caution, this is the first study to identify such potential risk factors. Larger longitudinal studies are necessary to confirm our results.Clinical Trial Protocol Record: 2013-A00663-42.

Can microcalcifications' characteristics predict the risk of breast cancer metastasis to bone?

Can microcalcifications' characteristics predict the risk of breast cancer metastasis to bone?

Abstract PD9-09: Proteomic analysis of extracellular matrix helps define drivers of metastatic progression

Abstract Tumor microenvironment (TME) plays a central role in the development of distant metastasis. However, given its complexity, successful targeting of TME will require a detailed understanding of its composition. Cancer-associated fibroblasts represent a large component of TME and are a major contributor to the extracellular matrix (ECM). Previously, we demonstrated the presence of two fibroblast sub-populations (CDCP1pos vs CD146pos fibroblasts) in breast cancer TME, which determine therapeutic response in estrogen receptor (ER) positive breast cancer. We hypothesized that the same fibroblast subtypes would also influence ECM composition and alter the metastatic potential of breast cancer cells. Here, we present the development of a novel fibroblast driven orthotopic model of ER+ breast cancer metastasis, which we combined with an innovative proteomics approach to precisely quantify extracellular matrix proteins. Results: 1.CDCP1pos fibroblasts promote increased breast cancer cell mobility, invasion and metastasis compared to CD146pos fibroblasts.We demonstrate that CDCP1pos fibroblasts significantly increase the invasion potential of breast cancer cells, when compared to CD146pos fibroblasts. Orthotopic co-implantation of ER+ tumor cells with CDCP1pos fibroblasts into the mammary fat pad of mice more frequently drives distant organ metastases to lung and brain when compared with tumors implanted with CD146pos fibroblasts. 2. Proteomic analysis of ER+ tumors influenced by CDCP1pos fibroblasts revealed known and novel drivers of breast cancer metastasis. Breast cancer cells mixed with CDCP1pos fibroblasts produce a non-uniform collagen orientation to the tumor boarder. Our novel proteomic analysis of TME specific proteins revealed that tumors influenced by CDCP1pos fibroblasts have high expression of many ECM proteins linked to increased risk of breast cancer metastasis, including TNC, FN1, COL5A3, and FBN1 among others. Derived proteomic TME signature accurately predicted lymph node involvement in patients who presented with early stage (T0 and T1) tumors in a cohort of 1,009 breast cancer patients from Cancer Genome Atlas Database. 3. Inhibiting fibroblast production of Tenascin C (TNC) results in decreased breast cancer cell invasion. Our in vitro mixed co-culture models, which contain ER+ breast cancer cells with both fibroblast subtypes, demonstrate that only CDCP1pos fibroblasts produce TNC. Furthermore, in spheroid assays with CDCP1posfibroblasts and breast cancer tumor cells, invasion is inhibited by TNC knockdown. The invasion phenotype can be rescued by addition of EGF, which suggests TNC promotes invasion via EGFR signaling. Conclusion: Metastatic spread of cancer cells relies heavily on TME alterations and makeup of the extracellular matrix. Our data suggest that fibroblast composition directly influences ECM properties and metastatic potential in breast cancer. Taken together, we believe that a better understanding of ECM composition will lead to a more personalized approach to breast cancer treatment. Citation Format: Brechbuhl HM, Barrett AS, Kopin E, Gillen A, Hagen J, Johnson L, Finaly-Shultz J, Sartorius CA, Hansen KC, Kabos P. Proteomic analysis of extracellular matrix helps define drivers of metastatic progression [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD9-09.

Abstract # 3145 Rage-associated S100a8/A9 levels associated with circulating proinflammatory cytokines and social well-being in patients with breast cancer over the first 12 months of treatment

Inflammation and social adversity have been linked to poorer long-term outcomes in breast cancer (BCa) patients, whereas greater social and family well-being (sfwb) relates to less inflammation and longer disease-free interval in BCa. S100A8A9 is a RAGE-related ligand that can up-regulate inflammatory and pro-metastatic processes and has been associated with increased risk of BCa metastasis. Here we examined associations between sfwb, s100A8A9 levels, and pro-inflammatory cytokines in 50 (M age = 53.65) distressed BCa patients in the weeks post-surgery (T1) and at 12-months (T2). Regression analyses controlling for age and disease stage showed lower S100A8A9 levels related to greater FACTsfwb at T1 ( β = -.353, p = .001) and T2 ( β = -.398, p =.01). S100A8A9 significantly related to T1 levels of IL6 ( β =.412, p = .008) and IL1 β ( β =.588, p = .001), and T2 levels of IL6 ( β =.486, p = .001) and IL-1 β ( β =.548, p = .001). Additionally, 12-month change in S100A8A9 was significantly related to 12-month change in IL6 ( β =.279, p = .001) and IL1 β ( β = .164, p =.001). These findings demonstrate that greater social and family well-being during BCa primary treatment is associated with lower levels of upstream and downstream inflammatory markers and point to the potential value of psychosocial interventions that target enhancement of social and family well-being during this time. .

Current Evidence on the Association between Four Polymorphisms in the Matrix Metalloproteinases (MMP) Gene and Breast Cancer Metastasis

The matrix metalloproteinases (MMP) can degrade various components of the extracellular matrix and its functional genetic polymorphisms may be associated with cancer metastasis. However, this relationship remains inconclusive in breast cancer. To better understand the roles of MMP polymorphisms in breast cancer metastasis, we conducted this comprehensive meta-analysis encompassing 1,165 cases and 3,418 controls. Four polymorphisms including MMP1 (-1607 1G/ 2G), MMP2 (-1575 G.A), MMP3 (1612 5A/6A), and MMP9 (-1562 C.T) were studied in this meta-analysis. Crude odds ratios (OR) with 95% confidence intervals (CI) were used to assess the strength of association. We found significant association between MMP1 (-1607 1G/2G) and breast cancer metastasis (for the recessive model 2G/2G vs. 1G/2G+1G/1G: OR 1.81, 95% CI 1.32–2.48; for the dominant model 2G/2G+1G/2G vs. 1G/1G: OR 1.60, 95% CI 1.02– 2.50). For the MMP3, significant association between MMP3 (-1171 5A/6A) and breast cancer metastasis was also detected under the dominant model (6A/6A+5A/6A vs. 5A/5A: OR 0.55, 95% CI 0.37-0.80). In conclusion, our results demonstrate that MMP1 may contribute the risk of breast cancer metastasis, while MMP3 played a protective role in breast cancer metastasis. Further studies are needed to evaluate these associations with breast cancer metastasis.

Selective gene-expression profiling of migratory tumor cells in vivo predicts clinical outcome in breast cancer patients

IntroductionMetastasis of breast cancer is the main cause of death in patients. Previous genome-wide studies have identified gene-expression patterns correlated with cancer patient outcome. However, these were derived mostly from whole tissue without respect to cell heterogeneity. In reality, only a small subpopulation of invasive cells inside the primary tumor is responsible for escaping and initiating dissemination and metastasis. When whole tissue is used for molecular profiling, the expression pattern of these cells is masked by the majority of the noninvasive tumor cells. Therefore, little information is available about the crucial early steps of the metastatic cascade: migration, invasion, and entry of tumor cells into the systemic circulation.MethodsIn the past, we developed an in vivo invasion assay that can capture specifically the highly motile tumor cells in the act of migrating inside living tumors. Here, we used this assay in orthotopic xenografts of human MDA-MB-231 breast cancer cells to isolate selectively the migratory cell subpopulation of the primary tumor for gene-expression profiling. In this way, we derived a gene signature specific to breast cancer migration and invasion, which we call the Human Invasion Signature (HIS).ResultsUnsupervised analysis of the HIS shows that the most significant upregulated gene networks in the migratory breast tumor cells include genes regulating embryonic and tissue development, cellular movement, and DNA replication and repair. We confirmed that genes involved in these functions are upregulated in the migratory tumor cells with independent biological repeats. We also demonstrate that specific genes are functionally required for in vivo invasion and hematogenous dissemination in MDA-MB-231, as well as in patient-derived breast tumors. Finally, we used statistical analysis to show that the signature can significantly predict risk of breast cancer metastasis in large patient cohorts, independent of well-established prognostic parameters.ConclusionsOur data provide novel insights into, and reveal previously unknown mediators of, the metastatic steps of invasion and dissemination in human breast tumors in vivo. Because migration and invasion are the early steps of metastatic progression, the novel markers that we identified here might become valuable prognostic tools or therapeutic targets in breast cancer.