CCR7 Immune Cell Receptor Expression in Inflammatory Breast Cancer

https://openworks.mdanderson.org/sumexp22/1011/thumbnail.jp

Lipocalin 2 promotes inflammatory breast cancer tumorigenesis and skin invasion

Inflammatory breast cancer (IBC) is an aggressive form of primary breast cancer characterized by rapid onset and high risk of metastasis and poor clinical outcomes. The biological basis for the aggressiveness of IBC is still not well understood and no IBC-specific targeted therapies exist. In this study, we report that lipocalin 2 (LCN2), a small secreted glycoprotein belonging to the lipocalin superfamily, is expressed at significantly higher levels in IBC vs non-IBC tumors, independently of molecular subtype. LCN2 levels were also significantly higher in IBC cell lines and in their culture media than in non-IBC cell lines. High expression was associated with poor-prognosis features and shorter overall survival in IBC patients. Depletion of LCN2 in IBC cell lines reduced colony formation, migration, and cancer stem cell populations in vitro and inhibited tumor growth, skin invasion, and brain metastasis in mouse models of IBC. Analysis of our proteomics data showed reduced expression of proteins involved in cell cycle and DNA repair in LCN2-silenced IBC cells. Our findings support that LCN2 promotes IBC tumor aggressiveness and offer a new potential therapeutic target for IBC

Inflammatory breast cancer appearance at presentation is associated with overall survival

Background: Inflammatory breast cancer (IBC) is a clinical diagnosis. Here, we examined the association of a "classic" triad of clinical signs, swollen involved breast, nipple change, and diffuse skin change, with overall survival (OS). Method: Breast medical photographs from patients enrolled on a prospective IBC registry were scored by two independent reviewers as classic (triad above), not classic, and difficult to assign. Chi-squared test, Fisher's exact test, and Wilcoxon rank-sum test were used to assess differences between patient groups. Kaplan-Meier estimates and the log-rank test and Cox proportional hazard regression were used to assess the OS. Results: We analyzed 245 IBC patients with median age 54 (range 26-81), M0 versus M1 status (157 and 88 patients, respectively). The classic triad was significantly associated with smoking, post-menopausal status, and metastatic disease at presentation (p = 0.002, 0.013, and 0.035, respectively). Ten-year actuarial OS for not classic and difficult to assign were not significantly different and were grouped for further analyses. Ten-year OS was 29.7% among patients with the classic sign triad versus 57.2% for non-classic (p < 0.0001). The multivariate Cox regression model adjusting for clinical staging (p < 0.0001) and TNBC status (<0.0001) demonstrated classic presentation score significantly associated with poorer OS time (HR 2.6, 95% CI 1.7-3.9, p < 0.0001). Conclusions: A triad of classic IBC signs independently predicted OS in patients diagnosed with IBC. Further work is warranted to understand the biology related to clinical signs and further extend the understanding of physical examination findings in IBC

Risk-Factor induced changes in the Breast Microenvironment facilitate Inflammatory Breast Cancer Progression and Lymphovascular Invasion

Inflammatory breast cancer (IBC) is a rapidly progressing, rare and highly lethal form of breast cancer. IBC is a clinical diagnosis, requiring \u3e1/3 involvement on the affected breast and/or skin by erythema, and disease onset of \u3c 6 months. The clinical symptoms of IBC vary in severity and presentation, these include redness, warmth, skin thickening and bruised or pink/purple discoloration appearance and skin changes such as peau d’orange. These skin symptoms are not attributed to inflammation, rather IBC is characterized by florid lymphovascular tumor emboli clogging dermal lymphatics. This leads to “classic” symptoms of breast swelling and skin edema or discoloration. To date, unique genomic drivers which differentiate IBC from non-IBC invasive breast cancers have not been identified highlighting a role for the microenvironment. Several epidemiological studies have unveiled subtype-specific risk factors associated with IBC that are known to alter the microenvironment. Obesity is an established risk factor for all subtypes of IBC. Never-breastfeeding increases risk for developing the most aggressive, triple-negative IBC. Further, never breastfeeding is associated with later clinical stage and worse outcomes. We worked to model these overlapping risk factors to understand microenvironment changes that may lead to the lymphatic change’s indicative of IBC. First, we investigated the association of a “classic” triad of clinical IBC signs with overall survival among patients to demonstrate the most overt clinical findings of lymphatic involvement were impacting prognosis. We evaluated a triad of IBC signs, including swollen involved breast, nipple change, and diffuse skin change, using breast medical photographs from patients enrolled on a prospective IBC registry. We reported that the ten-year OS was 29.7% among patients with the classic sign triad versus 57.2% for non-classic (P \u3c .0001). We determined that a triad of classic IBC signs independently predicted OS in patients diagnosed with IBC suggesting the clinical outcome of diffuse lymphatic invasion by IBC is prognostic, and potentially that targeting the mechanisms that promote lymphatic function and tumor invasion may improve outcomes. Our prior work implied the changes in the breast that permit florid lymphatic involvement may even occur prior to tumor initiation. Noting a limitation of existing studies of breast cancer risk factors is the experimental isolation of risk factors, which fails to model the patient experience. Thus, we modeled synergistic effects of IBC risk factors, obesity and weaning timing, on in vivo lymphatic function pre- and post-tumor initiation, IBC tumor growth, and the mammary gland microenvironment. We hypothesized that weaning status (duration of breast feeding) and high fat diet (HFD) would synergize to induce pro-lymphatic changes in the microenvironment before tumor initiation and promote IBC tumor growth. We found that HFD increased lymphatic contractile activity prior to tumor initiation. This HFD-induced increase in lymphatic function correlated to increased SUM149 tumor growth and increased inflammatory cells in the mammary gland. Tumors increased lymphatic function to a similar extent. Increased CCL21+ cells, a ligand for lymphatic traffic homing, was correlated to pulsing. Thus, the relationship between lymphatic pulsing, tumor growth and CCR7-CCL21 related tumor trafficking warrant further investigation. CCR7 is an immune cell receptor that mediates immune cell trafficking into lymphatics that can be expressed on tumor cells. We examined the expression of CCR7 in IBC and non-IBC cell lines, and IBC patient tumors to determine the prevalence of this receptor in IBC tumor cells. We found that CCR7 gene expression is increased in IBC versus non-IBC tumors, and was present across tumor subtypes in IBC cell lines and in both ER+ and ER- IBC patient tumors. Further, CCR7 was expressed on 23/24 IBC patient samples assessed by immunohistochemistry. This highlights the potential direction for developing novel therapies targeting CCR7, to improve therapeutic options and outcomes for IBC patients

Lymphatic Endothelial Cell Secretion Effect On Inflammatory Breast Cancer Cell Growth

https://openworks.mdanderson.org/sumexp21/1094/thumbnail.jp

Lipocalin 2 promotes inflammatory breast cancer tumorigenesis and skin invasion

International audienceInflammatory breast cancer (IBC) is an aggressive form of primary breast cancer characterized by rapid onset and high risk of metastasis and poor clinical outcomes. The biological basis for the aggressiveness of IBC is still not well understood and no IBC-specific targeted therapies exist. In this study, we report that lipocalin 2 (LCN2), a small secreted glycoprotein belonging to the lipocalin superfamily, is expressed at significantly higher levels in IBC vs non-IBC tumors, independently of molecular subtype. LCN2 levels were also significantly higher in IBC cell lines and in their culture media than in non-IBC cell lines. High expression was associated with poor-prognosis features and shorter overall survival in IBC patients. Depletion of LCN2 in IBC cell lines reduced colony formation, migration, and cancer stem cell populations in vitro and inhibited tumor growth, skin invasion, and brain metastasis in mouse models of IBC. Analysis of our proteomics data showed reduced expression of proteins involved in cell cycle and DNA repair in LCN2-silenced IBC cells. Our findings support that LCN2 promotes IBC tumor aggressiveness and offer a new potential therapeutic target for IBC