VAV3 mediates resistance to breast cancer endocrine therapy.

INTRODUCTION: Endocrine therapies targeting cell proliferation and survival mediated by estrogen receptor α (ERα) are among the most effective systemic treatments for ERα-positive breast cancer. However, most tumors initially responsive to these therapies acquire resistance through mechanisms that involve ERα transcriptional regulatory plasticity. Herein we identify VAV3 as a critical component in this process. METHODS: A cell-based chemical compound screen was carried out to identify therapeutic strategies against resistance to endocrine therapy. Binding to ERα was evaluated by molecular docking analyses, an agonist fluoligand assay and short hairpin (sh)RNA-mediated protein depletion. Microarray analyses were performed to identify altered gene expression. Western blot analysis of signaling and proliferation markers, and shRNA-mediated protein depletion in viability and clonogenic assays, were performed to delineate the role of VAV3. Genetic variation in VAV3 was assessed for association with the response to tamoxifen. Immunohistochemical analyses of VAV3 were carried out to determine its association with therapeutic response and different tumor markers. An analysis of gene expression association with drug sensitivity was carried out to identify a potential therapeutic approach based on differential VAV3 expression. RESULTS: The compound YC-1 was found to comparatively reduce the viability of cell models of acquired resistance. This effect was probably not due to activation of its canonical target (soluble guanylyl cyclase), but instead was likely a result of binding to ERα. VAV3 was selectively reduced upon exposure to YC-1 or ERα depletion, and, accordingly, VAV3 depletion comparatively reduced the viability of cell models of acquired resistance. In the clinical scenario, germline variation in VAV3 was associated with the response to tamoxifen in Japanese breast cancer patients (rs10494071 combined P value = 8.4 × 10-4). The allele association combined with gene expression analyses indicated that low VAV3 expression predicts better clinical outcome. Conversely, high nuclear VAV3 expression in tumor cells was associated with poorer endocrine therapy response. Based on VAV3 expression levels and the response to erlotinib in cancer cell lines, targeting EGFR signaling may be a promising therapeutic strategy. CONCLUSIONS: This study proposes VAV3 as a biomarker and a rationale for its use as a signaling target to prevent and/or overcome resistance to endocrine therapy in breast cancer.

Evidence for a link between TNFRSF11A and risk of breast cancer.

Intracellular signaling mediated by the receptor activator of nuclear factor-κB [Rank, encoded by the tumor necrosis factor receptor superfamily, member 11a (Tnfrsf11a) gene] is fundamental for mammary gland development in mice, regulating the expansion of stem and progenitor cell compartments. Conversely, Rank overexpression in mice promotes abnormal proliferation and impairs differentiation, leading to an increased incidence of tumorigenesis. Here, we show that a common genetic variant near the 5'-end of TNFRSF11A, rs7226991, is associated with breast cancer risk in the general population and among carriers of mutations in the breast cancer 2, early onset (BRCA2) gene. Akin to the results of the Cancer and Genetics Markers of Susceptibility initiative, combined analysis of rs7226991 in two Spanish case-control studies (1,365 controls and 1,323 cases in total) revealed a significant association with risk: odds ratio (OR) = 0.88, 95% confidence interval (CI) 0.78-0.98, P (trend) = 0.025. Subsequent examination of BRCA1 (n = 1,017) and BRCA2 (n = 885) mutation carriers revealed a consistent association in the latter group: weighted hazard ratio ((w)HR) = 0.70; 95% CI 0.55-0.88; and P (trend) = 0.003; compared to BRCA1 mutation carriers, (w)HR = 0.91; 95% CI 0.76-1.10; and P (trend) = 0.33. The results of this study need to be replicated in other populations and with larger numbers of BRCA1/2 mutation carriers.

Hospital admissions for bleeding events associated with treatment with apixaban, dabigatran and rivaroxaban.

OBJECTIVES: To analyse the hospital admissions for bleeding events associated with treatment with direct oral anticoagulants (DOACs). To describe the characteristics and outcomes of those patients. METHODS: A retrospective observational study was carried out in the framework of an integral risk management plan of drugs and proactive pharmacovigilance of hospital admissions for bleeding associated with apixaban, dabigatran and rivaroxaban from April 2015 through December 2016. Cases were identified using the information management tool of Orion Clinic (hospital electronic medical history) and by reviewing the hospital discharge reports. Various biometric, clinical and pharmacotherapeutic variables of each patient were registered. RESULTS: 37 hospitalisation episodes for DOAC-induced bleeding in 32 patients (15 received rivaroxaban, 9 apixaban and 8 dabigatran) were detected, representing an incidence rate of 3.44 per 100 person-years (95% CI 2.35 to 4.86). The most common bleeding site was gastrointestinal (27 cases, 73.0%). Intracranial bleeding was rare (three cases, 8.1%). Four patients (12.5%) were receiving DOACs at full doses and had a 'dose reduction indication'. The mean (SD) length of stay was 8.4 (5.2) days. Three patients (8.1%) died during the hospitalisation. Among bleeding episodes without fatal outcome, DOACs were stopped in 14 cases, continued in 14 cases, switched for another DOAC in two cases and the dose was reduced in four cases. CONCLUSIONS: DOACs are associated with serious bleeding events that require hospitalisation. The risk/benefit ratio assessment considering patient preferences and an individualised follow-up, especially in patients who are elderly, polymedicated or have impaired renal function, can help to reinforce the safe use of DOACs.

Cross-sectional analysis of retrospective case series of hospitalisations for gastropathy caused by non-steroidal anti-inflammatory treatment: risk factors and gastroprotection use.

OBJECTIVES: To analyse the risk factors of gastropathy caused by using non-steroidal anti-inflammatory drugs (NSAIDs) in detected hospital admissions and to analyse the use of gastroprotective treatment concerning these risk factors. METHODS: A retrospective observational study was carried out in the framework of an integral risk management plan of drugs and proactive pharmacovigilance of hospital admissions for NSAID-induced gastropathy occurring between 2011 and 2015. Cases were identified after reviewing the ICD-9 codes related to NSAID-induced gastropathy in hospital discharge reports. Various biometric, clinical and pharmacotherapeutic variables of each patient were registered. The gastroprotective criteria set out in the therapeutic decision algorithm of the Valencian Health System were followed. RESULTS: 62 hospital admissions for NSAID-induced gastropathy were detected. The mean length of stay was 5.3±3.8 days. Ibuprofen was the most prevalent NSAID (28 cases, 45.2%). 24 cases (38.7%) took NSAIDs in the week before hospitalisation. The prevalence of relevant risk factors for gastropathy were age >60 years (37 cases, 59.7%), concomitant medication (24 cases, 38.7%) and a history of peptic ulcer (9 cases, 14.5%). 41 patients (66.1%) met gastroprotective major criteria, 18 of whom (43.9%) were using a proton pump inhibitor following a prevention plan. CONCLUSIONS: In this study all relevant gastroprotective criteria were associated with the use of gastroprotection in detected hospital admissions for NSAID-induced gastropathy. However, a lack of gastroprotection was observed in a large number of detected cases with the criteria to use it. The feedback of our results to health area agents can serve to reinforce the safe use of NSAIDs.

Do primary mammary osteosarcoma and chondrosarcoma exist? A review of a large multi-institutional series of malignant matrix-producing breast tumours.

UNLABELLED: The existing literature describing the clinicopathological features and behaviour of matrix-producing (MP) malignant breast tumours presents conflicting results. As a consequence it remains uncertain whether these tumours should be treated as sarcoma and managed by a specialist sarcoma team or treated using the same principles as conventional ductal carcinoma, a dilemma that prompted this study. Improved understanding of the clinicopathological characteristics of primary mammary MP-sarcomas, namely osteosarcoma and chondrosarcoma, is required. METHODS: In this large international multicenter series of malignant MP-tumours of the breast (no = 101) with follow-up information has been assessed and their outcome is compared to other subtypes of metaplastic breast carcinoma (MBC) (no = 253) and to grade, lymph node and hormone receptor-matched ductal breast carcinomas (no = 258). RESULTS: The majority of MP-cancers were associated with epithelial features, which supports the concept that the majority of, if not almost all, primary MP breast sarcomas are of epithelial in origin (MBC). 21% showed nodal metastasis and the distribution of distant metastases resembled conventional mammary carcinoma. The prognosis of MP-MBC is comparable to matched ductal breast carcinoma and slightly better than other subtypes of MBC. Advanced stage (T3&T4) and development of recurrences were predictors of shorter survival in MP-MBC while grade and vascular invasion were not. CONCLUSION: Most malignant MP breast tumours are variants of MBC. MP-MBC with predominant mesenchymal components behaves similar to ductal carcinomas and although data on their response to systemic therapy is limited, there is no evidence that they should be managed differently from other forms of triple negative breast cancer.

RANK induces epithelial-mesenchymal transition and stemness in human mammary epithelial cells and promotes tumorigenesis and metastasis.

Paracrine signaling through receptor activator of NF-κB (RANK) pathway mediates the expansion of mammary epithelia that occurs during pregnancy, and activation of RANK pathway promotes mammary tumorigenesis in mice. In this study we extend these previous data to human cells and show that the RANK pathway promotes the development of mammary stem cells and breast cancer. Overexpression of RANK (FL-RANK) in a panel of tumoral and normal human mammary cells induces the expression of breast cancer stem and basal/stem cell markers. High levels of RANK in untransformed MCF10A cells induce changes associated with both stemness and transformation, including mammary gland reconstitution, epithelial-mesenchymal transition (EMT), increased migration, and anchorage-independent growth. In addition, spheroids of RANK overexpressing MCF10A cells display disrupted acinar formation, impair growth arrest and polarization, and luminal filling. RANK overexpression in tumor cells with nonfunctional BRCA1 enhances invasiveness in acinar cultures and increases tumorigenesis and metastasis in immunodeficient mice. High levels of RANK were found in human primary breast adenocarcinomas that lack expression of the hormone receptors, estrogen and progesterone, and in tumors with high pathologic grade and proliferation index; high RANK/RANKL expression was significantly associated with metastatic tumors. Together, our findings show that RANK promotes tumor initiation, progression, and metastasis in human mammary epithelial cells by increasing the population of CD44(+)CD24(-) cells, inducing stemness and EMT. These results suggest that RANK expression in primary breast cancer associates with poor prognosis.

Estudio comparativo entre el método One step nucleic acid amplification y el método convencional en la estadificación en cáncer de mama: un aumento en la detección de micrometástasis / Comparative study of the One step nucleic acid amplification method and the conventional method in axillary staging in breast cancer: an increase in the detection of micrometastases

Objetivos. El método One step nucleic acid amplification (OSNA) se ha incorporado para el estudio del ganglio centinela (GC) en cáncer de mama como alternativa al estudio convencional histológico (MC). El propósito de nuestro estudio fue comparar la estadificación por ganglio centinela (EGC) obtenida por el método OSNA con la obtenida mediante MC. Material y métodos. Se seleccionaron pacientes con cáncer de mama y EGC recogidas durante los años 2009-2010 y 2012-2013, estudiadas con MC y método OSNA. Se analizaron diferentes parámetros clínico-patológicos. Resultados. Se incluyó a 1.124 pacientes, 590 estudiadas por MC y 534 por método OSNA. La EGC inicial fue: pN0: MC 349 (59,2%) y OSNA 335 (62,7%); pN0(i+): MC 74 (12,5%) y OSNA 14 (2,6%); pN1mi: MC 59 (10%) y OSNA 77 (14,4%); pN1: MC 108 (18,3%) y OSNA 108 (20,3%). Se encontraron diferencias estadísticamente significativas entre la EGC por método OSNA y MC (p<0,001), a expensas de las tasas de pN1mi y pN0(i+). Se seleccionó a 224 pacientes con EGC pN1mi y pN0(i+) para determinar si las diferencias encontradas podrían atribuirse a distintas características clínico-patológicas. El método OSNA detecta el doble de micrometástasis (84,6%). Conclusiones. En nuestra casuística, por el método OSNA se observa un incremento significativo de pN1mi (84,6% vs. 44,4%) y una disminución de pN0(i+) respecto al estudio convencional, diferencias que no están condicionadas por los parámetros clínico-patológicos. El 75% de casos con pN1mi por OSNA muestra un número de copias inferior a 1.000 (AU)

Objetives. The One Step Nucleic Acid Amplification (OSNA) method has been incorporated in the study of the sentinel lymph node (SLN) in breast cancer as an alternative to conventional histological study. The aim of our study was to compare sentinel lymph node staging (SLNS) obtained by the OSNA method with that obtained by the conventional method (CM). Material and methods. We identified patients with breast cancer and SLN study during the periods 2009-2010 and 2012-2013, who underwent the CM and by OSNA. We analysed different clinicopathological parameters. Results. A total of 1124 patients were studied, 590 by CM and 534 by OSNA. SLNS was: pN0: CM 349 (59.2%) and OSNA 335 (62.7%); pN0(i+): CM 74 (12.5%) and OSNA 14 (2.6%); pN1mi: CM 59 (10%) and OSNA 77 (14.4%); pN1: CM 108 (18.3%) and OSNA 108 (20.3%). Statistically significant differences were found between the SLNS by OSNA and CM (p <0.001), due to the rates of pN1mi and pN0(i+). To determine whether this statistical significance could be attributed to different clinicopathological features, 224 patients were selected from the initial series with SLN pN1mi and pN0(i+). In this subgroup, the OSNA method detected twice as many micrometastases (pN1mi) (84.6%). Conclusions. In our series, the OSNA method resulted in a significant increase in pN1mi (84.6% vs 44.4%) and a decrease in pN0(i+) compared with the conventional method. Those differences were not affected by clinicopathological parameters. Most cases (75%) with pN1mi by OSNA showed less than 1000 copies (AU)

[Diverticular disease of the cecal appendix]. / Enfermedad diverticular del apéndice cecal.

INTRODUCTION: Appendiceal diverticula are uncommon, with an incidence of less than 1% in surgical specimens. We report a series of 14 patients with diverticular disease of the cecal appendix. PATIENTS AND METHOD: A total of 547 patients with a clinical diagnosis of acute appendicitis underwent surgery over 4 years. Of these, 11 patients showed acute appendiceal diverticulitis at histological examination, and three patients showed diverticulosis associated with appendicitis. Clinical features were compared between the group of patients with diverticular disease and the group with acute appendicitis. Statistical analysis was performed using Students t-test and the chi-squared test. RESULTS: The overall incidence of appendiceal diverticula was 2.6%, and 2% of cases had acute diverticulitis. In the group with diverticular disease, the mean age and the percentage of patients under clinical observation before the decision to perform surgery was made were significantly higher. There was a nonsignificant predominance of male over female patients and no differences were found in mean white cell count. No radiological investigations were performed in the diverticular group. CONCLUSIONS: The incidence of appendiceal diverticula was much higher in our series than that reported in the literature. We found no clinical or perioperative data that would serve to differentiate acute diverticulitis from acute appendicitis.

Enfermedad diverticular del apéndice cecal / Diverticular disease of the cecal appendix

Introducción. Los divertículos apendiculares constituyen una rareza y su incidencia es < 1%. Se presenta una serie de 14 pacientes con este diagnóstico. Pacientes y método. De un total de 547 pacientes intervenidos por apendicitis aguda durante un período de 4 años, 11 presentaban una diverticulitis apendicular aguda y 3 una diverticulosis asociada a apendicitis. Se comparan las características clínicas entre el grupo de pacientes con enfermedad diverticular del apéndice y el grupo de apendicitis aguda. El estudio estadístico se realiza mediante el test de la t de Student y el test de la (..) (AU)

Introduction. Appendiceal diverticula are uncommon, with an incidence of less than 1% in surgical specimens. We report a series of 14 patients with diverticular disease of the cecal appendix. Patients and method. A total of 547 patients with a clinical diagnosis of acute appendicitis underwent surgery over 4 years. Of these, 11 patients showed acute appendiceal diverticulitis at histological examination, and three patients showed diverticulosis associated with appendicitis. Clinical features were compared between the group of patients with diverticular disease and the group with acute appendicitis. Statistical analysis was performed using Student’s t-test and the chi-squared test. Results. The overall incidence of appendiceal diverticula was 2.6%, and 2% of cases had acute diverticulitis. In the group with diverticular disease, the mean age and the percentage of patients under clinical observation before the decision to perform surgery was made were significantly higher. There was a nonsignificant predominance of male over female patients and no differences were found in mean white cell count. No radiological investigations were performed in the diverticular group. Conclusions. The incidence of appendiceal diverticula was much higher in our series than that reported in the literature. We found no clinical or perioperative data that would serve to differentiate acute diverticulitis from acute appendicitis (AU)

Metástasis cutánea de carcinoma de vejiga urinaria productor de gonadotrofina coriónica humana / Skin metastasis from urinary bladder carcinoma producing human chorionic gonadotropin

Las metástasis cutáneas de carcinomas originados en órganos internos son infrecuentes. Entre ellas, las metástasis originadas en un carcinoma de vejiga urinaria son excepcionales. Presentamos el caso de un paciente de 74 años con un carcinoma de células transicionales de la vejiga urinaria al que se practicó una resección endoscópica; con posterioridad recibió tratamiento quimioterápico. En el curso evolutivo de la enfermedad, el paciente desarrolló una lesión exofítica en la piel preesternal. Tras la exéresis quirúrgica de la lesión cutánea, mediante estudio inmunohistoquímico pudo demostrarse la presencia de gonadotrofina coriónica humana, tanto en la tumoración de la piel como en el carcinoma de células transicionales, estableciéndose así el diagnóstico de metástasis cutánea. La transformación parcial del tejido del carcinoma de células transicionales de vejiga urinaria en tejido sincitiotrofoblástico causó la producción y secreción de gonadotrofina coriónica humana. Asimismo, se determinaron las concentraciones plasmáticas de hormona, que resultaron muy elevadas, lo que parece asociarse con una mayor agresividad tumoral y un peor pronóstico (AU)