Laura Yébenes

One-step nucleic acid amplification (OSNA) for the detection of sentinel lymph node metastasis in endometrial cancer

OBJECTIVE To evaluate the efficacy of one-step nucleic acid amplification (OSNA) for the diagnosis of sentinel lymph node (SLN) metastasis compared with histopathological examination in patients with endometrial carcinoma. METHODS A total of 94 SLNs from 34 patients with endometrial carcinoma were enrolled. The central 1-mm portion of each node was subjected to semi-serial sectioning, sliced at 200-μm intervals and examined by hematoxylin and eosin and cytokeratin 19 (CK19) immunohistochemical staining, and the remaining tissue was analysed by OSNA using CK19 mRNA. The accuracy of the OSNA assay was evaluated based on histopathological diagnosis. RESULTS Histologically, 89 SLNs were determined to be metastasis negative, and the remaining five SLNs were metastasis positive. Using the breast cancer cutoff value for detecting lymph node metastasis (OSNA criteria for breast cancer, >250copies/μl) the sensitivity of the OSNA assay was 100%; specificity was 87.6%; diagnostic accuracy was 88.3%. Discordant results were recorded for 11 of 94 SLNs. In all 11 cases, a positive result was given by the OSNA assay but not by histopathological examination. In two SLNs from the same patient, histopathological examination revealed the presence of benign epithelial inclusions that were CK19 positive; both SLNs yielded a positive result in the OSNA assay (true-false positive). All remaining nine histologically-negative/OSNA-positive SLNs were classified as micrometastasis (+) by the OSNA assay. CONCLUSION The OSNA assay shows high sensitivity and specificity, which suggests its utility as a novel tool for the molecular detection of SLN metastasis in patients with endometrial carcinoma.

Síndrome de DRESS y nefritis tubulointersticial aguda tras tratamiento con vancomicina y betalactámicos.: Descripción de un caso y revisión de la literatura

On the clinical analysis leukocytosis is observed with important eosinophilia and acute renal failure with creatinine: 6.1mg/dl, urea: 156mg/dl and vancomycin levels at 36.19μg/ml (normal levels: 5-10μg/ml). Vancomycin is immediately suspended and a catheter is placed the right femoral artery. Haemodialysis session begins. In the immunological study antinuclear antibodies, antineutrophil cytoplasmic antibodies, anti-GBM antibodies and protein electrophoresis were all within the normal range. Sterile blood and urine cultures are collected. Serology is extracted and tested for hepatitis B and C, human immunodeficiency virus, herpes virus, herpes 6, Epstein-Barr virus, Chlamydia and Mycoplasma, all with negative results. Suspecting immunoallergic acute renal failure, corticosteroids treatment is implemented on 3 daily doses of 250mg of methylprednisolone, followed by an intravenous dose of prednisone 1mg/kg. Later on, when the general state of the patient allows it, it is decided to carry out a renal biopsy. The biopsy includes 18 glomeruli, 6 of them with global sclerosis of the glomerular capillary. On the 12 preserved glomeruli, there are no significant intra-capillary lesions. A diffuse moderate tubulointerstitial lesion is detected, with inflammatory infiltrates made up of polymorphs formed by small lymphocytes, plasma cells and abundant eosinophils (Figure 1), and numerous images of tubulitis with lymphocyte infiltrates at the level of the tubular epithelium. On small arteries and small intralobular arteries there were no lesions. All the findings are compatible with the diagnosis of acute tubulointerstitial nephritis with eosinophils suggesting immunoallergic nephritis.

Tumor de células de Leydig de ovario: una causa infrecuente de virilización en la mujer posmenopáusica

Resumen El tumor de celulas de Leydig de ovario es un tipo de neoplasia muy raro que suele presentarse en mujeres posmenopausicas. Presentamos el caso de una mujer de 72 anos que consulto por virilizacion. Las determinaciones hormonales mostraron una elevacion muy marcada de la testosterona (12.038 pg/ml), con valores normales del resto de hormonas sexuales. En la ecografia se observo una tumoracion de 10 mm en el ovario izquierdo. Se realizo histerectomia y doble anexectomia. El estudio histologico demostro la existencia de un tumor de celulas de Leydig en el ovario izquierdo. Tras la intervencion, los valores plasmaticos de testosterona se normalizaron, y la paciente mostro una lenta regresion de los sintomas clinicos.

Comparison of risk classification between EndoPredict and MammaPrint in ER-positive/HER2-negative primary invasive breast cancer

Purpose To compare the concordance in risk classification between the EndoPredict and the MammaPrint scores obtained for the same cancer samples on 40 estrogen-receptor positive/HER2-negative breast carcinomas. Methods Formalin-fixed, paraffin-embedded invasive breast carcinoma tissues that were previously analyzed with MammaPrint as part of routine care of the patients, and were classified as high-risk (20 patients) and low-risk (20 patients), were selected to be analyzed by the EndoPredict assay, a second generation gene expression test that combines expression of 8 genes (EP score) with two clinicopathological variables (tumor size and nodal status, EPclin score). Results The EP score classified 15 patients as low-risk and 25 patients as high-risk. EPclin re-classified 5 of the 25 EP high-risk patients into low-risk, resulting in a total of 20 high-risk and 20 low-risk tumors. EP score and MammaPrint score were significantly correlated (p = 0.008). Twelve of 20 samples classified as low-risk by MammaPrint were also low-risk by EP score (60%). 17 of 20 MammaPrint high-risk tumors were also high-risk by EP score. The overall concordance between EP score and MammaPrint was 72.5% (κ = 0.45, (95% CI, 0.182 to 0.718)). EPclin score also correlated with MammaPrint results (p = 0.004). Discrepancies between both tests occurred in 10 cases: 5 MammaPrint low-risk patients were classified as EPclin high-risk and 5 high-risk MammaPrint were classified as low-risk by EPclin and overall concordance of 75% (κ = 0.5, (95% CI, 0.232 to 0.768)). Conclusions This pilot study demonstrates a limited concordance between MammaPrint and EndoPredict. Differences in results could be explained by the inclusion of different gene sets in each platform, the use of different methodology, and the inclusion of clinicopathological parameters, such as tumor size and nodal status, in the EndoPredict test.

High-throughput 3-dimensional culture of epithelial ovarian cancer cells as preclinical model of disease

Background Recent reports have identified distinct genomic patterns in ovarian carcinoma, including proliferative and mesenchymal-like groups, with worse outcome. The exact mechanisms driving the onset and progression of these tumors are still poorly understood. Additionally, researchers are concerned about the correct subtype stratification of the available cell line models, and the exploration of alternatives to monolayer culture. Identification of biomarkers to stratify cell lines, characterization of important processes as epithelial-mesenchymal transition (EMT), and the use of three-dimensional (3D) cultures as alternative models could be useful for cell line classification. Methods and Results In this work, we present a descriptive analysis of 16 commonly used ovarian cancer cell lines. We have studied their morphology in 2- and 3D culture, and their response to cisplatin, observing in the majority of them an increased resistance in 3D. We have also performed an immunohistochemical analysis for proliferation marker Ki-67, and EMT related markers to establish phenotypes. Epithelial cells tend to show higher proliferative rates, and mesenchymal cells show an increase in EMT related markers, especially when cultured in 3D conditions. Conclusions We have stated the complex heterogeneity of ovarian cancer models, resembling primary tumors, agreeing with the argument that the cell line model for in vitro experiments must be carefully chosen. Our results also support that tridimensional culture could be a very helpful alternative in ovarian cancer research. Regarding EMT, a very important process for the development of this disease, some related biomarkers might be further characterized for their role in this disease development.

Predicting Response to Standard First-line Treatment in High-grade Serous Ovarian Carcinoma by Angiogenesis-related Genes

Background/Aim: Predicting response to treatment in high-grade serous ovarian carcinoma (HGSOC) still remains a clinical challenge. The standard-of-care for first-line chemotherapy, based on a combination of carboplatin and paclitaxel, achieves a high response rate. However, the development of drug resistance is one of the major limitations to efficacy. Therefore, identification of biomarkers able to predict response to chemotherapy in patients with HGSOC is a critical step for prognosis and treatment of the disease. Several studies suggest that angiogenesis is an important process in the development of ovarian carcinoma and chemoresistance. The aim of this study was to identify a profile of angiogenesis-related genes as a biomarker for response to first-line chemotherapy in HGSOC. Materials and Methods: Formalin-fixed paraffin-embedded samples from 39 patients with HGSOC who underwent surgical cytoreduction and received a first-line chemotherapy with carboplatin and paclitaxel were included in this study. Expression levels of 82 angiogenesis-related genes were measured by quantitative real-time polymerase chain reaction using TaqMan low-density arrays. Results: Univariate analysis identified five genes [angiopoietin 1 (ANGPT1), aryl hydrocarbon receptor nuclear translocator (ARNT), CD34, epidermal growth factor (EGF) and matrix metallopeptidase 3 (MMP3)] as being statistically associated with response to treatment. Multivariable analysis by Lasso-penalized Cox regression generated a model with the combined expression of seven genes [angiotensinogen (AGT), CD34, EGF, erythropoietin receptor (EPOR), interleukin 8 (IL8), MMP3 and MMP7)]. The area under the receiver operating characteristics curve (0.679) and cross-validated Kaplan–Meier survival curves were used to estimate the accuracy of these predictors. Conclusion: An angiogenesis-related gene expression profile useful for response prediction in HGSOC was identified, supporting the important role of angiogenesis in HGSOC.

Primary Leiomyosarcoma of the Ovarian Vein: Case Report and Literature Review:

Primary leiomyosarcoma arising from the ovarian vein is extremely rare, with only 10 cases reported in the literature. We report on a case of leiomyosarcoma of the left ovarian vein in a 67-year-old woman who presented with abdominal discomfort. Pelvic ultrasound revealed a large, solid, irregular mass in close relation to the left ovary. The patient subsequently underwent a total hysterectomy with bilateral salpingo-oophorectomy. Histologically, the tumor was composed of interlacing fascicles of spindle cells with abundant eosinophilic cytoplasm, hyperchromatic nuclei, and prominent nucleoli. Mitotic activity was high, with 24 mitoses in 10 high-power fields. Areas of necrosis and hemorrhage were present within the tumor. Immunohistochemically, the tumor cells showed diffuse immunoreactivity for vimentin, muscle-specific actin, desmin, and caldesmon. The patient received chemotherapy postoperatively but subsequently developed disseminated metastatic disease (lung, liver, iliac lymph nodes, and peritoneum). Primary leiomyosarcomas arising from the ovarian vein are aggressive neoplasms, and the prognosis correlates with stage.

Predictive value of angiogenesis-related gene profiling in patients with HER2-negative metastatic breast cancer treated with bevacizumab and weekly paclitaxel

Bevacizumab plus weekly paclitaxel improves progression-free survival (PFS) in HER2-negative metastatic breast cancer (mBC), but its use has been questioned due to the absence of a predictive biomarker, lack of benefit in overall survival (OS) and increased toxicity. We examined the baseline tumor angiogenic-related gene expression of 60 patients with mBC with the aim of finding a signature that predicts benefit from this drug. Multivariate analysis by Lasso-penalized Cox regression generated two predictive models: one, named G-model, including 11 genes, and the other one, named GC-model, including 13 genes plus 5 clinical covariates. Both models identified patients with improved PFS (HR (Hazard Ratio) 2.57 and 4.04, respectively) and OS (HR 3.29 and 3.43, respectively). The G-model distinguished low and high risk patients in the first 6 months, whereas the GC-model maintained significance over time.

907PASSOCIATION BETWEEN ANGIOGENESIS-RELATED GENES AND THE RESPONSE TO MULTIMODAL THERAPY IN HIGH GRADE SEROUS ADVANCED OVARIAN CARCINOMA (AOC)

ABSTRACT Aim: Cytoreductive surgery plus chemotherapy is the standard of care in high grade serous AOC. To date, the outcome prediction after multimodal therapy remains one of the most important challenges for patients (pts) stratification according to effective treatments. Angiogenesis has maintained its prominent role in the ovarian cancer research field for years, being one of the most studied processes for the identification of alternative therapies in AOC. Previous studies identified several genes involved in angiogenesis as prognostic markers in this tumour. Methods: We included 39 pts with stages III or IV high grade serous ovarian cancer. These pts underwent surgical cytoreduction and received a carboplatin plus paclitaxel chemotherapy regimen. RNAs were collected from formalin-fixed paraffin-embedded samples. Expression levels of 82 angiogenesis-related genes were measured using quantitative real-time polymerase chain reaction (qRT-PCR). Results: 74% of pts (29 out of 39) achieved a complete response after surgical and chemotherapy treatments. The only clinical factor associated to treatment complete response was the remaining tumour after debulking surgery (HR 18.9, 95% CI (2.0 - 2458.5)). Regarding the genetic variables analyzed, we found that the expression of ANGPT1, ARNT, CD34, EGF and MMP3 genes were related to response in the univariate analysis. We tried also to generate a genetic model for response prediction, being the most interesting one composed of the combined expression of 7 genes (AGT, CD34, EGF, EPOR, IL8, MMP3 and MMP7). After leave one out cross validation (LOOCV), we obtained an AUC of 0.67 and the accuracy assigned was 0.74. The only factor that remains statistically significant in the multivariate analysis (including clinical and genetic factors) was the residual tumour after surgery. Conclusions: Remaining tumour after debulking surgery remains the most important factor to date to successfully achieve a complete response after the multimodal treatment in high-grade serous AOC pts. The specific roles of genes identified in this study will need to be further studied. Disclosure: All authors have declared no conflicts of interest.