SOX2 Gene Amplification and Overexpression is Linked to HPV-positive Vulvar Carcinomas.

SOX2 (SRY-related HMG-box 2) belongs to the SOX gene family of high-mobility transcription factors indispensably involved in gene regulation in pluripotent stem cells and neural differentiation. SOX2 copy number increases have been frequently reported in various types of squamous cell cancer. To better understand the effect of SOX2 aberrations on vulvar cancer phenotype and patient prognosis, we analyzed SOX2 copy number changes using fluorescence in situ hybridization and SOX2 expression by immunohistochemistry in 55 squamous cell carcinomas of the vulva. SOX2 amplification was found in 20.8% of tumors; 27.3% of vulvar carcinomas showed SOX2 protein overexpression. SOX2 amplification was correlated with SOX2 overexpression in our data set (P<0.01). Amplification of the SOX2 locus was associated with high tumor grade (P<0.05) and human papillomavirus (HPV) positivity (P<0.01). SOX2-amplified tumors showed more frequently a basaloid phenotype than nonamplified carcinomas. SOX2 protein overexpression was also correlated with basaloid phenotype and positive HPV status of vulvar carcinomas (P<0.05, each). SOX2 amplification and expression were not associated with patient overall survival. In conclusion, SOX2 copy number increases are detectable in a substantial proportion of high-grade HPV-positive vulvar carcinomas with basaloid differentiation. Our study provides further evidence for different molecular alterations in HPV-positive and HPV-negative vulvar carcinomas.

Regulatory role of G protein-coupled estrogen receptor for vascular function and obesity.

We found that the selective stimulation of the intracellular, transmembrane G protein-coupled estrogen receptor (GPER), also known as GPR30, acutely lowers blood pressure after infusion in normotensive rats and dilates both rodent and human arterial blood vessels. Stimulation of GPER blocks vasoconstrictor-induced changes in intracellular calcium concentrations and vascular tone, as well as serum-stimulated cell proliferation of human vascular smooth muscle cells. Deletion of the GPER gene in mice abrogates vascular effects of GPER activation and is associated with visceral obesity. These findings suggest novel roles for GPER in protecting from cardiovascular disease and obesity.

Deep learning for the standardized classification of Ki-67 in vulva carcinoma: A feasibility study.

BACKGROUND: The aim of this study is to demonstrate the feasibility of automatic classification of Ki-67 histological immunostainings in patients with squamous cell carcinoma of the vulva using a deep convolutional neural network (dCNN). MATERIAL AND METHODS: For evaluation of the dCNN, we used 55 well characterized squamous cell carcinomas of the vulva in a tissue microarray (TMA) format in this retrospective study. The tumor specimens were classified in 3 different categories C1 (0-2%), C2 (2-20%) and C3 (>20%), representing the relation of the number of KI-67 positive tumor cells to all cancer cells on the TMA spot. Representative areas of the spots were manually labeled by extracting images of 351 × 280 pixels. A dCNN with 13 convolutional layers was used for the evaluation. Two independent pathologists classified 45 labeled images in order to compare the dCNN's results to human readouts. RESULTS: Using a small labeled dataset with 1020 images with equal distribution among classes, the dCNN reached an accuracy of 90.9% (93%) for the training (validation) data. Applying a larger dataset with additional 1017 labeled images resulted in an accuracy of 96.1% (91.4%) for the training (validation) dataset. For the human readout, there were no significant differences between the pathologists and the dCNN in Ki-67 classification results. CONCLUSION: The dCNN is capable of a standardized classification of Ki-67 staining in vulva carcinoma; therefore, it may be suitable for quality control and standardization in the assessment of tumor grading.

The Paris System for reporting urinary cytology in daily practice with emphasis on ancillary testing by multiprobe FISH.

AIMS: The Paris System (TPS) was introduced in the diagnostic routine with the goal to simplify and standardise diagnostic reporting of urinary cytology. The diagnostic categories of TPS are based on defined cytological criteria, with a focus on high-grade urothelial carcinoma (HGUC). While the categories 'negative for HGUC (NHGUC)' and 'HGUC' are straightforward, the categories 'atypical urothelial cells (AUC)' and 'suspicious of HGUC (SHGUC)' remain inconclusive. In this study, we evaluated the feasibility of TPS in daily practice with special emphasis on ancillary fluorescence in situ hybridisation (FISH) testing in the setting of TPS categories. METHODS: In a 19-month period, TPS was prospectively applied in the routine diagnostic setting on 3900 urinary cytology cases comprising bladder and upper urinary tract washings and voided urine specimens. Additionally, we analysed the results of the FISH assay UroVysion prospectively performed on a cohort of 128 cases enriched for AUC and SHGUC categories. RESULTS: The most frequently reported category was NHGUC (n=3496, 89.7%), followed by AUC (n=178, 4.6%), HGUC (n=155, 4%), SHGUC (n=61, 1.6%), low-grade urothelial neoplasia (n=6, 0.1%) and other malignancies (n=4, 0.1%). In the FISH cohort, 40/90 (44%) cases within the AUC category were FISH positive, consistent with urothelial neoplasia. In the SHGUC category, 16/21 (76%) cases were FISH positive. CONCLUSIONS: When prospectively applying TPS in urinary cytology, inconclusive atypia accounts only for a small subset of cases. FISH additionally improves the stratification between reactive and malignant cells in the indeterminate AUC and SHGUC categories.