Vol 18, No 3 (2020)

Congenital adrenal hyperplasia (CAH) is a group of disorders caused by mutations in genes that encode steroidogenic enzymes. The most frequent form of CAH is caused by defects in CYP21A2 gene leading to 21-hydroxylase deficiency (21-OHD). There are 3 clinical forms of CAH: classic (salt-wasting (SW), simple virilizing (SV)), and nonclassic (NCCAH). Classic forms of 21-OHD can be specifically detected by the measurement of 17-hydroxyprogesterone in blood. Nonclassic form is often characterized by equivocal level of 17-hydroxyprogesterone and non-specific symptoms, so the genotyping is essential for the diagnosis. Moreover, molecular analysis of CYP21A2 mutations is useful for predicting the severity of disease and important for genetic counseling. We discuss the structure of CYP21A2 gene, types of mutations, mechanisms of clinical manifestation of disorder, including clinical features of heterozygote carriers. We present results of the study of 85 patients with hyperandrogenemia that were genotyped by multiplex test detecting 15 most common mutations in CYP21A2.

Introduction. Uveal melanoma is a malignant neoplasm of neuroectodermal origin, prone to early metastasis. The tumor microenvironment, including «inflammatory cells» and connective tissue cells, affects its progression, angiogenesis and metastasis. Purpose. To carry out clinical and morphological correlations of the cellular microenvironment of uveal melanoma with the detection of predictors of bad prognosis. Methods. 43 enucleated eyes with uveal melanoma (258 histological preparation) of adult patients were examined. Pathological type of tumors: epithelioid cell (n=9), spindle cell type AB (n=15), mixed cell (n=19). Tumor thickness - 4,7±1,3 mm, bazal diameter - 13,5±3,3 mm. T1N0M0 tumors in 6 eyes were detected, which were located juxtapapillary with involvement of the optic nerve head, which required enucleation. The presence and localization of lymphocytes, macrophages, mast cells, plasma cells, myeloid-derived suppressor cells (promyelocytes, neutrophils, eosinophils), fibroblasts and degree of tumor tissue infiltration were analyzed. Statistical methods: Microsoft Excel, Statistica 10.1, Spearman’s rank correlation coefficient - rs. Results. Analyzing the obtained results, we revealed higher correlations of the epithelium-cell type with the presence of macrophages, myeloid-derived suppressor cells (promyelocytes, neutrophils, eosinophils). The high degree of pigmentation correlated mainly with the presence of mast cells, myeloid-derived suppressor cells (promyelocytes, eosinophils), fibroblasts. The high degree of tumor vascularization correlated mainly with the presence of mast cells. At the same time, the moderate correlation between growth of tumor cells by emissaries with mast cells, myeloid-derived suppressor cells (promyelocytes, eosinophils), fibroblasts was noted. Conclusion. Thus, identification of certain cells in uveal melanoma microenvironment may be a predictor of an unfavorable tumor course and may serve as a pretext for the development of accompanying target therapy aimed to eliminating or reprogramming altered immunity cells and connective tissue.

Background. Lung cancer is the most frequent cause of cancer mortality. The prevailing type is non-small cell lung cancer (NSCLC), in the treatment of which EGFR tyrosine kinase inhibitors are actively used, as well as a combination of BRAF and MEK inhibitors. The indication for the targeted therapy is the genotyping of tumor samples for the detection of predictive markers, such as mutations in the EGFR, KRAS, BRAF and HER2 genes. One approach to the rapid and complex identification of aberrations in the genes of interest is the use of methods with high sensitivity: multi-target single-base elongation (MSE) and fragment analysis (FA). Objective. Determination of clinicopathological characteristics of patients with NSCLC with the presence of driver mutations in the EGFR, KRAS, BRAF and HER2 genes. Materials and methods. A cryobank of biological specimens taken from patients with NSCLC was organized at Pavlov First Saint Petersburg State Medical University. For genotyping of the central sites of tumor samples from 60 patients with NSCLC were used multi-target single-base elongation and fragment analysis. Results. EGFR mutations were found exclusively in patients with lung adenocarcinoma (p=0,0008) with a frequency of 31%. These aberrations are significantly associated with female gender (p=0,0002) and the absence of smoking status (p=0,0007). The ratio of exon 19 deletion to L858R was 2,5:1. The prevalence of minor mutations in EGFR-positive patients was 22,2%. Thus, the coexistence of minor mutations G719S and S768I was revealed in one tumor sample. The cause of acquired resistance to the first-generation EGFR TKI in a patient with exon 19 deletion was determined - T790M in the EGFR gene. KRAS mutations were found in 10% of male patients with NSCLC who was a current or former smoker. V600E in the BRAF gene and insertions in the HER2 gene were not detected. Conclusion. An integrated methodological approach allows to accurately determine clinically significant aberrations in the EGFR, KRAS, BRAF and HER2 genes in NSCLC patients according to the recommendations.