BACKGROUND: The search for effective biomarkers for ovarian cancer (OC) early diagnosis is an urgent task of modern oncogynecology. Metabolic profiling by ultra-high performance liquid chromatography and mass spectrometry (UHPLC-MS) provides information on the totality of all low molecular weight metabolites of patient's biological fluids sample, reflecting the processes occurring in the body. The aim of the study was to research blood plasma and urine metabolomic profile of patients with serous ovarian adenocarcinoma by UHPLC-MS. MATERIAL AND METHODS: To perform metabolomic analysis, 60 blood plasma samples and 60 urine samples of patients diagnosed with serous ovarian carcinoma and 20 samples of apparently healthy volunteers were taken. Chromatographic separation was performed on a Vanquish Flex UHPLC System chromatograph (Thermo Scientific, Germany). Mass spectrometric analysis was performed on an Orbitrap Exploris 480 (Thermo Scientific, Germany) equipped with an electrospray ionization source. Bioinformatic analysis was performed using Compound Discoverer Software (Thermo Fisher Scientific, USA), statistical data analysis was performed in the Python programming language using the SciPy library. RESULTS: Using UHPLC-MS, 1,049 metabolites of various classes were identified in blood plasma. In patients with OC, 8 metabolites had a significantly lower concentration (P < 0.01) compared with conditionally healthy donors, while the content of 19 compounds, on the contrary, increased (P < 0.01). During the metabolomic profiling of urine samples, 417 metabolites were identified: 12 compounds had a significantly lower concentration compared to apparently healthy individuals, the content of 14 compounds increased (P < 0.01). In patients with ovary serous adenocarcinoma, a significant change in the metabolome of blood plasma and urine was found, expressed in abnormal concentrations of lipids and their derivatives, fatty acids and their derivatives, acylcarnitines, phospholipids, amino acids and their derivatives, derivatives of nitrogenous bases and steroids. At the same time, kynurenine, myristic acid, lysophosphatidylcholine and L-octanoylcarnitine are the most promising markers of this disease. CONCLUSION: The revealed changes in the metabolome can become the basis for improving approaches to the diagnosis of serous ovarian adenocarcinoma.

• Klíčová slova
• Urine, blood plasma, metabolomic profile, serous ovarian adenocarcinoma, ultra-high performance liquid chromatography and mass spectrometry, urine,
• MeSH
• hmotnostní spektrometrie MeSH
• lidé středního věku MeSH
• lidé MeSH
• metabolom MeSH
• metabolomika * metody   MeSH
• nádorové biomarkery krev   moč   MeSH
• nádory vaječníků * metabolismus   krev   moč   diagnóza   MeSH
• serózní cystadenokarcinom metabolismus   diagnóza   krev   moč   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• nádorové biomarkery MeSH

High-grade serous carcinoma of the ovary, fallopian tube and peritoneum (HGSC), the most common type of ovarian cancer, ranks among the deadliest malignancies. Many HGSC patients have excess fluid in the peritoneum called ascites. Ascites is a tumour microenvironment (TME) containing various cells, proteins and extracellular vesicles (EVs). We isolated EVs from patients' ascites by orthogonal methods and analyzed them by mass spectrometry. We identified not only a set of 'core ascitic EV-associated proteins' but also defined their subset unique to HGSC ascites. Using single-cell RNA sequencing data, we mapped the origin of HGSC-specific EVs to different types of cells present in ascites. Surprisingly, EVs did not come predominantly from tumour cells but from non-malignant cell types such as macrophages and fibroblasts. Flow cytometry of ascitic cells in combination with analysis of EV protein composition in matched samples showed that analysis of cell type-specific EV markers in HGSC has more substantial prognostic potential than analysis of ascitic cells. To conclude, we provide evidence that proteomic analysis of EVs can define the cellular composition of HGSC TME. This finding opens numerous avenues both for a better understanding of EV's role in tumour promotion/prevention and for improved HGSC diagnostics.

• Klíčová slova
• ascites, extracellular vesicles (EV), fallopian tube and peritoneum (HGSC), high-grade serous carcinoma of the ovary, macrophage, ovarian cancer (OC), tandem mass spectrometry (MS/MS), tumour microenvironment (TME),
• MeSH
• ascites metabolismus   patologie   MeSH
• extracelulární vezikuly * metabolismus   MeSH
• lidé MeSH
• nádorové mikroprostředí MeSH
• nádory vaječníků * diagnóza   MeSH
• proteomika MeSH
• serózní cystadenokarcinom * diagnóza   genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: IMP2 and IMP3 are mRNA binding proteins involved in carcinogenesis. We examined a large cohort of ovarian tumors with the aim to assess the value of IMP2 and IMP3 for differential diagnosis, and to assess their prognostic significance. METHODS: Immunohistochemical analyses with antibodies against IMP2 and IMP3 were performed on 554 primary ovarian tumors including 114 high grade serous carcinomas, 100 low grade serous carcinomas, 124 clear cell carcinomas, 54 endometrioid carcinomas, 34 mucinous carcinomas, 75 mucinous borderline tumors, and 41 serous borderline tumors (micropapillary variant). The associations of overall positivity with clinicopathological characteristics were evaluated using the chi-squared test or Fisher's Exact test. RESULTS: We found IMP2 expression (in more than 5% of tumor cells) in nearly all cases of all tumor types, so the prognostic meaning could not be analyzed. The positive IMP3 expression (in more than 5% of tumor cells) was most common in mucinous carcinomas (82%) and mucinous borderline tumors (81%), followed by high grade serous (67%) and clear cell carcinomas (67%). The expression was less frequent in endometrioid carcinomas (39%), low grade serous carcinomas (23%), and micropapillary variant of serous borderline tumors (20%). Prognostic significance of IMP3 could be evaluated only in low grade serous carcinomas in the case of relapse-free survival, where negative cases showed better RFS (p = 0.033). CONCLUSION: Concerning differential diagnosis our results imply that despite the differences in expression in the different ovarian tumor types, the practical value for diagnostic purposes is limited. Contrary to other solid tumors, we did not find prognostic significance of IMP3 in ovarian cancer, with the exception of RFS in low grade serous carcinomas. However, the high expression of IMP2 and IMP3 could be of predictive value in ovarian carcinomas since IMP proteins are potential therapeutical targets.

• Klíčová slova
• IMP2, IMP3, Immunohistochemistry, Ovarian carcinoma,
• MeSH
• endometroidní karcinom * patologie   MeSH
• lidé MeSH
• lokální recidiva nádoru MeSH
• mucinózní adenokarcinom * diagnóza   patologie   MeSH
• nádorové biomarkery analýza   MeSH
• nádory vaječníků * patologie   MeSH
• peritoneální nádory * MeSH
• prekancerózy * MeSH
• serózní cystadenokarcinom * metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• IGF2BP2 protein, human MeSH Prohlížeč
• IMP3 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH

PURPOSE: In multiple oncological settings, expression of the coinhibitory ligand PD-L1 by malignant cells and tumor infiltration by immune cells expressing coinhibitory receptors such as PD-1, CTLA4, LAG-3, or TIM-3 conveys prognostic or predictive information. Conversely, the impact of these features of the tumor microenvironment on disease outcome among high-grade serous carcinoma (HGSC) patients remains controversial. EXPERIMENTAL DESIGN: We harnessed a retrospective cohort of 80 chemotherapy-naïve HGSC patients to investigate PD-L1 expression and tumor infiltration by CD8+ T cells, CD20+ B cells, DC-LAMP+ dendritic cells as well as by PD-1+, CTLA4+, LAG-3+, and TIM-3+ cells in relation with prognosis and function orientation of the tumor microenvironment. IHC data were complemented with transcriptomic and functional studies on a second prospective cohort of freshly resected HGSC samples. In silico analysis of publicly available RNA expression data from 308 HGSC samples was used as a confirmatory approach. RESULTS: High levels of PD-L1 and high densities of PD-1+ cells in the microenvironment of HGSCs were strongly associated with an immune contexture characterized by a robust TH1 polarization and cytotoxic orientation that enabled superior clinical benefits. Moreover, PD-1+TIM-3+CD8+ T cells presented all features of functional exhaustion and correlated with poor disease outcome. However, although PD-L1 levels and tumor infiltration by TIM-3+ cells improved patient stratification based on the intratumoral abundance of CD8+ T cells, the amount of PD-1+ cells failed to do so. CONCLUSIONS: Our data indicate that PD-L1 and TIM-3 constitute prognostically relevant biomarkers of active and suppressed immune responses against HGSC, respectively.

• MeSH
• antigen CTLA-4 imunologie   metabolismus   MeSH
• antigeny CD274 imunologie   metabolismus   MeSH
• buněčný receptor 2 viru hepatitidy A imunologie   metabolismus   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dospělí MeSH
• epiteliální ovariální karcinom imunologie   metabolismus   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny asociované s lyzozomy imunologie   metabolismus   MeSH
• míra přežití MeSH
• nádorové biomarkery imunologie   metabolismus   MeSH
• nádorové proteiny imunologie   metabolismus   MeSH
• prognóza MeSH
• regulace genové exprese u nádorů * MeSH
• retrospektivní studie MeSH
• senioři MeSH
• serózní cystadenokarcinom imunologie   metabolismus   patologie   MeSH
• tumor infiltrující lymfocyty imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• antigen CTLA-4 MeSH
• antigeny CD274 MeSH
• buněčný receptor 2 viru hepatitidy A MeSH
• CD274 protein, human MeSH Prohlížeč
• CTLA4 protein, human MeSH Prohlížeč
• HAVCR2 protein, human MeSH Prohlížeč
• LAMP3 protein, human MeSH Prohlížeč
• membránové glykoproteiny asociované s lyzozomy MeSH
• nádorové biomarkery MeSH
• nádorové proteiny MeSH

OBJECTIVE: The objectives of this study were to assess the immunohistochemical expression of p53, bcl-2, c-erbB-2, Ki-67, estrogen (ER) and progesterone (PR) receptors, matrix metalloproteinase-7 and -26 (MMP-7 and MMP-26) in endometrial cancer patients and to assess the relation between steroid receptor positivity and other markers. DESIGN: Experimental prospective study. SETTING: Department of Obstetrics and Gynecology, Department of Genetics, Department of Pathology, Palacký University Medical School and University Hospital Olomouc. METHODS: We studied 144 cases of primary untreated endometrial carcinoma in which the p53, bcl-2, c-erbB-2, Ki-67, ER, PR, MMP-7, and MMP-26 antigens were investigated with the use of immunohistochemical methods. We evaluated the correlations among immunohistochemical staining and the age, International Federation of Gynecology and Obstetrics stage, grading, depth of invasion, and metastatic spread to lymph nodes. RESULTS: Mean age was 65.7 years (range, 34-90 years). p53, bcl-2, c-erbB-2, Ki-67, ER, and PR were positive in 35 (24.3%), 100 (69.4%), 41 (28.4%), 65 (45.1%), 115 (79.8%), and 127 (88.1%) cases, respectively. Matrix metalloproteinases were evaluated in a group of 70 patients, wherein MMP-7 was positive in 33 patients (47.1%) and MMP-26 was positive in 40 patients (57.1%). The expression of MMP-7 decreased with higher patient age. p53 and Ki-67 overexpression was found to be related to poor differentiation. Immunostaining for bcl-2 correlated with the positivity of steroid receptors status, whereas immunostaining for c-erbB-2 correlated inversely with ER-positive group of cases. CONCLUSIONS: The overexpression of p53 and Ki-67 seems to indicate a more malignant phenotype, whereas bcl-2 expression in dependence of steroid receptor positivity could contribute to the identification of high-risk tumors.

• MeSH
• antigen Ki-67 metabolismus   MeSH
• dospělí MeSH
• imunoenzymatické techniky MeSH
• invazivní růst nádoru MeSH
• lidé středního věku MeSH
• lidé MeSH
• matrixová metaloproteinasa 7 metabolismus   MeSH
• metaloproteinasy secernované do matrix metabolismus   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorový supresorový protein p53 metabolismus   MeSH
• nádory endometria metabolismus   patologie   MeSH
• papilární karcinom metabolismus   patologie   MeSH
• prognóza MeSH
• prospektivní studie MeSH
• protoonkogenní proteiny c-bcl-2 metabolismus   MeSH
• receptor erbB-2 metabolismus   MeSH
• receptory pro estrogeny metabolismus   MeSH
• receptory progesteronu metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• serózní cystadenokarcinom metabolismus   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigen Ki-67 MeSH
• matrixová metaloproteinasa 7 MeSH
• metaloproteinasy secernované do matrix MeSH
• MMP26 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• nádorový supresorový protein p53 MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• receptor erbB-2 MeSH
• receptory pro estrogeny MeSH
• receptory progesteronu MeSH
• TP53 protein, human MeSH Prohlížeč