Testicular germ cell tumors (TGCTs) belong to the most chemosensitive solid tumors; however, a small proportion of patients fail to be cured with cisplatin-based chemotherapy. Inhibitors of ...PD-1/PD-L1 pathways represent a new class of promising drugs in anticancer therapy. The aim of this study was to evaluate expression and prognostic value of PD-1 and PD-L1 in TGCTs.
Surgical specimens from 140 patients with TGCTs (131 with primary testicular tumor and 9 with extragonadal GCTs) were included into the translational study. PD-1 and PD-L1 expression was detected in the tumor tissue by immunohistochemistry using monoclonal antibodies, scored by the multiplicative quickscore (QS) method, compared with their expression in normal testicular tissue and correlated with clinicopathological characteristics and clinical outcome.
None of the GCTs exhibited PD-1 protein, although expression of PD-L1 was significantly higher in GCTs in comparison with normal testicular tissue (mean QS = 5.29 versus 0.32, P < 0.0001). Choriocarcinomas exhibit the highest level of PD-L1 with decreasing positivity in embryonal carcinoma, teratoma, yolk sac tumor and seminoma. PD-L1 expression was associated with poor prognostic features, including ≥3 metastatic sites, increased serum tumor markers and/or non-pulmonary visceral metastases. Patients with low PD-L1 expression had significantly better progression-free survival hazard ratio (HR) = 0.40, 95% confidence interval (CI) 0.16–1.01, P = 0.008 and overall survival (HR = 0.43, 95% CI 0.15–1.23, P = 0.040) compared with patients with high PD-L1 expression.
In this translational study, we showed, for the first time, the prognostic value of PD-L1 expression in TGCTs and our data imply that the PD-1/PD-L1 pathway could be a novel therapeutic target in TGCTs.
Cytokines are involved in cancer invasion and metastasis. Circulating tumor cells (CTCs) play key role in tumor dissemination and are an independent survival predictor in breast cancer patients. The ...aim of this study was to assess correlation between CTCs and plasma cytokines in primary breast cancer (PBC) patients.
This study included 147 chemotherapy naïve PBC patients. Peripheral blood mononuclear cells (PBMC) were depleted of hematopoetic cells using RossetteSep™ negative selection kit. RNA extracted from CD45-depleted PBMC was interrogated for expression of EMT (Twist1, Snail1, Slug, Zeb1) and epithelial (Ck19) gene transcripts by qRT-PCR. The concentrations of 51 plasma cytokines were measured using multiplex bead arrays.
CTCs were detected in 25.2% patients. CTCs exhibiting only epithelial markers (CTC_EP) and only EMT markers (CTC_EMT) were present evenly in 11.6% patients, while CTCs co-expressing both markers were detected in 2.0% patients. Patients with presence of CTC_EP in peripheral blood had significantly elevated levels of plasma IFN-α2, IL-3, MCP-3, β-NGF, SCF, SCGF-β, TNF-β and SDF-1 compared to patients without CTC_EP. CTC_EP exhibited overexpression of SDF-1 receptor and CXCR4, but not other corresponding cytokine receptor, and in multivariate analysis SDF-1 was independently associated with CTC_EP. There was an inverse correlation between CTC_EMT and plasma cytokines CTACK, β-NGF and TRAIL, while presence of either subtype of CTCs was associated with increased level of TGF-β2.
Using cytokine profiling, we identified cytokines associated with CTCs subpopulations in peripheral blood of PBC. Our data suggest that CXCR4-SDF-1 axis is involved in mobilization and trafficking of epithelial CTCs.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Circulating tumor cells (CTCs) play a crucial role in tumor dissemination and are an independent survival predictor in breast cancer (BC) patients. Epithelial to mesenchymal transition (EMT) is ...involved in cancer invasion and metastasis. The aim of this study was to assess correlation between CTCs and expression of EMT transcription factors TWIST1 and SLUG in breast tumor tissue.
This study included 102 early BC patients treated by primary surgery. Peripheral blood mononuclear cells (PBMC) were depleted of hematopoietic cells using RossetteSep™ negative selection kit. RNA extracted from CD45-depleted PBMC was interrogated for expression of EMT (TWIST1, SNAIL1, SLUG, FOXC2 and ZEB1) and epithelial (KRT19) gene transcripts by qRT-PCR. Expression of TWIST1 and SLUG in surgical specimens was evaluated by immunohistochemistry and quantified by multiplicative score.
CTCs were detected in 24.5 % patients. CTCs exhibiting only epithelial markers were present in 8.8 % patients, whereas CTCs with only EMT markers were observed in 12.8 % of pts and CTCs co-expressing both markers were detected in 2.9 % pts. We observed lack of correlation between CTCs and expression of TWIST1 and SLUG in breast cancer cells or cancer associated stroma. Lack of correlation was observed for epithelial CTCs as well as for CTCs with EMT.
In this translational study, we showed a lack of association between CTCs and expression of EMT-inducing transcription factors, TWIST1 and SLUG, in breast tumor tissue. Despite the fact that EMT is involved in cancer invasion and metastasis our results suggest, that expression of EMT proteins in unselected tumor tissue is not surrogate marker of CTCs with either mesenchymal or epithelial features.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Germ cell neoplasia in situ (GCNIS), is preinvasive stage of testicular germ cell tumours (TGCTs). Fibrillins, which are integral components of microfibrils are suggested to be involved in cancer ...pathogenesis and maintenance of embryonic stem cells pluripotency. The aim of this study was to examine fibrillin-1 (FBN-1) expression in TGCTs patients.
Surgical specimens from 203 patients with TGCTs were included into the translational study. FBN-1 expression was evaluated in the tumour tissue, in GCNIS and in adjacent non-neoplastic testicular tissue in all available cases. Tissue samples were processed by the tissue microarray method. FBN-1 was detected by immunohistochemistry using goat polyclonal antibody and the expression was evaluated by the multiplicative quickscore (QS).
The highest FBN-1 positivity was detected in GCNIS (mean QS = 11.30), with overexpression of FBN-1 (QS >9) in the majority (77.1 %) of cases. Expression of FBN-1 in all subtypes of TGCTs was significantly lower in comparison to expression in GCNIS (all p <0.001). Seminoma had significantly higher expression compared to EC, ChC and TER (all p <0.05), but not to YST (p = 0.84). In non-neoplastic testicular tissue the FBN-1 positivity was very low (mean QS = 0.02). Sensitivity, specificity, positive and negative predictive value of FBN-1 expression for diagnosis of GCNIS were 97.1, 98.8, 98.6 and 97.7 %.
FBN-1 is overexpressed in TGCTs and especially in GCNIS when compared to non-neoplastic testicular tissue in patients with germ cell tumors and could be involved in germ cell neoplasia in situ development.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The purpose of this study was to monitor the association between single umbilical artery (SUA), chromosomal abnormalities and associated anomalies during the routine examination of spontaneous or ...induced miscarriages and premature births.
During 1992-2015 we morphologically and cytogenetically examined a series of 4098 samples. For 1330 cases the number of umbilical cord vessels could be reported.
The presence of single umbilical artery was identified in 67 fetuses of 1330 pregnancies (5.04 %); 36 of the 67 fetuses (53.7 %) had additional congenital malformations. The cultures were unsuccessful in 29 of 67 cases (43.3 %). 38 cases (56.7 %) were successfully karyotyped; 20 out of them had a normal karyotype and 18 had chromosomal anomalies including trisomy 18 (n = 4), trisomy 13 (n = 3), trisomy 21 (n = 2), trisomy 11 (n = 1), triploidy (n = 3), monosomy X (n = 3) and structural chromosomal aberrations (n = 2).
Isolated SUA is not at increased risk of chromosomal abnormalities and generally does not endanger pregnancy. All chromosomally abnormal embryos and fetuses had associated congenital anomalies. The most frequently associated congenital anomalies were in the musculoskeletal system, central nervous system and genitourinary tract (Tab. 4, Ref. 44).
To analyze a rare triad of intracranial fetal pathologies and clinical study of the novel defined sequence pathogenesis based on prenatal and postmortem findings.
Complex multidisciplinary clinical ...analysis and review of up-to-date literature.
In an 18-gestational-week fetus the screening ultrasound scan resembled the semilobar type of holoprosencephaly and oral tumor. After the indicated termination of pregnancy, the histopathology results confirmed another pathologies - oral meningoencephalocele, teratoma of the sellar area and large arachnoidal cyst of the anterior cerebral fossa. The surprising final results were evaluated by specialists in prenatal diagnosis, histopathology, genetics, neurology, and radiology.
We defined the final diagnosed triad oral meningoencephalocele - intracranial sellar teratoma- arachnoidal cyst as a novel sequence defect malformation. In the detailed sequence pathogenesis, the intracranial sellar teratoma created an aperture for meningoencephalocele in the cranial base and the arachnoidal cyst facilitated, by its growth and pressure, the protrusion of the brain tissues (Fig. 4, Ref. 10). Text in PDF www.elis.sk.
IntroductionCirculating tumour cells (CTCs) are associated with metastatic potential in a variety of human cancers. Although DNA methylation of cancer-related genes is a promising diagnostic and ...prognostic tool, it was rarely studied in relation to the presence of CTCs. ADAM23 gene, member of ADAM (a disintegrin and metalloproteinase domain) family, is involved in various biological events such as cell adhesion, fusion, migration, membrane protein shedding and proteolysis. The aberrant DNA methylation of ADAM23 was associated with expression silencing in breast, gastric, brain and other tumours. In our study we investigated the role of ADAM23 methylation in hematogenous spread of breast cancer (BC).Material and methodsTo detect CTCs, we analysed expression levels of KRT19, SLUG and TWIST genes in the CD45- enriched peripheral blood samples of 203 primary BC patients. The methylation levels of ADAM23 were quantified by pyrosequencing in formalin-fixed, paraffin-embedded tumour tissues. The hypermethylation cut-off was defined as mean ±2 standard deviation of DNA methylation in healthy breast tissues. Protein expression was evaluated in the same material by ImmunoReactive Score (German IRS system), based on the proportion of positive cells and the staining intensity of the nuclei or cytoplasm. A logistic regression was used to determine the effect of studied variables on presence of CTCs in peripheral blood.Results and discussionsCTC positivity (epithelial or mesenchymal markers) was identified in 23% of patients (16% mesenchymal, 9% epithelial and 2% both). ADAM23 hypermethylation (DNA methylation ≥10%) did not occur in mesenchymal CTC positive patients, while it was present in 28.8% of mesenchymal CTC negative patients (p=0.001). Among studied variables, the multivariate analysis identified low ADAM23 methylation level (p=0.004) and high ki67 proliferation index (p=0.005) as two most significant predictors of mesenchymal CTCs. It was shown recently that although ADAM23 inhibition enhanced invasion in breast cancer, it was not sufficient to trigger metastasis (Costa et al., 2015).ConclusionOur data indicate possible involvement of ADAM23 in hematogenous spread of BC cells. Better understanding of reversible epigenetic regulations hold promise for development of novel therapeutic approaches.This study was supported by the grants APVV-0076–10, APVV-16–0010, ITMS 26240220058 and VEGA grants no. 2/0092/15, 2/0102/17 and 1/0271/17.
The fifth edition of World Health Organization classification of urinary and male genital tumours also brought news regarding testicular tumours. In contrast to the previous editions' radical ...alterations, the adjustments in the fifth edition are subtle and mostly impact the terminology, categorization of some of the rare tumours and diagnostic criteria.
Acquainting with current terms and tumor classification, which is necessary for good clinical practice.
Abstract
Background: Cancer is a risk factor for venous thromboembolism (VTE) and plasma D-dimer (DD) and tissue factor (TF) are established blood markers associated with VTE. CTCs are an independent ...predictor of survival in early and metastatic breast cancer (BC) patients (pts) and CTCs are associated with the risk of VTE in metastatic BC pts. In this study, we hypothesized correlation between plasma DD and TF with presence of CTCs. Moreover, we hypothesized, that activation of urokinase plasminogen activator system (uPA) could be involved in CTCs released into peripheral blood (PB) as well as coagulation activation.
Methods: This prospective study included 119 early BC patients treated by primary surgery followed by systemic therapies when indicated, at the NCI in Slovakia from March to December 2012. Isolated peripheral blood mononuclear cells (PBMC) were depleted of hematopoietic cells (CD45+) using RossetteSepTM negative selection kit. RNA extracted from CD45-depleted PBMC was interrogated for expression of EMT-inducing transcription factors (TWIST1, SNAIL1, SLUG, ZEB1) and epithelial (CK19) gene transcripts by qRT-PCR. Expressions of gene transcripts in CD45- PBMC of pts and healthy donors (HD) were compared. Patient samples with higher epithelial and/or mesenchymal gene transcripts than those of HD (n = 60) were considered as CTC positive. Plasma DD, TF, uPA (urokinase-type plasminogen activator) and PAI-1 (plasminogen activator inhibitor) levels were detected by ELISA, while expressions of TF and uPA system in surgical specimens were evaluated by immunohistochemistry.
Results: CTCs were detected in 27.7% pts. The majority of CTCs exhibited either epithelial differentiation (13.5% pts) or mesenchymal phenotype (EMT markers in 15.1% of pts). A small fraction of CTCs showed co-expression of both markers (0.8%). Pts with any CTCs in PB had significantly higher mean ± SEM plasma DD levels (ng/mL) than those of pts without CTCs (632.4 ± 75.9 vs. 365.4 ± 47.0, p = 0.00002). This association was observed for both, epithelial CTCs (688.4 ± 110.4 vs. 400.7 ± 43.5 (p = 0.026) and mesenchymal CTCs (573.1 ± 105.8 vs. 415.6 ± 44.7, p = 0.0007). There was no association between plasma TF levels or breast tumor TF expression and CTCs. Patients with any CTCs had higher mean ± SEM plasma uPA (ng/mL) (374.6 ± 32.6 vs. 307.1 ± 20.3, p = 0.02) and PAI-1 (pg/mL) levels (5.3 ± 0.7 vs. 4.4 ± 0.4, p = 0.28), than those of patients without CTCs, however, there was no correlation between plasma uPA system activation and DD or TF levels. In multivariate analysis CTCs, patients age and tumor grade were independent factors associated with plasma DD levels.
Conclusion: This prospective study showed for the first time a positive association between plasma D-dimer levels and CTCs indicating a potential role for activation of the coagulation cascade in the early metastatic process. We hypothesize that CTCs could be directly involved in coagulation activation in breast cancer patients, or alternatively an increased CTCs count could be a marker of more aggressive disease and increased risk of VTE. Future studies will need to address the prognostic implications of these observations with the potential for therapeutic interventions for the metastatic process.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-04-02.
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