The type of distribution of PD-L1 positive immune cells and PD-L1 expression in tumor cells correlate with the development of non-classical differentiation in urinary bladder cancer

Background: The basic diagnostic tool of urinary bladder cancer is the histopathological assessment.However, it is insufficient to accurately predict the progression of this disease. There is a need to lookfor new prognostic factors that will make the therapeutic process more effective. The aim of this study isto evaluate the effect of activation of a PD1 – PD-L1 immune checkpoint in immune effector cells (IECs)and tumor cells, on the development of malignancy in the form of non-classic differentiation in urinarybladder cancer.Materials and methods: 110 patients with stage pT1-pT4 urothelial bladder carcinoma who underwent radicalcystectomy/cystoprostatectomy between 2011 and 2014 were included in the study. Tumor advancement(pT stage), grade (G), as well as, non-classic differentiation frequency and number were evaluated pathologically.In each case, the area of the tumor containing PD-L1+ IECs was analyzed. The distribution ofPD-L1+ immune effector cells within the tumor was also assessed as dispersed or aggregated.Results: The frequency of non-classic differentiation was significantly lower in urothelial bladder cancertumors with a dispersed pattern of distribution of PD-L1+ IECs. A correlation between the extent of PD-L1expression in tumor cells and the non-classic differentiation number in UBC was identified.Conclusions: The distribution of cells expressing the immune checkpoint biomarker PD-L1 constitutes anew prognostic factor and may play a key role in the selection of individualized immunotherapy. In addition,the evaluation of non-classic differentiation in the tumor may complement the assessment of PD-L1expression due to its capacity to characterize the current malignant potential of the tumor, whereas theassessment of extent and distribution of PD-L1+ in tumor-associated immune cells indicates the functionalstatus of the immune system

Expression of PD-L1 in tumor and immune system cells affects the survival of patients with urinary bladder cancer

Background: The prediction of tumor malignancy is still one of the most demanding diagnostic tasksin urinary bladder cancer because of its clinicopathological heterogeneity. The aim of this study was toevaluate the expression of PD-L1 in tumor cells (TCs) and immune effector cells (IECs) as well as thepattern of distribution of PD-L1+ IECs within the tumor (dispersed or aggregated) and their associationwith survival of patients with pT1-pT4 urinary bladder cancer.Materials and methods: 110 patients with stage pT1-pT4 urothelial bladder carcinoma who underwentradical cystectomy/cystoprostatectomy between 2011 and 2014 were included in the study. Paraffin blocksmost representative of the tumor were selected for H&E staining as well as immunostaining with the useof rabbit anti-PD-L1 (Ventana clone SP142, Roche). In each sample, the area of the tumor containing PDL1+IECs, as well as, the pattern of distribution (dispersed or aggregated) of PD-L1+ immune effectorcells within the tumor were analyzed. In addition, the expression of PD-L1 in TCs was also assessed.Results: Patients had a shorter survival time in pT2-pT4 cases without TCs expressing PD-L1 (p = 0.007)and/or when PD-L1+ IECs displayed a predominantly dispersed pattern of distribution (p = 0.013).Conclusions: The expression of PD-L1 on TCs and IECs is a prognostic factor which allows for stratificationof patient survival in UBC. The predominance of dispersed or aggregated pattern of distribution ofPD-L1+ IECs in the tumor may be considered as a new prognostic factor in pT1-T4 UBC and indicate thefunctional status of the immune system

The effect of RORa expression on the development of biological malignancy of urinary bladder cancer

Background: Morbidity and mortality relating to urinary bladder cancer have remained largely unchanged for many years. Similarly, the five-year survival rate in this disease has not improved considerably. New developments in individualized therapy necessitate the search for novel factors that could predict the development of malignancy in UBC. In this study, we provide the first evidence that the expression of ROR alpha transcription factor influences the development of malignancy in UBC. Materials and methods: 105 patients with stage pT1-pT4 urothelial bladder carcinoma who underwent cystectomy were included in the study. 4 μm tissue samples were stained immunohistochemically with a polyclonal anti-RORa antibody. The expression of RORa by the tumor cells (TCs) was assessed by counting TCs with a cytoplasmic and/or nuclear staining for RORa per 1000 TCs. The association between the extent of RORa expression and non-classic differentiation, tumor advancement (pT), grade (G) and regional lymph node spread was analyzed. Results: The cytoplasmic expression of RORa was detected in near all analyzed tumor samples (104/105). The extent of RORa expression was significantly higher in tumors which were more malignant with more propensity for non-classic differentiation and lymph node metastasis. We noted a lower percentage of TCs expressing RORa in poorly differentiated tumors (G3), compared to tumors moderately and higher differentiated (G1/G2). Conclusions: Our results suggest that RORa may play a significant role in the progression of urinary bladder cancer. RORa has a broad spectrum of regulatory activity relating to cell and tissue differentiation the mechanism of which is not fully understood. This study represents another step in the process of understanding the mechanisms of RORa regulation and highlights its potential role as a therapeutic target in urothelial bladder cancer

Differential relationship between two hypoxia markers: HIF-1α and GLUT1 and classic prognostic factors in invasive breast carcinoma

Background: Tumor hypoxia is an adverse prognostic factor which promotes cancer aggressiveness and limits its radio- and chemosensitivity. The aim of our study was to explore the relationship between endogenous hypoxia markers and classic prognostic factors, including clinical stage and the expression of ER, PR, and HER2 in primary untreated breast carcinoma. Methods: A retrospective immunohistochemical analysis of archived tissue blocks collected from 153 women, who underwent total mastectomy and lymph node dissection, included the expression of two hypoxia-related proteins: HIF-1α and GLUT1. Results: GLUT1 labelling index (LI) showed a positive correlation with T stage (R = 0.18, p = 0.026) and HER2 status (R = 0.25, p = 0.002), and a negative correlation with the expression of ER (R = −0.19, p = 0.017) and PR (R = −0.17, p = 0.032). HIF-1α LI showed a positive correlation with ER expression (R = 0.16, p = 0.045). In the multivariate regression analysis, a different relationship between classic prognostic factors and the two tested hypoxia proteins was proven. Higher GLUT1 expression correlated with ER and PR negativity (p = 0.02 and p = 0.01, respectively) as well as with higher expression of HER2 (p = 0.04). HIF-1α showed no association with PR and HER2, but a positive correlation with ER (p = 0.02). Neither of the hypoxia proteins was associated with a tumor grade. Only one clinical feature, T stage, correlated with both of the hypoxia markers: positively with GLUT1 (p = 0.049) and negatively with HIF-1α (p = 0.01) expression. Conclusions: In breast cancer, GLUT1 expression may be considered an additional prognostic factor which correlates with an adverse status of HER2 and hormonal receptors, and indicates a more hypoxic, radio- and chemotherapy refractory profile of carcinoma

Impact of specific KRAS Mutation in Exon 2 on clinical outcome of chemotherapy- and radiotherapy-treated colorectal adenocarcinoma patients

BACKGROUND AND OBJECTIVES: Knowledge obtained via high-throughput technologies, used for tumor genome sequencing or identifying gene expression and methylation signatures, is clinically applicable thanks to molecular characterization in the context of tumor development and progression. This study was conducted to assess the impact of specific KRAS mutation in codons 12 and 13 on clinical outcome of chemotherapy and radiotherapy in colorectal cancer patients. METHODS: A total of 239 samples of colorectal adenocarcinoma underwent histological evaluation and DNA isolation. RESULTS AND CONCLUSIONS: Patients with a mutation in KRAS codon 13 experienced worse outcome than those with a mutation in KRAS codon 12. Moreover, the cases of mutations in KRAS codons 12 or 13 were associated with a significantly higher mortality than the cases of wild-type KRAS, and some patients with KRAS mutated in codon 12 had an exceptionally long overall survival. Finally, primary preoperative radiation therapy followed by surgery significantly increased overall survival more efficiently than surgery followed by chemotherapy. This should be investigated in further studies. The fact that all patients treated with radiotherapy + surgery were alive, again focused our attention on the effect of preoperative radiation therapy on the prognosis for colorectal cancer patients. However, the number of patients in this subgroup is too small to allow any specific explanation for this observation. We should, rather, point out a problem for further investigation

The evaluation of metallothionein expression in nasal polyps with respect to immune cell presence and activity

<p>Abstract</p> <p>Background</p> <p>The expression of metallothionein (MT) is involved in acquiring resistance to immune-mediated apoptosis; it is also a negative regulator of the immune response. Nasal polyps are typified by a resistance to immune-mediated apoptosis as well as by excessive immune cell infiltration. RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is a membrane protein capable of inducing the apoptosis of CTLs and NK cells. The aim of the present study has been to explore the expression of metallothionein with respect to immune cell presence and immune cell activity. In our study, we identified immune cells using CD4 and CD68 antigen expression and evaluated their activity using CD25 antigen expression. We then analyzed metallothionein, RCAS1, CD25, CD4, and CD68 in a sampling of 50 nasal polyps using the immunohistochemistry method. We were able to divide the nasal polyps into three main groups according to their predominant immune cell infiltration: eosinophilic nasal polyps (21 cases), lymphocytic nasal polyps (17 cases), and neutrophilic nasal polyps (12 cases).</p> <p>Results</p> <p>In the present study, statistically significant differences between the MT expression in the epithelium and that in the stroma of the nasal polyps along with the accompanying alterations in activation markers on immune cells were found and the number of macrophages in both the eosinophilic and the lymphocytic nasal polyps was assessed. RCAS1-expressing macrophages were found only in the eosinophilic nasal polyps.</p> <p>Conclusion</p> <p>MT expression seems to favor the survival of nasal polyp epithelial cells in the adjacent area of increasingly cytotoxic immune activity. RCAS1-expressing macrophages seem to participate in creating the immune suppressive microenvironment and so help to sustain local inflammation.</p

Changes in Immunogenicity during the Development of Urinary Bladder Cancer: A Preliminary Study

In the present study, we evaluated tumor-infiltrating lymphocytes (TILs) and blood regulatory T lymphocyte (Tregs, CD4+/CD25+/FoxP3+) expression in bladder cancer patients. The number of CD4+, CD8+, CD25+, FoxP3+ and CD20+ TILs was analyzed in association with clinico-pathomorphological features. In more advanced metastasizing tumors, showing non-classic differentiation (ND) and a more aggressive tissue invasion type (TIT), the number of TILs decreased. A low number of CD4+ TILs was associated with poor prognosis. Similarly, Treg frequency before surgery and after surgical treatment was significantly lower in more advanced tumors. The changes in TILs, as well as of local and systemic Tregs, were accompanied by changes in the histological phenotype of urothelial carcinoma regarding pT stage, NDs, TIT, and clinical outcomes. The number of TILs and the frequency of blood Tregs (indicators of antitumor response) may be essential for choosing an immunotherapy that is adjusted to the immune status according to the phase of tumor growth. Moreover, a significant reduction in the number of CD4+ and CD8+ TILs with the development of NDs in more advanced tumors may be associated with lower tumor immunogenicity, resulting in immune tolerance towards tumor tissue. These observations and the tendency of urothelial bladder carcinoma to undergo NDs in a heterogeneous manner during tumor progression suggest complex interactions between bladder cancer immunogenicity and stages of tumor progression