Prognostic significance of PD-L1 protein expression and copy number gains in locally advanced cervical cancer.

BACKGROUND: Although immune checkpoint inhibitors against programmed death-1 (PD-1) and its ligand (PD-L1) have demonstrated promising results in several solid malignancies, including cervical cancer, there are some limitations to using PD-L1 immunohistochemical expression as a predictive biomarker for selecting patients who may benefit from such therapy. OBJECTIVE: To examine the protein expression and genetic status of PD-L1 with clinical outcomes in locally advanced cer- vical cancer. METHODS: We investigated the PD-L1 gene copy number gains assessed by fluorescence in situ hybridization (FISH) and PD-L1 expression using immunohistochemistry in 123 patients with locally advanced cervical cancers between December 2008 and December 2016. RESULTS: The prevalence of PD-L1 immunohistochemical expression was detected in 103/123(83%) cases. PD-L1 gene am- plification and polysomy were detected in 7% and 40% of cases, respectively. PD-L1 gene amplification and polysomy were associated with positive PD-L1 immunostaining (score 1+ to 3+) in 88% and 68% of cases, respectively. Clinically, PD-L1 immunopositivity was associated with parametrial invasion at diagnosis. In contrast, PD-L1 polysomy was associated with parametrial invasion and FIGO stages III-IV, whereas PD-L1 amplification was associated with nodal metastasis. In multi- variate analysis, PD-L1 amplification was predictive of worse RFS (HR, 5.68; 95%CI, 1.98-16.28; p = 0.001), whereas PD-L1 polysomy was predictive of worse LRR (HR, 4.13; 95%CI, 1.63-10.49; p = 0.003). PD-L1 immunohistochemical expression was not associated with worse outcomes in Cox models. CONCLUSIONS: Our results showed that an increase in PD-L1 gene copy number could be a novel prognostic and possible predictive biomarker for anti-PD-1/PD-L1 therapy in locally advanced cervical cancer.

Anaplastic lymphoma kinase (ALK) translocation in paediatric malignant peritoneal mesothelioma: a case report of novel ALK-related tumour spectrum.

AIMS: To report a case of paediatric malignant peritoneal mesothelioma (MPM) with evidence of anaplastic lymphoma kinase (ALK) translocation. METHODS AND RESULTS: We describe a 10-year-old girl who presented with abdominal pain and progressive abdominal distension. She had no history of asbestos exposure. Histopathological, immunohistochemical and ultrastructural analyses were performed and showed a biphasic malignant mesothelioma. In addition, we also studied on a selected set of immunomarkers which may be the potential therapeutic molecular targets including ALK, c-kit (CD117), epidermal growth factor receptor (EGFR) and human epidermal growth factor 2 (HER2)/neu, as well as corresponding molecular analysis. Consequently, we identified ALK expression by immunohistochemistry, together with evidence of ALK translocation by fluorescent in-situ hybridization (FISH) analysis. CONCLUSIONS: Paediatric MPM is associated with ALK translocation in our case. The results may open up a new avenue for the study of molecular genesis of paediatric malignant mesothelioma in the future and help to determine whether patients MMs with ALK translocation would benefit from ALK inhibitor treatment.

Huge peritoneal malignant mesothelioma mimicking primary ovarian carcinoma.

Peritoneal malignant mesothelioma (PMM) is less commonly found in female than male. The most important differential diagnosis of PMM in female patient is primary ovarian carcinoma because of their similar symptoms e.g. dyspepsia, abdominal discomfort from ascites, palpable abdominal mass, etc. However common clinical presentation of PMM is diffuse spread of peritoneal lesions without dominating tumor mass while primary ovarian tumor usually presents with large pelvic mass and smaller exta-ovarian metastatic lesions. The surgeon may make a provisional intraoperative diagnosis of PMM if both ovaries are clearly identified Unfortunately, both conditions frequently elicit fibrosis and adhesion that the exact location or the origin of tumor cannot be clearly stated. Histopathologic diagnosis of PMM is also difficult because it has three patterns of histopathology as biphasic tumors composed of epithelial and sarcomatous components or it may be monophasic of either type. When only the epithelial component is found, serous ovarian carcinoma is the important differential diagnosis while the biphasic mesothelioma must be differentiated from malignant mesodermal mixed tumor or carcinosarcoma of the ovary. The pathologist generally requires immunohistochemical study to achieve a correct diagnosis. The clinical feature and detailed histopathologic findings of the patient with PMM will be discussed

PD-L1 protein expression and copy number gains in HIV-positive locally advanced cervical cancer.

BACKGROUND: The programmed death-1/programmed death-ligand-1 (PD-1/PD-L1) axis may represent a target for cervical cancer; however, it is poorly understood in human immunodeficiency virus (HIV)-infected patients. METHODS: We evaluated HIV-positive (n = 42) and HIV-negative (n = 110) women with locally advanced cervical cancer regarding their PD-L1 expression, determined by combined positive score (CPS) ⩾ 1 and tumor proportion score (TPS) ⩾ 25%, and PD-L1 copy number alterations, assessed by fluorescence in situ hybridization. RESULTS: Regardless of HIV status, 84.9% and 44.8% of cases were PD-L1-positive according to CPS ⩾ 1 and TPS ⩾ 25%. Per CPS ⩾ 1, PD-L1 positive rate was similar between HIV-positive and HIV-negative women, whereas a significant difference was seen per TPS ⩾ 25%. Tumor size and parametrial invasion were correlated with PD-L1 positivity in HIV-negative women, whereas anti-retroviral therapy (ART) was correlated with TPS < 25%. Low CD4-positive cell counts were associated with CPS < 1 in HIV-positive women. No significant difference was observed in PD-L1 copy number status between HIV-positive and HIV-negative women. PD-L1 amplification and polysomy were independently associated with TPS ⩾ 25%, whereas the presence of parametrial invasion was independently associated with CPS ⩾ 1. Cancer stage and PD-L1 amplification were identified as independent predictors of recurrence-free survival [hazard ratio (HR) = 2.40 (1.32-4.36) and HR = 5.33 (1.94-14.61)] and cancer-specific survival [HR = 13.62 (5.1-36.38) and HR = 3.53 (1.43-8.69)]. PD-L1 polysomy was an independent predictor of locoregional recurrence-free survival [HR = 3.27 (1.27-8.41)]. HIV status and PD-L1 expression (CPS ⩾ 1 or TPS ⩾ 25%) were not associated with poor patient outcomes. CONCLUSION: PD-L1 amplification and polysomy are the strongest drivers of PD-L1 expression in cervical cancer, and could represent prognostic biomarkers for anti-PD-1/PD-L1 therapy. Cervical cancer biology may be modulated by HIV infection, CD4-positive cells, and HIV treatments.

Phosphohistone H3 (PHH3) as a surrogate of mitotic figure count for grading in meningiomas: a comparison of PHH3 (S10) versus PHH3 (S28) antibodies.

Mitotic figure (MF) counting is important in the evaluation of meningioma grading. Nevertheless, mitosis assessment on hematoxylin and eosin (H&E)-stained slides may be problematic because of technical factors and pathologist's experience. Phosphohistone H3 (PHH3) is a mitosis-specific antibody that has proven to facilitate mitotic count in various tumors. However, the antibody performance between PHH3 serine10 (S10) and serine28 (S28) has never been compared in these tumors before. In this study, 48 cases of meningioma (28 grade I, 14 grade II, 6 grade III) were evaluated using immunohistochemical stains for four commercially available PHH3 (S10) and S28 antibodies to identify MFs and validate PHH3 intra- and interobserver reproducibility and agreement. Two pathologists counted MFs on both H&E- and PHH3-stained slides. H&E and PHH3 MFs were highly correlated (Spearman's rho = 0.96 for PHH3 (S10)-Biocare, 0.96 for PHH3 (S10)-CST, 0.91 for PHH3 (S28)-Abcam, and 0.89 for PHH3 (S28)-Santa Cruz. The mean difference between an H&E and PHH3 mitotic count is 0.81 for PHH3 (S10)-Biocare, 0.95 for PHH3 (S10)-CST, - 0.97 for PHH3 (S28)-Abcam, and - 0.97 for PHH3 (S28)-Santa Cruz. For comparison among four PHH3 antibodies, PHH3 mitotic counts had both a good intra- and interobserver reproducibility (p > 0.05). Regarding to World Health Organization (WHO) grade, there was not a significant discrepancy in the stratification of tumor grades for all four PHH3 antibodies in terms of interobserver agreement. The Cohen's kappa coefficient (K) was 0.93 for PHH3 (S10)-Biocare, 0.82 for PHH3 (S10)-CST, 0.76 for PHH3 (S28)-Abcam, and 0.80 for PHH3 (S28)-Santa Cruz. Considering survival analyses, all five proliferation indices were univariately associated with recurrences. Increased PHH3 mitotic indices (MIs) were significantly associated with recurrence-free survival in univariate Cox proportional hazards regression analysis (p < 0.001) and remained an independent predictor in multivariate analysis (p < 0.05). The appropriate prognostic cutoff values for recurrence prediction were 5 or more per 10 high-power fields (HPFs) for PHH3 (S10) and 3 or more per 10 HPFs for PHH3 (S28).

Impact of antiretroviral drugs on PD-L1 expression and copy number gains with clinical outcomes in HIV-positive and -negative locally advanced cervical cancers.

Cervical cancer has become a leading cause of death in both HIV-infected and uninfected women. Previous studies have revealed that antiretroviral therapy (ART) possesses anti-human papillomavirus (HPV) and antitumour properties, potentially serving as an anticancer agent and improving functional immunity in HIV-positive individuals. However, to the best of our knowledge, no studies have examined the association between ART and the clinical outcome of patients with pre-existing invasive cervical cancer. The current study analysed 48 HIV-positive and 123 HIV-negative patients with locally advanced stage IB2-IVA cervical cancer between December 2008 and December 2016. Tumours were categorized based on programmed cell death-ligand 1 (PD-L1) immunoreactivity and copy number alterations in the PD-L1 gene, as determined by fluorescence in situ hybridization. The results revealed that ART-treated patients exhibited a lower prevalence of PD-L1 immunopositivity, PD-L1 amplification and polysomy compared with patients that did not receive ART and those that were HIV-negative. Furthermore, ART-treated patients with PD-L1 immunonegativity exhibited an improved recurrence-free survival (RFS) compared with patients that did not receive ART and HIV-negative individuals with PD-L1 immunopositivity (P=0.041 vs. P=0.030). Additionally, ART-exposed patients with PD-L1 disomy demonstrated improved locoregional recurrence-free survival (LRR) when compared with HIV-negative patients with PD-L1 amplification and polysomy (P=0.039 vs. P=0.007), RFS (P<0.001 vs. P=0.006) and cancer-specific survival (CSS) (P=0.021 vs. P=0.025). ART-exposed patients with PD-L1 disomy also exhibited improved RFS (P<0.001) and CSS (P<0.001) compared with HIV-negative patients with PD-L1 amplification. Improved LRRs were demonstrated in ART-exposed patients with PD-L1 disomy (P=0.028) compared with non-HIV patients with polysomy. Following multivariate analysis, International Federation of Gynaecology and Obstetrics stage and PD-L1 amplification were determined to be predictors of poor a RFS [hazard ratio (HR), 2.43; 95% confidence interval (CI), 1.37-4.30; P=0.002 vs. HR, 7.03; 95% CI, 2.79-17.74; P<0.001) and CSS (HR, 11.47; 95% CI, 4.70-27.99; P<0.001 vs. HR, 4.05; 95% CI, 1.64-9.98; P=0.002). However, only PD-L1 polysomy was determined to be a predictor of poor LRR (HR, 2.50; 95% CI, 1.11-5.63; P=0.027). HIV status was not associated with poor outcomes, as determined using Cox models. The results of the current study indicated that ART may be used for the treatment of cervical cancer in both HIV-infected and uninfected patients. However, additional research is required to further elucidate these results.

Expression of the p16 and Ki67 in Cervical Squamous Intraepithelial Lesions and Cancer.

PURPOSE: To evaluate the expression of p16 and Ki67 in cervical intraepithelial neoplasia (CIN) and cancer. MATERIALS AND METHODS: We performed a immunohistochemical study of p16 and Ki67 in 243 cervical tissues 53 nondysplastic lesions, 106 CIN1, 61 CIN2/3 and 23 squamous cell carcinomas. The expression of p16 and Ki67 was interpreted independently by 2 researchers and the sensitivity and specificity to detect clinically significant lesions (≥ CIN2) were determined. RESULTS: The overall agreement results of positive or negative immunostaining of intrainter observer variability were 0.659 for p16 and 0.808 for Ki67. p16 expression was demonstrated in 91.3% of invasive carcinomas, 78.7% of CIN2/3, 10.4% of CIN1 and 9.4% of nondysplasic lesions. The corresponding Ki67 expression was: 100% of all invasive carcinomas, 75.4% of CIN2/3, 22.6% of CIN1, and 11.3% with nondysplasia. The expression was significantly different between CIN2/3 vs CIN1 for both p16 and Ki67 (pvalues <0.001 both), and cancer vs CIN2/3 for Ki67 (pvalue 0.008). The differences were not significant between CIN1 vs nondysplasia (pvalues 1.000 for p16 and 0.130 of Ki67), and cancer vs CIN2/3 for p16 (p value 0.219). The sensitivity and specificity to detect > CIN2 were 84.5% and 90.5% by p16 and 82.1% and 88.6% by Ki67. CONCLUSIONS: The rates for 16 and Ki67 expression were directly associated with the severity of cervical lesions. Significant differences in these markers expression may be useful in cases with equivocal histologic features among cervical intraepithelial lesions, but not between CIN1 and nondysplastic lesions. The two markers had high sensitivity and specificity in determining >CIN2.