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Objective Around 7–10% of endometrial carcinomas are characterized by polymerase-e-(POLE) exonuclease-domain-mutations, an ultra-mutated-phenotype and a favorable prognosis. It is currently unknown whether POLE ultra-mutated-tumors are more immunogenic when compared to the other groups of endometrial cancers. Methods We used autologous-dendritic-cells (DC) pulsed with whole-tumor-extracts to assess the level of CD8+ and CD4+ T-cell-activation induced by POLE-ultramutated (+) and POLE wild-type (−) endometrial cancer cells in vitro. T-lymphocyte-proliferations were evaluated using CFSE and/or [3H] thymidine-incorporation-assays while the ability to specifically kill autologous-tumor-cells by cytotoxic-T-lymphocyte (CTL) was tested in standard 4-h- 51 Cr-cytotoxicity-assays. In order to correlate cytotoxic activity and proliferation by CD4+ and CD8+ T-lymphocytes, respectively, with a particular lymphoid subset, two-color-flow-cytometric analysis of intracellular-cytokine-expression (IFN-γ vs IL-4) at the single cell level was also performed. Results DC-pulsed with tumor extracts were able to induce CTL-responses against autologous-tumor-cells in both POLE (+) and POLE (−) cancer patients (P=0.305). These CD8+ T-cell-populations were cytotoxic against tumor-cells but they did not lyse PHA-stimulated-autologous-lymphocytes or autologous-EBV-transformed-lymphoblastoid-control-cell-lines. In contrast, only POLE (+) tumor-lysate-pulsed-DC were able to induce significant proliferation and high IFN-γ expression (i.e., Th1-cytokine-bias) in autologous in vitro DC-stimulated CD4+ T-cells as well as naive CD4+ and CD8+ T-cells from patients-peripheral-blood (P Conclusions POLE ultra-mutated-tumors are significantly more immunogenic when compared to POLE (−) tumors, in particular to the helper arm of the immune system. These data lend support to the hypothesis that the better prognosis of patients with POLE (+) tumors may at least in part be linked to their enhanced immunogenicity.

Objectives: We evaluated the role of PIK3CA-mutations as mechanism of resistance to trastuzumab in primary HER2/neu-amplified uterine-serous-carcinoma (USC) cell lines. Methods: Fifteen whole-exome-sequenced USC cell lines were tested for HER2/neu-amplification and PIK3CA-mutations. Four HER2/neu-amplified USC (2-harbouring wild-type-PIK3CA-genes and 2-harbouring oncogenic-PIK3CA-mutations) were evaluated in in vitro dose-titration-proliferation-assays, cell-viability and HER2 and S6-protein-phosphorylation after exposure to trastuzumab. USC harbouring wild-type-PIK3CA were transfected with plasmids encoding oncogenic PIK3CA-mutations (i.e., H1047R/R93Q) and exposed to trastuzumab. Finally, trastuzumab efficacy was tested by using two USC xenograft mouse models. Results: Seven out of fifteen (46%) of the USC cell lines were HER2/neu-amplified by fluorescence in situ hybridisation. Within these tumours four out of seven (57%) were found to harbour oncogenic PIK3CA-mutations vs two out of eight (25%) of the HER2/neu not amplified cell lines (P=0.01). HER2/neu-amplified/PIK3CA-mutated USC were highly resistant to trastuzumab when compared with HER2/neu-amplified/wild-type-PIK3CA cell lines (P=0.02). HER2/neu-amplified/PIK3CA wild-type cell lines transfected with oncogenic PIK3CA-mutations increased their resistance to trastuzumab (P

(OS) and progression-free survival (PFS) were estimated using the Kaplan–Meier method. The associations of smoking and of smoking + treatment (i.e., bevacizumab vs. no bevacizumab) with PFS and OS were evaluated with the Cox proportional hazards model. Results: Smoking questionnaires were available for analysis in 99% of study participants. Two hundred eighty-six women (64%) were active or former smokers. In the entire study population, this group had significantly worse OS than never smokers (14.2 vs. 16.8 months, HR 1.29, 95% CI 1.03–1.61, P= 0.026). Smokers who received chemotherapy plus bevacizumab (n= 148, 52%) experienced significantly longer PFS (median 7.9 vs. 5.7 months, log-rank P b 0.01) and significantly longer OS (median 16.7 vs. 12.0 months, log rank P= 0.027) than smokers who received chemotherapy alone (n= 138, 48%). Overall response rate among smokers was 49.3% (chemotherapy plus bevacizumab) vs. 29% (chemotherapy alone) (P= 0.002). The median number of cycles of therapy was 6 for smokers and nonsmokers (interquartile range, 4–10, smokers vs. 2–9, nonsmokers) (P=ns). Smokers receiving bevacizumab experienced the following adverse events: grade 2+ fistula (16.2%), grade 3+ venous thromboembolism (9.5%), grade 2+hypertension (27%), and grade 2+proteinuria (2.7%). Cervical cancer was the reported cause of death in 74.5% of smokers. Conclusions: Smoking is common and associated with a 29% increased risk of death among women with advanced or recurrent cervical cancer. The survival benefits conferred by antiangiogenesis therapy are sustained, even among smokers in this population. Smokers with CxCA should be informed of these results and efforts at smoking cessation strongly encouraged.