Clinical characteristics and management of a Greek female patient cohort with breast conserving treatment.

PURPOSE: The purpose of this retrospective single-center study was to examine histopathological characteristics and treatment options in a cohort of Greek female patients treated with breast conserving surgery (BCS) and to evaluate potential predictive factors of breast cancer (BC) local recurrence. METHODS: The clinic's medical records from 1995 up to the end of July 2016 were scanned in order to identify female patients treated with BCS. We recognized 1175 patients who underwent BCS, representing 35.8% of the entire sample (3281 patients). RESULTS: The mean age of the patients enrolled in this study was 54.7 years, with a median follow-up period of 58.5 months. Nine deaths (0.8%) were registered with 5-year overall survival (OS) rate being 100%. Regarding adjuvant therapies, radiotherapy (RT) was assigned to 94.4% of the patients, endocrine therapy to 86.0%, chemotherapy to 51.3%, while all therapies were given simultaneously to 38.3% of the sample patients. Herceptin was administered to 14.1% of the patients. Twenty-eight recurrences (2.4%) with 3 deaths (10.7%) were confirmed. Over all traditional parameters studied, only lymph node status appeared to be statistically correlated with local recurrence (p=0.005). T3 stage can be considered as a hint that this tumor size might be a risk factor of local recurrence. Age at diagnosis seems to be an independent factor of BC local recurrence. CONCLUSIONS: A very low rate of local recurrence (2.4%) was validated which can be attributed to patients being appropriately selected for BCS, who were then enrolled in this study, and to the uniformity of the RT plan.

Impact of Oncotype DX on chemotherapy assignment: a retrospective single-center study on female breast cancer patients.

PURPOSE: This study was designed to determine the Recurrence Score (RS) distribution and its associated risk assessments based on traditional clinicopathologic characteristics in a single-center breast cancer (BC) deriving cohort in Greece, and to evaluate the impact of the RS results on adjuvant treatment decisions applied in this cohort. METHODS: This was a retrospective, single-center study regarding Greek female patients with early-stage breast cancer (ESBC). From 2009 to 2015, 114 cases lacking unanimity in the multidisciplinary breast meeting (MDM) fulfilled the inclusion criteria. The RS of the Oncotype DX (ODX) assay was the main outcome. RESULTS: The mean RS in the sample was 16.38 (SD=6.87). RS was positively correlated with Ki-67 (p=0.008). A negative progesterone receptor (PR) was associated with a higher RS (p<0.05). RS was higher for cases of chemotherapy assignment (p<0.001). According to the oncologists' pre-DX assay recommendations, 62.8% of the patients would have been 'wrongly' assigned to chemotherapy, while 14.3% of patients would have not been recommended this treatment even though they should have. The overall chemotherapy recommendation was significantly altered after the ODX RS assay was carried out (p=0.008) and, in the sample, it diminished by 39.5%. CONCLUSIONS: The distribution of the ODX RS in the specific cohort of Greek women is similar to that reported in other geographic regions of the world. Knowledge of the RS resulted in a shift in treatment recommendations towards lower-intensity regimens and in a greatly reduced proportion of chemotherapy recommendations.

Clinical characteristics and management of a Greek female patient cohort with breast ductal carcinoma in situ.

PURPOSE: The purpose of this retrospective single-centre study was to examine the histopathological characteristics of breast ductal carcinoma in situ (DCIS) lesions in a cohort of Greek female patients and describe our experience regarding the clinical management of the disease. METHODS: The medical records from 1995 up to mid-2014 were scanned in order to trace DCIS cases. One hundred and seventy two patients (6.8% of all breast cancer cases) were diagnosed with pure DCIS and no invasive components; 32.0% underwent a second surgery, mainly due to first surgery positive margins. RESULTS: Age at first surgery ranged from 27 to 79 years (mean±SD 50±11) and median tumor size was 10mm (interquartile range/IQR7equals;12mm). Comedo necrosis (CN) was identified in 28.5% of the cases. The detection of CN was significantly associated with older age at diagnosis, larger tumor size and lower probability of highly differentiated tumors. Radiotherapy (RT) and hormonotherapy (HT) were applied to 44.8% and 63.4% of the patients, respectively. CONCLUSIONS: We implemented international practices (surgery, radiotherapy and prophylactic hormonal therapy) to patients diagnosed with DCIS and have observed only two relapses. It is our belief that DCIS requires a multidisciplinary approach and patient-tailored therapy which can potentially contribute to minimization of the local recurrence risk.

Interleukin-13 displaying retargeted oncolytic measles virus strains have significant activity against gliomas with improved specificity.

The majority of glioblastoma multiforme (GBM) tumors (80%) overexpress interleukin-13 receptor alpha2 (IL-13Ralpha2), but there is no expression of IL-13Ralpha2 in normal brain. Vaccine strains of measles virus have significant antitumor activity against gliomas. We tested the hypothesis that measles virus entry could be retargeted via the IL-13Ralpha2. MV-GFP-H(AA)-IL-13 was generated from the Edmonston-NSe vaccine strain, by displaying human IL-13 at the C-terminus of the H protein, and introducing CD46 and signaling lymphocyte activation molecule (SLAM)-ablating mutations in H. The IL-13 retargeted virus showed significant cytopathic effect (CPE) against IL-13Ralpha2 overexpressing glioma lines, and lack of CPE/viral replication in normal human astrocytes and normal human fibroblasts not expressing IL-13Ralpha2. In vivo treatment of orthotopically implanted GBM12 xenografts demonstrated significant prolongation of survival in mice treated with the retargeted strain (P < 0.0001), and comparable activity between the IL-13R retargeted strain and MV-GFP (P = 0.6377). In contrast to MV-GFP-treated mice, administration of the retargeted strain in the central nervous system of measles replication-permissive Ifnar(ko) CD46 Ge mice resulted in lack of neurotoxicity. Strains of measles virus retargeted against the glioma-specific IL-13Ralpha2 receptor have comparable therapeutic efficacy, and improved specificity as compared with the unmodified measles virus strain MV-GFP in vitro and in vivo.

Oncolytic measles virus strains in the treatment of gliomas.

BACKGROUND: Recurrent gliomas have a dismal outcome despite use of multimodality treatment including surgery, radiation therapy and chemotherapy. OBJECTIVE: In this article the authors discuss potential applications of oncolytic measles virus strains as novel antitumor agents in the treatment of gliomas. METHODS: Important aspects of measles virus development as an anticancer therapeutic agent including engineering, retargeting and combination studies with other therapeutic modalities are discussed. The translational process that led to the first clinical trial of an engineered measles virus derivative in patients with recurrent glioblastoma multiforme is also described. RESULTS/CONCLUSIONS: Oncolytic measles virus strains hold promise as novel antitumor agents in the treatment of gliomas.

Interleukin-13 Displaying Retargeted Oncolytic Measles Virus Strains Have Significant Activity Against Gliomas With Improved Specificity.

The majority of glioblastoma multiforme (GBM) tumors (80%) overexpress interleukin-13 receptor α2 (IL-13Rα2), but there is no expression of IL-13Rα2 in normal brain. Vaccine strains of measles virus have significant antitumor activity against gliomas. We tested the hypothesis that measles virus entry could be retargeted via the IL-13Rα2. MV-GFP-HAA-IL-13 was generated from the Edmonston-NSe vaccine strain, by displaying human IL-13 at the C-terminus of the H protein, and introducing CD46 and signaling lymphocyte activation molecule (SLAM)-ablating mutations in H. The IL-13 retargeted virus showed significant cytopathic effect (CPE) against IL-13Rα2 overexpressing glioma lines, and lack of CPE/viral replication in normal human astrocytes and normal human fibroblasts not expressing IL-13Rα2. In vivo treatment of orthotopically implanted GBM12 xenografts demonstrated significant prolongation of survival in mice treated with the retargeted strain (P < 0.0001), and comparable activity between the IL-13R retargeted strain and MV-GFP (P = 0.6377). In contrast to MV-GFP-treated mice, administration of the retargeted strain in the central nervous system of measles replication-permissive Ifnarko CD46 Ge mice resulted in lack of neurotoxicity. Strains of measles virus retargeted against the glioma-specific IL-13Rα2 receptor have comparable therapeutic efficacy, and improved specificity as compared with the unmodified measles virus strain MV-GFP in vitro and in vivo.

Combination of measles virus virotherapy and radiation therapy has synergistic activity in the treatment of glioblastoma multiforme.

PURPOSE: Glioblastoma multiforme is the most frequent primary brain tumor in adults and represents one of the most lethal malignancies with a median survival of 12-16 months. We have previously shown that an oncolytic measles virus derivative expressing soluble human carcinoembryonic antigen (MV-CEA) has significant antitumor activity against glioblastoma multiforme cell lines and xenografts. Radiation therapy (RT) represents one of the mainstays of glioma treatment. Here we tested the hypothesis that the combination of RT with MV-CEA would have synergistic activity against gliomas. EXPERIMENTAL DESIGN: 3-(4,5-Dimethyl-thiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and clonogenic assays were used to test cytoxicity of the combination treatment in vivo. To examine the mechanism of synergy, one-step viral growth curves, terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays, and Western blot analyses were performed. In vivo assessment of synergistic antitumor activity was conducted in a U87 glioma model. RESULTS: MTS and clonogenic assays showed a strong synergistic interaction between MV-CEA and RT in glioblastoma multiforme cells including both primary and established glioma lines. Furthermore, significant antitumor efficacy was observed in vivo in a subcuteneous U87 xenograph model. There was significant prolongation of survival (P = 0.001) in the combination treatment group as compared with single modality- or control-treated animals. One-step viral growth curves showed increased viral burst size by up to 2 log in MV/RT combination-treated cells, as compared with single agent MV-CEA-treated glioma cells. Changes in CEA levels and expression of viral N and H protein were also consistent with increased viral production. Furthermore, TUNEL assays and Western blot analysis showed increase in apoptosis in MV/RT combination-treated cells. The pan-caspase inhibitor Z-VAD-FMK and the caspase-8 inhibitor Z-IETD-FMK, but not the caspase-9 inhibitor Z-IEHD-FMK, protected glioma cells from MV-CEA/RT-induced cleavage of poly(ADP-ribose) polymerase (PARP), indicating that the apoptotic death in combination-treated cells is mostly mediated via the extrinsic caspase pathway. The Fas/Fas ligand interaction blocking antibody NOK-1 blocked MV/RT-induced PARP cleavage whereas the Fas agonistic antibody CH11 increased PARP cleavage in MV/RT combination-treated cells. Reverse transcription-PCR, fluorescence-activated cell sorting analysis and immunohistochemistry showed up-regulation of Fas in combination-treated tumor in vitro and in vivo cells. CONCLUSIONS: There is synergy between MV-CEA and RT in vitro and in vivo. The synergistic effect of the combination seems to be due to increase in viral burst size and increase in apoptotic cell death. This latter effect is mostly mediated via the extrinsic caspase-8 pathway, activated via increased signaling through the Fas death receptor pathway. These results could have translational implications in glioma therapy.

Epidermal growth factor receptor (EGFR)-retargeted measles virus strains effectively target EGFR- or EGFRvIII expressing gliomas.

A retargeted measles virus strain MV-GFP-H(AA)-scEGFR was generated by engineering the MV-NSe Edmonston vaccine strain to incorporate both CD46 (Y481A) and signaling lymphocyte activation molecule (SLAM) (R533A) ablating mutations in the hemagglutinin protein in combination with the display of a single-chain antibody against epidermal growth factor receptor (EGFR) at the C terminus of hemagglutinin. The unmodified MV-GFP virus was used as a positive control. Specificity of the EGFR retargeted virus was demonstrated in non-permissive Chinese hamster ovary (CHO) cells stably transfected to express either the natural receptors CD46 or SLAM or the target receptors EGFR and EGFRvIII. In vitro, the retargeted virus had potent antitumor activity against EGFR- or EGFRvIII-overexpressing primary glioblastoma multi-forme (GBM) cell lines that was comparable to the activity of the unmodified MV-GFP virus. Intratumoral administration of MV-GFP-H(AA)-scEGFRvIII in orthotopic GBM12 xenografts resulted in tumor regression, as demonstrated by bioluminescence imaging and significant prolongation of survival, that was comparable to the effect of the unmodified strain. In contrast to MV-GFP, central nervous system administration of the targeted MV-GFP-H(AA)-scEGFR virus in measles replication-permissive Ifnar(ko) CD46 transgenic mice resulted in no neurotoxicity. In conclusion, EGFR-retargeted measles virus strains have comparable therapeutic efficacy to the unmodified virus in glioma cells overexpressing EGFR or EGFRvIII in vivo and in vitro, and improved therapeutic index, a finding with potential translational implications in glioma virotherapy.