Longer OS Research Articles

P2.05-057 Baseline Inflammatory and Immunological Profile Predict the Survival of NSCLC Patients Undergone Palliative Radiotherapy: Topic: Toxicities

Dose-fractioned cisplatin and oral etoposide (mPE) +/- bevacizumab (mPEBev) is a metronomic treatment showed anti-tumor anti-angiogenic and immunological activity in non-small-cell-lung-cancer (mNSCLC) patients enrolled in BEV2007 trial. These effects could altogether contribute to final antitumor activity. Recent findings suggested that radiotherapy may induce immunological effects on this bases we investigated whether palliative radiotherapy could affect the survival of patients enrolled in BEVA trial. We therefore, carried out a retrospective analysis in the subset of 47 who received palliative radiation therapy after four courses of mPE +/- bevacizumab. All of the patients had received chemotherapy with cisplatin (30mg/sqm, days 1-3q21) and oral etoposide (50mg, days 1-15q21) (mPE) while thirty-five also received bevacizumab at the dosage of 5mg/kg on the day 3q21 (mPEBev regimen). Radiation therapy was delivered with a palliative intent to different target sites including bones (19 patients), brain (Whole brain) (18 patients), lung parenchymal lesions and nodes (7 patients), stereo-tactic ablative radiation therapy (3 patients). Our statistical analysis found that the use of RT was associated to a much longer survival (RT vs no RT: 23.26 vs 16.05 months, P=0.003) months, with no difference in term of PFS 1.65 vs 13.12 P= 0.135). We found no differences with treatment (+/- bevacizumab), sex, grading, stage (IIIB versus IV). Log-rank tests revealed a much longer OS in those patients presenting serum levels of IL17 (p:0.046), c-reactive-protein (p:0.056) and ESR nome per esteso (p:0.014) lower than median value, after Mpe +/- bevacizumab and prior irradiation. We finally observed a longer survival in patients showing a CD4+/CD8+ T cell ratio higher than median value (p:0.050). These results suggest that palliative radiation therapy delivered after our metronomic regimen in mNSCLC is associated to a longer survival with a mechanism presumably driven by immunological effectors. These results represent a solid rationale to test our metronomic regimen and RT in sequential combination with immune-checkpoint inhibitors in mNSCLC patients.

Clinical outcome of neoadjuvant chemotherapy for advanced ovarian cancer

ObjectiveTo evaluate clinical outcome in patients selected to receive neoadjuvant chemotherapy (NACT) compared to primary debulking surgery (PDS). MethodsRetrospective study including all consecutive patients diagnosed and treated for advanced (stages III-IV) ovarian cancers between the years 2003–2015. Results263 women were included in the study, of these, 127 patients were selected to receive NACT and 136 were treated with PDS followed by adjuvant chemotherapy.PDS was associated with longer OS in stage IIIc disease (median OS: 60.2 vs. 48.8months; p-value 0.039) compared with NACT.Patients achieved higher rates of complete cytoreduction in the NACT group compared to the PDS group (65.9% vs. 40.2%; p=0.001). Patients attaining complete cytoreduction after PDS had the best survival, (median OS 106months) followed by those with complete cytoreduction after NACT (median OS 71months), followed by those with residual disease after PDS (median OS 55months). Patients with residual disease following interval debulking after NACT had the worst outcome (median OS 36months).Platinum sensitivity following first line and second line chemotherapy was similar whether patients received neoadjuvant chemotherapy or not. ConclusionPDS was associated with improved outcome. NACT appears to improve survival outcome in patients that would have had residual disease after PDS, and attain complete cytoreduction at the time of interval cytoreduction. This treatment option can be used in selected patients that are not candidates for complete cytoreduction at PDS.

Comparison of immunophenotypes of primary breast carcinomas and multiple corresponding distant metastases: an autopsy study of 25 patients.

Phenotypical change in metastatic breast carcinoma has widely been accepted as an inherent biological feature rather than technical fault. We analyzed the immunohistochemical phenotype and histopathological features of 25 primary breast carcinomas and 90 corresponding distant metastases in 23 organs retrospectively. Histological slides were reviewed for prognostic and predictive factors. Overall, metastases were more similar to each other and often differed from the primary tumor. We created a 3-step grouping system based on the localization of metastases. Regions: tumors metastasizing to the abdominal region were likely to lose ER (p=0.002); we detected loss of PR in metastases to the thorax (p=0.039) and abdomen (p<0.001). Organ systems: loss of ER and PR was observed in metastases to the gastrointestinal system (p=0.026 and p=0.001, respectively), in the respiratory system only the loss of PR was significant (p=0.05). Individual organs: the primaries were likely to lose the hormone receptors in liver metastases (ER p=0.026; PR p=0.004). In lung metastases only loss of PR was apparent (p=0.049). We did not observe significant change in HER2 status, regarding Ki67 change occurred only in bone metastases compared to the primary (p=0.048). 7/25 patients' distant metastases had heterogeneous immunoprofiles. The later the metastasis was discovered the more likely it had a differing IHC profile compared to the primary tumor, patients who had longer OS had a higher chance to develop a discordant metastasis. Immunoprofile of metastases may differ from primary breast cancer and from each other, probably resulting in different response to therapy.

1565P - Impact of a salvage chemotherapy with carboplatin plus docetaxel on testosterone levels in metastatic castration - and docetaxel-resistant prostate cancer (mDRPC)

Recent data suggest that carboplatin plus docetaxel (DC) may be effective in mDRPC. Platinum(II)-complexes interfere with steroid biosynthesis by inhibiting the cholesterol side chain cleavage enzyme (CYP11A1), 3&bgr;-hydroxysteroid dehydrogenase (HSD3B1,2) and 17&agr; hydroxylase/C17,20-lyase (CYP17A1). Docetaxel failure/resistance was defined according to the Prostate Cancer Working Group (PCWG2 2007) criteria. Treatment consisted of at least two cycles of carboplatin AUC5 iv for 30 min on day 1 every 4 weeks, docetaxel at a dose of 35 mg/m2 iv for one hour on days 1, 8, 15 plus prednisone 2x5mg/day orally after receiving informed consent until disease progression or occurrence of intolerable adverse effects. Efficacy measures were done following PCWG2 recommendations. Testosterone/androgene levels were measured before (n = 73) and during carboplatin/docetaxel chemotherapy (n = 63). Of the 96 pts. treated since February 2005, 95.8% had bone, 42.7% lymph node, 26.0% liver and 19.8% lung involvement. At the current analysis, the median follow-up time was 14.6 months, 86 pts. had died and 90 had progressive disease. The objective response rate was 22/59 (37.3%). Response of prostate-specific antigen (≥50%) was observed in 48/96 (50%) patients. Median progression-free survival (PFS) was 7.2 months (CI 95% 6.3, 8.1) and median OS was 15.6 months (CI 95% 11.6, 19.7). The most common reversible grade 3/4 toxicity was leukopenia/ neutropenia (39.6/33.3%). Median free testosterone levels were 0.63 pg/ml before and <0.18 pg/ml during carboplatin/docetaxel treatment (nadir levels, p < 0.001; detection limit <0.18 pg/ml). Median serum androgene levels (T + DHT) were 0.1 ng/ml before and below the detection limit of <0.05 ng/ml during DC treatment. In multivariate analyses, LDH, PSA response, free testosterone nadir levels below the detection limit (<0.18 pg/mL) during DC treatment were associated with longer OS (p < 0.05). These data suggest that carboplatin plus weekly docetaxel may be an important second-line treatment option for DRPC patients by inhibiting the testosterone biosynthesis.

High expression of miR-16 and miR-451 predicating better prognosis in patients with gastric cancer.

To investigate the expression pattern of miR-16 and miR-451 and evaluate their prognostic value in 180 GC patients undergoing surgery. In our previous study, a panel of five circulating miRNAs (miR-16, miR-25, miR-92a, miR-451 and miR-486-5p) can be used as a potential biomarker for detecting of early-stage gastric carcinoma (GC). Tissue microarrays were constructed from 180 patients with GC after surgery. MiR-16 and miR-451 expression was detected by miRNA-locked nucleic acid in situ hybridization, and their relationship with clinicopathological parameters and overall survival was analyzed. MiR-16 expression was decreased in 30.6% (55/180) of GC, increased in 54.4% (98/180) and unchanged in 15.0% (27/180), compared with paracancerous normal tissue (P<0.001). MiR-451 expression was decreased in 17.8% (32/180), increased in 62.8% (113/180) and unchanged in 19.4% (35/180) of GC, compared with paracancerous normal tissue (P<0.001).Univariate analysis indicated that low miR-16 and miR-451 expression, tumor stage, tumor status, node status and tumor size were significant negative prognostic predictors for overall survival in patients with GC (P<0.001, P<0.001, P=0.002, P<0.001 and P=0.001, respectively). Multivariate regression analysis demonstrated that stage [hazard ratio (HR) 1.80; 95% confidence interval (CI) 1.0-3.26; P=0.05], low expression of miR-16 (HR 2.26; 95% CI 1.51-3.40; P<0.001) and miR-451 (HR 2.01; 95% CI 1.36-2.96; P<0.001) predicted shorter OS, while tumor status (HR 1.59; 95% CI 0.73-3.48 P=0.242), lymph node metastasis (HR 1.41; 95% CI 0.71-2.82; P=0.326) and tumor size (HR 1.53; 95% CI 0.92-2.55; P=0.099) were not. Moreover, patients with both miR-16 and miR-451 high expression have better OS than those with two miRNAs unchanged or low expression in GC tissues. Patients with both miR-16 and miR-451 high have better OS than patients with single miR-451 high expression. High expression of miR-16 and miR-451 was associated with longer OS in GC patients. Especially patients with miR-16 and miR-451 double high expression will predict better OS. MiR-16 and miR-451 may be used as novel makers to evaluate prognosis and provide a new treatment target in GC.

D05 - FOLFOXIRI plus bevacizumab (bev) as upfront treatment for metastatic colorectal cancer (mCRC) patients (pts) with initially unresectable liver-limited disease (LLD)

Background: Surgical resection of colorectal liver metastases is a potentially curative option for a subgroup of initially unresectable mCRC pts. Active upfront regimens may increase the percentage of pts converted to resection and positively affect their survival. The aim of the present study was to assess the activity and efficacy of FOLFOXIRI plus bev in mCRC pts with LLD. Patients and Methods: Pts with unresectable LLD treated with FOLFOXIRI plus bev in FOIB, TRIBE and MOMA studies were selected. Baseline clinical and molecular characteristics and outcome parameters were collected. Results: 205 pts with LLD were identified. 184 (90%) pts presented with synchronous disease, 119 (61%) had = 4 metastases, liver metastases were bilobar in 140 (79%) cases, and primary tumor was unresected in 74 (36%). 137 (67%) pts responded, and 124 (60%) achieved early response. 91 (44%) pts underwent resection: R0, R1 and R2 resections were performed in 63 (69%), 11 (12%) and 17 (19%) cases, respectively. Having less than 6 involved segments (p = 0.014), achieving RECIST response (p < 0.001), early response (p = 0.002), and deeper response (p = 0.063) were associated with higher probability to undergo resection in the multivariable model. As compared to unresected pts, those achieving resection had longer PFS (median PFS: 18.1 vs 10.6 mos, HR: 0.47 [0.34-0.65], p < 0.001) and OS (median OS: 44.3 vs 22.5 mos, HR: 0.29 [0.20-0.44], p < 0.001). At a median follow up of 36.5 mos, the estimated 5ys-PFS and 5ys-OS in resected pts were 10% and 38%, respectively. Neither number and size of liver metastases, nor their distribution, nor number of involved segments, nodal status, CEA levels, Fong and Nordlinger score predicted RFS and OS of resected pts. Notably, BRAF mut pts resected after FOLFOXIRI plus bev achieved survival results comparable with RAS/BRAF wt and RAS mut. Radiological (p = 0.047) and pathological response (TRG 1-2 vs 3-4, p = 0.088) was associated with longer OS. Conclusion: FOLFOXIRI plus bev allowed to obtain high resection rate and remarkable survival results in a population of mCRC pts with extensive LLD, not selected for a conversion intent. Radiological and pathological response to FOLFOXIRI plus bev predict survival in resected pts, differently from traditional clinical risk scores and RAS/BRAF status.

Abstract 4159: Characterization of the T-cell receptor (TCR) repertoire in extensive disease small cell lung cancer (ED SCLC)

Abstract Background: The current study explores T-cell (TC) clonality and molecular factors associated with this metric. Tumors employ multiple mechanisms to evade antitumor immune responses. One process involves TC inhibition via upregulation of immune checkpoint (IC) ligands in the tumor microenvironment (TME). Gradual upregulation of inhibitory receptor at their cellular surface results in a decreased capacity to proliferate and activate cytotoxic pathways against tumor cells presenting antigenic peptides.1 In melanoma, this subset of exhausted TCs has been described as highly clonal, where the majority of PD-1-expressing TC population shares the same TCR sequence specific against the same antigenic fragment.2 Previous studies have demonstrated that a clonal TCR repertoire appears to be associated in part with therapeutic responses during IC blockade.3 Methods: We performed TCR sequencing on 82 blood and 73 archival tumor tissue samples collected from ED SCLC patients (pts) in an ongoing longitudinal cohort study in US community oncology practices. Of these, 82 blood and 48 tissue samples had sufficient material available to quantify a clonality metric. To quantify TC abundance as a fraction of total nucleated cells, 82 blood and 58 tissue samples had sufficient material available. Results: Within the subset of 48 tumor samples, a more clonal (ie, less diverse) TCR repertoire was associated with less necrosis (P≤0.012) and lower levels of inflammatory cell infiltration in the local TME (P≤0.021). When pts were divided into 2 equal groups according to the median clonality level, pts with a less clonal TCR repertoire (n = 24/48) who were treated with non-immune-targeted therapy trended toward a longer overall survival (OS; 446 vs 301 days; P≤0.039). In contrast, the percentage of TCs in the TME did not correlate with improved survival (P≤0.412), necrosis (P≤0.131), and inflammation (P≤0.615). This observation differs from results in melanoma describing the impact of IC blockade where pts responding to therapy were associated with a more clonal TME TC population and increased CD8 TCs in the tumor compartment and at the invasive margin.3 In blood, while a less clonal TCR repertoire was associated with a similar but non-significant trend toward longer survival (P≤0.148), pts with increased TC abundance had longer OS (P≤0.025). No association was observed between clonality in TME and clonality (P≤0.571) or TC abundance (P≤0.965) in blood. Conclusion: We hypothesize that a diverse TCR repertoire in the TME and increased peripheral TC abundance are 2 predictors of longer OS in ED SCLC. To further explore factors that may influence TC responses in ED SCLC, the current TCR sequencing results will be integrated with transcriptome and whole genome sequencing analyses.

Abstract 1403: Correlations between overall survival and patient regulatory T-cell levels at initiation of Folfiri therapy in colorectal cancer

Abstract Twenty-three patients with metastatic colon or rectal carcinoma were enrolled in a first-line study of Folfiri therapy consisting of irinotecan (180 mg/m2 day 1), levo-leucovorin (200 mg/m2 day 1), 5-FU (400 mg/m2 bolus day 1 and 2400 mg/m2 continuous infusion over 48h), and bevacizumab (5 mg/kg day 1). This treatment schedule was repeated every 2 weeks. Peripheral blood samples were collected prior to the start of cycle I, and after 30 days, and assayed by 4-color flow cytometry and regulatory T-cell (Treg) suppression assay. Cytofluorometric analysis of PBMCs of the entire group of patients revealed no statistical differences between the PBMCs collected at baseline and post 30 days of therapy in terms of PBMC amount, percent of CD4+ T cells, CD8+ T cells, or Tregs, or the ratios of CD4+ T cells or CD8+ T cells vs. Tregs. Lower levels of Tregs (&amp;lt; 2.5% of PBMC) at baseline, i.e., prior to chemotherapy, were associated with a better PFS by log-rank analysis. The median PFS for patients with &amp;lt; 2.5% Tregs at baseline was 23.3 weeks, and for patients with &amp;gt;2.5% Tregs at baseline PFS was 10.7 weeks (p = 0.037, n = 23). Furthermore, a forward step-wise regression analysis model demonstrated that lower Tregs at baseline (regression coefficient = 0.467, p = 0.004) and the decrease in Tregs (as% of PBMC) at 30 days of chemotherapy (regression coefficient = 0.454, p = 0.006) were both independent predictors of better OS, independent of age, gender, ALP, LDH, mucinous vs. non-mucinous phenotype, serum CEA, serum 19.9, and number of metastatic sites. Of the 23 patients on study, 14 demonstrated clinical responses by RECIST criteria and nine did not. Eight of the 14 (57%) responders by RECIST criteria displayed Treg levels &amp;lt; 2.5% of PBMC at baseline, whereas only two of the seven (22%) non-responders demonstrated &amp;lt; 2.5% Tregs at baseline. In addition, nine of the responders by RECIST criteria (64%) showed a trend in the decrease of Tregs as% of PBMC at 30 days of therapy, whereas only 22% of the non-responders had a decrease in Tregs (p = 0.049, Chi2 = 3.9 with Pearson test). In conclusion, we found lower frequencies of Tregs at baseline in responders than non-responders, which was associated with longer PFS. In addition, lower Tregs at baseline and a decrease in Treg frequency after 1 month of Folfiri therapy were both independent predictors of longer OS. Citation Format: Caroline Jochems, Vittore Cereda, Romaine I. Fernando, Jacob Richards, Italia Grenga, Patrizia Ferroni, Fiorella Guadagni, Jeffrey Schlom, Mario Roselli. Correlations between overall survival and patient regulatory T-cell levels at initiation of Folfiri therapy in colorectal cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1403.

Abstract 2779: A CpG island methylator phenotype in acute myeloid leukemia independent of IDH mutations and associated with a favorable outcome

Abstract Background: Acute myeloid leukemia (AML) causes the most leukemia-related deaths in the United States, and has frequent epigenetic aberrations, including a CpG island methylator phenotype (CIMP). CIMP defines unique molecular subtypes of other cancers and has been linked to mutations in IDH1/2, however the clinical consequences of CIMP and the role of IDH1/2 mutations in AML remain unclear. Methods: To measure genome-wide CpG methylation we used Digital Restriction Enzyme Analysis of Methylation (DREAM) on AML bone marrow samples and normal peripheral blood controls. For validation we used methylation data from patient samples from The Cancer Genome Atlas (TCGA) on the Illumina Infinium HumanMethylation450 platform. We also used RNA-seq data from TCGA, and microarray data from GEO (GSE6891). Statistical analysis was done using R. Results: Genome-wide analysis of variably methylated CpG sites in 96 AML bone marrow samples using DREAM revealed two distinct CpG island methylator phenotypes by hierarchical clustering: IDH-CIMP (I-CIMP) in which 7/10 cases had oncogenic IDH1/2 mutations, and AML-CIMP (A-CIMP), which lacked any mutations in IDH1/2. At median follow-up of 6.16 years, A-CIMP cases, but not I-CIMP cases were associated with longer overall survival (median OS, years: A-CIMP = Not reached, P = 0.08; I-CIMP = 3.35, P = 0.50; CIMP-negative = 1.17). We validated and extended these findings using TCGA data. In this cohort A-CIMP cases also had significantly longer OS compared to CIMP-negative (median OS, years: A-CIMP = 2.34, P = 0.01; I-CIMP = 1.25, P = 0.89; CIMP-negative = 1.00). Aberrant hypermethylation in A-CIMP occurred preferentially at CpG islands by a factor greater than 3, while I-CIMP cases demonstrated a slight preference for hypermethylation at sites outside CpG islands. Interestingly, A-CIMP was enriched in CEBPA (19%) and WT1 mutations (14%), but inversely correlated with IDH, TET2, and NPM1 mutations. Functional pathway analysis revealed that genes hypermethylated in A-CIMP are associated with pluripotency maintenance - including PAX6, GBX2, and HOXA9 - and RNA-seq data largely, but not entirely, recapitulated methylation-based patterns. There was a strong correlation between promoter CpG island methylation and gene expression for many A-CIMP genes. Finally, the transcriptional program associated with A-CIMP was found to correlate with outcome and genetic backgrounds in both TCGA and additional independent datasets. Conclusions: Taken together, our data suggest that CIMP in AML is complex, multifactorial and cannot be explained solely by coding gene mutations (e.g. IDH1/2, TET2). There is an association between A-CIMP and curability in multiple AML datasets that cannot be recapitulated by mutational data alone and that may be worth validating in prospective studies. Citation Format: Andrew D. Kelly, Heike Kroeger, Jumpei Yamazaki, Rodolphe Taby, Frank Neumann, Sijia Yu, Justin T. Lee, Rong He, Shoudan Liang, Yue Lu, Matteo Cesaroni, Sherry A. Pierce, Steven M. Kornblau, Carlos E. Bueso-Ramos, Farhad Ravandi, Hagop M. Kantarjain, Jaroslav Jelinek, Jean-Pierre J. Issa. A CpG island methylator phenotype in acute myeloid leukemia independent of IDH mutations and associated with a favorable outcome. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2779.

Expression of CD30 in Diffuse Large B Cell Lymphoma and Its Clinical Significance

To evaluate the expression and clinical significance of tumor necrosis factor receptor (TNFR) superfamily protein CD30 in diffuse large B cell lymhoma (DLBCL). The CD30 expression, clinical characteristics and prognosis of 63 patients with DLBCL, NOS out of 149 patients with DLBCL admitted in our hospital between January 2008 and December 2012 were analyzed retrospectively. no significant relationship existed between CD30 expression and clinical features, such as age, sex, B symptoms, staging, ECOG PS, LDH level, extranodal site involvement, IPI, GCB or non GCB type, bone marrow involvement. By univariate analysis, the clinical factors associated with general OS and EFS, included CD30, ECOG PS, B symptoms, extranodal site involvement, LDH level, IPI, bone marrow involvement and rituximab. Univariate analysis in GCB DLBCL indicated that CD30 had no significant effect on OS and EFS. However, univariate analysis in non-GCB DLBCL indicated CD30 was associated with longer OS (P=0.037) and showed a tendency of better EFS (P=0.067). In multivariate analysis, IPI and CD30 were independent prognostic factors for OS (IPI: P=0.000, 95%CI 0.042-0.374, CD30: P=0.044, 95%CI 1.055-60.613), and IPI also was independent prognostic factors for EFS (P=0.000, 95%CI 0.040-0.360). CD30+ and DLBCL have a tendency of better EFS (P=0.050, 95%CI 0.996-56.501). CD30 expression level correlates with the prognosis of DLBCL and has a certain clinical value, which may be a new prognostic index of DLBCL.

The impact of brain metastasis on quality of life, resource utilization and survival in patients with non-small-cell lung cancer.

This systematic review aims to improve understanding of the burden of disease associated with brain metastases from non-small-cell lung cancer (NSCLC) in terms of survival, quality of life (QoL) and economic impact. PubMed/MEDLINE, Cochrane collaboration and EMBASE databases were searched for articles published in English from 2000 to 2014. Of 3288 abstracts retrieved, 3156 were eliminated without a full-text review. Of the 132 articles that received a full-text review, a final set of 93 articles was included in an initial literature analysis. In order to homogenize the patient populations evaluated, we included entries that were either entirely composed of NSCLC patients or that had >50% of NSCLC patients in the total study population. From the studies identified in this systematic review, median OS and PFS varied based on the type of treatment received, although whole-brain radiotherapy (WBRT) was associated with the shortest OS and PFS durations. Regimens incorporating targeted therapy in molecularly selected patients were associated with the longest OS and PFS durations. QoL findings varied among studies, generally WBRT resulted in stable or worsening QoL scores rather than improvements. Healthcare costs were increased following diagnosis of brain metastases regardless of treatment. The findings from this review highlight the need for more effective treatments of brain metastases from NSCLC that improve survival function, QoL and potentially decrease costs.

Abstract P4-04-07: Association between tumor infiltrating immune cells and disseminated tumor cells in the bone marrow of patients with primary breast cancer: A ten year follow-up

Abstract Background: Disseminated tumor cells (DTCs) in the bone marrow (BM) of breast cancer patients (pts) were shown to be an independent prognostic factor with regard to reduced PFS and OS. Although this negative prognostic impact has been related to chemotherapy resistant and stem cell-like cells, the microenvironment of the primary tumor might also influence tumor cell spread and thus, outcome of the disease. After a ten year follow-up, we here analyzed the prognostic significance of infiltrating leukocytes in the tumor (TILs) and stroma (SILs) with regard to PFS and OS as well as their association with DTCs in 163 pts with primary breast cancer, diagnosed between 2001 and 2004. Patients and Methods: Tissue microarrays (TMA) were stamped from paraffin embedded specimens and 5µm thick sections were subjected to immunohistochemistry using antibodies against CD3 (DCS, Hamburg, Germany), CD68, CD20, CD8 (all Dakocytomation, Glostrup, Denmark), CD4 (Zytomed Systems, Berlin, Germany) and CD45R0 (Labvision, Fremont, USA), respectively. Each TMA has been evaluated for TILs and SILs assessing intraepithelial and stromal tumor areas occupied by respective immune cells separately: score 0: no staining; score 1: ≤ 10%; score 2: 11-60%; score 3: ≥61%, respectively, using light microscopy at magnifications of x100-200. Statistical analysis was conducted with the R programming language (version 3.20) and Cox proportional hazard regression and the Kruskal-Wallis test. DTCs were studied at the time of primary diagnosis using immunocytochemistry, applying the pan cytokeratin (CK) antibody A45-B/B3. Results: Univariate analyses showed significant associations between prolonged OS and high rates of CD8TIL (p=0.01) and CD4TIL (p=0.028). In the subgroup of triple-negative carcinomas, CD3 TIL and CD8SIL were positively correlated with OS (p=0.038 and p=0.046, respectively), whereas CD8TIL was predictive in HR+/HER2- tumors (p=0.029). In a multivariate model, only CD4TIL was significantly associated with longer OS (p=0.049) in the total group of 163 pts, whereas in the triple-negative subgroup, CD3TIL was significantly correlated with favorable outcome (p=0.044). In contrast, no significant associations were found between TILs/SILs and PFS. The results for DTCs were available for 92/163 pts resulting in a positivity rate of 53%. Intriguingly, despite failing overall associations between DTCs and TILs/SILs, an unfavorable CD4SIL/CD8SIL ratio in the tumors of DTC-positive pts predicted significantly shorter PFS and OS (p=0.009 and p=0.018, respectively). In the DTC-negative subgroup, high CD4SILs were a prognostic marker for shorter OS. Notably, no significant associations were found between CD20, CD45R0 and CD68 and any parameter tested. Conclusion: Analyzing a variety of leukocyte subtypes, we here demonstrate that especially the distribution of CD4 and CD8 in the primary tumor and their ratio seems to be involved in the prognosis of DTC-positive as well as DTC-negative primary breast carcinomas. Since the results for DTCs were only available in a subgroup of pts, further studies will have to prove these findings. Citation Format: Kasimir-Bauer S, Spiekermann A, Friedrich CM, Hoffmann O, Schmid KW, Kimmig R, Bankfalvi A. Association between tumor infiltrating immune cells and disseminated tumor cells in the bone marrow of patients with primary breast cancer: A ten year follow-up. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-04-07.

A phase II study of Chinese patients (pts) treated with nab-paclitaxel (nab-P) plus gemcitabine (Gem) for metastatic pancreatic cancer (MPC).

327 Background: nab-P + Gem demonstrated significantly better overall survival (OS; median: 8.7 vs 6.6 mo; HR 0.72; P &lt; 0.001) than Gem alone as first-line treatment for pts with MPC in the MPACT study. This phase II bridging study evaluated efficacy and safety of nab-P + Gem in Chinese pts with MPC. Methods: Efficacy and safety of nab-P 125 mg/m2 + Gem 1000 mg/m2 on days 1, 8, and 15 every 4 weeks was evaluated in a 3-part sequential study. In part 1, safety was examined. In part 2, efficacy was evaluated by the Simon optimal 2-stage design. If there were &gt; 2 responses in 28 pts in stage 1 of part 2, an additional 54 pts would be treated in stage 2. The study would be completed if &gt; 9 responses were observed. If there was an insufficient number of responses in either stage of part 2, part 3 would be triggered to compare nab-P + Gem vs Gem alone. The primary endpoint was overall response rate (ORR). Secondary endpoints included duration of response (DOR), OS, and safety. Correlation of OS and baseline neutrophil-to-lymphocyte ratio (NLR) was also analyzed. Results: Eighty-three pts were treated. Median age was 57.0 y; 19% were aged &gt; 65 y; baseline Karnofsky performance status was 90 - 100 in 70% of pts and 70 - 80 in 30%. Combining results for stages 1 and 2, ORR was 35% by independent assessment, median DOR was 8.9 mo (95% CI, 6.01 - 8.94), median OS and PFS were 9.2 mo (95% CI, 7.60 - 11.10) and 5.5 mo (95% CI, 5.29 - 7.16), respectively (Table). Baseline NLR ≤ 5 was associated with a nonsignificant trend toward longer OS vs NLR &gt; 5 (median, 10.0 vs 8.3 mo; HR 0.62; P= 0.148). The most common grade ≥ 3 adverse events included neutropenia (37%), leukopenia (31%), and fatigue (14%). Grade ≥ 3 peripheral neuropathy occurred in 7% of pts. Part 3 was not triggered per study design. Conclusions: nab-P + Gem demonstrated efficacy in Chinese pts with MPC. The OS and ORR were numerically better than those of the phase III MPACT trial (Table). A trend toward longer OS in pts with baseline NLR ≤ 5 vs &gt; 5 confirmed previous findings from the MPACT study. No new safety signals were identified. Clinical trial information: NCT02135822. [Table: see text]

Molecular characterization of colorectal tumors in young patients compared with older patients and impact on outcome.

505 Background: The incidence of colorectal cancer (CRC) among young individuals is rising. The effect of an individual’s age on their tumor molecular profile is unknown. Methods: Molecular profiles of 4,821 tumors obtained from younger and older CRC patients (pts) were reviewed and correlated with pt outcome. Protein expression (IHC), gene amplification (ISH), sequencing (NGS and Sanger), and fragment analysis were performed. T and Fisher’s exact tests determined differences between age groups; Kaplan-Meier methodology estimated survival. Results: Tumors from 1,277 younger (median age 40; range 15-45 yr) and 3,544 older (72; 65-98) pts were studied. Most frequently mutated genes included TP53, APC, KRAS, PIK3CA, SMAD4, and BRCA1/2. Mutation rates for BRAF (14.4% vs. 4.8%, p &lt; 0.001), APC (62% vs. 54%, p = 0.0034), FBXW7 (9.6% vs. 5.5%, p = 0.01), and KRAS (45% vs. 41%, p = 0.02) were higher in older pts; NRAS mutation rates (4.5% vs. 3.9%) were similar in both groups. Younger pts had higher overexpression rates of HER-2/neu (3.2% vs. 1.8%, p = 0.017) and MGMT (64% vs. 58%, p = 0.001). Microsatellite instability (MSI), determined by IHC (MLH1, MSH2, MLH6 and PMS2) and fragment analysis, was similar between cohorts (10.3% vs. 8.1%), but somatic MSI high (determined by concurrent BRAF mutation) was higher in older pts (6% vs. 0%, p &lt; 0.0001). There was no difference in TS (37% vs. 34%), ERCC1 (25% vs. 26%) or TOPO1 (49% vs. 46%) expression between age groups. Older pts showed a trend toward higher PD-L1 expression (2.9% vs. 0.7%, p = 0.0512) but there was no difference in the frequency of PD-1 expression on tumor-infiltrating lymphocytes (39% vs. 43%). Outcome was evaluable for 82 pts (younger, 47; older, 35). Most pts received FOLFOX + Bev as first line Rx. Median OS was worse in younger pts (p = 0.03). Low ERCC1 expression was associated with prolonged survival in older (p &lt; 0.01) but not in younger (p = 0.3) pts. There was no association between TOPO1 expression and OS (p = 0.8). Pts with low TS expression showed a trend toward longer OS (p = 0.08). Conclusions: Young pts with CRC may carry genetic alterations that are distinct from older pts. A better understanding of disease biology may help to identify therapeutic targets for younger pts.

Cytotoxic Chemotherapy as First-Line Therapy for Advanced Non-Small-Cell Lung Cancer in Taiwan: Daily Practice.

Aim: Cytotoxic chemotherapy is the standard first-line therapy for patients with advanced non-small cell lung cancer (NSCLC) without specific gene alterations. This study examined the prescription pattern and the survival outcome of cytotoxic chemotherapy regimens in daily practice in Taiwan.Methods:We established a population-based cohort of patients diagnosed with advanced NSCLC between 2005 and 2009 using the databases of Taiwan Cancer Registry and National Health Insurance in Taiwan. We then analyzed chemotherapy prescriptions and the survival outcomes of patients.Results:A total of 25,008 patients with advanced NSCLC were identified, 17,443 (70.0%) of which received first-line chemotherapy and were therefore included in this study. Among them, 11,551 (66.2%) patients had adenocarcinoma and 3,292 (18.9%) patients had squamous cell carcinoma (SCC). Approximately 70% of the patients were diagnosed with NSCLC in medical centers. Platinum-based doublet chemotherapy was administered to 66.9% of the patients. Among all chemotherapy regimens, platinum with gemcitabine (33.8%) was the most common, irrespective of geographic region. The second and third most common regimens were vinorelbine alone (13.0%) and platinum with docetaxel (11.6%). The prevalence of platinum-based doublet chemotherapy regimens decreased from 71.4% in 2005 to 64.1% in 2009. Among patients with adenocarcinoma histology, those who received platinum with pemetrexed had longer OS than did patients who received other platinum-based regimens (p < 0.001).Conclusion: Our findings reaffirm that in real-world practice, treatment plans of advanced NSCLC should be drawn up according to histology type.

High-grade glioma in children and adolescents: a single-center experience.

The aim of this study was to report the outcome in children with high-grade astrocytoma outside the brain stem and spinal cord that were treated at a single center. The study included 26 patients with anaplastic astrocytoma and 37 patients with glioblastoma; all patients were aged ≤18 years. At initial diagnosis, 18 of the patients with glioblastoma received only temozolomide (TMZ), 14 received other chemotherapies, and 5 did not receive any chemotherapy. Among the patients with anaplastic astrocytoma, 9 received TMZ, 9 received other chemotherapy regimens, and 8 patients did not receive any chemotherapy. The median radiotherapy dose in all patients was 60 Gy. Median age of the patients was 12.5 years. Median overall survival was 20 months and mean progression-free survival was 4.7-11.3 months (median: 8 months) in all patients. Patients with a Karnofsky performance score (KPS) ≥70 had median overall survival of 32 months, versus 7 months in those with a KPS < 70. Patients aged <15 years had median survival of 38 months, versus 16 months in those aged 15-18 years. Patients with anaplastic astrocytoma that received TMZ, other chemotherapy regimens, and no chemotherapy had median survival of 21 months, 132 months, and 11 months, respectively. Patients with glioblastoma that received TMZ, other chemotherapy regimens, and no chemotherapy had median survival of 32 months, 12 months, and 8 months, respectively. In the present study, patients with anaplastic astrocytoma treated with chemotherapy protocols other than TMZ had the longest OS; however, in the glioblastoma group, OS was 32 months in those treated with standard TMZ and 12 months in those treated with other protocols (P = 0.493). Although TMZ is less toxic than PCV, it was not shown to be superior.

FOXO1 Activation Is an Effector of SYK and AKT Inhibition in Tonic BCR Signal-Dependent Diffuse Large B-Cell Lymphomas

Introduction: In normal B lymphocytes, B-cell receptor (BCR)-induced activation of PI3K-AKT kinases and subsequent inactivation of FOXO1 is a critical pro-survival component of tonic BCR signaling. In murine models, conditional deletion of FOXO1 protected quiescent peripheral B cells from apoptosis mediated by inducible loss of the BCR, demonstrating that PI3K-AKT-FOXO1 axis plays a central role in B-cell homeostasis. Disruption of the BCR signaling by SYK inhibitor leads also to the apoptosis of BCR-dependent DLBCLs, at least in part via a mechanism involving decreased activity of PI3K/AKT axis. Herein, we investigated the role of FOXO1 in toxicity of BCR pathway/SYK inhibition in human BCR-dependent lymphomas.Methods: BCR-dependent DLBCL cell lines (DHL4, DHL6, Ly1, Ly7) were incubated with SYK inhibitor R406 (4 µM) and AKT phosphorylation, FOXO1 activation and transcriptional activity were assessed by phospho-specific flow cytometry, Western Blot and qPCR. Toxicity of active FOXO1 was assessed by transducing DLBCL cells with a constitutively nuclear FOXO1 mutant. FOXO1 knock-down in DLBCL cell was achieved with shRNA. DREAM cleavage and HRK expression were assessed by Western Blot and qPCR, respectively, in the absence or presence of caspase inhibitors. DLBCL cell viability and apoptosis after incubation with SYK and/or AKT inhibitors R406 and MK2206, were assessed by MTS assay and Annexin V/PI staining. Drug interactions were quantitated by CompuSyn software. The expression of p-SYK and FOXO1 in DLBCL samples was assessed by immunohistochemistry (IHC) in 60 DLBCL patients.Results: Since FOXO1 is a major effector of tonic BCR signaling, we assessed the activity of FOXO1 in DLBCL cells after SYK inhibition. In all tested cell lines, AKT and FOXO1 phosphorylations decreased after incubation with SYK inhibitor. Diminished FOXO1 phosphorylation resulted in its nuclear relocalization and induction of FOXO1-dependent expression of p27, BIM, TRAIL and GADD45A. To assess whether the increased activity of FOXO1 is sufficient to induce apoptosis of DLBCL cells, we transduced DHL4 cells with a constitutively nuclear and transcriptionally active FOXO1-3A mutant. The mutant induced G1/S cell cycle arrest and triggered apoptosis, whereas wild-type FOXO1 did not change proliferation or cellular viability, demonstrating that FOXO1 activation is sufficient to induce apoptosis of DLBCL cells. Next, we assessed the toxicity of SYK inhibitor in cells lacking FOXO1. Cells with depleted FOXO1 exhibited 70% lower sensitivity to SYK inhibitor than control cells (p<0.0001). Since in previous studies expression of the proapoptotic member of BCL2 family, HRK, was required for SYK-inhibitor induced cell death in DLBCL cells, we determined the role of FOXO1 activation in HRK expression. HRK expression was dramatically increased in SYK-inhibitor treated control cells, but not in FOXO1-deficient cells. Consistent with this, SYK inhibitor triggered cleavage of HRK transcriptional repressor DREAM only in control cells, but not in FOXO1-depleted cells. HRK induction was blocked by caspase inhibitors. Since AKT is major kinase regulating both FOXO1 activity and HRK induction, we assessed the combined effects of the AKT inhibitor MK-2206 with R406 and found markedly synergistic toxicity (combination index [CI] 0.5-0.8). Combination of inhibitors in FOXO1-depleted cells did not trigger cell death, highlighting the critical effector role of FOXO1. FOXO1 expression was present in 80% of primary DLBCL samples and correlated with SYK activity (p=0.009). High levels of FOXO1 protein expression were associated with longer OS (log rank, p=0.04).Conclusions: Taken together, our results demonstrate that targeting the BCR signaling at multiple levels is a rational therapeutic strategy. Proapoptotic activity of FOXO1 is an important effector of SYK and AKT inhibition in DLBCLs and its expression is required for SYK-and AKT inhibitor-induced toxicity. The underlying mechanism linking FOXO1 activation and cell death involves caspase-dependent cleavage of transcriptional repressor DREAM and subsequent induction of a proapoptotic BCL2 family member, HRK. DisclosuresNo relevant conflicts of interest to declare.

Lung cancer and other second neoplasms after treatment of Hodgkin lymphoma.

To evaluate the risk factors associated with lung cancer (LC) and other second neoplasms (SN) in Hodgkin lymphoma (HL) survivors. We retrospectively analyzed the clinical characteristics and outcomes of 604 patients treated in our institution between 1968 and 2012. 90 out of 604 patients developed SN: 27 LC and 63 other SN. The median time elapsed until LC and other SN was 16.5 and 11.8 years, respectively (p = 0.003). In the LC group, 85.5 % of patients were male and 84.6 % smokers (HR 7, 95 % CI 2.4-20.7, p < 0.001). Radiotherapy (RT) doses applied were higher in the SN group with an increased risk of LC (HR: 4.0 95 % CI 1.1-11.6, p = 0.010) and other SN (HR: 3.3 95 % CI 1.6-6.7 p = 0.001) with doses higher than 42 Gy. No association was found between alkylating agents and development of SN. In LC, the most frequent histology was adenocarcinoma with an elapsed time after HL of 13.2 years in early stages and 21.3 in advanced (p = 0.02). Median OS after a diagnosis of LC was 12.6 months ranging from 5.9 (in cases presenting due to symptoms) to 49.1 (incidentally diagnosed cases) (p = 0.005). RT treatment, especially with doses higher than 42 Gy, and smoking increase the risk of SN after HL. In this series, LC patients with early stages had a shorter elapsed time from HL diagnosis and longer OS, therefore the role of LC screening in HL survivors should be prospectively evaluated and smoking cessation counseling ought to be a key aspect during follow-up.

Therapeutic effect and prognostic factors for patients with mucoepidermoid carcinoma of parotid gland

Objective To study the therapeutic effect of adjuvant chemotherapy and radiotherapy after operation in patients with mucoepidermoid carcinoma of parotid gland, and to screen the indicators ralated to the prognosis of tumor. Methods Eighty patients with mucoepidermoid carcinoma of parotid gland in First People′ Hospital of Yibin of Sichuan Province from January 2005 to December 2009 were analysed retrospectively in our research. We studied the survival of patients who were treated wtih simple operation(30 cases) or postoperative adjuvant therapy(50 cases). Then we further analyzed the relationships between the prognosis of the patients and some variables (age, gender, smoking, alcohol drinking, lymph node metastasis, distant organ metastasis, treatment method, differentiation degree and T grading). Results Kaplan-Meier survival curves showed that patients with postoperative adjuvant therapy had longer PFS and OS than those without adjuvant therapy (94.4 months vs 69.3 months; 114.9 months vs 96.7 months), with statistical significance (χ2=11.246, P=0.001; χ2=15.803, P=0.001). COX univariate analysis showed that gender (χ2=22.346, P=0.000), smoking (χ2=7.891, P=0.041), lymph node metastasis (χ2=12.371, P=0.005), distant organ metastasis (χ2=9.813, P=0.002), treatment method (χ2=25.261, P=0.000), differentiation degree (χ2=4.361, P=0.006) and T grading (χ2=5.336, P=0.014) were related to the PFS of patients. COX multivariate analysis showed that lymph node metastasis (χ2=11.003, RR=2.827, 95%CI: 1.965-3.851, P=0.011), distant organ metastasis (χ2=7.611, RR=0.472, 95%CI: 0.240-0.775, P=0.016), treatment method (χ2=24.542, RR=5.390, 95%CI: 3.585-9.602, P=0.000), degree of differentiation (χ2=3.221, RR=2.118, 95%CI: 1.845-4.719, P=0.009) and T grading (χ2=4.336, RR=0.804, 95%CI: 0.681-0.916, P=0.024) were related to the PFS of patients. COX univariate analysis showed that smoking (χ2=4.551, P=0.008), alcohol drinking (χ2=11.742, P=0.048), lymph node metastasis (χ2=14.886, P=0.009), distant organ metastasis (χ2=6.713, P=0.005), treatment method (χ2=22.411, P=0.000), degree of differentiation (χ2=8.116, P=0.012) and T grading (χ2=14.443, P=0.035) were related to the OS of patients. COX multivariate analysis showed that lymph node metastasis (χ2=11.711, RR=2.985, 95%CI: 1.521-3.999, P=0.005), distant organ metastasis (χ2=5.390, RR=0.400, 95%CI: 0.201-0.793, P=0.009), treatment method (χ2=19.327, RR=5.086, 95%CI: 3.241-8.006, P=0.000), degree of differentiation (χ2=7.084, RR=2.301, 95%CI: 1.908-4.503, P=0.001) and T grading (χ2=13.229, RR=0.561, 95%CI: 0.348-0.867, P=0.040) were related to the OS of patients. Conclusion Adjuvant radiation and chemotherapy can obviously prolong the PFS and OS for the patients with mucoepidermoid carcinoma of parotid gland. Lymph node metastasis, distant organ metastasis, treatment method, differentiation degree and T grading can greatly influence the prognosis of patients with mucoepidermoid carcinoma of parotid gland, which can be used as independent prognostic indicators for the patients with mucoepidermoid carcinoma of parotid gland. Key words: Mucoepidermoid tumor; Treatment outcome; Prognosis

Use of proton therapy for re-irradiation in pediatric intracranial ependymoma

Background and purposeTo report disease control, survival and treatment-associated toxicity with the use of proton therapy (PRT) for re-irradiation of intracranial ependymoma. Materials and methodsTwenty patients underwent 33 PRT re-irradiation courses for recurrent or metastatic lesions between June 2004 and February 2015 at Massachusetts General Hospital. ResultsThe majority of patients were female (60%), with infratentorial tumors (90%), anaplastic histology (55%), and initially received 55.8 GyRBE (52.2–59.4) involved field (IF) PRT. First failure was local (55%), distant (30%) or both (15%) at a median time of 23.9months (9.9–98.5) from first treatment. Salvage therapy included re-resection (75%), chemotherapy (60%) and IFPRT (70%) to a median dose 50.4 GyRBE (35–55.8) in the majority of patients. The median follow-up was 37.8months (5.5–138.0). Three year OS and PFS are 78.6% (95% CI 67.6–89.6) and 28.1% (95% CI 15.6–40.6), respectively. Longer OS was significantly associated with surgical resection of recurrent disease (HR 9.19, 95% CI 1.27–66.44, p=0.028). The pattern of second failure after re-irradiation was directly related to the pattern of first failure (p<0.01). Three of 14 patients (21.4%) locally re-treated experienced grade 2 radiation-associated treatment change. ConclusionsProton therapy appears safe and efficacious for the re-treatment of recurrent intracranial ependymoma.