Independent Prognostic Value For Overall Survival Research Articles

Prognostic significance of high NPC2 expression in gastric cancer

Gastric cancer is one of the most common malignancies worldwide, and the third leading cause of cancer-related death. The identification of novel biomarkers and therapeutic targets is critical to improve the prognosis. A total of 380 patients with primary gastric cancer from the TCGA database were analyzed. The receiver operating characteristic curves were plotted. We further evaluated the independent prognostic ability of NPC2 expression for overall survival (OS) and relapse-free survival (RFS) through the Kaplan–Meier curve and Cox analysis. The NPC2 expression was significantly higher (P < 0.001) in gastric cancer. High NPC2 expression was significantly (P < 0.0001) associated with poor OS and poor RFS. The age, stage, radiation therapy, residual tumor, and NPC2 expression showed independent prognostic value for OS. The gender and NPC2 expression showed independent prognostic value for RFS. The higher NPC2 expression was observed in gastric cancer, compared with adjacent normal tissue (P < 0.001), confirmed by the IHC staining. The CCK-8 assay showed that NPC2 knockdown inhibits cell proliferation while NPC2 overexpression promotes cell proliferation (P < 0.05). NPC2 expression may serve as a promising prognostic biomarker for patients with gastric cancer.

The development and validation of a m6A-lncRNAs based prognostic model for overall survival in lung squamous cell carcinoma.

No biomarkers have been identified for the prognosis of lung squamous cell carcinoma (LUSC). Risk models based on m6A-lncRNAs help to predict survival in some cancers. However, very few studies have reported m6A-lncRNA risk models in LUSC. We aimed to construct a prognostic model based on m6A-lncRNAs in LUSC. The clinical and RNA-sequencing information of 504 LUSC patients were downloaded from The Cancer Genome Atlas (TCGA) database. Prognostic m6A-lncRNAs were identified by a Pearson correlation analysis and univariate Cox regression analysis. The ConsensusClusterPlus algorithm was used to cluster the prognostic m6A-lncRNAs. The overall survival (OS) and clinicopathological characteristics of the 2 clusters were compared. A gene set enrichment analysis (GSEA) analysis was performed to analyze the genes enriched in the 2 clusters. A least absolute shrinkage and selection operator (LASSO) Cox regression analysis was used to construct the risk-score model. Two hundred and forty eight patients were randomly chosen from TCGA-LUSC cohort for the training set. The receiver operating characteristic (ROC) curve analysis was used to assess the predictive ability of the model. The clinical characteristics and OS in the high- and low-risk groups were compared. The independent prognostic value of the model was tested by Cox regression analyses. Thirteen m6A-lncRNAs were identified as prognostic lncRNAs and classified into cluster A and cluster B. The OS of patients in cluster A was better than that of patients in cluster B (P<0.001). Patients in cluster B had higher expressions of immune checkpoints. Immune score, stromal score, and ESTIMATE score were higher in cluster B (P<0.001). Seven of the 13 lncRNAs were used to construct the risk-score model. Patients in the high-risk group had a worse OS. ROC curves showed a under the curve (AUC) of 0.639 in the training set and 0.624 in the validation set. A high risk was associated with cluster B, a high immune score, and stage III-IV disease. Patients in the high-risk group had increased expressions of immune checkpoints. The Cox regression analyses showed that the risk-score model had independent prognostic value for OS. The risk-score model retained its prognostic value in different subgroups. The m6A-lncRNA risk-score model is an independent prognostic factor for OS in LUSC patients. However, the risk-score model need to be further tested clinically.

Real-life long-term outcomes of upfront surgery in patients with resectable stage I-IIIA non-small cell lung cancer.

BackgroundTreatment of early-stage non-small cell lung cancer (NSCLC) is rapidly evolving. When introducing novelties, real-life data on effectiveness of currently used treatment strategies are needed. The present study evaluated outcomes of stage I–IIIA NSCLC patients treated with upfront radical surgery in everyday clinical practice, between 2010–2017.Patients and methodsData of 539 consecutive patients were retrieved from a prospective hospital-based registry. All diagnostic, treatment and follow-up procedures were performed at the same thoracic oncology centre according to the valid guidelines. The primary outcome was overall survival (OS) analysed by clinical(c) and pathological(p) TNM (tumour, node, metastases) stage. The impact of clinicopathological characteristics on OS was evaluated using univariable (UVA) and multivariable regression analysis (MVA).ResultsWith a median follow-up of 53.9 months, median OS and 5-year OS rate in the overall population were 90.4 months and 64.4%. Five-year OS rates by pTNM stage I, II and IIIA were 70.2%, 60.21%, and 49.9%, respectively. Both cTNM and pTNM stages were associated with OS; but only pTNM retained its independent prognostic value (p = 0.003) in MVA. Agreement between cTNM and pTNM was 69.0%. Next to pTNM, age (p = 0.001) and gender (p = 0.004) retained their independent prognostic value for OS.ConclusionsThe study showed favourable outcomes of resectable stage I–IIIA NSCLC treated with upfront surgery in real-life. Relatively low agreement between cTNM and pTNM stages and independent prognostic value of only pTNM, observed in real-life data, suggest that surgery remains the most accurate provider of the anatomical stage of disease and important upfront therapy.

Benefits of neoadjuvant therapy compared with adjuvant chemotherapy for the survival of patients with HER2-positive breast cancer: A retrospective cohort study at FUSCC

PurposeNeoadjuvant therapy (NAT) is considered the standard of care for patients with HER2-positive breast cancer (BC). However, there is no proven survival benefit of NAT compared to adjuvant therapy for the survival of patients with early-stage HER2-positive BC. This study aimed to compare the prognosis of HER2-positive BC patients treated with NAT to that of patients treated with adjuvant therapy. MethodsThis was a single-center real-world retrospective study. This study analyzed the disease-free survival (DFS) and overall survival (OS) of 538 HER2-positive BC patients treated with neoadjuvant therapy and 2684 patients treated with adjuvant therapy at Fudan University Shanghai Cancer Center (FUSCC) between 2012 and 2016. Patients with a clinical tumor size (cT) ≤5 cm or >5 cm were matched using the propensity score matching (PSM) method to prevent selection bias. ResultsAfter PSM, among patients with cT ≤ 5 cm, there was no significant difference in DFS (P = 0.08) or OS (P = 0.11) between the two groups. The analysis of survival outcomes of patients treated with neoadjuvant and adjuvant therapy in the different chemotherapy subgroups yielded consistent results. According to multivariate analysis, lymph node status and response to NAT showed independent prognostic value for OS and DFS. Among patients with cT > 5 cm, the DFS (P = 0.25) and OS (P = 0.57) of patients treated with NAT were similar to those of patients treated with adjuvant therapy after PSM. ConclusionWe confirmed the equivalent effects of adjuvant therapy and NAT in HER2-positive BC patients. Neoadjuvant therapy should be used for patients with HER2-positive BC.

Downregulated mRNA Expression of ZNF385B Is an Independent Predictor of Breast Cancer.

Background ZNF385B, a zinc finger protein, has been known as a potential biomarker in some neurological and hematological studies recently. Although numerous studies have demonstrated the potential function of zinc finger proteins in tumor progression, the effects of ZNF385B in breast cancer (BC) are less studied. Methods The Oncomine database and “ESurv” tool were used to explore the differential expression of ZNF385B in pan-cancer. Furthermore, data of patients with BC were downloaded from The Cancer Genome Atlas (TCGA). The receiver operating characteristic (ROC) curve of ZNF385B expression was established to explore the diagnostic value of ZNF385B and to obtain the cut-off value of high or low ZNF385B expression in BC. The chi-square test as well as Fisher exact test was used for identification of the relationships between clinical features and ZNF385B expression. Furthermore, the effects of ZNF385B on BC patients' survival were evaluated by the Kaplan-Meier and Cox regression. Data from the Gene Expression Omnibus (GEO) database were employed to validate the results of TCGA. Protein expression of ZNF385B in BC patient specimens was detected by immunohistochemistry (IHC) staining. Results ZNF385B expression was downregulated in most types of cancer including BC. Low ZNF385B expression was related with survival status, overall survival (OS), and recurrence of BC. ZNF385B had modest diagnostic value, which is indicated by the area under the ROC curve (AUC = 0.671). Patients with lower ZNF385B expression had shorter OS and RFS (relapse-free survival). It had been demonstrated that low ZNF385B expression represented independent prognostic value for OS and RFS by multivariate survival analysis. The similar results were verified by datasets from the GEO database as well. The protein expression of ZNF385B was decreased in patients' samples compared with adjacent tissues by IHC. Conclusions Low ZNF385B expression was an independent predictor for worse prognosis of BC patients.

Prognostic Value of the Albumin-to-γ-glutamyltransferase Ratio for Gallbladder Cancer Patients and Establishing a Nomogram for Overall Survival.

Purpose: The albumin-to-γ-glutamyltransferase ratio (AGR), a novel inflammation-related index, has been reported to have prognostic importance in several malignancies but not yet in gallbladder cancer (GBC). This study intended to assess the prognostic value of AGR in GBC and to develop a nomogram based on AGR for predicting overall survival (OS) in GBC patients after surgery.Methods: Medical records of 140 qualified GBC patients between July 2003 and June 2017 were retrospectively analyzed. The function “surv_cutpoint” in the R package “survminer” was implemented to discover the optimal cut-off value of AGR. A nomogram on the fundamental of Cox model was established in the training cohort and was internally validated using calibration curves, Harrell's concordance index, time-dependent AUC plots and decisive curve analyses.Results: The optimal AGR cut-off value concerning overall survival was 2.050. Univariate and multivariate analyses demonstrated that AGR (HR=0.354, P=0.004), T stage (HR=3.114, P=0.004), R0 resection (HR=0.448, P=0.003), BMI (HR=0.470, P=0.002) and CA19-9 (HR=1.704, P=0.048) were independent predictors for OS. The nomogram combining these prognostic factors showed considerable prognostic performance in term of consistency, discrimination and net benefit.Conclusion: AGR has independent prognostic value for OS in GBC patients receiving surgery. A nomogram incorporating AGR, T stage, R0 resection, CA19-9 and BMI achieved enhanced prognostic ability.

Impact of Aspirin on clinical outcome in advanced HCC patients receiving sorafenib and regorafenib

Background and aimThe aim of our retrospective study is to evaluate the prognostic significance of aspirin in patients with advanced HCC treated with sorafenib. Methods304 patients with HCC,consecutively treated with sorafenib from May 2007 to September 2018, were included in the clinical study. Of Them 93 patients token aspirin. Progression-free survival (PFS)and overall survival (OS)were estimated with the Kaplan–Meier method and compared with the log-rank test. ResultsThe concomitant use of sorafenib and aspirin was associated with a median OS of 18.3 months compared to 8.8 months of patients who did not receive aspirin (HR 0.57; P < 0.0001). The concomitant use of sorafenib and aspirin was associated with a median PFS of 7.3 months compared to 3.0 months of patients who did not receive aspirin (HR 0.61; P = 0.0003). In the multivariate analysis, the use of aspirin maintained an independent prognostic value for OS(HR 0.61; P = 0.0013). In second line the concomitant use of regorafenib and aspirin was associated with a median OS of 16.9 months compared to 8.0 months of patients who did not receive aspirin (HR 0.30; P = 0.02). ConclusionGlobally, our data seem to suggest that aspirin use may improve the clinical outcome of patients with advanced hepatocellular carcinoma receiving sorafenib and regorafenib.

Combination of CD47 and signal-regulatory protein-α constituting the "don't eat me signal" is a prognostic factor in diffuse large B-cell lymphoma.

The interaction between CD47 and signal‐regulatory protein‐α (SIRPα) inhibits phagocytosis, thus affecting the clinical outcomes of neoplastic diseases. Although CD47 upregulation is associated with poor prognosis in several malignancies, the effect of SIRPα expression and its coexpression with CD47 remains unclear. This study aimed to investigate the clinicopathologic effect of CD47 and SIRPα expression in diffuse large B‐cell lymphoma (DLBCL). Immunostaining of 120 biopsy samples showed that CD47 is primarily expressed in tumor cells, whereas SIRPα is expressed in nonneoplastic stromal cells, mostly macrophages. CD47high cases showed higher MYC protein expression and lower MYC translocation. The SIRPαhigh cases presented significantly shorter overall survival (OS) and progression‐free survival (PFS) than SIRPαlow cases in the activated B‐cell (ABC) subtype of DLBCL (P = .04 and P = .02, respectively). Both CD47high and SIRPαhigh presented significantly shorter OS and PFS than other cases among all DLBCL patients (P = .01 and P = .004, respectively), and the ABC type (P = .04 and P = .008, respectively) but not the germinal center B‐cell type. Both CD47high and SIRPαhigh yielded a constant independent prognostic value for OS and PFS in multivariate analysis (hazard ratio [HR], 2.93; 95% confidence interval [CI], 1.20‐7.43; P = .02; and HR, 2.87; 95% CI, 1.42‐5.85; P = .003, respectively). To the best of our knowledge, this is the first study to report that combinatorial CD47 and SIRPα expression is a potential independent prognostic factor for DLBCL. Evaluation of CD47 and SIRPα expression could be useful before CD47 blockade therapy.

LTBK-12. EORTC 26951, RANDOMIZED STUDY OF ADJUVANT PCV AFTER 59.4 GY RADIOTHERAPY: VERY LONG TERM FOLLOW-UP

Abstract BACKGROUND Between 1995 and 2002 the EORTC Brain Tumor Group conducted a prospective phase III study on adjuvant procarbazine, CCNU and vincristine (PCV) chemotherapy in anaplastic oligodendroglioma (AOD). A mature follow-up presented in 2012 showed survival benefit of the addition of PCV, in particular in 1p/19q co-deleted tumors and tumors with MGMT promoter methylation. We now present very long term follow-up. MATERIALS AND METHODS Patients were eligible if locally diagnosed with a newly diagnosed AOD. They were randomized between radiotherapy (RT, 33 x 1.8 Gy) and the same RT followed by 6 cycles PCV (RT/PCV). Primary endpoints were overall survival (OS) and progression free survival (PFS). 1p/19q status (FISH) was determined in 300 patient. Kaplan- Meier technique and Cox modeling were used for long term survival analysis. Primary analyses were adjusted for known prognostic factors. For other analyses no adjustment was performed. RESULTS With 368 patients included, a median follow-up of 18.4 years and 307 (83%) survival events, median and 20-year survival after RT/PCV versus RT alone were 42.3 mo and 16.8% vs 30.6 months and 10.1% (HR 0.78; 95% CI (0.63, 0.98), adjusted p=0.06). Eighty patients were 1p/19q codel of which 26 (33%) were still alive, in this subgroup median and 20-year survival after RT/PCV versus RT alone were 14 years and 37.1% versus 9.3 years and 13.6% (HR 0.60, 95% CI (0.35, 1.03), unadjusted p=0.06). Twenty year PFS in 1p/19q codel was 31.3% in RT/PCV treated patients and 10.8% in RT only treated patients (HR 0.49, 95% CI (0.29, 0.83), unadjusted p=0.007). In the 1p/19q codel subgroup age, WHO PS and necrosis at pathology were identified to be of independent prognostic value for OS. CONCLUSION This long term analysis confirms the earlier conclusions and provides data on long term survival in this patient group. In 1p/19q codel patients treated with RT/PCV, the 20-year PFS and OS rates are 31% and 37% respectively.

Elevated pre-existing lymphocytic infiltrates in tumour stroma predict poor prognosis in resectable urothelial carcinoma of the bladder.

Lymphocytic infiltrates are predominantly distributed in the tumour stroma, and represents the tumour-related immune response. The aim of this study was to elucidate the prognostic value of stromal lymphocytic infiltrates (SLI) in resectable urothelial carcinoma of the bladder (UCB). The prognostic significance of SLI in UCB was assessed in a discovery cohort (n=226; 60 deaths) and in a validation cohort (n=417; 103 deaths). SLI was categorised into intense (≥50% SLI) and non-intense (<50% SLI). A multivariable Cox model was used to analyse the associations of SLI score with overall survival (OS) and disease-free survival. Immunofluorescence staining was used to examine the composition and phenotypes of SLI. The median follow-up times were 58.1 and 64.9months in the discovery and validation cohorts, respectively. SLI was intense in 38.1% of patients in the discovery cohort and in 20.9% of patients in the validation cohort (P<0.001). SLI score had independent prognostic value for OS [hazard ratio (HR) 2.132; P=0.016] and disease-specific survival (DSS) (HR 1.952; P=0.04) in the discovery cohort, which was confirmed in the validation cohort (OS: HR 1.636; P=0.023; DSS: HR 1.627; P=0.029). SLI score was positively associated with histological grade, tumour stage and lymph node status in both cohorts. Moreover, in the stroma, SLI displayed a broad spectrum of inhibitory immune cells, by expressing several major immune checkpoint molecules, i.e. programmed cell death protein 1, programmed death-ligand 1, indoleamine 2,3-dioxygenase, and T-cell immunoglobulin and mucin domain 3. Intense pre-existing SLI was validated as a reliable marker of poorer prognosis for survival in UCB patients, which may add to the prognostic significance of the TNM classification.

Reduced protocadherin17 expression in leukemia stem cells: the clinical and biological effect in acute myeloid leukemia

BackgroundLeukemia stem cell (LSC)-enriched genes have been shown to be highly prognostic in acute myeloid leukemia (AML). However, the prognostic value of tumor suppressor genes (TSGs) that are repressed early in LSC remains largely unknown.MethodsWe compared the public available expression/methylation profiling data of LSCs with that of hematopoietic stem cells (HSCs), in order to identify potential tumor suppressor genes in LSC. The prognostic relevance of PCDH17 was analyzed on a cohort of 173 AML patients from The Cancer Genome Atlas (TCGA), and further validated in three independent cohorts (n = 339).ResultsWe identified protocadherin17 (PCDH17) and demonstrated that it was significantly down-regulated and hypermethylated in LSCs compared with HSCs. Our analyses of primary AML patient samples also confirmed these deregulations. Clinically, low PCDH17 expression was associated with female sex (P = 0.01), higher WBC (P < 0.0001), higher percentages of blasts in bone marrow (BM) and peripheral blood (PB) (P = 0.04 and < 0.001, respectively), presence of FLT3-internal tandem duplications (P = 0.002), mutated NPM1 (P = 0.02), and wild-type TP53 (P = 0.005). Moreover, low PCDH17 expression predicted worse overall survival (OS) in four independent cohorts as well as in the molecularly defined subgroups of AML patients. In multivariable analyses, low PCDH17 expression retained independent prognostic value for OS. Biologically, PCDH17 expression-associated gene signatures were characterized by deregulations of EMT- and Wnt pathway-related genes.ConclusionsPCDH17 gene was silenced by DNA methylation in AML. Low PCDH17 expression is associated with distinct clinical and biological features and improves risk stratification in patients with AML.

Pre-Treatment Derived Neutrophil to Lymphocyte Ratio As a Prognostic Factor in Extranodal Natural Killer/T-Cell Lymphoma

Introduction: Extranodal natural killer/T-cell lymphoma (ENKTL) is an aggressive type of non-Hodgkin lymphoma (NHL), which is relatively more common in Asia. Accumulating evidence suggested that tumor microenvironment and inflammatory response played important roles in determining the clinical course and outcome of human malignancies. Recently, the derived neutrophil to lymphocyte ratio (dNLR) was demonstrated to act as prognostic factor in several malignancies. Nevertheless, the significance of dNLR in ENKTL has not been elucidated to date. The aim of this study is to investigate the prognostic value of dNLR in ENKTL.Methods: We included patients with pathological diagnosis of ENKTL from February 2002 to June 2018 at Shandong Provincial Hospital Affiliated to Shandong University. All the diagnoses were based on the criteria of the WHO 2016 Classifications of the Tumors and Hematopoietic and Lymphoid Tissues. The pretreatment clinical-pathological data, such as age, gender, stage, performance status, serum lactate dehydrogenase (LDH) levels, bone marrow involvement, and neutrophil count and leukocyte count were collected. The dNLR was calculated by absolute neutrophil count divided by (leukocyte count - neutrophil count).The primary end point was overall survival (OS). The secondary end point was progression-free survival (PFS). Survival analyses were performed using the Kaplan-Meier method and log-rank test. Multivariate analyses were performed using Cox proportional-hazards regression model. This study was approved by the Medical Ethical Committee of Shandong Provincial Hospital Affiliated to Shandong University. P<0.05 was considered statistically significant.Results: A total of 33 patients with pathological diagnosis of ENKTL were included in this study, including 26 (78.8%) male patients and 7 (21.1%) female patients. The median age at diagnosis was 50 years (range 13-75 years), and 6 (18.2%) of them were more than 60 years old. Among them, 13 (39.4%) patients were diagnosed at stage I/II with mean dNLR of 1.801 ± 0.832, versus 20 (60.6%) at stage III/IV with mean dNLR of 2.834 ± 1.962 (P=0.047). In addition, elevated serum LDH (P=0.026), distant lympho-node involvement (P=0.013), high β2-microglobulin (β2-MG, P=0.035) and high International Prognostic Index (IPI) score (P=0.044) were also significantly associated with high dNLR at diagnosis of ENKTL (Table 1).To determine the clinical significance of dNLR, cutoff value of 3.6 for dNLR, which was determined by receiver operating characteristic (ROC) analysis, was selected as the best discriminate between patients' survival and death (AUC 0.714, 95%CI 0.524-0.904, P=0.043, Figure 1). Patients were divided into dNLR<3.6 and dNLR≥3.6 groups. Patients with elevated dNLR (≥3.6) presented adverse pre-treatment factors, including elevated serum LDH (P<0.001), high Eastern Cooperative Oncology Group performance status (ECOG PS) score (P=0.004), increased β2-MG (P=0.008) and high IPI score (P=0.018). Kaplan-Meier curve revealed that high dNLR (≥3.6) was related to shorter OS (P=0.001) and PFS (P=0.047) of ENKTL patients (Figure 2-3).We next explored the correlation between clinical characteristics and survival of ENKTL patients by univariate and multivariate analysis. Univariate Cox proportional analysis revealed that high ECOG PS score (≥2, P=0.004), elevated β2-MG (P=0.014), high IPI score (≥2, P=0.007) and elevated dNLR (>3.6, P=0.001) as prognostic factors of poor outcome of patients. To further assess whether dNLR could act as an independent prognostic value for OS, multivariate analysis with Cox proportional hazard model was performed. In the multivariate analysis, which included all independent parameters significantly associated with clinical outcome in univariate analysis, we identified that high dNLR (≥3.6) as an independent prognostic factor for OS in patients with ENKTL (P=0.004).Conclusions: In this study, we validated for the first time the prognostic value of dNLR in ENKTL patients. High dNLR was associated with poor survival of ENKTL. The dNLR may serve as a novel and convenient prognostic factor for untreated ENKTL patients. Further validation of these results in larger cohorts of patients with ENKTL is warranted. DisclosuresNo relevant conflicts of interest to declare.

MTOR and mTOR phosphorylation status in primary and metastatic renal cell carcinoma tissue: differential expression and clinical relevance.

Impaired regulation of the Akt/mammalian target of rapamycin (mTOR) pathway has been implicated in mechanisms related to neoplastic transformation in renal cell cancer (RCC) through enhancement of cell proliferation and survival and mTOR activation has been reported to occur due to phosphorylation of mTOR. To further determine the relevance of mTOR expression and activation and to analyze their putative role as a biomarker for systemic treatment in metastatic RCC, we investigated the expression of mTOR and phospho(p)-mTOR in primary RCC and metastases and correlated levels with pathological variables and clinical outcome. Tissue microarrays (TMA) from paraffin-embedded tissue from 342 patients with primary clear cell renal cell carcinoma and 90 patients undergoing surgical resection for metastases were immunohistochemically stained for mTOR and p-mTOR and expression was quantified with immunoreactivity scores. Clinical patient characteristics and follow-up were recorded. Comparative evaluation of protein expression levels and association of expression with clinical variables and survival was performed. mTOR staining revealed differential expression in benign, primary and RCC metastasis (average staining score: 1.64, 0.78, and 1.44, respectively). Average staining of p-mTOR was 0.99 in benign kidney tissue, 0.73 in primary RCC and 1.14 in RCC metastasis tissue. Elevated mTOR expression in primary RCC tissue was associated with the presence of tumor necrosis, while a high level of p-mTOR was significantly correlated with advanced T-stage, high Fuhrman grade, the presence of tumor necrosis and sarcomatoid features. An elevated ratio of p-mTOR/mTOR was significantly correlated with advanced stage and sarcomatoid histology. mTOR expression was not predictive of overall survival (OS), while high p-mTOR levels were associated with impaired OS (p = 0.0046) and cancer-specific survival (p = 0.0067). In univariate analysis, advanced stage (HR 3.78), high Fuhrman grade (HR 4.0), the presence of tumor necrosis (HR 1.99), and sarcomatoid features (HR 5.12) were significant predictors of OS. Moreover, elevated levels of p-mTOR (HR 1.67) and an elevated ratio of p-mTOR/mTOR ratio (HR 1.73) were significantly predictive of OS. In the multivariate regression model only the presence of locally advanced tumors (HR 2.44) was of independent prognostic value for OS, while there was a trend for impaired OS for patients with a high p-mTOR (HR 1.27, p = 0.21). Phosphorylated mTOR is differentially expressed in localized RCC and metastasis. Elevated phosphorylation of mTOR is associated with aggressive pathologic features and unfavorable outcome. Whether these findings portend to relevance for mTOR inhibition treatment for metastatic RCC should be objective of further investigations.

Prognostic value of cyclooxygenase-2 gene polymorphisms in advanced non-small cell lung cancer patients treated with first-line platinum-based chemotherapy.

Cyclooxygenase-2 (COX-2) plays an important role in cell apoptosis, angiogenesis and tumor invasion, and over-expression of COX-2 is associated with tumor development and occurrence. The aim of this study is to investigate the association between COX-2 polymorphisms and clinical outcomes in advanced non-small cell lung cancer (NSCLC) patients treated with first-line platinum-based chemotherapy. A total of 190 patients with IIIB or IV NSCLC who received platinum-based chemotherapy were recruited in this study. Four functional COX-2 polymorphisms, including rs689465, rs689466, rs3218625 and rs20417, were genotyped by PCR-based restriction fragment length polymorphism methods. Kaplan-Meier methods were used to compare survival by different genotypes. Cox proportional hazard models were used to identify independently significant variables. The rs689465 AA genotype was significantly associated with longer overall survival (OS) (13.0 months vs 8.8 months, P = 0.019 for log-rank test; hazard ratio [HR] 0.624; 95% confidence internal [CI] 0.418-0.931) and progression-free survival (5.3 months vs 4.0 months, P = 0.018 for log-rank test; HR 0.627; 95% CI 0.421-0.934) compared with AG or GG genotype, especially in patients with adenocarcinoma (P = 0.002), performance status of 1 (P = 0.009) and stage IV disease (P = 0.012), and treated with gemcitabine-based chemotherapy (P = 0.012). Multivariate regression analysis showed that COX-2 rs689465 polymorphism had a significantly independent prognostic value for OS (P = 0.017, HR = 1.637, 95% CI = 1.093-2.453). Our study suggested that rs689465 polymorphism could be a prognostic biomarker for advanced NSCLC patients treated with first-line platinum-based chemotherapy.

Initial Characteristics, Treatment and Prognosis Of Elderly (≥ 70 years) Patients With Chronic Lymphocytic Leukemia (CLL): An Analysis Of a Series Of 367 Cases

AbstractIntroduction Although CLL predominates in the elderly, there are few studies focusing on the characteristics, treatment, and prognosis of elderly patients with CLL. Patients and MethodsRetrospective single center study of 949 patients diagnosed with CLL between 1990 and 2012. We defined two age-groups:<70 years (“young”, n=582) and ≥ 70 (“elderly”, n=367). Statistical analysis was performed with SPSS and R. ResultsBaseline characteristics of patients are shown in the Table. Median follow-up time was 7.8 years (0.1-23.1) for the overall series. Elderly patients presented with more advanced clinical stage (p<.001) and higher B2M (p<.001) than younger ones. However, elderly patients presented with a lower WBC count (19.6 vs. 23.6 x 109/L; p<.001) and a longer LDT (p=.004). In contrast to younger patients, both sexes were equally distributed within the elderly (p<.001). There were no significant differences regarding ZAP70, FISH cytogenetics, IGHV mutational status and mutations of NOTCH1 and SF3B1. Elderly patients were treated less frequently (39% vs. 62%; p<.001; TTFT 23.8% vs. 41.9% at 3 years; p<.001) and the type of treatment given to elderly vs. younger patients was: alkylating agents (70% vs. 36%), purine analogs (10% vs. 32%), chemoimmunotherapy (5% vs. 25%), other (15% vs. 7%) (p<.001). The response rate was lower in elderly patients (49 % with 14% CR vs. 69% with 31% CR; p<.001). The overall survival (OS) was shorter in the elderly (6.6 vs. 13.3 years; p<.001) in whom a higher CLL-unrelated mortality (34.9% vs. 6.9% at 10 years; p<.001) was observed. The univariate analysis showed a correlation between advanced clinical stage (p<.001), increased B2M (p<.001), 11q- and 17p- (p=.016), high ZAP70 (p<.001), unmutated IGHV (p=.002) and NOTCH1 (p=.002) and SF3B1 mutations (p<.001) and shorter OS. A high co-morbidity index at diagnosis (CIRS-D >6) also correlated with OS (p=.064). With the only exception of cytogenetics by FISH all variables included in the multivariate analysis retained independent prognostic value (clinical stage: HR 1.68, p=.012; ZAP-70: HR 1.64, p=.01; CIRS-D: HR 1.42, p=.035 and B2M: HR 1.09, p=.021). IGHV mutational status, NOTCH1 and SF3B1 mutations were not included in the analysis due to missing data. In elderly patients requiring treatment only co-morbidity burden at treatment initiation (CIRS-T>4: HR 2.13, p=.009) and response to therapy (PR vs. CR: HR 2.03, p=.039; and failure vs. CR: HR 4.33, p<.001) had independent prognostic value for OS. ConclusionsPatients above 70 years were diagnosed at more advanced phases of the disease than younger ones but no significant differences in the biology of the disease were observed. Importantly, once patients required treatment only co-morbidity and response to therapy were prognostically relevant. Unfortunately, however, only a small proportion of elderly patients could receive effective therapy as currently conceived. In addition, an important fraction of patients died because of causes not related to CLL. These findings underscore the importance of developing active and well tolerated treatments, as well as that of a comprehensive medical care for elderly patients with CLL.TablePrinciple baseline characteristics of the 949 CLL patients separated by age< 70 years (n=582)≥ 70 years (n=367)p-valueMedian age57 (28-69)77 (70-97)-Median follow-up9.5 (0.4-23.2)5.2 (0.1-20.3)-Gender, male (%)365 (63)190 (52).001ECOG > 1 (%)10/543 (2)35/341 (10)<.001Rai III/IV (%)29/566 (5)43/358 (12)<.001Binet C (%)26/566 (5)35/358 (10)<.001WBC x 109/L32.3± 44.426.7±37.002Lymphocytes x 109/L23.6 ± 35.219.6± 33.4.002Hb g/dl13.8 ± 1.713.1± 1.9<.001Platelets x 109/L197 ± 65200 ± 82NSLDT<12 m (%)79/444 (18)31/287 (10).004LDH increased (%)58/540 (11)48/347 (14)NSB2M mg/ml2.41 ± 1.463.41 ± 1.98<.001ZAP-70 increased (%)162/436 (37)68/227 (30)NSCD 38 increased (%)131/405 (32)61/205 (30)NSIGHV unmutated (%)174/375 (46)68/157 (43)NSNOTCH1 mutated (%)35/350 (10)22/151 (15)NSSF3B1 mutated (%)27/301 (9)8/111 (7)NSFISH (number, %)NS13q-147/427 (30)85/214 (40)Normal126/427 (30)62/214 (29)+ 1258/427 (14)29/214 (14)17p-56/427 (13)23/214 (11)11q-40/427 (9)15/214 (7)NS: not significant; ZAP-70 increased: ≥ 20% of CLL cells positive; CD38 increased: >30% of CLL cells positive; IGHV unmutated: ≥ 98% identity with germline; CIRS-D: Cumulative Illness Rating Scale at diagnosis Disclosures:No relevant conflicts of interest to declare.

Bendamustine with or without rituximab in the treatment of relapsed chronic lymphocytic leukaemia: an Italian retrospective study

To retrospectively assess the efficacy of bendamustine alone and with rituximab (R-B), 109 patients with relapsed chronic lymphocytic leukaemia (CLL) were enrolled in 24 Italian centres. The median age was 66 years (range 39-85). Forty-three percent of patients had relapsed and 57% were resistant (median previous therapies = 3; range 1-8). Twenty-two patients received bendamustine alone and 87 patients received R-B (median B dosage: 100 mg/m(2) per day, range 90-130 mg/m(2) per day). The overall response rate was 69·6% (complete response 28·6%; partial response 41%), and was significantly higher in patients treated with R-B (P = 0·014) and in those responsive to the previous treatment (P=0·04). After a median follow-up of 7·9 months (range 1-148), the median progression-free survival was 16 months and the median duration of response was 13 months. Median overall survival (OS) was 16·8 months for the whole cohort; patients not responding to the treatment had a significantly worse outcome than those who attained a response (P = 0·0001). In multivariate analysis, only resistant disease status at start of bendamustine treatment (HR 3·2, 95% CI 1·4-7·3, P = 0·006) had an independent prognostic value for OS. Toxicity was manageable and mostly haematological. In conclusion, in our experience R-B was an effective and well-tolerated treatment for relapsed/refractory CLL patients, producing a remarkable high CR rate and mild toxicity.

High serum lactate dehydrogenase adds prognostic value to the international myeloma staging system even in the era of novel agents

High serum lactate dehydrogenase (LDH) is associated with features of advanced disease and inferior survival in multiple myeloma. It is however unclear whether LDH adds to the prognostic value of International Staging System (ISS) and whether it retains its prognostic significance in patients who are exposed to novel agent-based therapies. To address these issues we analyzed 996 consecutive symptomatic patients who were included in the database of the Greek Myeloma Study Group and received frontline treatment between January 1, 1995 and December 31, 2008. The median overall survival (OS) of all patients was 40 months with a clear improvement in those who started treatment after January 1, 2000 (49 vs. 31 months; P < 0.01). A multivariate model showed that LDH, ISS, performance status, age and platelet counts had an independent prognostic value for OS (P < 0.001 for all parameters). The median OS of patients with high (11% of patients) and normal LDH was 15 vs. 44 months (P < 0.001). High LDH was associated with inferior OS within all ISS groups: 22 vs. 76 months for high and normal LDH groups, respectively, in ISS-1 (P < 0.01); 11 vs. 40 months in ISS-2 (P < 0.001) and 17 vs. 27 months in ISS-3 (P < 0.01). The median OS of high and normal LDH groups among patients who received novel agents was 21 vs. 51 months, respectively (P < 0.001). Lactate dehydrogenase is a readily available and inexpensive variable, which has a major impact on the survival of myeloma patients even when they belong to a low or intermediate ISS subgroup and even when they receive novel agent-based therapies.

High Microvascular Density Correlates with Poor Outcome in Patients with Diffuse Large B-Cell Lymphoma (DLBCL) Treated with Rituximab Plus Chemotherapy (R-CT).

Abstract 1948Poster Board I-971Survival after treatment of diffuse large B-cell lymphoma (DLBCL) is influenced by differences in the tumor microenvironment. Gene expression profiling (GEP) studies have shown that the angiogenesis-related signature (stromal-2 signature) is prognostically unfavorable. However, the clinical and biological significance of angiogenesis quantified in tumor tissue sections of DLBCL from patients treated with rituximab plus chemotherapy (R-CT) is not yet fully explored. CD31, the platelet adhesion molecule PECAM1, is one of the genes included in the “stromal-2 signature” reported in the GEP studies. The objective of this study was to determine whether the microvessel density (MVD) and microvessel number (MVN) in DLBCL patients treated with R-CT were associated with the clinicopathological features of the tumors and related to the outcome of the patients. The MVD and MVN were assessed in a series of 160 patients with DLBCL from the Leukemia Lymphoma Molecular Profiling Project consortium (LLMPP) 86M /74F; median age 64 yrs. The GEP was investigated in 116 of these including 50 germinal center B (GCB), 55 activated B-cell (ABC) and 11 unclassifiable cases. An independent series of 129 patients from the Catalan Lymphoma-Study Group (GELCAB) (67M/62F; median age 64 yrs) was used to validate the results. Front-line treatment was R-CT in all cases of both series. Tissue Microarrays (TMA) were constructed from pretreatment biopsy specimens of de novo DLBCL. High grade B cell lymphoma otherwise unclassifiable, primary mediastinal B cell lymphoma, T-cell-rich B cell lymphoma, and tumors associated with immunodeficiency were excluded. All cases were stained in an automated immunostainer with an antibody against CD31 (DAKO). The MVD and MVN were quantified using digitalized images of the tumor using Olympus Cell B Basic Imaging Software. Microvessel areas were defined as vascular areas delineated by CD31+ staining. The MVD was calculated as the sum of all microvessel areas divided by the total area analyzed. The MVN was the sum of all identified individual vessels, divided by the total area analyzed. TMAs were independently scored by two observers and discrepancies were resolved over a double-headed microscope. To determine whether the angiogenic values scored using the TMA were representative of the tumor sample, whole tissue sections and TMA cores from the same tumor were evaluated in 40 cases and compared by a linear regression analysis. MVD and MVN were grouped in quartiles when necessary and considered high or low when above or below the 50th percentile, respectively. Linear correlation analysis between the CD31 (+) MVD results on TMA cores and on the corresponding whole tissue sections in 40 cases showed a good correlation (R2=0.81). In the LLMPP cohort, DLBCL with an ABC profile showed higher MVD than those with GCB profile (p=0.05). In addition, higher MVD was observed in patients with advanced stage (p<0.01), but there was no significant correlation with other clinical features. 5-yr overall survival (OS) according to CD31(+) MVD was 74% vs. 47% for patients with low and high MVD respectively (p=0.0015). Both the International prognostic index (IPI) (relative risk 3.3; p=0.001) and MVD (relative risk 2.2; p<0.001) showed independent prognostic value for OS in a Cox model. In addition, MVD and GEP type (GCB vs. ABC) were also independent predictors of OS. MVN showed no meaningful relation with initial features or with OS. In the validation cohort from the GELCAB, all the above mentioned results were confirmed, including the influence of MVD on OS (5-yr OS 78% vs. 50% for low and high MVD, respectively; p=0.02) that was also independent of IPI in a Cox model. In conclusion, increased MVD is able to discriminate poor-risk patients in DLBCL treated with R-CT independently of the IPI risk groups. This finding highlights the relevance of angiogenesis in the behavior of these tumors and suggests that it may be an important parameter when assessing the impact of new therapies, particularly anti-angiogenic drugs. Disclosures:Gascoyne:Roche Canada: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Connors:Roche Canada: Research Funding. Rimsza:High Throughput Genomics: .

High syndecan-1 expression in breast carcinoma is related to an aggressive phenotype and to poorer prognosis.

Syndecan-1 is a transmembrane heparan sulphate proteoglycan that is involved in cell-cell adhesion, organization of cell-matrix adhesion, and regulation of growth factor signaling. Specimens from 254 consecutive breast carcinoma (BC) cases (110 N0, 144 N1/2) with long-term follow-up (median, 95 months) were immunostained for syndecan-1, estrogen receptor (ER), progesterone receptor (PgR), and p53; in 154 cases, c-erbB-2 status was known. Syndecan-1 mRNA and protein expression also were evaluated in 20 breast tissue samples (10 normal and tumor pairs). Syndecan-1 was expressed at high levels in 106 (42%) BCs; syndecan-1 up-regulation was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) studies. High syndecan-1 expression was associated with high histologic grade, large tumor size, high mitotic count, c-erbB-2 overexpression, and ER and PgR negative status. At univariate survival analysis syndecan overexpression was related to poor prognosis (P < 0.01 for both overall survival (OS) and disease-free survival). Bivariate survival analysis showed an additive adverse effect for syndecan-1 and c-erbB-2 overexpression. At multivariate analysis, syndecan-1 overexpression was independently associated with poor OS (hazard ratio [HR], 1.71; 95% confidence interval [CI], 1.08-2.69). High syndecan-1 expression also was of independent prognostic value for OS in the group of 102 ER-negative patients (HR, 2.42; 95% CI, 1.21-4.82). Stratifying patients on the basis of the type of adjuvant therapy given, high syndecan-1 expression was associated with a higher risk of death only in patients treated with the cyclophosphamide-methotrexate-fluorouracil regimen (HR, 1.9; P = 0.09); at multivariate analysis for OS, this association proved to be of independent statistical significance (P = 0.03; HR, 2.15). Syndecan-1 is expressed at high levels in a significant percentage of breast carcinomas and is related to an aggressive phenotype and poor clinical behavior.