Prognostic Marker For Overall Survival Research Articles

N-cadherin is a therapeutic target in t(4;14)-positive multiple myeloma

e238 Patients and Methods: CXCL9 serum levels were studied by ELISA in newly-diagnosed (ND) MM (n1⁄4105) and MGUS patients (n1⁄417), and healthy donors (n1⁄437). CXCL9 expression was analyzed by qPCR in MM cell lines (HMCLs), MM-BMMNCs, MM-PBMCs and in a publically available GEP-dataset (GSE2658). Concurrent analysis of CXCL9-11 serum levels was performed by FACS-CBA array. Results:Median CXCL9 levels (161 pg/ml; range: 9-1966) were significantly elevated in MM patients compared to MGUS (93; 6-1303) and healthy donors (106; 51e391). CXCL9 levels significantly correlated with B2M, ISS stage, calcium, albumin, LDH, hemoglobin and age. High CXCL9 levels predicted adverse survival (17.0 months vs. not reached, P<0.001), which was upheld when age-adjusted cut-off levels were used. Similarly, in MM patients (n1⁄4559) treated within the TT2 and 3 protocol (GSE2658) shorter OS was found in patients with a present compared to those with an absent detection call for CXCL9 (P1⁄40.004). Expression of CXCL9 was only detected in 1 of 7 HMCLs and 12% of previously published MM cell samples. Hence, increased serum levels of CXCL9 are likely derived from other cell types. We observed increased expression of CXCL9 in PBMCs compared to BMMNCs of MM patients, suggesting that elevated secretion of CXCL9 might result from aberrant cytokine production in cells of the PB. CXCL9 serum levels were also analyzed in conjunction with CXCL10 and 11 in 65 MM patients by FACS-CBA. Again, we observed a significant correlation with B2M for CXCL9-11. Survival was significantly worse in patients with high compared to low CXCL9 (median OS 25.3 months vs. not reached, P1⁄40.003) and CXCL10 (19.97 months vs. not reached, P1⁄40.0006). In multivariate analysis, CXCL9 (P1⁄40.03) and CXCL10 (P1⁄40.013) were revealed as independent predictors of survival. Conclusion: Our findings depict CXCL9 as novel prognostic marker for overall survival in ND-MM patients. High levels of CXCL9 in the PB probably increase TH1/TH2 ratios and may shield myeloma cells from immune attack by CD8+ cytotoxic T cells as previously shown for T cell lymphomas. Further experiments will aim to confirm these results and define the functional role of CXCL9 in MM.

RETRACTED ARTICLE: Downregulation of miR-133b/miR-503 acts as efficient prognostic and diagnostic factors in patients with osteosarcoma and these predictor biomarkers are correlated with overall survival.

It has been suggested that microRNAs can act as prognostic markers, diagnostic markers, and potential thera- peutic targets. Therefore, it can be useful to evaluate the ex- pression level of miR-133b/miR-503 in clinical samples. Our study was aimed to evaluate miR-133b/miR-503 expression levels in patient's tissues suffering from osteosarcoma. The association of miR-133/miR-503 expressions with path- ological factors was evaluated. Total RNA was purified from patients with osteosarcoma and noncancerous bone tissues, and real-time PCR was used to assess the expression rates of miR-133b and miR-503. Our results indicated that the miR-133b expression was strongly lower in osteosarcoma bone tissue compared with noncancerous bone tissues. Furthermore, expression level of miR-503 was decreased in osteosarcoma bone tissue than those corresponding noncan- cerous bone tissues. Our findings showed that low expression of miR-133b was remarkably correlated to large size of tumor (P=0.031), advanced TNM stage (P=0.003), and metastasis or recurrence (P=0.012). In addition, low expression of miR-503 were strongly linked to large tumor size (P=0.027), advanced TNM stage (P=0.006), and metastasis or recurrence (P=0.25). Furthermore, Kaplan-Meier survival analysis and log-rank test revealed that the low expressions of miR-133b/miR-503 were correlated with shorter overall sur- vival(log-ranktestP=0.019;P=0.028).ThemultivariateCox proportional hazards model suggested that decreased expres- sions of miR-133b/miR-503 (P=0.027; P=0.021), TNM stage (P=0.021; P=0.001), metastasis (P=0.001; P=0.037), and large tumor size (P=0.035; P=0.023) were independent prognostic markers of overall survival of patients. Our result indicated that miR-133b/miR-503 expression levels in tissues may be useful as efficient prognostic factors in patients suf- fering from osteosarcoma, and that downregulation of miR-133b/miR-503 are correlated with overall survival.

Prognostic value of abnormally expressed long non-coding RNAs in patients with osteosarcoma: a meta-analysis

// Fashuai Wu 1, * , Deyao Shi 1, * , Shidai Mu 2 , Yu Huang 3 , Feng Gao 1 , Xiangcheng Qing 1 and Zengwu Shao 1 1 Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 2 Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 3 Department of Otorhinolaryngology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China * These authors contributed equally to this work Correspondence to: Zengwu Shao, email: profszw1962@163.com Keywords: lncRNA; osteosarcoma; prognosis; biomarker; meta-analysis Received: May 23, 2017 Accepted: December 26, 2017 Published: January 12, 2018 ABSTRACT Long non-coding RNAs (lncRNAs) contribute to progression of various cancers including osteosarcoma through abnormal regulation of cancer-related cellular processes. Recent researches have shown that various lncRNAs are abnormally expressed in osteosarcoma and associated with the prognosis. With the aim of gaining a better insight into the association between expression level of lncRNAs and prognosis of osteosarcoma, multiple databases including PubMed, Embase, Cochrane Library and Web of Science were carefully searched for available studies up to March 14, 2017. Finally, 19 publications with 1298 patients matched our inclusion criteria and were evaluated in our meta-analysis. The quality of each study was scored using the Newcastle-Ottawa Scale and studies not reaching a minimum threshold were excluded. Results of the meta-analysis demonstrated that abnormal expression level of lncRNAs predicted poor overall survival (pooled HR 3.064, 95% CI: 2.487–3.775) and event-free survival (pooled HR 2.642, 95% CI 1.759–3.970) in osteosarcoma and subgroup analysis showed consistent prognostic value. Furthermore, combining data of Cox multivariable analysis indicated that abnormal expression level of lncRNAs was an independent prognostic marker for overall survival (pooled HR 2.864, 95% CI: 2.246–3.651) in osteosarcoma patients. The clinicopathological parameters analysis further showed that abnormal expression level of lncRNAs was correlated with tumor size, tumor stage, metastasis and differentiation grade of osteosarcoma. Limitations of the meta-analysis included variation of cut-off value definition, Chinese provenance of most studies, publication bias and so on. In conclusion, this meta-analysis suggested that abnormal expression level of lncRNAs has a promising future for predicting the prognosis of patients with osteosarcoma.

Mucin 1-specific B cell immune responses and their impact on overall survival in breast cancer patients

Considering the diverse functions of B cells, responses to tumor-associated antigens (TAA) have been thought to be the main source of B cell-mediated antitumor immunity. Polymorphic epithelial mucin (MUC1) is considered one of the most specific TAA in patients with breast cancer. The present study aims to dissect the level and subclasses of naturally occurring anti-MUC1 antibodies in regard to tumor biologic parameters, clinical characteristics and overall survival. In 288 primary, non-metastatic breast cancer patients, pretreatment serum levels of anti-MUC1 immunoglobulin G (IgG) and its subclasses G1–4 as well as immunoglobulin M (IgM) were analyzed via ELISA. With respect to overall survival (Kaplan–Meier analysis), tumor biologic parameters as hormone receptor status, human epidermal growth factor receptor 2 (Her2), Ki-67 expression and tumor grading have been correlated as well as clinical characteristics as nodal involvement, tumor stage and patients' age at the time of diagnosis. Median follow-up time was 148 mo (IQR: 73.1–158.5 mo). A significant increase in IgG antibody titers was correlated highly significantly with an improved overall survival of patients. In multivariate analysis, total IgG proved to be an independent prognostic marker for overall survival (p = 0.002). IgG subclass analysis did not reveal any correlation of IgG1, IgG3 and IgG4 levels with overall survival, while increased immunoglobulin G2 (IgG2) values, although statistically not significant, tended to correlate with prolonged patient survival. MUC1-specific IgM antibodies were shown not to be predictive of overall survival. Altogether, humoral immune responses appear to play a crucial part in the tumor immunity of breast cancer patients. The present data confirms the positive impact of tumor-specific IgG on prolonged overall survival in breast cancer patients. MUC1-antibody testing might be a useful tool to identify high-risk patients who may need adjuvant therapy and potentially might benefit from MUC1-directed immunotherapy.

Matrix metalloproteinase-12 expression is increased in cutaneous melanoma and associated with tumor aggressiveness.

Cutaneous melanoma is the most malignant form of skin cancer characterized by aggressive invasion. Matrix metalloproteinases play essential roles in tumor invasion due to their ECM degrading capacity. However, the clinical significance of matrix metalloproteinasis (MMP)-12 in human cutaneous melanoma has not been addressed yet. In the present study, we investigated MMP-12 expression level in 298 patients with cutaneous melanoma and 60 normal skin tissue specimens by immunohistochemistry assay. Appropriate statistical analysis was utilized to determine the association of MMP-12 with clinical features and prognosis of melanoma. Results showed that MMP-12 expression was increased in cutaneous melanoma compared with that in normal skin. It was also found that MMP-12 expression in melanoma was significantly associated with tumor invasion and metastasis. Univariate survival analysis indicated that patients with melanoma of high MMP-12 expression had unfavorable overall survival compared with those of low MMP-12 expression. Cox's proportional hazards analysis showed that MMP-12 expression was an independent prognostic marker of overall survival for patients with cutaneous melanoma. These results proved that MMP-12 expression was increased in cutaneous melanoma and associated with tumor progression. It also provided the first evidence that MMP-12 level could be an independent prognostic marker for patients with cutaneous melanoma.

Prognostic Value of Dynamic Susceptibility Contrast-Enhanced and Diffusion-Weighted MR Imaging in Patients with Glioblastomas.

Prediction of survival in patients with glioblastomas is important for individualized treatment planning. This study aimed to assess the prognostic utility of presurgical dynamic susceptibility contrast and diffusion-weighted imaging for overall survival in patients with glioblastoma. MR imaging data from pathologically proved glioblastomas between June 2006 to December 2013 in 58 patients (mean age, 62.7 years; age range, 22-89 years) were included in this retrospective study. Patients were divided into long survival (≥15 months) and short survival (<15 months) groups, depending on overall survival time. Patients underwent dynamic susceptibility contrast perfusion and DWI before surgery and were treated with chemotherapy and radiation therapy. The maximum relative cerebral blood volume and minimum mean diffusivity values were measured from the enhancing part of the tumor. Maximum relative cerebral blood volume values in patients with short survival were significantly higher compared with those who demonstrated long survival (P < .05). No significant difference was observed in the minimum mean diffusivity between short and long survivors. Receiver operator curve analysis demonstrated that a maximum relative cerebral blood volume cutoff value of 5.79 differentiated patients with low and high survival with an area under the curve of 0.93, sensitivity of 0.89, and specificity of 0.90 (P < .001), while a minimum mean diffusivity cutoff value of 8.35 × 10(-4)mm(2)/s had an area under the curve of 0.55, sensitivity of 0.71, and specificity of 0.47 (P > .05) in separating the 2 groups. Maximum relative cerebral blood volume may be used as a prognostic marker of overall survival in patients with glioblastomas.

High p53 protein expression in therapy-related myeloid neoplasms is associated with adverse karyotype and poor outcome

Identification of p53-positive cells by immunohistochemistry in bone marrow from primary myelodysplastic syndrome patients correlates with the presence of TP53 mutations and poor prognosis. Mutations in the tumor suppressor gene TP53 are more frequent in therapy-related acute myeloid leukemia and myelodysplastic syndrome than in de novo disease, but the role of p53 immunohistochemistry in the therapy-related setting has not been specifically investigated. We studied p53 protein immunoreactivity in bone marrow biopsies of therapy-related myeloid neoplasms and correlated protein expression with TP53 mutation status, clinicopathologic features and outcome. We first studied 32 patients with therapy-related acute myeloid leukemia and 63 patients with therapy-related myelodysplastic syndrome/chronic myelomonocytic leukemia from one institution and then validated our results in a separate group of 32 patients with therapy-related acute myeloid leukemia and 56 patients with therapy-related myelodysplastic syndrome from a different institution. Strong p53 immunostaining in ≥1% of bone marrow cells was highly predictive of a TP53 gene mutation (P<0.0001) and was strongly associated with a high-risk karyotype (P<0.0001). The presence of ≥1% p53 strongly positive cells was associated with poorer overall and disease-specific survival, particularly in the subset of patients treated with stem-cell transplantation. In a multivariable Cox regression model, the presence of ≥1% p53 strongly expressing cells was an independent prognostic marker for overall survival in both cohorts, with hazard ratios of 3.434 (CI: 1.751–6.735, P<0.0001) and 3.156 (CI: 1.502–6.628, P=0.002). Our data indicate that p53 protein expression, evaluated in bone marrow biopsies by a widely available immunohistochemical method, prognostically stratifies patients with therapy-related myeloid neoplasms independent of other risk factors. p53 immunostaining thus represents an easily applicable method to assess risk in therapy-related acute myeloid leukemia/myelodysplastic syndrome patients.

An integrated approach of gene expression and DNA-methylation profiles of WNT signaling genes uncovers novel prognostic markers in acute myeloid leukemia.

BackgroundThe wingless-Int (WNT) pathway has an essential role in cell regulation of hematopoietic stem cells (HSC). For Acute Myeloid Leukemia (AML), the malignant counterpart of HSC, currently only a selective number of genes of the WNT pathway are analyzed by using either gene expression or DNA-methylation profiles for the identification of prognostic markers and potential candidate targets for drug therapy. It is known that mRNA expression is controlled by DNA-methylation and that specific patterns can infer the ability to differentiate biological differences, thus a combined analysis using all WNT annotated genes could provide more insight in the WNT signaling.ApproachWe created a computational approach that integrates gene expression and DNA promoter methylation profiles. The approach represents the continuous gene expression and promoter methylation profiles with nine discrete mutually exclusive scenarios. The scenario representation allows for a refinement of patient groups by a more powerful statistical analysis, and the construction of a co-expression network. We focused on 268 WNT annotated signaling genes that are derived from the molecular signature database.ResultsUsing the scenarios we identified seven prognostic markers for overall survival and event-free survival. Three genes are novel prognostic markers; two with favorable outcome (PSMD2, PPARD) and one with unfavorable outcome (XPNPEP). The remaining four genes (LEF1, SFRP2, RUNX1, and AXIN2) were previously identified but we could refine the patient groups. Three AML risk groups were further analyzed and the co-expression network showed that only the good risk group harbors frequent promoter hypermethylation and significantly correlated interactions with proteasome family members.ConclusionOur results provide novel insights in WNT signaling in AML, we discovered new and previously identified prognostic markers and a refinement of the patient groups.

Fluorodeoxyglucose positron emission tomography/computerized tomography in differentiated thyroid cancer management: Importance of clinical justification and value in predicting survival.

The purpose of this study was to evaluate the added value of follow-up fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) to clinical assessment and predicting survival outcome in patients with differentiated thyroid cancers. This is an institutional review board approved, retrospective study of 202 biopsy-proven thyroid cancer patients at a single tertiary centre. A total of 327 follow-up or surveillance PET/CT scans done 6 or more months from initial treatment completion were included in this study. Median follow-up from completion of primary treatment was 94 months (range, 6.17-534.1 months). Overall survival benefit was measured using Kaplan-Meier plots with a Mantel-Cox log-rank test. Multivariate Cox regression model is provided with clinical covariates. Of the 327 PET/CT scans from 202 patients, 161 were positive and 166 as negative for recurrence or metastasis. A total of 23 patients died during the study period. Patients with a positive PET/CT scan had shorter overall survival than those who had a negative scan (P < 0.0001, hazard ratio 6.1 (3.0-14.3) ). In the context of clinical assessment, PET/CT identified recurrence in 50% (25/50) of scans without prior clinical suspicion and ruled out recurrence in 36.8% (102/277) of scans with prior clinical suspicion. In a multivariate Cox regression model, factors associated with overall survival were stage (P < 0.0001), time to scan (P = 0.0005) and PET/CT result (P < 0.0001). FDG PET/CT performed in follow-up more than 6 months from primary treatment completion adds value to clinical judgment and a prognostic marker of overall survival in thyroid cancer patients.

Down-regulation of C/EBP homologous protein (CHOP) expression in gastric cardia adenocarcinoma: Their relationship with clinicopathological parameters and prognostic significance

C/EBP homologous protein (CHOP) is a multi-functional protein involved in the apoptosis pathway of endoplasmic reticulum stress (ERS) and is related to cancer progression. The purpose of this study was to assess CHOP expression as a prognostic biomarker in gastric cardia adenocarcinoma (GCA). The levels of CHOP mRNA and protein in GCA and matched adjacent non-cancerous tissues were evaluated by quantitative real-time-polymerase chain reaction (qRT-PCR) and western blot. Furthermore, the CHOP protein expression and localization were examined by immunohistochemistry in GCA and corresponding adjacent non-cancerous tissues, gastritis and normal cardiac tissues. The association of CHOP expression with clinical pathological parameters and prognosis of GCA patients was statistically analyzed. Compared with adjacent non-cancerous tissues, the CHOP was down-regulated at mRNA and protein levels in GCA (P<0.01). In addition, immunohistochemistry analysis showed that CHOP positivity was lower in GCA than that in paired adjacent non-cancerous tissues, gastritis and normal tissues (P<0.01). CHOP expression rate gradually decreased with an increase in clinical stage, tumor differentiation and lymph node metastasis of GCA (P<0.05). Kaplan-Meier survival analysis revealed that low expression of CHOP correlated with poor prognosis of GCA patients. Moreover, univariate and multivariate analyses showed that CHOP was an independent prognostic marker for overall survival of GCA patients. Our results suggest that low CHOP expression predicts poor prognosis of GCA patients, and CHOP may be potentially a prognostic biomarker for GCA.

Circulating Tumor Cells (CTCs): From Detection to Dissection

Summary �䀆e breast cancer represents one of the most prevalent cancers in the world. �䀆e outlook for patients diagnosed with breast cancer has been changing for the better over time, with several sgnficant advances coming through in the past 10 years; among them, and a�怇er highlighting the importance of prevention and early diagnosis, relevant advances as a better understanding of the process through gene expression analysis, at the ability to use specfic therapeutic targets, and a more advanced hormonal therapy played a more than sgnficant positive role. Nevertheless, and despite the above mentioned improvements in treatment and prevention, breast cancer remains one of the leading causes of cancer mortality in women. Approximately 90% of cancer mortality is due to the development of metastatic disease. �䀆e metastatic process involves the discharge of tumor cells from the primary tumor into peripheral blood (circulating tumor cells, CTCs) to a target organ (1). Recently, the clinical implication of detection of CTCs in breast cancer has been recognized and accepted (2). In fact, the presence of CTCs has been correlated with a worse prognosis in early and metastatic breast cancer. As a result of these investigations, it has been reported that the amount of CTCs in the blood of these patients is an independent prognostic marker for overall survival and progression-free survival (OS and PFS) (3). However, despite the accepted role of CTCs in the prognosis of breast cancer, the detection of CTCs alone does not provide enough information. Although the changes in CTC's amount per blood unit might indicate sensitivity or resistance to an anticancer therapy and provides information about the prognosis of the disease, this information is in many cases not enough (4). �䀆e question we need to ask is, how can we eliminate these CTCs in order to combat the disease �䀆e annihilation of CTCs, involves the knowledge of their biology. �䀆e molecular and phenotypic characterization of these CTCs could be the key to understanding the mechanism of resistance to administered therapies and the door to the development of novel cancer treatments that specficalltarget circulating tumor cells. In this short communication, we would like to brefldiscuss the relevance of the molecular characterization of CTCs. Firstly, as new tools to potential clinical use in the monitoring of breast cancer, and secondly as potential biomarkers to development of new therapeutic targets. Similar but not identical

TP53 Mutation Status Divides MDS Patients with Complex Karyotypes into Distinct Prognostic Risk Groups: Analysis of Combined Datasets from the International Working Group for MDS-Molecular Prognosis Committee

Background:Complex karyotypes in patients with myelodysplastic syndromes (MDS) are defined by the presence of 3 or more cytogenetic abnormalities and are considered indicators of a poor prognosis by nearly all models in clinical use. There is evidence that the prognostic significance of complex karyotypes in MDS can be refined by considering the number of cytogenetic abnormalities or the presence of specific lesions, the presence of monosomy, and the mutation status of the TP53 gene. Mutations of TP53 are strongly associated with adverse clinical features including complex karyotypes, yet still have independent prognostic significance. Whether these mutations or other elements associated with complex karyotypes carry the greatest prognostic significance is unknown and has been the subject of recent contradictory findings. Here we examine the relationship between TP53 mutations, the number and type of chromosomal abnormalities, and clinical features including overall survival in a combined analysis of MDS patients with complex karyotypes.Methods:Data on 258 complex karyotype MDS patients from the US and Europe gathered by members of the International Working Group for Prognosis in MDS-Molecular Committee were combined for this analysis. Patients gave informed consent to have their data and clinical samples collected at their respective institutions in accordance with the Declaration of Helsinki. Tumor samples were examined for TP53 mutations primarily with next generation sequencing techniques. Categorical variables were compared using a Fisher exact test or Kruskal-Wallis test as appropriate, while continuous variables were compared using a Wilcoxon rank-sum test. All p-values reported are two-sided, unadjusted for multiple testing, and considered significant at the 0.05 level.Results:Of the 258 complex karyotype MDS samples examined, TP53 mutation status was determined in 223, of which 133 (60%) carried a TP53 mutation. Patients with TP53 mutations were slightly older than unmutated patients (median age 70.8 vs. 65.5 years, p=0.055), were more thrombocytopenic (median platelet count 42 vs. 78 x109/L, p=0.002), and had a greater percentage of bone marrow blasts (7.75% vs 4%, p=0.03). Mutated patients were also more likely to have a monosomal karyotype (85% vs. 59%, p<0.0001), have 5 or more chromosomal abnormalities (85% vs. 51%, p<0.0001), and have abnormalities of chromosome 17 (38% vs. 23%, p=0.03).Patients with TP53 mutation had a significantly shorter median overall survival of 7.7 months compared to 23.4 months for patients with unmutated TP53 (p<0.0001, see figure). Multivariate analysis that considered potential confounders such as monosomal karyotypes, number of abnormalities, blast proportions and platelet count identified TP53 mutation status as the most significant prognostic marker for overall survival (hazard ration [HR] 2.12, p<0.0001). The presence of only 3 or only 4 karyotype abnormalities was protective compared with 5 or more such lesions (HR 0.385, p=0.0005 for 3 lesions and HR 0.505, p=0.0083 for 4 lesions).Discussion:Despite strong associations with adverse clinical and cytogenetic abnormalities incorporated into existing prognostic scoring systems, TP53 mutations carry significant independent prognostic value for patients with MDS. In our multivariable analyses, TP53 mutation appears to dominate over monosomal karyotype as an adverse prognostic marker in MDS patients with complex cytogenetics. Many of the TP53 mutations identified had a small variant allele fraction suggesting that they were present in a disease subclone. The small number of mutant cells in these cases may be insufficient to alter clinical measures, but could still indicate a poor prognosis if they represent a clone capable of treatment resistance and rapid disease progression.This combined analysis is a work in progress that will include data from additional groups and explore mutation status of other MDS-related genes, TP53 variant allele fraction, clinical risk scores, and data from patients without complex karyotypes.Conclusions:Somatic TP53 mutations are highly enriched in MDS patients with complex karyotypes and identify a group with significantly shortened overall survival. Sequencing of TP53 can improve the prediction of prognosis in MDS patients with 3 or more chromosomal abnormalities. [Display omitted] DisclosuresBejar:Genoptix Medical Laboratory: Consultancy, Honoraria, Licensed IP, no royalties Patents & Royalties, Membership on an entity’s Board of Directors or advisory committees; Celgene: Membership on an entity’s Board of Directors or advisory committees. Haferlach:MLL Munich Leukemia Laboratory: Other. Garcia-Manero:Celgene: Research Funding. Sekeres:Celgene: Membership on an entity’s Board of Directors or advisory committees; Amgen Corp: Membership on an entity’s Board of Directors or advisory committees; Boehringer-Ingelheim Corp: Membership on an entity’s Board of Directors or advisory committees. Campbell:14M Genomics Limited: Consultancy, Equity Ownership.

Prognostic Role of Serum Albumin Level in DLBCL before and during the Rituximab Era. Retrospective GISL Study over 738 Cases

Introduction.Serum albumin (SA) has been shown to be a prognostic marker in many hematological malignancies, and in diffuse large B-cell lymphoma (DLBCL) before the introduction of rituximab. SA may be a surrogate for age, comorbid status, and disease severity. Here, we aimed to assess the role of SA as prognostic marker for overall survival (OS) in patients with newly diagnosed DLBCL treated with and without rituximab (R) containing regimens (CHOP and CHOP-like).Patients and methodWe collected 738 patients complete for IPI factors [age, LDH, extra nodal sites, stage and performance status (PS)] and SA recorded in the GISL database from 1998 to 2012. OS was estimated using the Kaplan-Meier method, with Cox proportional hazard model used to identify potential risk factors for time-to-event data. Optimal cut-off for SA was calculated by means of ROC curve at 5 years of follow-up. Stability of the cut-point was analyzed using bootstrap techniques. The role of SA was explored interacting SA with R and adjusting by IPI. The strength of SA as prognostic factor was evaluated counting the bootstrap frequency of inclusion (BIF, 1000 re-samples) of SA, adjusted by the IPI factors, in Cox PH regression.ResultsOf the 738 patients included in the study 322 (44%) were treated with R-CHOP and 416 (56%) were treated without regimens containing R. The median age at diagnosis was 60 years (range 21-89) and 45%, 25% and 30% were categorized in IPI 0-1, 2 and 3/5 respectively; and the median SA was 3.8 g/dL (range 1.0-6.4). Patients treated with R showed a greater % of high IPI 3/5 (43%) compared to those not treated with R (20%, P<0.001) and a lower SA level (3.7 vs 3.9 g/dL, P=0.001). SA showed correlation with IPI (4.0, 3.8 and 3.5 g/dL in IPI 0-1, 2 and 3/5, respectively, P=0.0001).Overall, the median follow-up was 64 months (range 1-157) with 241 dead. The 5-ys OS was 66% (95 CI 62-70) and the best cut-off for SA was 3.7 g/dL with ROC=68.5%. OS at 5-yrs in patients not treated with R was 74% and 52% for SA ≥3.7 and <3.7 g/dL, respectively (P<0.001). In patients treated with R the 5-yrs OS was 78% and 50% for SA ≥3.7 and <3.7 g/dL (P<0.001). The SA <3.7 g/dL, interacted by R and adjusted by IPI, had HR = 1.71 (95 CI 1.22-2.40, P=0.002) and HR = 1.99 (95 CI 1.31-3.01, P=0.001) in cases not treated and treated with R, and the HRs did not vary between treatments (P=0.582). The BIF of SA<3.7 g/dL adjusted by IPI factors was 88% and 79% in patients without and with R chemotherapy.ConclusionThe SA is a simple variable collected from the clinical-chemistry, that showed a good prognostic property in DLBCL. Although correlated with the IPI, SA has maintained a strong prognostic value both before or during the rituximab era. The biologic role of SA over the survival is unclear. It has been hypothesized that low level of SA may be associated with 1) increased inflammatory response to the tumor, 2) increased cytokine release and 3) poor nutritional status, and due to this condition the patients would not be able to receive the full cycles of chemotherapy. DisclosuresNo relevant conflicts of interest to declare.

The Ki67/CD138 Ratio Independently Predicts Overall Survival in the Upfront Treatment of Newly Diagnosed Multiple Myeloma

Background: Assessment of malignant plasma cell cycling via plasma cell labeling index (PCLI) has been a validated prognostic tool in multiple myeloma (MM) but the test requires specialized technical expertise and is not widely available. Ki67 is a well-known protein marker of cellular proliferation on immunohistochemical (IHC) staining with prognostic utility in other malignancies. In an effort to develop a simpler system to provide analogous information to PCLI, we used a novel IHC co-staining technique for CD138 and Ki67 to quantify plasma cells in active cycling. We then performed a retrospective analysis of the ratio of Ki67/CD138 (Ki67%) in newly diagnosed patients with multiple myeloma receiving 1st-line therapy to correlate with clinical outcomes.Methods: A retrospective cohort study of patients (pts) with treated symptomatic MM was performed by interrogation of the clinical database at the Weill Cornell Medical College / New York Presbyterian Hospital. For inclusion in the analysis, subjects must have started first-line treatment in the period of 2005-2010, and had available bone marrow biopsies. Double-staining with Ki67 and CD138 was performed by IHC. The Ki67% was calculated as the percent of plasma cells expressing CD138 that were also found to express Ki67. Treatment outcomes were stratified and compared based on %Ki67. Response was determined by monthly serum protein electrophoresis / immunofixation (IFX) with free light chain analysis according to International Multiple Myeloma Working Group (IMWG) guidelines. Pts who were IFX negative but had no subsequent bone marrow biopsy were classified as being in unconfirmed complete remission.Results: We identified 151 patients with newly diagnosed MM and available %Ki67 expression who received first-line therapy over the period of 2005-2010. Patient were subdivided into two groups based on %Ki67: Low: %ki67 <= 5%, n = 87; and High: %Ki67 >5, n=64, to allow for comparison of treatment response and survival analysis. Specific therapeutic agent exposure history did not differ significantly between patients. Both groups had similar depth of response rates (ORR) to front-line therapy, Table 1. Median progression-free survival for the high versus low %Ki67 groups approached statistical significance at 54 months (95% CI 30.8,67.4) versus 26.9 months (95% CI 21.6,40.2), respectively (P = 0.083). At data cut-off, there were 30 deaths in the low %Ki67 group (1-yr OS 93%, 5-yr OS 71%) and 36 deaths in the high %Ki67 group (1-yr OS 94%, 5-yr OS 62%). Median overall survival (OS) was not reached for Ki67% <= 5% (95% CI 97.3,NR) vs. 78.9 months (95% CI 55.9,93.1) for Ki67% > 5%, (P = 0.0434), Figure 1. Multivariate cox regression for factors with influence on OS showed that only high-risk cytogenetics (HR 2.05, 95% CI 1.17, 2.92, P = 0.027), ISS (HR 1.835, 95% CI 1.33, 3.60, P = 0.000), and %Ki67 group status had an independent effect on survival outcome. Low (<=5%) versus high (>5%) %Ki67 influenced overall survival with a hazard ratio of 1.76 (CI 1.07,2.92, P = 0.027). Survival after ASCT was significantly longer in the low %Ki67 group with median OS not reached (95%CI, 97.3, NR) versus 86.9 months (95% CI 43.9, NR) for high %Ki67 group (P = 0.04).Discussion: The ratio of IHC double positive Ki67 and CD138 of > 5% is an independent prognostic marker for overall survival in newly diagnosed MM undergoing 1st line therapy. The %Ki67 serves as a simpler and widely available analog to PCLI that can be presently performed in most hematopathology laboratories.Table 1:First Line Treatment and Best Response (modified IMWG Criteria)Ki67% <= 5(N = 87)n (%)Ki67% > 5(N = 64)n (%)PTreatment Exposure*Lenalidomide59 (67.8)48 (75)0.34Thalidomide30 (34.5)14 (21.9)0.09Bortezomib25 (28.7)14 (21.9)0.34Alkylating agent11 (12.6)4 (6.3)0.19ASCT27 (31)22 (34.4)0.66Best ResponseOverall Response(>= Partial response)77 (88.4)57 (89.1)0.41Complete response15 (17.2)22 (34.4)Unconfirmed complete response**14 (16.1)8 (12.5)Very good partial response23 (26.4)15 (23.4)Partial response25 (28.7)12 (18.8)Stable disease9 (10.3)5 (7.8)Progressive disease1 (1.2)2 (3.1)* Percentages do not add to 100% due to instances of concurrent therapy use** Unconfirmed complete response: immunofixation negative, but no confirmatory bone marrow biopsy available [Display omitted] DisclosuresMark:Celgene: Membership on an entity’s Board of Directors or advisory committees, Research Funding, Speakers Bureau; Millennium: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Onyx: Research Funding, Speakers Bureau. Rossi:Celgene: Speakers Bureau. Pekle:Celgene: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Millennium: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau. Perry:Celgene: Speakers Bureau. Coleman:Celgene: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Millennium: Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Onyx: Honoraria. Niesvizky:Celgene: Membership on an entity’s Board of Directors or advisory committees, Research Funding, Speakers Bureau; Millennium: Membership on an entity’s Board of Directors or advisory committees, Research Funding, Speakers Bureau; Onyx: Membership on an entity’s Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Prognostic significance of VEGF-C, semaphorin 3F, and neuropilin-2 expression in oral squamous cell carcinomas and their relationship with lymphangiogenesis.

Neuropilin-2 (NRP2), a receptor for vascular endothelial growth factor C (VEGF-C) and semaphorin 3 F (SEMA3F), is a possible regulator of tumor progression and angiogenesis. However, little evidence of a correlation between NRP2 expression and lymphangiogenesis has been reported. SEMA3F might suppress lymphangiogenesis by competing with VEGF-C for binding to NRP2. We evaluated lymphatic vessel density (LVD), lymphatic vessel invasion (LVI), the expression levels of VEGF-C, SEMA3F, and NRP2 by immunohistochemistry in 80 cases of oral squamous cell carcinomas (OSCCs). In these tumors, the expression of NRP2 was positively associated with T stage classification, lymph node metastasis, LVD, LVI, and the expression of VEGF-C. In contrast, low expression of SEMA3F was significantly related to poor differentiation and higher incidence of lymph node metastasis. Patients expressing high levels of VEGF-C or NRP2, or low levels of SEMA3F had a higher risk of recurrence and shorter overall survival. Multivariate analysis showed that VEGF-C and NRP2 were independent prognostic markers for overall survival. Results indicate SEMA3F is an inhibitor of tumor progression and provide evidence for an association between NRP2 and VEGF-C, lymphangiogenesis, lymph node metastasis. This suggests the prognostic significance and potential therapeutic value of the VEGF-C/SEMA3F/NRP2 axis for OSCCs.

PPM1D overexpression predicts poor prognosis in non-small cell lung cancer.

It has been reported that protein phosphatase, Mg(2+)/Mn(2+) dependent, 1D (PPM1D) plays an important role in cancer tumorigenesis. However, the clinical and functional significance of PPM1D expression has not been characterized previously in non-small cell lung cancer (NSCLC). The purpose of this study was to assess PPM1D expression and to explore its contribution to NSCLC. We examined PPM1D messenger RNA (mRNA) expression in 53 NSCLC tissues and matched adjacent noncancerous tissues by quantitative reverse transcription PCR (qRT-PCR). Furthermore, the PPM1D protein expression was analyzed by immunohistochemistry in 157 NSCLC samples. The relationship between PPM1D expression and clinicopathological features was analyzed by appropriate statistics. Kaplan-Meier analysis and Cox proportional hazards regression models were used to investigate the correlation between PPM1D expression and prognosis of NSCLC patients. The relative mRNA expression of PPM1D was significantly elevated in NSCLC tissues as compared with adjacent noncancerous tissues (P < 0.001). The high expression of PPM1D in NSCLC tissues was significantly correlated with tumor grade (P = 0.006), tumor size (P = 0.017), clinical stage (P = 0.001), and lymph node metastases (P = 0.002). Kaplan-Meier survival analysis revealed that high PPM1D expression correlated with poor prognosis of NSCLC patients (P < 0.001). Multivariate analysis showed that PPM1D expression was an independent prognostic marker for overall survival of NSCLC patients. In conclusion, PPM1D plays an important role in the progression of NSCLC. PPM1D may potentially be used as an independent biomarker for the prognostic evaluation of NSCLC.

Abstract 4155: Genome-wide association study of survival in early stage non-small cell lung cancer

Abstract Lung cancer, especially non-small cell lung cancer (NSCLC), is the leading cause of cancer-related deaths all over the world. Studies have indicated that molecular biomarkers including genetic variants may provide additional values for the targeted treatments and clinical outcomes of NSCLC patients. In this study, we evaluated the associations between genome-wide genetic variants and the survival of NSCLC in a cohort of 365 early-stage NSCLC patients with surgery, and replicated sixteen promising loci in another independent cohort with 327 early-stage NSCLC patients in Chinese population. The results showed that rs10023113 in CAMK2D was consistently associated with survival of early-stage NSCLC in the GWAS scan and the replication cohort [GWAS scan: hazard ratio (HR) = 2.84; 95% confidence interval (CI) = 1.90-4.23, P = 1.29×10-6; replication cohort: HR = 2.19, 95% CI = 1.15-4.21, P = 1.80×10-2]. When combining all the patients, the results showed that the variant allele of rs10023113 was significantly associated with poor prognosis of NSCLC with P value reaching 3.40×10-7 (HR = 2.30, 95% CI = 1.67-3.17). These findings suggest that CAMK2D rs10023113 may be a potentially prognostic marker for overall survival of early-stage NSCLC patients in Chinese population. Citation Format: Hongxia Ma, Guangfu Jin, Zhibin Hu, Hongbing Shen. Genome-wide association study of survival in early stage non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4155. doi:10.1158/1538-7445.AM2014-4155

Abstract 4718: ERO1L, a novel prognostic marker of gastric cancer patient survival, mediates cancer cell invasion and chemoresistance

Abstract Gastric cancer is the second cause of cancer-related deaths worldwide. Even though within same stage, gastric cancer patients present diverse clinical manifestations and prognosis. Molecular markers will be important in predicting patients' outcomes and tailoring personalized treatments according to individual biology. In this study, we analyzed the gene expression profile of human gastric cancer (published on CCR) to identify potential biomarkers. We found that TXN family genes and ERO1L were significantly overexpressed and related to prognosis. We evaluated ERO1L significantly overexpressed in gastric cancer and ERO1L was very highly expressed in hypoxic condition. The other study has identified ERO1L as included in the small group of eight genes predicting poor survival of patients with pulmonary adenocarcinoma. We show that ERO1L is a prognostic marker for overall survival among patients with gastric cancer. To investigate the function of ERO1L gene in gastric cancer cell line (AGS and MKN1), we tested the effect of ERO1L expression on gastric cancer cells. To determine the biologic role of ERO1L in regulating cancer cell proliferation, stable transfection of shRNA and expression vector for ERO1L in gastric cancer cells. Our results showed that shERO1L decrease cell proliferation. Next, we tested whether or not the ERO1L gene is involved in progression to metastatic disease in gastric cancer, especially in tumorigenesis, including migration and invasion. In Functional studies, ERO1L silencing decrease gastric cancer cell migaration and invasion, whereas EROL expression significantly increase cell migration and invasiveness. We examined whether ERO1L plays a role in chemoresistance in gastric cancer cells. After silencing or overexpressing ERO1L, we carried out cell viability assays in gastric cancer cells with a panel of chemotherapy agents used to treat gastric cancer patients: a microtubule stabilizer (paclitaxel) and an antimetabilite (5-FU). Surprisingly, Silencing ERO1L expression led gastric cancer cells to become more sensitive to paclitaxel and 5-FU. In conclusion, our findings show that a prognostic molecular signature that can predict the poor progression of gastric cancer tumors. Furthermore, unequal distribution of expression patterns reflecting activation of ERO1L with different survival rates supports a personalized target therapy in gastric cancer with biomarker gene signature driven patient selection. Citation Format: So-Young Seol, Jae Yun Lim, Sun Och Yoon, Soon Won Hong, Jong Won Kim, Seung Ho Choi, Jae Yong Cho. ERO1L, a novel prognostic marker of gastric cancer patient survival, mediates cancer cell invasion and chemoresistance. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4718. doi:10.1158/1538-7445.AM2014-4718

Abstract 4723: Prognostic significance of EGFR expression in localized gastric cancer

Abstract Background: The role of epidermal growth factor receptor (EGFR) expression as predictive or prognostic biomarker in gastric cancer is still in controversy. The aim of this study is to find prognostic role of tumor EGFR expression in gastric cancer patients after curative resection. Methods: From January 2000 to September 2003, gastric cancer patients who received curative gastrectomy were included in this study. We retrospectively collected clinical characteristics and survival data of the patients, and analyzed EGFR expression on tissue microarrays from formalin-fixed paraffin-embedded specimens of primary tumor tissues by immunochemical staining (IHC). EGFR expression was scored to 0, 1+, 2+ or 3+ based on intensity of reactivity. Results: Of total 937 patients, 295 (31.5%) patients, 101 (10.8%) patients and 36 (3.8%) patients showed EGFR 1+, 2+, and 3+ expression by IHC, respectively. EGFR overexpression (2+/3+) is more frequently found in intestinal type than diffuse or mixed type (24.2% vs. 5.1%, P&amp;lt;0.001). In stage III disease according to AJCC 7th edition, unlike other stages, the patients with EGFR overexpression (2+/3+) had longer disease free survival (hazard ratio [HR], 0.639; 95% confidence interval [CI], 0.434-0.938; P=0.022) and longer overall survival (HR, 0.630; 95% CI, 0.445-0.890, P=0.009) than other patients in univariate analysis. Additionally, in 279 patients who had stage III gastric cancer with diffuse type histology and received 5-fluorouracil (5-FU) based adjuvant chemotherapy, EGFR overexpression was favorable prognostic marker for overall survival (HR, 0.365; 95% CI, 0.161-0.827; P=0.016) in multivariate analysis, while old age and lymphovascular invasion were poor prognostic factors. Conclusion: EGFR overexpression was potentially favorable prognostic biomarker for stage III gastric cancer, especially in the patients who received 5-FU based adjuvant chemotherapy. Citation Format: Ji Soo Park, Hyo Song Kim, Yoon Sung Bae, Sung Hoon Noh, Hyunki Kim. Prognostic significance of EGFR expression in localized gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4723. doi:10.1158/1538-7445.AM2014-4723

Lewis Lung Cancer Tumor Growth Inhibition by Disruption of Transforming Growth Factor Beta (TGF-β1) Pathway in Murine Model

Results: The mean pre-diagnosis NLR was significantly associated with lung cancer patient overall survival. Compared to patients with a low NLR level, those with elevated NLR exhibited a significant unfavorable overall survival with a hazard ratio (HR) of 1.50 (95% confidence interval [CI]: 1.30-1.75, p Z 7.2 x 10) after adjusting for other covariates. The association was internally validated with bootstrap (p < 0.01 at 99% resampling). The effect of NLR significantly interacted with race (p interaction Z 0.005), and smoking status (p interaction Z 0.02). The association was more pronounced in nonwhite patients (HR Z 2.16, 95% CI Z 1.61-2.89, p Z 2.62 x 10) than in white patients (HR Z 1.31, 95% CI Z 1.10-1.57, P Z 0.002) (p interaction Z 0.005), and in never smokers or patients with unknown smoking status (HRZ 2.60, 95%CIZ 1.61-4.19, pZ 9.1 x 10) than in past smokers (HR Z 1.55, 95% CI Z 1.24-1.93, p Z 0.0001) or current smokers (HR Z 1.32, 95% CI Z 1.04-1.68, p Z 0.02). Flexible parametric survival model demonstrated that the increased risk for death associated with elevated pre-diagnosis NLR persisted over 18 months after diagnosis. Conclusions: Our data suggests that pre-diagnosis NLR might be used as an independent prognostic marker for overall survival for lung cancer patients. The effect of NLR on overall survival seems to be more pronounced for those patients with races other than white, and for nonsmokers when compared with the past and current smokers. We plan to further analyze the effect of the pre-diagnosis NLR on overall survival in the context of nutritional status and performance status of patients with lung cancer. Author Disclosure: V. Bar Ad: None. Y. Lai: None. B. Lu: None. J. Palmer: None. R. Myers: None. Z. Ye: None. C. Wang: None. R. Axelrod: None. B. Campling: None. M. Werner-Wasik: None. S. Cowan: None. N. Evans: None. R. Kumar: None. B. Hehn: None. C. Solomides: None. H. Yang: None.