Predictive Marker In Patients Research Articles

MicroRNA-21 in Colorectal Cancer: “Just Another Brick in the Wall”?

MicroRNAs (miRNAs)—small, noncoding RNA strands consisting of 18 to 25 base pairs—have been investigated extensively during the last decade. Initially seen as superfluous material, it has become clear that they play a critical role in translational and posttranscriptional regulation of protein processing. In cancer cells those processes are altered and cancer cells therefore display an irregular miRNA configuration. Many solid tumors have specific patterns of miRNA expression levels, which are used in several commercially available, and, in some cases, Food and Drug Administration approved, diagnostic tests. It is possible to determine tumor origin and, in part, the histologic subtype of solid tumors by its miRNA signature. Evaluating miRNA in tissue and body fluids is therefore an emerging field of biomarker development. MicroRNA 21 (miR-21) is found to be upregulated in a wide range of solid tumors including lung, breast, gastric, colon, and glioblastoma (1), and therefore as a single marker is not specific for colon cancer. Its reported RNA targets include transforming growth factor s (TGF-s), phosphatase and tensin homolog (PTEN), programmed cell death protein 4 (PDCD4), protein sprouty 1 (Spry-1), phosphatase of regenerating liver 3 (PRL-3), and nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB). Therefore, miR-21 may be involved in the modulation of critical steps in carcinogenesis such as proliferation, apoptosis, and migration (2). It has further been shown that colon cancer cell lines with higher expression levels of miR-21 have an enhanced ability of motility and invasion (3). Slaby and others demonstrated that tissue miR-21 expression is rising with higher Union for International Cancer Control stage (4,5) and therefore represents a marker for clinicopathologic features of the disease. First data have been published to state a role of miR-21 in chemoresistance to gemcitabine in cholangiocellular cancer cell lines (6) and 5-fluorouracil in colon cancer cells (7). Higher miR-21 tissue levels could further be linked to a decreased pathologic tumor response in patients treated with FOLFOX in a neoadjuvant setting for locally advanced colorectal cancer (CRC) (8). Owing to its capability to be secreted by cancer cells, miR-21 has been the first miRNA to be detected in the plasma in patients with diffuse large B-cell lymphoma in 2008 (9). These specifics taken together, miR21 represents an almost ideal candidate for a diagnostic tool in the diagnosis and treatment monitoring of CRC. The article by Toiyama et al. (10) in this issue of the Journal confirms previous data showing that miR-21 serum levels are higher in patients with CRC than in healthy controls (2,11). The authors showed that miR-21 levels increased with the stage of CRC. They suggested that miR-21 levels in the serum may be a useful diagnostic tool. MiR-21 serum levels were measured in 186 patients with CRC, 43 patients with advanced adenomas, and 53 healthy controls. The authors showed that miR-21 serum levels were statistically significantly higher in patients presenting with advanced adenomas or CRC compared to healthy controls. A differentiation between advanced adenomas and CRC was not achieved. Within the cohort of patients with CRC, serum levels of miR-21 were statistically significantly higher in metastatic (stage IV) disease than in stage I and II disease. Using a cutoff value determined by the receiver operating characteristic procedure, miR-21 levels had diagnostic and prognostic value. This association remained statistically significant after multivariate analysis adjusting for age, sex, TNM stage, lymph node metastasis, carcinoembryonic antigen level, distant metastasis, and pathologic differentiation. Several questions remain about the clinical use of miR-21 as a biomarker for CRC. All published miRNA data have been generated from retrospective analyses; therefore, prospective testing in independent control studies is critical for validation. Those studies should further evaluate whether miR-21 can serve as a prognostic or predictive marker in patients with CRC. It is essential to establish standardized techniques to measure serum miRNA levels. The authors tried to address this problem by

Cytidine deaminase status as a predictive marker in patients with hematologic malignancies treated with azacytidine or cytarabine.

7117 Background: Cytarabine and azacytidine are mainstays for treating haematological malignancies. As most nucleosidic analogs, both azacytidine and cytarabine are metabolized in the liver by an exclusive enzymatic step driven by cytidine deaminase (CDA). CDA is highly polymorphic and dysregulations have been repeatedly associated with poor clinical outcome with gemcitabine. Methods: We have used a test to determine, on a phenotype basis, CDA status in patients. This test was used prospectively in a subset of 39 adult patients (16F, 23M, mean age 77 years), all treated for various haematological malignancies (i.e., CML, AML, lymphomas, myelodysplastic syndromes) with either aza-cytidine or a cytarabine-containing regimen. Response and treatment-related toxicities were monitored following current standards. In addition, impact of CDA status on azacytidine tolerance was evaluated in mice with or without CDA deficiency, as a proof of concept for the actual implication of metabolic deregulations in the toxicities observed in patients. Results: In patients, mean CDA activity was 3.7 +/-2.8 U/mg (min: 1, max: 14.8 U/mg). Ten out of 39 patients (i.e., 25%) showed low CDA activities and were considered as PM. Conversely, 8 patients (i.e., 20%) displayed CDA activities particurlary elevated (i.e., > 6U/mg) and were considered as UM patients. Of note, PM patients all showed severe toxicities, including two toxic-deaths. Conversely, UM patients showed little efficacy when treated with either azacytidine or cytarabine. In mice with CDA-deficiency, standard azacytidine led to profound and long-lasting neutropenia, as compared with normal mice. Drug monitoring confirmed that individuals with low CDA activity and toxicities showed higher concentrations of azacytidine as compared with normal individuals. Conclusions: Overall this pilot study strongly suggests that CDA status could be a relevant marker for predicting clinical outcome in patients treated with either azacytidine or cytarabine. CDA status could be further used as a covariate to tailor drug dosage so as to ensure an optimal efficacy/toxicity balance in patients with haematological malignancies.

The vascular endothelial growth factor as a predictive and prognostic factor in patients with advanced gastric cancer treated with docetaxel, cisplatin, and 5-florouracil (DCF) regimen.

e15191 Background: Vascular endothelial growth factor (VEGF) is a key element stimulating the angiogenesis in solid tumors. In the current study, we aimed to assess the role of plasma VEGF levels as a prognostic and predictive marker for advanced gastric cancer patients treated with a modified docetaxel, cisplatinum and 5-florouracil (DCF) regimen. Methods: Thirty consecutive patients treated with a modified DCF regimen were included in the study. The plasma VEGF levels of the patients before treatment and following two cycles of chemotherapy were assayed. Results: The modified DCF regimen was well tolerated and yielded a median overall survival (OS) time of 9,0 months and 2-year OS rate of 20%. Good performance status, lower pre-treatment VEGF levels,and decrease in VEGF levels following chemotherapy were found to be independent prognostic factors for OS. Conclusions: The results of the current study suggest that the corrected plasma levels of VEGF could be used as a prognostic factor for advanced gastric cancer patients and a decrease in VEGF levels following 2 cycles of chemotherapy as a predictive marker for patients receiving DCF chemotherapy regimen.

Abstract 2372: Predictive markers in patients with non-small cell lung cancer with pemetrexed treatment.

Abstract Background: This study is desiged to evaluate biomarkers that are correlated with the efficacy of pemetrexed in patients with non-small-cell lung cancer (NSCLC). Methods: Patients with stage III or IV NSCLC who received pemetrexed monotherapy after failure of one or more chemotherapy regimen between January 2003 and August 2011 were included. Polymorphism of reduced folate carrier gene (SLC19A, rs1051298), 5,10-methylenetetrahydroflate reductase (MTHFR, rs1801133) and thymidylate synthase (TS, rs45445694, rs16430) gene were analyzed by PCR and direct sequencing method using peripheral blood. Results: Total 123 patients were included. The median age was 61 years (range 35-82). Fifty eight (47.2%) patients were female. Predominant histology was adenocarcinoma (92.7%). Metastatic diseases were 114 (80.5%) patients. Patients received one or more lines of systemic therapy before pemetrexed therapy, one line in 53 (43.1%) patients, two in 51(41.5%), three in 14 (11.02%), and four in 4 (3.3%). Median 4 cycles of chemotherapy were given (range 1-44). Response rate was 22.7% (CR, n=2, PR, n=26) and stable disease was detected in 54 patients (43.9%). Median follow up duration was 28.7months (range 4.07-84.33months). Median progression free survival (PFS) was 4.2 months (95% CI: 2.9-5.6) and median overall survival (OS) was 18.9 months (95% CI: 13.3-24.6). Clinical factors like, sex, histology of tumor (adenocarcinoma vs, nonadenocarcinoma), differentiation, smoking status and EGFR mutation status (active mutation vs. other mutation vs. wild type) did not have any significant difference on response and PFS. Only polymorphisms of TS promoter enhancer region (TSER) were related with PFS. 2R/2R repeat in the TSER (2R/2R vs 2R/3R, 3R/3R 1.0m vs 4.3m, p<0.01) and CC homozygote variant in second repeat of the 3R allele (CC vs GC, GG 2.8m vs 5.0m, p=0.04) had poor PFS. All polymorphism did not have effect on response rate and overall survival. Conclusions: This study suggests that polymorphisms of TSER associated PFS in pemetrexed treatment. Further prospective studies will be needed to validate these polymorphism as a useful biomarker of pemetrexed treatment in NSCLC patients. Citation Format: Hiun S. Chae, Yoon Ho Ko, Eun Kyoung Jeon, Sook Hee Hong, Jeong-Oh Kim, Seung Joon Kim, Kyo Young Lee, Young Kyoon Kim, HoonKyo Kim, Seok young Park, In Sook Woo, Jin Hyoung Kang. Predictive markers in patients with non-small cell lung cancer with pemetrexed treatment. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2372. doi:10.1158/1538-7445.AM2013-2372

Thymidylate synthase and ERCC1 as predictive markers in patients with pulmonary adenocarcinoma treated with pemetrexed and cisplatin

Increased expression of thymidylate synthase (TS) is thought to be associated with resistance to antifolate drugs such as pemetrexed. Excision repair cross-complementation group 1 (ERCC1) is a predictive marker for platinum-based chemotherapy. This study evaluated whether the expression of TS and ERCC1 proteins is associated with clinical outcomes of the patients with pulmonary adenocarcinoma who were treated with pemetrexed/cisplatin as first-line chemotherapy. The expressions of TS and ERCC1 were evaluated by immunohistochemistry in biopsy specimens obtained from patients with pulmonary adenocarcinoma who had received pemetrexed/cisplatin as first-line treatment. Patients were categorized according to median H-score. Response rate (RR), progression-free survival (PFS) and overall survival (OS) were analyzed retrospectively. Both low TS and ERCC1 expressions were significantly associated with better RR (p=0.037 and p=0.015, respectively) and longer PFS (p<0.001 and p=0.004, respectively). Low ERCC1 expression was also associated with longer OS (p=0.003) while TS only showed a trend (p=0.105). TS expression was independent predictor for the better PFS in multivariate analysis (hazard ratio [HR]=0.32, 95% confidence interval [CI]: 0.14–0.76). Combining the two markers, the low TS/low ERCC1 group showed significantly longer PFS (HR=0.48, 95% CI: 0.26–0.75) and OS (HR=0.57, 95% CI: 0.36–0.89) compared with high TS/high ERCC1 group.Protein expressions of TS and ERCC1 were associated with clinical outcomes in patients with pulmonary adenocarcinoma who were treated with pemetrexed/cisplatin as first-line chemotherapy. TS and ERCC1 protein expressions can be potential predictive markers in this setting.

Predictive and prognostic values of Tau and BubR1 protein in prostate cancer and their relationship to the Gleason score

The aim of this study is to detect the expression levels of spindle assembly checkpoint protein-BubR1 and microtubule-associated protein-Tau in human prostate cancer tissues of different Gleason score, and to test whether there is a relationship between their expression levels and clinicopathologic parameters including response to docetaxel treatment, Gleason score, and overall survival (OS). Moreover, to test whether Tau protein expressed in the cancerous prostate tissue is phosphorylated. Thirty patients who received at least three cycles docetaxel for metastatic castrate-resistant prostate cancer were included into the trial. The patients' formalin-fixed and paraffin-embedded prostate tissue specimens were retrospectively obtained from the pathology department archives of Ege University School of Medicine. The expression status of BubR1 protein was defined by immunohistochemical (IHC) using the anti-BubR1 antibody. The expression status of Tau protein was defined by IHC using the two types of Tau antibodies: anti-Tau-1 antibody (that recognizes Tau only in its dephosphorylated form) and anti-PHF-Tau antibody (that recognizes all isoforms of human Tau proteins independent of its phosphorylation status). The BubR1 and Tau were overexpressed in about 63 and 23 % of the study group, respectively. Tau overexpression was significantly associated with lower Gleason score. There was no significant association between the expression levels of BubR1 and Tau proteins, and docetaxel response. Reduced BubR1 expression was strongly associated with longer survival (P = 0.008), whereas Tau expression status did not effect survival. Moreover, the Tau expression of cancerous prostate tissue was highly dephosphorylated. In this clinicopathological study, our findings did not confirm the preclinical observations that low BubR1 and Tau expression confer selective sensitivity to microtubulisin drugs. Our data imply that reduced BubR1 expression was a predictor for longer OS, and the possibility that high Tau expression may be involved in better prognosis due to its relationship to the Gleason score. Furthermore, our data suggest that both Tau and BubR1 may be a promising prognostic marker rather than predictive marker in patients with prostate cancer.

Can Serum be Used for Analyzing the EGFR Mutation Status in Patients with Advanced Non-small Cell Lung Cancer?

Epidermal growth factor receptor (EGFR) mutations as prognostic or predictive marker in patients with non-small cell lung cancer (NSCLC) have been used widely. However, it may be difficult to get tumor tissue for analyzing the status of EGFR mutation status in large proportion of patients with advanced disease. We obtained pairs of tumor and serum samples from 57 patients with advanced NSCLC, between March 2006 and January 2009. EGFR mutation status from tumor samples was analyzed by genomic polymerase chain reaction and direct sequence and EGFR mutation status from serum samples was determined by the peptide nucleic acid locked nucleic acid polymerase chain reaction clamp. EGFR mutations were detected in the serum samples of 11 patients and in the tumor samples of 12 patients. EGFR mutation status in the serum and tumor samples was consistent in 50 of the 57 pairs (87.7%). There was a high correlation between the mutations detected in serum sample and the mutations detected in the matched tumor sample (correlation index 0.62; P<0.001). Twenty-two of 57 patients (38.5%) received EGFR-tyrosine kinase inhibitors as any line therapy. The response for EGFR-tyrosine kinase inhibitors was significantly associated with EGFR mutations in both tumor samples and serum samples (P<0.05). There was no significant differences in overall survival according to the status of EGFR mutations in both serum and tumor samples (P>0.05). Serum sample might be alternatively used in the difficult time of getting tumor tissue for analyzing the status of EGFR mutation status in patients with advanced NSCLC.

The association of ventricular tachycardia and endothelial dysfunction in the setting of acute myocardial infarction with ST elevation

BackgroundVentricular tachycardia (VT) is frequently seen in ischemic settings like acute myocardial infarction with ST segment elevation (STEMI). Endothelial dysfunction (ED) represents inflammation and the loss of all protective features of the endothelium. We aimed to examine the association between VT and ED in patients with STEMI.Material/MethodsThe study included 90 subjects (30 with VT and acute STEMI, 30 with STEMI without VT, and 30 controls). Sera of all subjects were tested on ED markers by enzyme immunoassay: sICAM-1 (intracellular adhesive molecule-1), sVCAM-1 (vascular adhesive molecule-1), P- and E-selectins, and VEGF (vascular endothelial growth factor). In addition, CRP (C-reactive protein) was detected.ResultsSignificantly increased values of low-density lipoprotein, triglycerides, leukocytes, creatinine, and the number of cigarettes smoked were observed among patients with VT+STEMI in comparison to controls. The levels of E-selectin were significantly lower in the VT+STEMI group than in the other groups, while the levels of VCAM-1 were significantly higher in the groups with STEMI and VT+STEMI compared to the controls. Lower levels of VEGF were recorded in STEMI and VT+STEMI groups compared to the control group. A significant correlation between CRP and VCAM-1 in patients with VT +STEMI was demonstrated.ConclusionsWe showed that ED may have a role in the immunopathogenesis of VT in patients with STEMI. The role of sE-selectin and correlation of sVCAM-1 with CRP as possible ED predictive markers in patients with VT+STEMI should be further investigated in a large cohort of patients.

Is ATP7B a Predictive Marker in Patients With Ovarian Carcinoma Treated With Platinum-Taxane Combination Chemotherapy?

This study examined the prognostic significance of copper-transporting P-type adenosine triphosphatase (ATP7B) expression in patients with ovarian carcinoma treated with platinum-taxane combination chemotherapy. Expression of ATP7B in ovarian carcinoma was assessed by immunohistochemistry and clinical data collected by retrospective review of medical charts. Overexpression of ATP7B was identified in 25 (29.1%) of 86 ovarian carcinomas. The frequency of ATP7B expression in clear cell carcinomas was significantly higher than that in serous high-grade carcinomas (P < 0.05). We observed no statistically significant correlations between high ATP7B protein expression and either disease-free survival (P = 0.722) or overall survival (P = 0.389). Our study is the first to demonstrate a lack of statistically significant differences between ATP7B positive and negative cases with respect to prognosis of patients with ovarian carcinoma treated with a platinum-taxane combination regimen. However, that ATP7B expression in clear cell carcinomas was significantly higher than that in serous carcinomas may partially explain the difference in chemotherapeutic response and prognosis between patients with these 2 types of carcinomas.

Correlation between BRCA1 expression and clinicopathological factors including brain metastases in patients with non-small-cell lung cancer

Previously identified as a breast and ovarian cancer susceptibility gene, BRCA1 has gained major scientific interest as a potential prognostic and/or predictive marker for various tumors, including non-small-cell lung cancer (NSCLC), the leading cause of cancer related mortality worldwide. BRCA1 plays a central role in DNA damage response (DDR. It undergoes phosphorylation by various DDR kinases at different serine residues, of which ser1524 is known to be specifically phosphorylated by ATM in response to genotoxic stress. We performed BRCA1 immunohistochemistry on several tissue microarrays (TMAs) of 113 early (I, II stage) and advanced (III, IV stage) NSCLCs, using MS110 antibody against the BRCA1 N-terminal and S1524 antibody against the phosphorylated form of BRCA1 protein at ser1524 (Abcam). Patients with III and IV stage disease were treated by adjuvant cisplatin-based chemotherapy. Staining results were correlated with overall survival (OS), disease free survival (DFS) and with the occurrence of brain metastases. BRCA1 S1524 nuclear positivity was significantly correlated with longer OS and DFS in stage I and II patients (P<0.05), while OS and DFS were shorter in S1524 positive stage III and IV patients (P<0.05). No significant correlation was found with brain metastases. The results show that BRCA1 phosphorylaton, at least in ser1524, differentiates the fate of early and advanced NSCLC as well as response to chemotherapy, but the underlying mechanisms are not completely understood. Detection of phosphorylated forms of BRCA1 might serve as a useful prognostic and predictive marker for patients with NSCLC.

Is Mean Platelet Volume a Predictive Marker in Patients With Venous Thrombosis?

>> boses have been considered as distinct physiopathological concepts through thrombus formation. Arterial thrombi are found in areas of high shear stress and consist mostly of platelets and fibrin. Venous thrombi usually develop in low flow vessels and consist predominantly of erythrocytes and fibrin. Although the data suggest the MPV may potentially be useful as a predictive marker, there are a number of concerns related to the technological limitations and variations in the measurement methods of the MPV that need to be considered. Mean platelet volume does not report a routine part of the complete blood count because of the EDTA induced changes over time. The EDTA-induced platelet shape changes result in a progressive increase in MPV with impedance technology. In general, the MPV increases up to 30% within 5 minutes of exposure and increases further by 10% to 15% over the next 2 hours. 3,4 However, some investigators have reported inconsistent increases in MPV with EDTA storage up to 50%. 3-5 The effect of EDTA on

High concordance between HercepTest immunohistochemistry and ERBB2 fluorescence in situ hybridization before and after implementation of American Society of Clinical Oncology/College of American Pathology 2007 guidelines

Human epidermal growth factor receptor 2 (HER2, ERBB2) is an important critical predictive marker in patients with invasive breast cancer. It is thus imperative to ensure accuracy and precision in HER2 and ERBB2 testing. In 2007, the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP) proposed new guidelines for immunohistochemistry and fluorescence in-situ hybridization scoring in an effort to improve accuracy and utility of these companion diagnostic tests. The goal of the 2007 guidelines was to improve concordance rates between the diagnostic tests and decrease the number of inconclusive cases. This study examines the impact in concordance rates and number of inconclusive cases based on the recent change in guidelines in a large study cohort. HER2 immunohistochemistry and ERBB2 fluorescence in-situ hybridization were performed on all specimens from our facility from years 2003 through 2010 (n=1437). Cases from 2003–2007 (n=1016) were scored using Food and Drug Administration guidelines, with immunohistochemical 3+ cases staining >10% of tumor cells and fluorescence in-situ hybridization amplification cutoff value of 2.0. The 2007 guidelines were implemented and scored accordingly for cases from 2008–2010 (n=421), with immunohistochemical 3+ cases staining >30% of tumor cells and fluorescence in-situ hybridization amplification cutoff value of 2.2. We compared concordance rates before and after 2007 guidelines. For the 2003–2007 study population, the concordance rate between the assays was 97.6% with a corresponding kappa coefficient (k) of 0.90. For the 2008–2010 study population, concordance rate was 97.6% with a corresponding k of 0.89. There was no significant difference in number of inconclusive rates before and after 2007 guidelines. In our study, implementation of the new ASCO/CAP 2007 HER2 guidelines did not show a significant difference in concordance rates and did not decrease the number of inconclusive cases.

Gastric Cancer: Basic Aspects

The worldwide incidence and mortality of gastric cancer (GC) remain high, and new concepts for diagnosis and treatment are needed. In this review, we summarize recent studies that applied next-generation sequencing approaches and also report the latest development in microRNA research. Two recently published studies identified somatic mutations in ARID1A gene in GC using exome sequencing. On the other hand, dysregulation of microRNA expression can alter processes such as proliferation, apoptosis, invasion, and metastasis. These novel markers may prove to be useful in earlier diagnosis and as prognostic or predictive markers in patients with GC .

Pair Box 8 (PAX8) protein expression in high grade, late stage (stages III and IV) ovarian serous carcinoma

ObjectivesPair-Box 8 (PAX8) is a transcription factor which has been found to be overexpressed in ovarian serous carcinoma (OSC). Silencing PAX8 by using shRNA led to a drop in cell viability in ovarian cancer cell lines, suggesting its use as a targeted therapeutic agent. The prognostic value of PAX8 in OSC is still widely unknown. The aim of this study was to evaluate PAX8 as a prognostic biomarker in patients with advanced stage OSC. MethodsPAX8 was evaluated using immunohistochemistry on a tissue microarray of 148 OSC and the expression was correlated to the following clinico-pathologic variables; age of diagnosis, tumor stage, optimal debulking, recurrence free survival (RFS) and overall survival (OS). ResultsWe found that PAX8 was expressed in 61% of cases. There was no association between PAX8 and tumor stage, optimal debulking and disease recurrence. In addition, PAX8 failed to have a predictive value in disease outcome. ConclusionDespite showing that PAX8 protein is not a useful predictive marker in patients with high grade, advanced stage OSC, its overexpression in a large number of these cases makes the inhibition of PAX8 a very attractive targeted therapy.

Expression of Vascular endothelial growth factor in Ewing's sarcoma

Vascular endothelial growth factor (VEGF) is the most potent stimulator of angiogenesis. The aim of this study was to evaluate the role of serum VEGF as a diagnostic, predictive and prognostic marker in Ewing's sarcoma. Patients with histopathologically proven diagnosis of Ewing's sarcoma without prior chemotherapy or radiotherapy were invited to take part in the study. Pre-chemotherapy, post-chemotherapy and post-surgery blood samples were collected for analysis of serum VEGF levels. Blood samples from ten sex- and age-matched healthy volunteers were collected for estimation of VEGF levels to act as control. Human VEGF Elisa kit (Bender Medsystem, Austria) was used to assess the serum VEGF levels. A total of nine cases of Ewing's sarcoma were included in the study. Mean age in the group was 12.44 years (range, seven to 18 years). Mean and median serums VEGF level in the study population were 4,547.78 pg/ml and 3,780.00 pg/ml, respectively. Ten age- and sex-matched healthy volunteers were selected as controls. No significant correlation was obtained between serum VEGF, age, sex and tumour size. Mean serum VEGF was significantly raised in the study group as compared to controls (p=0.001). We observed a significant decline in serum VEGF level following neoadjuvant chemotherapy (p = 0.008). No correlation could be established between serum VEGF level pulmonary metastasis and overall survival. Serum VEGF might have a role as a diagnostic and predictive marker in patients with Ewing's sarcoma.

Profiling of circulating tumor cells isolated from 105 metastatic gastric cancer patients revealed HER2 overexpression/activation for potential use in clinical setting.

10535 Background: Gastric cancer (GCA) is the second leading cause of cancer mortality in the world. Survival of patients with advanced GCA treated with chemotherapy remains low. New targeted therapies are urgently needed. There is mounting evidence of the role of HER2 overexpression in patients with GCA, and it has been highly correlated to poor outcomes with more aggressive disease. The ability to accurately determine HER2 status by testing circulating tumor cells (CTCs) may improve patient treatment by allowing ongoing assessment of HER2 status during treatment and/or identifying additional patients who could potentially benefit from HER2- targeted therapy. Methods: The Collaborative Enzyme Enhanced Reactive-immunoassay (CEER) was utilized to determine the expression and activation (phosphorylation) levels of HER2 in CTCs isolated from blood specimens obtained from 105 metastatic GCA patients. Results: Utilizing the CEER platform, the levels of HER2 expression and phosphorylation were determined for CTCs isolated from metastatic GCA patients. Evaluable CTCs were found in 33% (35/105) of enrolled patients. Out of 35 patients, 7 patients (20%) have high HER2 over expression, 6 patients (17%) have moderate HER2 expression and 11 patients (31%) have HER2 activation (phospho positive) with no HER2 over-expression. Conclusions: When CTCs were present, the CEER assay identified varying levels of HER2 involvements in 68% of metastatic GCA patients. HER2 positive CTCs could serve as a prognostic and/or predictive marker in patients with advanced GCA and CTC-HER2 profile shifts can be utilized to monitor the treatment efficacy.

IGFBP Ratio Confers Resistance to IGF Targeting and Correlates with Increased Invasion and Poor Outcome in Breast Tumors

To improve the significance of insulin-like growth factor-binding protein 5 (IGFBP-5) as a prognostic and potentially predictive marker in patients with breast cancer. Increased IGFBP-5 expression was identified in MCF-7 cells resistant (MCF-7R4) to the IGF-1R/insulin receptor (InsR) inhibitor BMS-536924 and its role examined by targeted knockdown and overexpression in multiple experimental models. Protein expression of IGFBP-5 was measured by immunohistochemistry in a cohort of 76 patients with breast cancer to examine correlative associations with invasive tumor fraction and outcome. The use of a combined IGFBP-5/IGFBP-4 (BPR) expression ratio was applied to predict anti-IGF-1R/InsR response in a panel of breast cancer lines and outcome in multiple breast tumor cohorts. IGFBP-5 knockdown decreased BMS-536924 resistance in MCF-7R4 cells, whereas IGFBP-5 overexpression in MCF-7 cells conferred resistance. When compared with pathologically normal reduction mammoplasty tissue, IGFBP-5 expression levels were upregulated in both invasive and histologically normal adjacent breast cancer tissue. In both univariate and multivariate modeling, metastasis-free survival, recurrence free survival (RFS), and overall survival (OS) were significantly associated with high IGFBP-5 expression. Prognostic power of IGFBP-5 was further increased with the addition of IGFBP-4 where tumors were ranked based upon IGFBP-5/IGFBP-4 expression ratio (BPR). Multiple breast cancer cohorts confirm that BPR (high vs. low) was a strong predictor of RFS and OS. IGFBP-5 expression is a marker of poor outcome in patients with breast cancer. An IGFBP-5/IGFBP-4 expression ratio may serve as a surrogate biomarker of IGF pathway activation and predict sensitivity to anti-IGF-1R targeting.

BCAR1 expression improves prediction of biochemical reccurence after radical prostatectomy

Because prostate cancer exhibits a great variability in clinical outcome, biomarkers that can be used in daily practice are needed to better stratify patients into prognostic groups. Since steroid hormones play a central role in the development and progression of prostate cancer, we aimed to analyze in a matched nested case-control study the value of molecules involved in steroid signaling, to predict recurrence after radical prostatectomy, independently from standard prognostic tools. Among 1,200 patients treated by radical prostatectomy with negative margins with at least 4 years follow-up, 121 prostate cancers with biochemical relapse were matched after pathological reassessment with 121 cancers with identical clinicopathological features but without relapse. Immunohistochemistry was performed on tissue microarrays, using antibodies directed against molecules involved in androgen and estrogen signaling, including hormone receptors, enzymes (such as the five alpha reductases 1,2 and 3, aromatase, alpha-keto reductase 1C3 and squalene epoxidase), the breast cancer antiestrogen resistance 1 (BCAR1), and the proliferation marker Ki67. The median follow-up for patients without recurrence was 7 years. Both cell proliferation and BCAR1 expression were significantly associated with biochemical relapse, in univariate and multivariate analysis. In subgroup analysis, the sole predictive marker in patients with well-differentiated prostate cancer was BCAR1 (P = 0.004), whereas only proliferation (P = 0.001) was significantly associated with relapse in less-differentiated prostate cancer patients. BCAR1 is an independent predictor of recurrence after radical prostatectomy for "low risk" prostate cancer. The use of this biomarker may enable more individualized treatment approaches.

P2-12-26: Impact of the Recurrence Score on Adjuvant Decision-Making in ER-Positive Early Breast Cancer – Results of a Large Prospective Multicentre Decision Impact Study in Node Negative and Node Positive Disease.

Abstract Background Oncotype DX® has become part of clinical routine in the diagnosis and decision-making progress in early breast cancer (EBC). Prospective data on its clinical use and impact on treatment decisions in ER-positive (ER+) node negative (N0) disease from various controlled clinical trials in different countries have been published recently. The Recurrence Score® (RS) has also been validated as a prognostic and predictive marker for patients with ER+ node positive (N+) disease. As of today no prospective data have been reported on its impact on decision making in these patients. We performed a large prospective study to evaluate RS-guided adjuvant therapy in N0 and N+ ER+ EBC. Material and Methods: Patients (pts) with ER+, HER2−negative N0 and N+ (1-3 positive lymph nodes) EBC and no contraindication for adjuvant chemotherapy were included in the study. Physicians’ adjuvant treatment recommendations and their confidence in these as well as patients’ decisional conflicts were assessed before and after knowledge of the results of the test using standardized questionnaires. Actual treatment data were collected to perform pharmacoeconomic analyses. Analyses were performed on the per-protocol population for whom a Recurrence Score result and treatment recommendations pre and post-test were available. Results: Overall 379 pts were recruited. In 11 pts Oncotype DX could not be performed and 2 pts dropped out leaving 366 pts in the per protocol population. Of these, 244 (66.7%) were N0 and 122 (33.3%) N+. Median age was 56 years (Range 25–85). Overall, 54.1% had low, 38.0% intermediate and 7.9% high RS values. For N0 disease, the distribution of RSs was 53.7%, 38.9%, 7.4% and for N+ disease 54.9%, 36.9% and 9.0%, respectively. Initial treatment recommendation changed in 33.1% of all cases; 30.3% in N0 and 38.5% in N+ disease, and in 36.4% for pts with low, 30.9% with intermediate and 20.7% with high RSs. In 21.6% of all pts a recommendation for adjuvant chemoendocrine therapy (CHT) was changed to endocrine therapy (HT), 10.7% of recommendations changed from HT to CHT. For N0 disease change rates were 18.4% from CHT to HT and 11.5% from HT to CHT. For N+ disease 27.9% of recommendations changed from CHT to HT and 9.0% of recommendations changed from HT to CHT. In 25% of all, 22% of N0 and 39% of N+ pts initially recommended HT the post-RS recommendation changed to CHT; in 38% of all, 39% of N0 and 37% of N+ pts initially recommended CHT the recommendation changed to HT. Overall, physicians’ confidence increased in 45.1% of all (p=0.047) and 44.7 of N0 and 45.9% of N+ cases, respectively. There was a moderate decrease of the decisional conflict score in all pts and subgroups that reached statistical significance for all pts (p=0.029) and the low RS subgroup (p=0.003). Conclusions: Results of this large prospective study show an impact of the RS on adjuvant treatment decision making in German clinical practice for patients with ER+ EBC. Recommendations were predominantly changed from chemoendocrine to endocrine adjuvant therapy resulting in a net reduction of chemotherapy usage. This effect was more pronounced for patients with 1–3 positive nodes. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-26.

Abstract B156: An epithelial to mesenchymal transition (EMT) gene expression signature predicts resistance to PI3K/Akt pathway inhibitors in non-small cell lung cancer.

Abstract Background: Epithelial/mesenchymal transition (EMT) is associated with loss of cell adhesion molecules such as E-cadherin and increased invasion, migration, and proliferation in epithelial cancers. In non-small cell lung cancer (NSCLC), EMT is associated with greater resistance to EGFR inhibitors. However, its potential to predict response to other targeted drugs or chemotherapy has not been well characterized. Using a previously-derived EMT gene signature, we tested the association between EMT and drug response in a panel of &amp;gt;50 NSCLC cell lines. Materials/Methods: NSCLC cell lines were classified as epithelial or mesenchymal by a previously described 76-gene EMT gene expression signature. IC50s for more than 30 drugs or drug combinations were determined by MTS assay. Differences in IC50 between epithelial and mesenchymal lines were assessed by t-test (p-values ≤ 0.05 were considered significant). Results: As expected, EGFR mutated cell lines were classified by the EMT signature as epithelial and were highly sensitive to EGFR inhibition by erlotinib. Among wild-type EGFR cell lines, those that had undergone EMT were significantly more resistance to erlotinib (p=0.029). NSCLC cell lines with mesenchymal signatures were also more resistant to drugs targeting the PI3K/Akt pathway such as GDC0941 (p=0.037) and 8-amino-adenosine (p=0.009). A trend towards greater resistance was also seen in mesenchymal cells treated with the Akt inhibitor MK2206 (p=0.12). There was no association between EMT and response to cytotoxic chemotherapies, including cisplatin, carboplatin, gemcitabine, pemetrexed, docetaxel, paclitaxel, and platinum-doublets (p-values ≥0.2). Conclusion: EMT is associated with greater resistance to PI3K/Akt pathway inhibitors and may be a useful predictive marker for patients treated with these drugs. The EMT signature is being investigated prospectively in patients treated with an Akt inhibitor in an ongoing clinical trial. Supported in part by the NIH Lung Cancer SPORE grant P50 CA70907. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr B156.