bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 280: Cancer risk in World Trade Center rescue and recovery workers
Authors: Boffetta Paolo , Solan Samara , Wallenstein Sylvan , Lucchini Roberto , Landrigan Philip
Published: May 2015

Objective: The purpose of this investigation was to evaluate cancer incidence in responders during the first 7 years after 11 September 2001.

Methods: Cancers among 20,984 consented participants in the WTC Health Program were identified through linkage to state tumor registries in New York, New Jersey, Connecticut, and Pennsylvania. The follow-up lasted until 2008. Standardized incidence ratios (SIRs) were calculated to compare cancers diagnosed in responders to predicted numbers for the general population. Multivariate regression models were used to estimate associations with degree of exposure.

Results: A total of 575 cancers were diagnosed in 552 individuals. Increases above registry-based expectations were noted for all cancer sites combined (SIR = 1.15; 95% CI: 1.06, 1.25), thyroid cancer (SIR = 2.39; 95% CI: 1.70, 3.27), prostate cancer (SIR = 1.21; 95% CI: 1.01, 1.44), combined hematopoietic and lymphoid cancers (SIR = 1.36; 95% CI: 1.07, 1.71), and soft tissue cancers (SIR = 2.26; 95% CI: 1.13, 4.05). When restricted to 302 cancers diagnosed ≥ 6 months after enrollment, the SIR for all cancers decreased to 1.06 (95% CI: 0.94, 1.18), but thyroid and prostate cancer diagnoses remained greater than expected. All cancers combined were increased in very highly exposed responders and among those exposed to significant amounts of dust, compared with responders who reported lower levels of exposure.

Conclusion: Estimates should be interpreted with caution given the short follow-up and long latency period for most cancers, the intensive medical surveillance of this cohort, and the small numbers of cancers at specific sites. However, our findings highlight the need for continued follow-up and surveillance of WTC responders. The follow-up of the cohort is currently being updated and efforts are being made to improve the assessment of exposure to carcinogens related to WTC experience.

Citation Format: Paolo Boffetta, Samara Solan, Sylvan Wallenstein, Roberto Lucchini, Philip Landrigan. Cancer risk in World Trade Center rescue and recovery workers. [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 280. doi:10.1158/1538-7445.AM2014-280
Abstract 2824: Circulating tumor cells (CTCs) can predict survival and response to treatment in non-metastatic colorectal cancer (CRC): Correlation between flowcytometry, quantitative real-time pcr (qPCR) and cytomorphology
Authors: Bahnassy Abeer , Zekri Abdel-Rahman N. , Salem Salem E. , Mohamad Marwa , Mohanad Marwa
Published: May 2015

Background: Circulating tumor cells (CTCs) were detected in patients with metastatic epithelial malignancies using different methodologies. The number of CTCs before and during treatment of advanced CRC is an independent predictor of poor response to treatment, progression-free (PFS) and overall survival (OS). We investigated 1) whether CTC count can predict outcome in patients with CRC and 2) the utility of FCM supplemented with PCR and/or cyto-morphology (CM) in enumerating CTCs.

Methodology: CTCs were enumerated in 56 CRC cases and 20 normal controls (NC) by flowcytometry (FCM), and quantitative real time PCR (qPCR) using CK19, MUC, CD44, CD133 and ALDH1after immune-magnetic separation. Results were correlated to cytomorphology, standard prognostic factors, overall and progression free survival (OS&PFS).

Results: CTCs < 3 were detected in 45% of the cases compared to 55% with ≥3 CTCs. CK19, MUC1, CD44, CD133 and ALDH1-RNA expression was detected in (55.4%, 44.6%, 53.6%, 43.6%, 46.4%; respectively). The concordance between FCM& qPCR, FCM&CM, qPCR& CM ranged from 82% to 96%. Significant correlations were reported between CTCs and tumor stage and grade (p<0.01). OS was significantly associated with pathological tumor type, T stage, lymph nodes involvement, CTC, CK, MUC1, CD44 and CD133. PFS was significantly associated with pathological tumor type, disease stage, N stage, CTC, CK, MUC1, CD44 and CD133 (p=).

Conclusions:1)enumeration of CTCs, pre- and after initial treatment, can predict outcome in CRC patients, 2)FCM is a sensitive, rapid and easy technique for enumeration of CTCs, especially if supplemented with qPCR for detection of CRC cells biomarkers (CK19), MUC, CD44, CD133 and ALDH1).

Citation Format: Abeer Bahnassy, Abdel-Rahman N. Zekri, Salem E. Salem, Marwa Mohamad, Marwa Mohanad. Circulating tumor cells (CTCs) can predict survival and response to treatment in non-metastatic colorectal cancer (CRC): Correlation between flowcytometry, quantitative real-time pcr (qPCR) and cytomorphology. [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 2824. doi:10.1158/1538-7445.AM2014-2824
Abstract 2848: Urinary prostaglandin E metabolite as a biomarker predicting response to EGFR therapy in patients with head and neck cancer
Authors: Kekatpure Vikram D. , Suresh Amritha , Rangarajan Bharath , Siddappa Gangotri , Kuriakose Moni Abraham
Published: May 2015

Background: Cross talk between the epidermal growth factor receptor (EGFR) and prostaglandin E2 pathways (PGE2) is well established. Activation of EGFR signaling can induce PGE2. Urinary PGE-M is a stable end metabolite of PGE2. The purpose of this study was to evaluate the role of urinary PGE-M as a biomarker predicting response to anti-EGFR monoclonal antibody therapy in patients with head and neck squamous cell carcinoma (HNSCC).

Patients and Methods: Ten HNSCC patients with recurrent and/or metastatic disease received Nimotuzumab, a humanized monoclonal antibody against EGFR. The response to Nimotuzumab was assessed 2 weeks following the fourth cycle using PERCIST criteria. Urine samples were collected before treatment and 2 weeks after the fourth cycle of therapy.

Results: Seven patients had progressive disease (PD) while two patients had stable disease (SD) and one patient had a partial response (PR) to therapy. The mean pretreatment value of urinary PGE-M in patients with SD or PR was 63 ng/mg Cr (range 48-78 ng/mg Cr). By comparison, patients with progressive disease had lower baseline levels of urinary PGE-M with mean pretreatment value of 37ng/mg Cr (range 15- 54ng/mg Cr). More than a 20% reduction in post treatment vs. pretreatment urinary PGE-M values was seen in patients with SD and PR. By comparison, patients with progressive disease had either a minimal reduction or an increase in the post treatment levels of urinary PGE-M.

Conclusion: Elevated pretreatment levels of urinary PGE-M may predict response to anti-EGFR therapy in patients with HNSCC. These findings need further validation.

Citation Format: Vikram D. Kekatpure, Amritha Suresh, Bharath Rangarajan, Gangotri Siddappa, Moni Abraham Kuriakose. Urinary prostaglandin E metabolite as a biomarker predicting response to EGFR therapy in patients with head and neck 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 2848. doi:10.1158/1538-7445.AM2014-2848
Abstract 2857: S100A14 plays an important role in cell growth and metastasis of epithelial ovarian cancer through PI3K-AKT pathway
Authors: Cho Hanbyoul , Lee Maria , Shin Ha-Yeon , Yang Wookyeom , Lee Eun-ju , Kwon Hyunja , Kim Sol , Kim Jae-Hoon
Published: May 2015

Background: S100A14 is an EF-hand containing calcium protein of the S100 protein family that exerts its biological effects on different types of cancers. The roles of S100A14 in tumorigenesis and its underlying mechanism have not been fully understood. This study aims to investigate the clinical significance and possible mechanisms of action of S100A14 in the growth and metastasis of epithelial ovarian cancer (EOC).

Method: S100A14 expression in ovarian cancer was studied using a total of 104 ovarian tissue specimens (13 normal ovarian tissues, 10 benign ovarian tumors, 10 borderline ovarian tumors, and 71 EOC tissues) via immunohistochemistry. We investigated the possible functions of S100A14 in EOC stable cell by lentivirus-mediated overexpression and short hairpin RNA (shRNA).

Results: S100A14 protein was significantly over-expressed in EOC cells and tissues. S100A14 expression was significantly associated with advanced stage (P<0.001), serous type (P=0.004), and poor tumor grade (P<0.001). Patients with S100A14 overexpression had shorter overall (Hazard Ratio = 4.53 [1.16-17.69], P=0.029) and disease-free survival (Hazard Ratio = 3.10 [1.22-7.89], P=0.017) in multivariate analysis. In vitro analysis of cell function revealed that cell proliferation, migration and invasion were regulated by S100A14. We investigated the underlying mechanism and found that the phosphorylation levels of AKT have increased after S100A14 overexpression. Inhibition of AKT by a specific AKT inhibitor (MK-2206) and PI3K inhibitor (wortmannin) at least partly reversed the AKT-phosphorylation events in S100A14-overexpressed cells.

Conclusions: The data revealed the aberrant expression of S100A14 in ovarian cancer, suggesting a potentially important role of S100A14 in ovarian carcinogenesis; S100A14 regulated AKT via phosphorylation of its ser473 site also implies that S100A14 could be used as a novel therapeutic target for ovarian cancer treatment.

Citation Format: Hanbyoul Cho, Maria Lee, Ha-Yeon Shin, Wookyeom Yang, Eun-ju Lee, Hyunja Kwon, Sol Kim, Jae-Hoon Kim. S100A14 plays an important role in cell growth and metastasis of epithelial ovarian cancer through PI3K-AKT pathway. [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 2857. doi:10.1158/1538-7445.AM2014-2857
Abstract 286: Epigenome-wide association study of normal lung tissues identifies hypomethylation of multiple CpG sites associated with cigarette smoking
Authors: Li Wen-Qing , Shi Jianxin , Wheeler William , Caporaso Neil , Laird-Offspringa Ite , Landi Maria Teresa
Published: May 2015

Background: DNA methylation due to tobacco smoking have been reported in studies of blood genomic DNA, indicating a possible role of methylation changes due to smoking in multiple adverse health outcomes. Since DNA methylation is known to be tissue-specific, we profiled methylation in normal human lung tissues and evaluated the association with cigarette smoking based on a large sample size.

Methods: We studied histologically normal lung tissues of 237 lung cancer patients (121 current cigarette smokers and 116 current nonsmokers) from the Environment And Genetics in Lung cancer Etiology study. DNA methylation was assayed using the Illumina Infinium HumanMethylation450 BeadChip arrays and methylation for 338,456 autosomal probes remained after quality control filters. We examined the association of log-transformed methylation with current smoking status, as well as with quantitative measures of smoking, using linear regression models adjusting for covariates. We sought to replicate the findings in The Cancer Genome Atlas normal lung samples (12 current smokers and 50 nonsmokers).

Results: Eight CpG sites were significantly hypomethylated in lung tissues of current smokers after Bonferroni-correction (P<1.48×10-7). Of them, six were associated with smoking duration, pack-years and years from quitting smoking, and were replicated in TCGA samples (P<0.10), including cg17113147 (NXN), cg05575921 (AHRR), cg07992500 (CDC42EP3), cg14120703 (NOTCH1), cg11152412 (EDC3), and cg03224163 (HIPK2). These loci have not been reported in prior studies, except for cg05575921, which was shown to be hypomethylated in smokers in studies of blood DNA.

Conclusions: Our study in lung tissues shows that smoking is associated with alterations in DNA methylation in genes involved in tobacco smoking metabolism or cancer progression. These findings suggest that smoking-associated epigenetic changes may have an important role in lung tumorigenesis.

Citation Format: Wen-Qing Li, Jianxin Shi, William Wheeler, Neil Caporaso, Ite Laird-Offspringa, Maria Teresa Landi. Epigenome-wide association study of normal lung tissues identifies hypomethylation of multiple CpG sites associated with cigarette smoking. [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 286. doi:10.1158/1538-7445.AM2014-286
Abstract 2865: High serum transforming growth factor β1 levels associated with poor survivals in patients with advanced hepatocellular carcinoma
Authors: Lin Tzu-Hsuan , Shao Yu-Yun , Chan Soa-Yu , Huang Chung-Yi , Cheng Ann-Lii , Hsu Chih-Hung
Published: May 2015

Background: TGF-β (Transforming growth factor β) signaling pathway plays a significant role in mediating progression, metastasis, and immune evasion of human malignancies. Previous studies have found that serum TGF-β1 levels were elevated in patients with hepatocellular carcinoma (HCC) compared to patients with chronic hepatitis or cirrhosis. However, the prognostic role of serum TGF-β1 levels in patients with advanced HCC is currently unknown.

Patients and Methods: We enrolled patients who had received sorafenib or sorafenib-based combination therapy for advanced HCC, defined as metastatic or locally advanced diseases not amenable to loco-regional therapies, at National Taiwan University Hospital, Taipei, Taiwan, between 2007 and 2012. The baseline blood samples were collected before sorafenib treatment with the consent of the patients. Serum TGF-β1 levels were measured by Human TGF-β1 Instant ELISA (eBioscience, Vienna, Austria), and were correlated with disease control rate (DCR), progression- free survival (PFS), and overall survival (OS) of the patients.

Results: The study enrolled 91 patients (M:F=82:9), with a median age of 56.9 years. Sixty-nine patients (75.8 %) had chronic hepatitis B virus (HBV) infection, 18 (19.7%) had chronic hepatitis C infection, 82 (90.1%) had Barcelona Clinic Liver Cancer stage C disease, 81(89.0%) had either extrahepatic metastasis or macrovascular invasion, and 88 (96.7%) had Eastern Cooperative Oncology Group performance status of ≤ 1. All patients had Child-Puch class A liver function reserve. Patients with high baseline serum TGF-β1 levels (higher than the median level of the whole patient cohort), compared to patients with low serum TGF-β1 levels, had significantly shorter PFS (median, 2.54 vs. 4.27 months, p = 0.022) and OS (median, 5.57 vs. 11.6 months, p = 0.029). The DCRs for patients with high and low TGF-β1 were not statistically different (54.3 % vs. 62.2%, p = 0.4463). High baseline TGF-β1 levels tended to associate with high serum alpha-fetal protein (AFP) levels and high Cancer of the Liver Italian Program (CLIP) scores, although not reaching statistical significance.

Conclusions: High baseline serum TGF-β1 levels were associated with poor PFS and OS in patients with advanced HCC. (This study was supported by grants NRPB-100CAP1020-2, NSC101-2314-B-002-141 & NSC102-2314-B-002 -120).

Citation Format: Tzu-Hsuan Lin, Yu-Yun Shao, Soa-Yu Chan, Chung-Yi Huang, Ann-Lii Cheng, Chih-Hung Hsu. High serum transforming growth factor β1 levels associated with poor survivals in patients with advanced hepatocellular carcinoma. [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 2865. doi:10.1158/1538-7445.AM2014-2865
Abstract 2879: Anti-cancer effects of REIC/Dkk-3-encoding adenoviral vector for the treatment of non-small cell lung cancer
Authors: Suzawa Ken , Toyooka Shinichi , Shien Kazuhiko , Tanaka Norimitsu , Watanabe Masami , Soh Junichi , Sakaguchi Masakiyo , Matsuo Keitaro , Yamamoto Hiromasa , Furukawa Masashi , Asano Hiroaki , Tsukuda Kazunori , Nasu Yasutomo , Huh Nam-Ho , Miyoshi Shinichiro , Kumon Hiromi
Published: May 2015

Objectives: REIC/Dkk-3 is down-regulated in a broad range of human cancer cells and is considered to function as a tumor suppressor. We previously reported that REIC/Dkk-3-expressing adenovirus vector (Ad-REIC) induced endoplasmic reticulum (ER) stress and cancer-specific apoptosis in human prostate cancer. In this study, we examined the therapeutic impact of Ad-REIC on non-small cell lung cancer (NSCLC).

Materials and Methods: We examined the anti-tumor effect of Ad-REIC on 25 NSCLC cell lines in vitro and A549 cells in vivo. Two of these cell lines were artificially established as EGFR-tyrosine kinase inhibitor (TKI) resistant sublines.

Results: Ad-REIC-treatment inhibited the cell viability by 40% or more in 13 (52%) of the 25 cell lines at multiplicity of infection (MOI) of 20 (20 MOI). These cell lines were regarded as being highly sensitive cells. The cell viability of a non-malignant immortalized cell line, OUMS-24, was not inhibited at 200 MOI of Ad-REIC. The effects of Ad-REIC on EGFR-TKI resistant sublines were equivalent to those in the parental cell lines. Here, we demonstrated that Ad-REIC treatment activated c-Jun N-terminal kinase (JNK) in NSCLC cell lines, indicating the induction of ER stress with GRP78/BiP (GRP78) up-regulation and resulting in apoptosis. A single intratumoral injection of Ad-REIC significantly inhibited the tumorigenic growth of A549 cells in vivo. As predictive factors of sensitivity for Ad-REIC treatment in NSCLC, we examined the expression status of GRP78 and coxsackievirus and adenovirus receptor (CAR). We found that the combination of the GRP78 and CAR expressional statuses may be used as a predictive factor for Ad-REIC sensitivity in NSCLC cells.

Conclusion: Ad-REIC induced JNK activation and subsequent apoptosis in NSCLC cells. Our study indicated that Ad-REIC has therapeutic potential against NSCLC and that the expression statuses of GRP78 and CAR may predict a potential therapeutic benefit of Ad-REIC.

Citation Format: Ken Suzawa, Shinichi Toyooka, Kazuhiko Shien, Norimitsu Tanaka, Masami Watanabe, Junichi Soh, Masakiyo Sakaguchi, Keitaro Matsuo, Hiromasa Yamamoto, Masashi Furukawa, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Nam-Ho Huh, Shinichiro Miyoshi, Hiromi Kumon. Anti-cancer effects of REIC/Dkk-3-encoding adenoviral vector for the treatment of 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 2879. doi:10.1158/1538-7445.AM2014-2879
Abstract 2887: Molecular farming and molecular engineering of cancer vaccines
Authors: Shukla Sourabh , Steinmetz Nicole F.
Published: May 2015

Human epidermal growth factor receptor 2 (HER2)+ cancers are aggressive, have poor prognoses, high rate of metastasis and relapse. Immunotherapy (e.g. trastuzumab) has shown improvement in outcome for HER2+ patients. Development of drug resistance and short half-lives limiting the duration of the therapy, are barriers in the development of effective therapeutic strategies. Furthermore, while passive immunotherapy provides treatment, it does not reduce the risk of recurrence of the disease. Therefore, we propose a HER2 vaccine to mount an active HER2-specific immune response, with long lasting immunological memory to overcome the limitations of passive immunotherapy and prevent tumor progression, metastasis, and relapse.

We hypothesize that integration of HER2 epitopes on the plant-produced vaccination platform potato virus X (PVX) will overcome immunological tolerance against HER2. We have shown that PVX naturally targets dendritic and B-cells stimulating Th1 biased immunity, important for successful tumor prevention. The carrier acts as an adjuvant, improves stability of epitopes, and increases circulation time, therefore ensuring a sustained stimulus post vaccination and production of HER2-specific antibodies that could act against HER2+ tumors via a variety of effector mechanisms. Immunizations of FBV/N mice resulted in the production of HER2-specific antibodies, as shown by ELISA and confocal microscopy using HER2+ human cancer cell lines. Vaccination studies using animals pre-disposed to HER2+ cancer are currently ongoing. Our long-term goal is to produce the vaccine through molecular farming in edible plant tissue and make it available globally, especially in underdeveloped countries where repeated administration of costly immunotherapies is not affordable or unavailable.

Citation Format: Sourabh Shukla, Nicole F. Steinmetz. Molecular farming and molecular engineering of cancer vaccines. [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 2887. doi:10.1158/1538-7445.AM2014-2887
Abstract 2909: A gene signature composed of 13 p53 target genes predicts for response to NVP-CGM097, a novel p53-Mdm2 inhibitor, in cell lines and in human primary tumor xenograft models
Authors: Jeay Sebastien , Gaulis Swann , Ferretti Stéphane , Albrecht Geneviève , Barys Louise , Guthy Daniel , Halilovic Ensar , Ito Moriko , Murakami Masato , Pornon Astrid , Ruetz Stephan , Venkatesan Kavitha , Yu Jianjun , Jensen Michael , Wiesmann Marion , Wuerthner Jens , Graus-Porta Diana
Published: May 2015

Patient selection biomarkers are essential for the successful and rapid development of emerging targeted anti-cancer therapeutics. In this study we have identified a novel patient selection strategy for NVP-CGM097, a p53-Mdm2 inhibitor currently in a Phase I clinical trial (NCT01760525). We have analyzed the sensitivity of over 500 cell lines from the “Cancer Cell Line Encyclopedia” to the p53-Mdm2 inhibitor in cell viability assays, and intersected response data with information on gene expression and genomic alterations. This analysis has led to the identification of a gene signature consisting of 13 genes, as a predictor for sensitivity to NVP-CGM097. Interestingly, these 13 genes are known p53 downstream target genes, supporting the hypothesis that the identified gene signature is reflective of the p53 pathway functionality in sensitive cell lines. We show the performance of the signature as ROC and Precision-Recall curves in cell lines as well as in a number of human primary tumor xenografts, both unselected as well as pre-selected for p53 wild-type status. Work is now ongoing to validate this gene signature using baseline tumor biopsy samples and RECIST-based efficacy readouts in the current Phase I clinical trial.

Citation Format: Sebastien Jeay, Swann Gaulis, Stéphane Ferretti, Geneviève Albrecht, Louise Barys, Daniel Guthy, Ensar Halilovic, Moriko Ito, Masato Murakami, Astrid Pornon, Stephan Ruetz, Kavitha Venkatesan, Jianjun Yu, Michael Jensen, Marion Wiesmann, Jens Wuerthner, Diana Graus-Porta. A gene signature composed of 13 p53 target genes predicts for response to NVP-CGM097, a novel p53-Mdm2 inhibitor, in cell lines and in human primary tumor xenograft models. [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 2909. doi:10.1158/1538-7445.AM2014-2909
Abstract 2926: PNA as novel targeted cancer therapy for BRAFV600E mutant melanoma
Authors: Rothman Jeffrey H. , Surriga Oliver , Vasudeva Shyamprasad D. , Ambrosini Grazia , Ouerfelli Ouathek , Schwartz Gary K.
Published: May 2015

Our aim is to target tumor cells specifically by directly suppressing their oncogenes with peptide nucleic acid (PNA) oligonucleotide analogues. PNA oligonucleotides bind to DNA over 1000-fold more avidly than its native complement, are completely resistant to intra and extra cellular enzymatic degradation, show no nonspecific toxicity at therapeutic levels, and when conjugated to delivery peptides can be made nuclear and cell membrane permeable. We have employed these PNA oligomers to target BRAFV600E which is prevalent in cutaneous melanoma in a sequence-specific complementary manner so as to disrupt transcription by strand invasion. For these studies, we have developed a novel delivery peptide conjugated to PNA modified to increase both cellular delivery and PNA stability towards its target. Our results indicate that exposure of the melanoma cell lines to a modified PNA-peptide conjugate selective for BRAFV600E results in a concentration-dependent inhibition of cell growth that is specific for the BRAFV600E mutant melanoma cell lines with an IC50 range of 250 to 500 nM. Moreover, there is no inhibition of BRAFWT cell growth at these concentrations. This is associated with suppression of BRAFV600E protein over time with no effect on BRAFWT protein levels. Furthermore, BRAFV600E protein expression was suppressed for up to 6 days following initial exposure proving the durability of this type of inhibition. Exposure to this modified PNA-peptide down-regulates BRAFV600E mRNA transcription exclusively in the mutant cell lines. Live cell imaging of BRAFV600E mutant cells confirms localization of fluorescein-labeled PNA-delivery peptide specific to BRAFV600E to the nucleus within 3 hours of treatment. Our results indicate that these PNA-peptide derivatives could represent a novel and promising new therapy for patients with BRAFV600E mutant melanoma, and this technology could be applied to a multitude of other cancers either with specific translocations or mutations differing from wild-type cells even by only a single base pair.

Citation Format: Jeffrey H. Rothman, Oliver Surriga, Shyamprasad D. Vasudeva, Grazia Ambrosini, Ouathek Ouerfelli, Gary K. Schwartz. PNA as novel targeted cancer therapy for BRAFV600E mutant melanoma. [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 2926. doi:10.1158/1538-7445.AM2014-2926
Abstract 2967: Metformin selectively targets tumor initiating cells in erbb-2 overexpressing breast cancer models
Authors: Zhu Pei , Davis Meghan , Blackwelder Amanda , Bachman Nora , Liu Bolin , Edgerton Susan , Williams Leonard L. , Thor Ann D. , Yang Xiaohe
Published: May 2015

Metformin is an oral biguanide used for type II diabetes. Epidemiologic studies suggest a link between metformin use and reduced risk of breast and other types of cancers. erbB-2 expressing breast cancer is a subgroup of tumors with poor prognosis. Previous studies demonstrated that metformin is a potent inhibitor of erbB-2 overexpressing breast cancer cells; metformin treatment extends the life span and impedes mammary tumor development in ErbB-2 transgenic mice in vivo. However, the mechanisms of metformin associated anti-tumor activity, especially in prevention models, remain unclear. In this study, we investigated the mechanisms of metformin associated prevention/inhibition of ErbB-2 mediated breast cancer development by focusing on the potential effect of metformin on tumor initiating cells (TICs)/cancer stem cells (CSCs) in the MMTV-erbB-2 transgenic mouse model and its context with existing in vitro models. We report here for the first time that systemic administration of metformin (250 mg/kg/day, ip, between 8- and 18-week of age) selectively inhibits CD61high/CD49fhigh subpopulation, a group of TICs of MMTV-ErbB-2 mammary tumors, in preneoplastic mammary glands. Metformin also inhibited CD61high/CD49fhigh subpopulation in MMTV-erbB-2 tumor-derived cells, which was correlated with their compromised tumor initiation/development in a syngeneic tumor graft model. Molecular analysis indicated that metformin induced downregulation of erbB-2 and EGFR expression and inhibited the phosphorylation of erbB family members, IGF1R, Akt, mTOR and Stat3 in vivo. In vitro data indicate that low doses (1 mM) of metformin inhibited the self-renewal/proliferation of CSCs/TICs in ErbB-2-overexpressing breast cancer cells. We further demonstrated that the expression and activation of ErbB-2 were preferentially increased in CSC/TIC-enriched tumorsphere cells, which promoted their self-renewal/proliferation and rendered them more sensitive to metformin. Our results, especially the in vivo data, provide fundamental support for developing metformin-mediated preventive strategies targeting erbB-2-associated carcinogenesis.

Citation Format: Pei Zhu, Meghan Davis, Amanda Blackwelder, Nora Bachman, Bolin Liu, Susan Edgerton, Leonard L. Williams, Ann D. Thor, Xiaohe Yang. Metformin selectively targets tumor initiating cells in erbb-2 overexpressing breast cancer models. [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 2967. doi:10.1158/1538-7445.AM2014-2967
Abstract 3050: Human rectal carcinoid tumors highly express DCLK1, a putative cancer stem cell marker
Authors: Ikezono Yu , Koga Hironori , Abe Mitsuhiko , Yoshida Takafumi , Nakamura Toru , Akiba Jun , Yano Hirohisa , Tsuruta Osamu , Torimura Takuji , Sata Michio
Published: May 2015

Background and Aim: Doublecortin-like kinase 1 (Dclk1) is known to regulate neuronal differentiation, migration, and neurogenesis (Shu et al., Neuron 2006). Accumulating evidence suggests that the protein is a putative marker for intestinal and pancreatic stem cells, including cancer stem cells (CSCs) of those organs (Gagliardi et al., Pathol Res 2012, Nakanishi et al., Nat Genet 2013, Bailey et al., Gastroenterology 2013). Since carcinoid tumor is defined as a neuroendocrine tumor, we assessed whether or not the Dclk1 protein, a marker of CSC, was expressed in human rectal carcinoid tumor tissues. Patients and Methods: Eighteen patients (9 male, 9 female; mean age, 51) with rectal carcinoid tumors were included in this study. Informed consent was obtained from all of the patients. The tumors were safely resected by endoscopic mucosal resection (EMR) method between 2003 and 2012 in the Kurume University Hospital. Mean diameter of the tumors was 5.4 mm (range, 2.0-8.0 mm). Immunohistochemical staining was performed to examine the Dclk1 protein expression, accompanied by the staining for the commonly-used neuroendocrine markers synaptophysin, chromogranin A, and CD56. The immune-reactivity was evaluated according to the following criteria: the intensity of staining was scored in each specimen on a scale of 0 to 3, in which 0 = negative staining, 1 = weakly positive staining, 2 = moderately positive staining, and 3 = strongly positive staining. The extent to which positive cells were seen in each specimen (‘‘extent of distribution’’ of positive cells) was estimated on a scale of 0 to 2, in which 0 = positive staining in 0% to 20% of cells; 1, in 21% to 60%; and 2, in 61% to 100%. Sum of the scores was calculated for each specimen. Results: The total score for Dclk1, synaptophysin, CD56, and chromogranin A was 87/90, 86/90, 60/90, and 34/90, respectively. Conclusions: Human rectal caricinoid tumor highly expressed Dclk1, and the high expression rate indicated that the protein expression was a novel feature for the neuroendocrine tumor. It is suggested that Dclk1 is a potential marker of not only CSCs but also non-epithelial tumors, such as carcinoid tumor, regulating crucial tumor cell behaviors.

Citation Format: Yu Ikezono, Hironori Koga, Mitsuhiko Abe, Takafumi Yoshida, Toru Nakamura, Jun Akiba, Hirohisa Yano, Osamu Tsuruta, Takuji Torimura, Michio Sata. Human rectal carcinoid tumors highly express DCLK1, a putative cancer stem cell marker. [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 3050. doi:10.1158/1538-7445.AM2014-3050
Abstract 3064: Importance of circulating tumor cells in newly diagnosed colorectal cancer
Authors: Dalum Guus van , Groot Marco R. de , Stam Gerrit-Jan , Scholten Loes F.A. , Mastboom Walter J.B. , Vermes Istvan , Tibbe Arjan G.J. , Terstappen Leon W.M.M.
Published: May 2015

Background: The presence of circulating tumor cells (CTC) is associated with poor prognosis in patients with metastatic colorectal cancer (CRC). This study was conducted to determine if the presence of CTC prior to surgery and during follow-up in patients with newly diagnosed non-metastatic CRC identifies patients who are at risk for disease recurrence.

Methods: In a prospective single center study 183 patients with newly diagnosed non-disseminated CRC scheduled for surgery were enrolled from 2003 till 2008 and followed up for a median of 5.1 years. CTC were enumerated with the CellSearch System in 4 aliquots of 7.5 ml of peripheral blood before and after surgery (1-26 weeks), after adjuvant therapy and 1, 2, 3 and 4 years after surgery.

Findings: ≥1 CTC/ 30ml of blood were detected in 44 (24%) patients before surgery. CTC frequency did not change significantly at the time points after surgery. Patients with CTC before surgery had a significant decrease in Recurrence Free Survival (RFS, logrank test p=0.014) and Colon Cancer Related Survival (CCRS, p=0.002). Five year RFS dropped from 75% to 61% and five year CCRS from 83% to 69% for patients with CTC before surgery. In a multivariate analysis of CTC, T-Stage and N-stage, the presence of CTC and N-stage remained as significant factors for RFS and CCRS. Surprisingly the presence of CTC after surgery was not significantly associated with RFS and CCRS whereas CTC 2-3 years after surgery was again significantly associated with RFS and CCRS.

Interpretation: The presence of CTC in patients with stage I-III CRC before surgery is associated with a significant reduction RFS and CCRS. Although similar amounts of CTC were detected within 3 months after surgery they were not associated with RFS or CCRS. In contrast CTC were again highly significant for RFS and CCRS 2-3 years after surgery. These findings suggest a role of CTC detection, to assess which patients need adjuvant treatment. To implement CTC detection in the non-metastatic setting a validated CTC detection technology is needed with increased sensitivity and specificity.

Citation Format: Guus van Dalum, Marco R. de Groot, Gerrit-Jan Stam, Loes F.A. Scholten, Walter J.B. Mastboom, Istvan Vermes, Arjan G.J. Tibbe, Leon W.M.M. Terstappen. Importance of circulating tumor cells in newly diagnosed colorectal 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 3064. doi:10.1158/1538-7445.AM2014-3064
Abstract 3080: Comparison of CellSearch with polymeric microfluidic devices for CTC isolation using EpCAM-negative tumor cell lines of malignant pleural mesothelioma
Authors: Chikaishi Yasuhiro , So Tomoko , Takenaka Masaru , Oka Soichi , Hirai Ayako , Iwanami Takashi , Shimokawa Hidehiko , Yoneda Kazue , Nagata Yoshika , Uramoto Hidetaka , Ohnaga Takeshi , Tanaka Fumihiro
Published: May 2015

Background. The CellSearch system is the most commonly used technique approved by the FDA to isolate and quantify circulation tumor cells (CTCs). However, in this system, the efficiency of CTC isolation is influenced by the epithelial cell adhesion molecule (EpCAM). Therefore, it is difficult to isolate CTCs in the setting of the epithelial to mesenchymal transition (EMT), malignant pleural mesothelioma and so on. We developed polymeric microfluidic devices for CTC isolation (CTC-chip) and compared the efficacy of CellSearch with CTC-chip using EpCAM-negative tumor cell lines of malignant pleural mesothelioma.

Methods. In order to evaluate the accuracy of the number of CTCs, we used PBS samples and peripheral blood samples (blood samples) collected from healthy donors to which spiking the tumor cell lines ACC-MESO1 and ACC-MESO4. Both tumor cell lines (obtained from ATCC) were derived from malignant pleural mesothelioma, positive for mesothelin and podoplanin and negative for EpCAM. The recovery rate was evaluated using two methods. CellSearch was performed according to the normal method, while CTC-chip was performed using a two coating pattern with anti-mesothelin antibody (chip1) and anti-podoplanin antibody (chip2).

Results. In the PBS samples, the recovery rate of captured ACC-MESO1 was 8.6%/98%/99% (CellSearch/chip 1/chip 2), while those of ACC-MESO4 was 21.1%/97%/99%, respectively. In the blood samples, the recovery rate of captured ACC-MESO1 was 1.2%/6.2%/13.3% (CellSearch/chip 1/chip 2), while that of ACC-MESO4 was 6.0%/3.7%/11.6%, respectively.

Conclusions. Our experiments showed that the recovery rate of each sample was higher using CTC-chip than that obtained with CellSearch. Moreover, CTC-chip coated anti-podoplanin antibody exhibited a higher recovery rate than coated anti-mesothelin antibody. Future studies using experiments of CTC isolation in cancer patients with malignant pleural mesothelioma are thus required.

Citation Format: Yasuhiro Chikaishi, Tomoko So, Masaru Takenaka, Soichi Oka, Ayako Hirai, Takashi Iwanami, Hidehiko Shimokawa, Kazue Yoneda, Yoshika Nagata, Hidetaka Uramoto, Takeshi Ohnaga, Fumihiro Tanaka. Comparison of CellSearch with polymeric microfluidic devices for CTC isolation using EpCAM-negative tumor cell lines of malignant pleural mesothelioma. [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 3080. doi:10.1158/1538-7445.AM2014-3080
Abstract 3951: Dual phosphoinositide 3-kinase/Mammalian target of rapamycin inhibitors, GSK2126458 and PKI-587, suppress tumor progression and increase radiosensitivity in nasopharyngeal carcinoma
Authors: Wang Wei , Li Qi , Liu Tongxin , Chen Longhua
Published: May 2015

Purpose

Although combined chemo-radiotherapy has provided considerable improvements for nasopharyngeal carcinoma (NPC), recurrence and metastasis are still frequent. The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway plays a critical role in tumor formation and tumor cell survival after radiation-induced DNA damage. In the present study, we evaluated whether inhibition of PI3K/mTOR by two novel dual inhibitors, GSK2126458 and PKI-587, could suppress tumor progression and sensitize NPC cells to radiation.

Experimental Design

Four NPC cell lines (CNE-1, CNE-2, 5-8F and 6-10B) were used to analyze the effects of GSK216458 and PKI-587 on cell proliferation, migration, invasion, clonogenic survival, amount of residual γ-H2AX foci, cell cycle and apoptosis after radiation. A 5-8F xenograft model was used to evaluate the in vivo effects of the two compounds in combination with ionizing radiation (IR).

Results

Both GSK216458 and PKI-587 effectively inhibited cell proliferation and motility in NPC cells and suppressed phosphorylation of Akt, mTOR, S6 and 4EBP1 proteins in a dose- and time-dependent manner. Moreover, both compounds sensitized NPC cells to ionizing radiation (IR) by increasing DNA damage, enhancing G2/M cell cycle delay and inducing apoptosis. In vivo, the combination of IR with GSK2126458 or PKI-587 significantly inhibited tumor growth. Tumor regression was correlated with induction of apoptosis and suppression of the phosphorylation of mTOR, Akt, and 4EBP1.

Conclusions

GSK2126458 and PKI-587 inhibit tumor progression and increase radiosensitivity by inhibiting PI3K/Akt/mTOR signaling in NPC. Combining IR with a dual PI3K/mTOR inhibitor, either GSK2126458 or PKI-587, might be a promising therapeutic strategy for NPC.

Citation Format: Wei Wang, Qi Li, Tongxin Liu, Longhua Chen. Dual phosphoinositide 3-kinase/Mammalian target of rapamycin inhibitors, GSK2126458 and PKI-587, suppress tumor progression and increase radiosensitivity in nasopharyngeal carcinoma. [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 3951. doi:10.1158/1538-7445.AM2014-3951
Abstract 3986: Characterization of prostaglandin signaling in primary neuroblastoma
Authors: Kock Anna , Larsson Karin , Ljungblad Linda , Idborg Helena , Korotkova Marina , Elfman Lotta , Johnsen John-Inge , Jakobsson Per-Johan , Kogner Per
Published: May 2015

Neuroblastoma (NB) is an embryonic childhood tumor of the sympathetic nervous system. Despite intensive treatment patients in the high-risk group of NB shows a poor prognosis.

We have shown the importance of inflammation in NB development and progression and that the pro-inflammatory lipid mediator prostaglandin E2 (PGE2) acts as a paracrine survival factor in NB. We have also shown that mPGES-1, essential for PGE2 synthesis, promotes growth and survival of several malignancies.

In this study we focus on the abundance of prostaglandins and their specific enzymes in primary NB of different biological and clinical subsets in the aim of finding a more specific and targeted NB treatment.

Levels of prostaglandins were assayed in 40 primary human NB tumors by mass spectrometry (MS) showing elevated levels of PGE2 in high risk 11q-deleted NB compared to low-risk NB whereas elevated levels of prostaglandin D2 (PGD2) could be coupled to NB with favorable clinical outcome.

The expression of COX-1, COX-2, mPGES-1, L-PGDS, H-PGDS and 15-PGDH were analyzed in tumors by immunohistochemistry (IHC). Expression of mPGES-1 and L-PGDS was found in all analyzed tumors.

Double stainings of mPGES-1 and the specific NB cell marker GD2 were performed on tumors with different biology using immunofluorescence. In low-risk tumors mPGES-1 is not expressed by the tumor cells whereas co-localization were found in 11q deleted high-risk tumors. To consider the level of inflammation and PGE2 signaling in these tumors we analyzed cell markers of both the innate and adaptive immune system together with mPGES-1. In all tumors mPGES-1 expressing cells are found in clusters closely together with T-cells, B-cells, dendritic cells and M1/M2 macrophages.

Together this suggests an involvement of inflammatory prostaglandins in neuroblastoma development and tumor progression opening up new strategies for targeted therapies.

Citation Format: Anna Kock, Karin Larsson, Linda Ljungblad, Helena Idborg, Marina Korotkova, Lotta Elfman, John-Inge Johnsen, Per-Johan Jakobsson, Per Kogner. Characterization of prostaglandin signaling in primary neuroblastoma. [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 3986. doi:10.1158/1538-7445.AM2014-3986
Abstract 3987: Evaluation of anti–malarial compounds as inhibitors of ezrin in osteosarcoma cells
Authors: Celik Haydar , Kallarakal Abraham , Colon-Lopez Daisy , Toretsky Jeffrey , Bosch Jurgen , Uren Aykut
Published: May 2015

Ezrin is a member of the ERM (Ezrin, Radixin, Moesin) family of proteins that functions as a cross-linker between the plasma membrane and actin cytoskeleton, thus allowing the cell to regulate cytoskeletal dynamics in response to external stimuli. Ezrin plays important roles in cell motility, membrane trafficking, morphogenesis, adhesion, survival and apoptosis. Ezrin also participates in crucial signal transduction pathways. Overexpression of ezrin contributes to the aggressive cancer behavior in various tumor types especially by enhancing the metastatic phenotype. Thus, ezrin represents a promising molecular target for cancer therapy. In our earlier studies, we have established that NSC305787 can directly bind to ezrin and inhibit its activity, which results in reduced metastasis. Since NSC305787 shares significant structural similarities with some anti-malarial drugs, we tested 400 anti-malarial compounds including 200 drug-like compounds and 200 probe-like compounds for their potential to directly bind to ezrin protein. We used surface plasmon resonance technology (Biacore) to screen direct binding. In this initial screen, 12 compounds demonstrated binding to ezrin over background values and selected as primary hits. Detailed SPR analysis of primary hits resulted in 7 compounds with comparable KD values to that obtained with our lead compound NSC305787. Primary hits were also evaluated for inhibition of the cell motility of murine osteosarcoma (OS) cells K7M2 and K12 that express high and low levels of ezrin, respectively. Cell motility and chemotaxis assays were conducted in parallel with viability assays to rule out general toxicity. Selected anti-malarial compounds were effective in inhibiting the motility of OS cells with high ezrin compared to cells with low ezrin. In conclusion, several anti-malarial drugs may have new indications in cancer therapy by inhibiting ezrin.

Citation Format: Haydar Celik, Abraham Kallarakal, Daisy Colon-Lopez, Jeffrey Toretsky, Jurgen Bosch, Aykut Uren. Evaluation of anti–malarial compounds as inhibitors of ezrin in osteosarcoma cells. [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 3987. doi:10.1158/1538-7445.AM2014-3987
Abstract 3988: The omega-3 fatty acids DHA and EPA inhibit medulloblastoma growth
Authors: Ljungblad Linda M. , Wickström Malin , Johnsen John-Inge , Kogner Per , Gleissman Helena
Published: May 2015

Background/Aim: Docosahexaenoic acid (DHA) and eicosapentaenic acid (EPA) are two essential omega-3 polyunsaturated fatty acid that have been shown to exert antitumoral effects in several cancer types, both in vitro and in vivo. DHA has also been shown to protect neural cells from apoptosis and is of importance in the maintenance of normal brain development and function.

Medulloblastoma (MB) is the most common malignant brain tumor in children with a poor 5-year survival of only 50-60%. It is a highly invasive embryonal neuroepithelial tumor that arises in the cerebellum and has a tendency to disseminate throughout the CNS early in its course. Multimodal treatment is always necessary in the form of surgery, radiotherapy and chemotherapy. However, treatment for this disease often results in significant endocrinological and intellectual sequelae. The neuroprotective and antitumoral properties of omega-3 fatty acids could therefore be of great benefit in this patient group. Here we aim to investigate the effect of DHA and EPA in medulloblastoma.

Methods: Cytotoxic activity of DHA and EPA was studied in cell viability assay in a panel of medulloblastoma cell lines. The molecular mechanisms were characterized using cell- and molecular biology techniques.

Results: By adding different concentrations of DHA and EPA we could demonstrate IC50s ranging between 1.9 and 46 µM for DHA and 3.8 and 68 µM for EPA in the six examined medulloblastoma cell lines. All cell lines showed greater sensitivity for DHA than EPA. Preliminary data showed that treatment with DHA inhibited the prostaglandin E2 production in medulloblastoma cells, indicating a possible mechanism of action.

Conclusions: Medulloblastoma cells are highly sensitive to omega-3 induced toxicity. Previous studies have showed that DHA can protect neurons from apoptosis. Therefore, omega-3 fatty acids are good candidates for improving medulloblastoma therapy by acting as both “sword and shield” by killing off cancer cells while protecting healthy neurons from therapy induced toxicity, thus possibly both inhibiting tumor growth and reducing the sequelae associated with other modes of brain tumor treatment.

Citation Format: Linda M. Ljungblad, Malin Wickström, John-Inge Johnsen, Per Kogner, Helena Gleissman. The omega-3 fatty acids DHA and EPA inhibit medulloblastoma growth. [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 3988. doi:10.1158/1538-7445.AM2014-3988