bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2248: Anti-leukemic activities of alcoholic extracts of two traditional Indian medicinal plants.
Authors: Malhotra Hemant , Malhotra Bharti , Bhargava Shipra , Rathode Om Singh , Sharma Pratibha , Mathur Ashwin , Jasuja Sandeep , Chiplunkar Shubha V.
Published: August 2014

Background: In recent years there has been a re-awakening of interest in the identification of Medicinal plants and their derivatives for the treatment of human diseases including cancer. The present work aims to investigate the in-vitro activity of two plants used in traditional Indian medicine - Zingiber Officinale Rose(Rhizome) and Nerium Oleander L(Leaves) against a human leukemic cell line.

Method: The plants were extracted with methanol and ethanol using soxalate apparatus. In-vitro anti-cancer activity was screened against leukemic cell line K562 using the SRB and MTT assay. Apoptosis and DNA fragmentation were characterized by PI staining flow cytometry and Gel electrophoresis, respectively.

Results: Notable cancer cell growth inhibition was observed for extracts from Zingiber Officinale Rose and Nerium Oleander L with IC50 values ranging between 10 -28 μg/ml. Furthermore, treatment of cells with both the extracts was shown to produce cell cycle arrest and DNA damage indicating that the observed cytotoxicity was mediated via apoptosis.

Conclusion: Extracts of Ginger and Oleander has anti-cancer activity against the K572 cell line in an in-vitro system. Based upon this initial screening work reported here, further studies aimed at the identification of active components of these extracts are planned. The activity of these agents against cell lines developed from actual patient samples are also planned.

Citation Format: Hemant Malhotra, Bharti Malhotra, Shipra Bhargava, Om Singh Rathode, Pratibha Sharma, Ashwin Mathur, Sandeep Jasuja, Shubha V. Chiplunkar. Anti-leukemic activities of alcoholic extracts of two traditional Indian medicinal plants. [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 2248. doi:10.1158/1538-7445.AM2013-2248
Abstract 2249: Induction of PIAS3 by curcumin as a novel treatment strategy in mesothelioma.
Authors: Dabir Snehal , Wildey Gary , Dowlati Afshin
Published: August 2014

Curcumin, a natural drug derived from the spice turmeric, has been extensively studied as an anti-cancer agent. The mechanism of action behind this promising bioactivity of curcumin, previously unclear, is now beginning to emerge. Curcumin has been shown to inhibit STAT3 phosphorylation and activity in melanoma cells, ovarian and endometrial cancer cells and small-cell lung cancer cells. In many cancers constitutively activated STAT3 (Signal Transducers and Activators of Transcription 3) is essential for cell proliferation. In this study we demonstrate that four mesothelioma cell lines demonstrate high levels of active STAT3. The addition of curcumin can suppress mesothelioma cell growth and proliferation, demonstrating the STAT3-dependence of these cells. Interestingly, the inhibitory effect of curcumin on STAT3 appears to be mediated through an upregulation of an endogenous inhibitor of STAT3 called PIAS3 (protein inhibitor of activated STAT3). Thus, the significance of our results is that curcumin not only targets an effector of mesothelioma cell proliferation, STAT3, but does this by increasing the expression of its endogenous inhibitor, PIAS3. This result prompted ongoing studies to determine if PIAS3 levels are deficient in mesothelioma tumor specimens obtained from the National Virtual Mesothelioma bank. Taken together, our data support the idea that mesothelioma is a STAT3-driven cancer caused by a lack of PIAS3 expression and that strategies to increase endogenous PIAS3 expression will have tumor suppressor effects. The ability of curcumin to increase PIAS3 expression will give us a jump start on pursuing this strategy because it represents a bioactive lead compound with many available structural analogs.

Citation Format: Snehal Dabir, Gary Wildey, Afshin Dowlati. Induction of PIAS3 by curcumin as a novel treatment strategy in mesothelioma. [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 2249. doi:10.1158/1538-7445.AM2013-2249
Abstract 2266: The characterization of anti-breast cancer compounds isolated from the Juniperus communis.
Authors: Robichaud Gilles A. , Picot Nadia , Jean Stéphanie , Carpenter Caitlyn , Gray Chris A.
Published: August 2014

Breast cancer is the most common form of cancer in the female population worldwide where approximately 30% of these women will succumb to their disease. The identification of novel, efficacious and safe anticancer drugs is thus essential to mitigate the morbidity and mortality associated with this disease. More recently, the study of anticancer properties from natural products has regained popularity as natural entities provide more biodiversity of chemical structures with biological and medicinal activity. Based on a documented library of the most common medicinal plants used by the indigenous people of North America, we have examined the anti-breast cancer properties of extracts from Juniperus communis (common juniper).

Using methanolic extracts from juniper needles and branches, we conducted bioassay-guided fractionation to assess growth inhibition of the highly malignant MB231 breast cancer cell line. The isolation and identification of active juniper constituents were performed through high performance liquid chromatography (HPLC) and standard spectroscopic analysis (1D- and 2D-nuclear magnetic resonance [NMR] and low-resolution and high-resolution mass spectrometry).

We characterized the diterpene isocupressic acid and the aryltetralin lignan deoxypodophyllotoxin as very potent inhibitors of cancer cell growth using dose-response and kinetic cell viability assays. Using a series of cellular and molecular tests, we also found that isocupressic acid and deoxypodophyllotoxin block MB231 cell cycling through the suppression of the ERK/MAPK and the NFκB signaling pathways. In addition to the blockade of the cell's survival cascade, both compounds concomitantly induced CASPASE-3-dependant apoptosis of MB231 cells within hours of treatment.

Our findings emphasize the potential and importance of natural products screening for new chemical entities with novel anticancer activities. Natural products research complemented with the wealth of information available through the ethnobotanical and ethnopharmacological knowledge of the indigenous natives will provide new candidate entities with desirable bioactivities to develop new cancer therapies.

Citation Format: Gilles A. Robichaud, Nadia Picot, Stéphanie Jean, Caitlyn Carpenter, Chris A. Gray. The characterization of anti-breast cancer compounds isolated from the Juniperus communis. [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 2266. doi:10.1158/1538-7445.AM2013-2266
Abstract 2267A: Red beetroot dye inhibits proliferation in squamous cell carcinoma of the head and neck.
Authors: Lew Daniel , Wrege Kay , Stoner Gary , Bock Jonathan M.
Published: August 2014

Head and Neck Squamous Cell Carcinoma (SCCHN) comprises more than 90% of all head and neck cancers. Despite the many recent advancements in cancer therapy, SCCHN survival has had little improvement with a 5-year survival rate of less than 50%. SCCHN is currently the 6th most common form of cancer in the United States and is rapidly becoming one of the most frequent cancers in developing nations. Red beetroot dye is a commonly used food additive to add a deep red color to commonly consumed foods like ice cream and yogurt. It also contains active anticancer chemical compounds such as betanins and betalains. These compounds are prepared by crushing the beetroots and mixing with water, ascorbic acid, citric acid, and dextrin. Previous work has shown that red beetroot dye can be effective in suppressing the development of multiorgan tumors in animal experiments, including up to 70% prevention of esophageal cancer in a rat chemoprevention study. No previous studies have evaluated the ability of red beetroot dye to inhibit the growth and proliferation of SCCHN. SCCHN cells were treated with increasing dose concentration of red beetroot dye and assessed for proliferation, apoptosis, cell cycle changes, and G1 protein expression changes. We observed dose dependent toxicity in SCCHN following red beetroot dye treatment with concomitant cell cycle arrest in the G1 and G2 cell cycle phases. Cell cycle arrest appeared to be more significant with p53 mutation. Decreased cell counts were induced by cells undergoing apoptosis and necrosis. UMSCC-17b (mutant p53) treated with increasing doses of red beetroot dye expressed lower levels of p53 and p21 by western blot analysis. Taken together, these results suggest that red beetroot dye may have significant potential as a chemopreventive or adjuvant treatment for SCCHN.

Citation Format: Daniel Lew, Kay Wrege, Gary Stoner, Jonathan M. Bock. Red beetroot dye inhibits proliferation in squamous cell carcinoma of the head and neck. [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 2267A. doi:10.1158/1538-7445.AM2013-2267A
Abstract 228: Curcumin attenuates tumor initiating stem-like property of head and neck cancer through miR145 axis-mediated paracrine signaling.
Authors: Chiou Shih-Hwa , Chiou Guang-Yuh , Lu Kai-Hsi
Published: August 2014

Recent reports have demonstrated that head and neck cancer derived tumor initiating cells (HNC-TICs) presented high tumorigenic, chemo-radioresistant, metastatic properties, and coupled with gain of epithelial-mesenchymal transition (EMT) characteristics. As curcumin, an active component of the spice turmeric, can exert cytotoxic effects on cancer cells without harming normal tissues. The aim of this study was to investigate the chemo-therapeutic effect and regulatory mechanisms of curcumin on HNC-TICs. We first observed that the treatment of curcumin significantly down-regulated the ALDH1 activity, CD44 positivity, self-renewal property, and side population of HNC cells in a dose dependent manner. Using miRNA/mRNA-microarray analysis, curcumin significantly increased expression of tumor suppressive miR-145. Further mechanistic studies showed that the re-expression of PU-PEI-mediated miR145 delivery led to decreased of TICs properties. Blocking of endogenous miR145 can dramatically enhance stemness and tumor-initiating properties in ALDH1-CD44- non-TICs HNC cells. Additionally, the repressive effect of miR-145 on TICs properties was mediated by regulating of EMT. Importantly, in vivo nude mice model showed that cucumin treatment by oral gavage or PU-PEI-mediated miR145 delivery to xenograft tumors reduced tumor growth and metastasis and prolonged the survival times of tumor-bearing mice. From these results, we conclude that the inhibition of tumor aggressiveness in HNC-TIC in by curcumin was in part was mediated by up-regulation of miR-145, suggesting that curcumin would be a valuable therapeutics clinically in treatment modalities for malignant head and neck cancers by elimination of tumor initiating stem-like, and EMT properties.

Citation Format: Shih-Hwa Chiou, Guang-Yuh Chiou, Kai-Hsi Lu. Curcumin attenuates tumor initiating stem-like property of head and neck cancer through miR145 axis-mediated paracrine signaling. [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 228. doi:10.1158/1538-7445.AM2013-228
Abstract 2283: Nano-scale proteomic profiles of response to targeted therapy in patients with RCC.
Authors: Fan Alice C. , Leppert John , Liliental Joanna E. , Xu Liwen , Metzner Thomas J. , Shroff Emelyn , Yaghi Alia , Yost Christine , Brooks James D. , Harshman Lauren C. , Sabatti Chiara , Srinivas Sandhya , Felsher Dean W.
Published: August 2014

Novel agents that inhibit targets in the hypoxia and MTOR pathways can achieve response rates of 30-60% in renal cell carcinoma (RCC); however, biomarkers have not yet been identified to measure degree of target inhibition and mechanism of biologic response in individual patients. We developed the use of a high-throughput, nanoscale immunoassay (NIA, Nanopro1000 Instrument, Protein Simple) to profile hypoxia pathways and downstream signaling in serially sampled clinical specimens from patients with RCC. NIA uses charge-based separation to distinguish multiple charged modifications of individual proteins, and measure relative ratios of individual unphosphorylated and phosphorylated isoforms. We first developed and optimized assays to measure therapeutic targets and proteins in hypoxia, proliferation and metabolic pathways (including CaIX, HIF, VEGFR, ERK, AKT, MYC, glutaminase) to analyze frozen surgical specimens, fine needle aspirates (FNA's) and blood PBMC specimens from patients with RCC. Next, we analyzed more than 200 FNA's from solid tumors, comparing RCC with paired adjacent non-tumor tissue and other epithelial malignancies. Basal MAPK signaling across the samples ranged across 3 logs for both normal and tumor tissue, demonstrating the difficulty in setting a threshold predictive of response based on a single pre-treatment measurement. To test whether analysis of serial specimens could detect changes, we prospectively collected and analyzed blood peripheral mononuclear cells from 20 RCC patients before and during treatment with standard targeted therapies including MTOR and tyrosine kinase inhibitors. We report for the first time that targeted inhibitors can preferentially inhibit or activate specific protein phospho-isoforms in patients with RCC. Our studies demonstrate that rapid and quantitative nanoproteomic profiling in very small amounts of serially sampled clinical specimen may accelerate translational studies for predictive proteomic biomarkers of response to targeted therapy.

Citation Format: Alice C. Fan, John Leppert, Joanna E. Liliental, Liwen Xu, Thomas J. Metzner, Emelyn Shroff, Alia Yaghi, Christine Yost, James D. Brooks, Lauren C. Harshman, Chiara Sabatti, Sandhya Srinivas, Dean W. Felsher. Nano-scale proteomic profiles of response to targeted therapy in patients with RCC. [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 2283. doi:10.1158/1538-7445.AM2013-2283
Abstract 231: Conformation-specific monoclonal antibodies against Notch1 target aberrant Notch1 signaling in cancer stem cells.
Authors: Sharma Ankur , Rangarajan Annapoorni , Dighe Rajan R.
Published: August 2014

Notch signaling pathway plays a key role in regulating various cell fates including those of stem cells, throughout the mammalian development and adult homeostasis. Here we report characterization of anti-Notch1 monoclonal antibodies (mAb's) with potential therapeutic applications. Antibodies against various domains of human-Notch1 receptor were generated and characterized, mAb's generated against primary ligand-binding domain (EGF like repeats 11-12) and activation domain NRR (Negative Regulatory region) showed most prominent therapeutic potential. The Notch1 specific mAb 602.101 against ligand-binding domain (EGF like repeats 11-12) prevents binding of Jagged and Delta-like ligands and subsequent Notch1 receptor activation in breast cancer stem cells. Further, antibody treatment leads to apoptosis, inhibition of epithelial to mesenchymal transitions as well as irreversible depletion of cancer stem cells. The conformation specific mAb's against Notch1 NRR can specifically distinguish between the mutant and the wild type human Notch1 receptor present on cancer cells. Anti-NRR mAb's 604.107 (mutant specific) and 604.132 (wild type specific) specifically inhibited Notch1 signaling in leukemic and breast cancer stem cells. These mAbs can serve as a potential tool to irreversibly combat therapy resistant cancer stem cell subpopulations and to inhibit the epithelial to mesenchymal transitions in various types of cancers.

Citation Format: Ankur Sharma, Annapoorni Rangarajan, Rajan R. Dighe. Conformation-specific monoclonal antibodies against Notch1 target aberrant Notch1 signaling in cancer stem cells. [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 231. doi:10.1158/1538-7445.AM2013-231
Abstract 2314: The chromatin structure regulates selective glucocorticoid receptor recruitment to the genome in intact tissue.
Authors: Grontved Lars , John Sam , Baek Songjoon , Liu Ying , Buckley John , Vinson Charles , Aguilera Greti , Hager Gordon L.
Published: August 2014

The dissection of regulatory pathways in physiological and pathophysiological states and during pharmacological intervention necessitates the application of biochemical and genomic approaches in intact target tissues. The hypothalamus-pituitary-adrenal axis is a highly conserved neuroendocrine pathway that results in the synthesis of glucocorticoid hormones from the adrenal gland to control reactions to stress and inflammatory processes. Exploiting a mouse model in which animals have been adrenalectomized to remove endogenous glucocorticoids, we monitor changes of the chromatin landscape in liver, following a single glucocorticoid injection, using ChIP-seq and DNase-seq methodologies.

We observe that GR exclusively occupies accessible chromatin, demonstrating that accessible chromatin can be highly predictive of transcription factor occupancy in tissue. Importantly, changes in RNA polymerase II recruitment to GR regulated genes are associated with proximal regions of induced and repressed chromatin accessibility. Underscoring the highly tissue-specific GR-dependent transcriptional response, GR binding sites in other cell types are occluded in liver through highly positioned nucleosomes and inaccessible chromatin. Remarkably, we demonstrate that sixty two percent of GR binding sites in liver are associated with C/EBPbeta occupancy. At the majority of these sites chromatin is pre-programmed, suggesting a priming function of C/EBPbeta in GR recruitment. However, at a subset of GR binding sites C/EBPbeta loading is assisted by GR recruitment in conjunction with de novo chromatin remodeling.

Disruption of C/EBPbeta binding to chromatin using a dominant negative C/EBP targeted to the liver results in an attenuation of pre-programmed chromatin accessibility, GR recruitment and GR induced chromatin remodeling only at sites occupied by GR and C/EBP. Our results demonstrate a highly cooperative mechanism by which C/EBP regulates selective GR binding to the genome in liver tissue and provides a mechanism for phenotypic similarities described for GR and C/EBP mouse models. We suggest that the selective targeting of GR in other tissues is likely mediated by the combined action of cell-specific priming proteins and chromatin remodelers.

Citation Format: Lars Grontved, Sam John, Songjoon Baek, Ying Liu, John Buckley, Charles Vinson, Greti Aguilera, Gordon L. Hager. The chromatin structure regulates selective glucocorticoid receptor recruitment to the genome in intact tissue. [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 2314. doi:10.1158/1538-7445.AM2013-2314
Abstract 2343: A novel ERK inhibitor is active in models of acquired resistance to BRAF and MEK inhibitors.
Authors: Samatar Ahmed A. , Morris Erick J. , Jha Sharda , Clifford Restaino R. , Luttrerbach Bart , Pelletier Marc , Philippar Ulrike , Jayaraman Lata , Zawel Leigh , Fawell Steve , Gilliland Gary
Published: August 2014

The high frequency of activating BRAFV600E mutations in melanoma (40-70%), thyroid (50%) and colorectal cancer (10%), or KRAS/NRAS mutations in melanoma (20%), pancreatic (90%), colorectal (50%) and non-small cell lung cancer (30%), provides strong rationale for targeting the MAPK pathway as a therapeutic strategy 1-6. Vemurafenib (PLX4032) and dabrafenib (GSK2118436), selective BRAF inhibitors, and trametinib (GSK1120212), an allosteric MEK inhibitor, have shown robust clinical efficacy in melanoma patients 7-10. However, the majority of responses are transient and cellular resistance is often associated with pathway reactivation involving the downstream extracellular-signal-regulated kinases 1 and 2 (ERK1/2) (reviewed in 11). We hypothesized that pathway blockade at ERK, the last signaling node prior to MAPK transcriptional programming, would not only be efficacious in MAPK-activated tumors but would also have utility in BRAF or MEK inhibitor resistant settings. We therefore sought to identify small molecule inhibitors of ERK. This report describes the identification and characterization of SCH772984, a potent and selective ATP competitive inhibitor of ERK1/2 which displays behaviors of both type I and type II kinase inhibitors. SCH772984 has nanomolar cellular potency on tumor cells with mutations in BRAF, NRAS, or KRAS and induces tumor regressions in xenograft models at tolerated doses. Importantly, SCH772984 effectively inhibited MAPK signaling and cell proliferation in BRAF or MEK inhibitor resistant models as well as in the context of BRAF/MEK combination resistance. Together these data support the clinical development of ERK inhibitors, not only in patients with MAPK activated tumors, but also in patients who have developed acquired resistance to BRAF or MEK inhibitors or resistance to the recently described combination of these agents.

Citation Format: Ahmed A. Samatar, Erick J. Morris, Sharda Jha, Restaino R. Clifford, Bart Luttrerbach, Marc Pelletier, Ulrike Philippar, Lata Jayaraman, Leigh Zawel, Steve Fawell, Gary Gilliland. A novel ERK inhibitor is active in models of acquired resistance to BRAF and MEK inhibitors. [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 2343. doi:10.1158/1538-7445.AM2013-2343
Abstract 2354: An E2F signature predicts benefit of adjuvant chemotherapy in early-stage non-small cell lung cancer.
Authors: Kurtyka Courtney A. , Chen Dung-Tsa , Chen Lu , Brazelle William , Welsh Eric A. , Berglund Anders E. , Eschrich Steven A. , Schabath Matthew B. , Haura Eric B. , Cress W. Douglas
Published: August 2014

Early-stage non-small cell lung cancer (NSCLC) is primarily treated by surgical resection. Unfortunately, after resection, one-third to one-half of early-stage patients will die of metastatic recurrence. Adjuvant chemotherapy (ACT) improves the survival of patients with early-stage disease and has become the standard treatment for patients with resected stage II-III NSCLC. However, the five-year survival advantage of ACT is only 4%-15% suggesting that many patients do not benefit. Given the morbidity associated with ACT, it is imperative to develop new prognostic tools to identify those patients with a high probability of relapse. Toward this end, we have used small inhibitory RNAs targeting multiple E2F pathway components to derive an E2F gene expression signature in vitro. This signature was refined by filtering for its components that were altered in non-small cell lung cancers compared to normal tissue. Principle component analysis was then used to represent the signature which was tested for correlation to overall survival in two large cohorts. The first of the two cohorts was the Molecular Classification of Lung Adenocarcinoma (MCLA) from the Director's Challenge Consortium and the second was a novel database on 444 lung adenocarcinomas treated as a part of Moffitt's Total Cancer Care Network. The E2F signature is strongly prognostic in both cohorts with P values <0.0001. Additionally, using data in the JBR.10 trial (GSE14814) for patients who either did or did not receive ACT we were able to determine that patients having a high E2F signature benefit from ACT (have increased overall survival), whereas patients with a low E2F signature do not. Overall, these results suggest that this approach could be optimized in the clinical setting to distinguish patients likely to benefit from ACT from those who will not.

Citation Format: Courtney A. Kurtyka, Dung-Tsa Chen, Lu Chen, William Brazelle, Eric A. Welsh, Anders E. Berglund, Steven A. Eschrich, Matthew B. Schabath, Eric B. Haura, W. Douglas Cress. An E2F signature predicts benefit of adjuvant chemotherapy in early-stage non-small cell lung cancer. [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 2354. doi:10.1158/1538-7445.AM2013-2354
Abstract 2365: Comparison of epidermal growth factor receptor mutation tests in patients with recurred non-small cell lung cancer after surgery.
Authors: Oh In-Jae , Kim Woo-Jin , Kang Hyunwook , Kim Kyu-Sik , Kim Young-Chul , Song Sang-Yun , Choi Yoo-Duk , Hwang Jun-Eul , Yang Sei Hoon
Published: August 2014

Background: Epidermal growth factor receptor (EGFR) mutations are predictive marker of EGFR-tyrosine kinase inhibitor (TKI) therapy in non-small cell lung cancer (NSCLC). We compared the agreements between three EGFR mutation tests with surgical specimens of recurred NSCLC.

Methods: We retrospectively analyzed paraffin-embedded tissues, which were obtained from 111 (76 adenocarcinoma, 17 squamous cell carcinoma, 18 others) cases of 108 patients treated by EGFR-TKIs after surgery between 2003 and 2011 in Chonnam National University Hwasun Hospital. EGFR mutations in exons 18-21 were analyzed by three different methods- Cobas® EGFR mutation test (Roche), PNA Clamp™ EGFR mutation test (Panagene) and direct sequencing- and compared with radiologic response to EGFR-TKIs. The degree of agreement was evaluated by Cohen's kappa value.

Results: The mean treatment duration of total 111 (64 gefitinib, 47 erlotinib) cases was 188.5±198.7 days. The response rate was 18.9%, and disease control rate was 63.1%. EGFR mutation rates were 38.7% in PNA clamping, 34.2% in Cobas test, and 31.5% in direct sequencing. The agreement of Cobas test and PNA clamping was 93.7% (k=0.864, p<0.001), that of Cobas test and sequencing was 84.7% (k=0.653, p<0.001), and that of PNA and sequencing was 78.4% (k=0.528, p<0.001). The prediction rates of EGFR-TKIs response were 39.5% of Cobas test, 37.2% of PNA clamping, and 34.3% of sequencing. The degree of agreement between mutation and clinical response was better in Cobas test (k=0.350, p<0.001) and PNA clamping (k=0.330, p<0.001) than sequencing (k=0.252, p=0.008).

Conclusion: The strength of agreement of Cobas test was very good with PNA clamping and good with direct sequencing. So Cobas test can be useful test for EGFR-TKI therapy in recurred NSCLC after surgery.

Citation Format: In-Jae Oh, Woo-Jin Kim, Hyunwook Kang, Kyu-Sik Kim, Young-Chul Kim, Sang-Yun Song, Yoo-Duk Choi, Jun-Eul Hwang, Sei Hoon Yang. Comparison of epidermal growth factor receptor mutation tests in patients with recurred non-small cell lung cancer after surgery. [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 2365. doi:10.1158/1538-7445.AM2013-2365
Abstract 2368: An online tool for the validation of survival-predicting biomarkers in non small-cell lung cancer using microarray data of 1,329 patients.
Authors: Gyorffy Balazs , Lanczky Andras
Published: August 2014

To decipher the molecular basis of lung cancer and to improve treatment strategies we have to identify genes correlated to therapy response and to survival. Here, we present the development of a new, freely available online tool suitable for the real-time meta-analysis of published lung cancer microarray datasets to validate expression based survival related biomarker candidates.

First we have searched the caBIG, GEO and TCGA repositories to identify datasets with published gene expression data and survival information. In this, three GEO platforms (GPL96, GPL570 and GPL3921) were considered as these possess 22,277 common probes, which were used for constructing the database. All together 1,329 NSCLC samples in 9 independent datasets were identified, 85% have overall survival and 45% progression free survival info, one-third adenocarcinomas and one-third squamous cell carcinomas, 57% male and the average overall survival is 45 months. Kaplan-Meier survival plot, and the hazard ratio with 95% confidence intervals and logrank P value are calculated and plotted in R using Bioconductor packages. To assess the prognostic value of a gene, each percentile (of expression) between the lower and upper quartiles are computed and the best performing threshold is used as the final cutoff in a Cox regression analysis. The complete meta-analysis tool can be accessed online at: www.kmplot.com/lung.

We demonstrate the application of this integrative analysis pipeline by validating 22 previously published survival associated biomarkers including VEGF, ADAM28, ANXA3, CADM1, CD24, CD82, CDK1, CEA, cyclin E, ERCC1, EZH2, HER2, IFNAR2, MMP9, OPN, P16, p53R2, RAD51, S100A4, survivin, XAF1 and XIAP. Of these, high significances (p<1E-16) were achieved by ANXA3 (HR=0.63), CADM1 (HR=1.7), IFNAR2 (HR=0.54), RAD51 (HR=1.9) and XAF1 (HR=0.58). Five additional genes were significant: S100A4 (HR=1.46, p=3.6E-4), CD82 (HR=1.34, p=0.018), OPN (HR=1.33, p=0.02), HER2 (HR=1.3, p=0.02) and CD24 (HR=0.75, p=0.3). The remaining genes did not achieved statistical significance.

In summary, we developed an online available meta-analysis tool for the validation of genes associated with survival in NSCLC.

Citation Format: Balazs Gyorffy, Andras Lanczky. An online tool for the validation of survival-predicting biomarkers in non small-cell lung cancer using microarray data of 1,329 patients. [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 2368. doi:10.1158/1538-7445.AM2013-2368
Abstract 2378: Upregulation of the ATR/CHK1 pathway and decreased therapeutic sensitivity in upper aerodigestive cancer cell lines with distal 11q loss.
Authors: Sankunny Madhav , Parikh Rahul A. , Gollin Susanne M.
Published: August 2014

Gemcitabine, a pyrimidine analog/antimetabolite, is commonly used to treat breast, lung, head and neck, pancreatic, ovarian, biliary, urinary bladder, and cervical carcinomas. Gemcitabine inhibits DNA synthesis by direct inhibition of ribonucleotide reductase by its metabolite, gemcitabine diphosphate, and the incorporation of its active form, gemcitabine triphosphate into the elongating DNA strand, resulting in chain termination. Gemcitabine-induced DNA damage leads to activation of the ATR/CHK1 pathway and S-phase cell cycle arrest. Our previous studies showed that carcinoma cells with our biomarker (distal 11q loss) overexpress ATR and CHK1, resulting in decreased sensitivity to ionizing radiation in biomarker-positive head and neck squamous cell carcinoma (HNSCC) and non-small cell lung carcinoma (NSCLC) cell lines. Several studies in the literature have shown that inhibition of CHK1 increases the sensitivity of tumor cell lines to replication inhibitors, including gemcitabine. In pancreatic cancer cell lines, small molecule inhibitor (SMI)-mediated disruption of CHK1 signaling has been reported to potentiate the cytotoxic effect of gemcitabine. Based on the literature and our previous findings, we now report that biomarker-positive HNSCC and NSCLC cell lines also show ATR/CHK1 upregulation and decreased sensitivity to gemcitabine, Further, we observed that siRNA knockdown of the ATR/CHK1 pathway and SMI inhibition of CHK1 signaling in HNSCC and NSCLC cell lines increase their sensitivity to treatment with gemcitabine. Our previous results showed increased efficacy of ionizing radiation combined with CHK1 inhibition in biomarker-positive cell lines; our new results show increased efficacy of gemcitabine combined with CHK1 inhibition in biomarker-positive HNSCC and NSCLC cell lines. These results provide further support for combining CHK1-targeted therapy with standard therapy in conjunction with an appropriate companion diagnostic biomarker, like distal 11q loss. As is the case with other targeted therapies, biomarker-based selection of the right drug for the right patient appears to increase therapeutic efficacy.

Citation Format: Madhav Sankunny, Rahul A. Parikh, Susanne M. Gollin. Upregulation of the ATR/CHK1 pathway and decreased therapeutic sensitivity in upper aerodigestive cancer cell lines with distal 11q loss. [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 2378. doi:10.1158/1538-7445.AM2013-2378
Abstract 2385: Identification of a molecular marker for cetuximab sensitivity and development of a novel combination therapy for non-small cell lung cancers.
Authors: Takata Miyako , Chikumi Hiroki , Yamaguchi Kosuke , Kurai Jun , Nakamoto Masaki , Burioka Naoto , Igishi Tadashi , Shimizu Eiji
Published: August 2014

The epidermal growth factor receptor (EGFR) is an important molecular target for non-small-cell lung cancers (NSCLCs). A recent phase III study (the FLEX study) revealed that the combination of the anti-EGFR antibody cetuximab with conventional chemotherapy improved the survival of patients with NSCLCs. However, the survival benefit was only about 5 weeks. Therefore, the identification of a molecular marker for cetuximab sensitivity is urgently needed. In this study, we investigated a new molecular marker for cetuximab sensitivity and developed a combination therapy to improve the effectiveness of cetuximab. First, we established a NSCLC cell line that showed resistance to cetuximab by culturing a parental sensitive NSCLC cell line in the presence of increasing concentrations of cetuximab. Next, we performed microarray analysis to compare the gene expression patterns of cetuximab-sensitive and -resistant cell lines. Using pathway analysis, we found that the Akt and Wnt pathways were significantly activated in the resistant cell line as compared to their activation in the sensitive cell line. We confirmed this result by performing western blot analysis, which showed Akt phosphorylation and beta-catenin dephosphorylation in the resistant cell line. Therefore, the activation status of either the Akt or Wnt pathway is the candidate pathway that influences cetuximab sensitivity. Finally, we focused on the Akt pathway and treated both resistant and parental sensitive cell lines with the PI3-K inhibitor LY294002 in combination with cetuximab. This combination therapy induced a synergistic growth-inhibiting effect in both cell lines. The data suggest that the activation status of the Akt pathway is one of the markers for cetuximab sensitivity and the combination of an Akt pathway inhibitor and cetuximab would be a promising anti-EGFR therapy for NSCLCs.

Citation Format: Miyako Takata, Hiroki Chikumi, Kosuke Yamaguchi, Jun Kurai, Masaki Nakamoto, Naoto Burioka, Tadashi Igishi, Eiji Shimizu. Identification of a molecular marker for cetuximab sensitivity and development of a novel combination therapy for non-small cell lung cancers. [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 2385. doi:10.1158/1538-7445.AM2013-2385

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2391: HPV status predicts therapy response in patients with locally advanced HNSCC receiving cetuximab-based radioimmunotherapy.
Authors: Stenzinger Albrecht , Wolf Janina , Jensen Alexandra , Endris Volker , Dietel Manfred , Weichert Wilko
Published: August 2014

HPV+ HNSCC show better therapy responses and improved survival when compared to HPV- HNSCC. No data exist on the prognostic value of HPV and combined HPV/EGFR status in patients with locally advanced HNSCC (LAHNSCC) treated with radiotherapy and cetuximab. The HPV status defined by p16INK4A immunohistochemistry and PCR-based chip analysis was assessed in a clinically well-characterized cohort of patients with LAHNSCC. Outcome was estimated using Kaplan-Meier analysis. Additionally, the EGFR expression levels and the effect of radioimmunotherapy in HPV+ and HPV- HNSCC cell lines were determined. 34% of the cases were p16INK4A/HPV positive. p16INK4A significantly correlated with male sex, younger patients and oropharnygeal localization and was a powerful predictor of OS (p=0.001) and PFS (p≤0.001). The HRs for HPV+ tumors were 0.17 (95%CI: 0.056-0.524, p=0.002) for OS and 0.12 (95%CI: 0.033-0.438, p=0.001) for PFS. P16INK4A correlated well with the HPV status (p≤0.001). The HPV+ UD-SCC2 cells had higher EGFR levels when compared to HPV- FaDu cells and the efficacy of combined radioimmunotherapy was stronger. This is the first study that demonstrates the prognostic value of HPV in patients with LAHNSCC receiving cetuximab-based radioimmunotherapy.

Citation Format: Albrecht Stenzinger, Janina Wolf, Alexandra Jensen, Volker Endris, Manfred Dietel, Wilko Weichert. HPV status predicts therapy response in patients with locally advanced HNSCC receiving cetuximab-based radioimmunotherapy. [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 2391. doi:10.1158/1538-7445.AM2013-2391
Abstract 2400: Differential phosphorylation of signaling targets as a function of RAS mutation status and tumor histology in non-small cell lung cancer.
Authors: Um Sang-Won , Chen Jingxiang , Lee Carrie , Sharpless Norman E. , Roberts Patrick J. , Hayes D. Neil
Published: August 2014

Background: While the importance of RAS and mTOR signaling pathways in non-small cell lung cancer (NSCLC) is well known, the activation status of key components in the pathway as a function of histology and oncogene mutation status has been not completely described. The purpose of this study is to investigate phosphorylation status of signal transduction pathway members and to elucidate their prognostic significance in relation to adenocarcinoma and squamous cell carcinoma.

Methods: Phosphorylated extracellular signal-regulated kinases 1 and 2 (pERK), MEK 1 and 2 (pMEK), and mammalian target of rapamycin (pmTOR) were immunohistochemically examined using tissue microarray samples of 269 surgically resected primary NSCLC. Thyroid transcription factor-1 (TTF-1) expression and mutation status of EGFR and KRAS were also evaluated. The relationships between 5-year progression free survival (PFS) and phospho-protein expression were investigated by univariate analyses of log-rank test and multivariate analyses using Cox regression model.

Results: A total of 98 adenocarcinomas and 134 squamous cell carcinomas were included in the analyses. In adenocarcinoma, pERK expression was significantly related to pMEK (P = 0.001), pmTOR (P < 0.001), and TTF-1 (P = 0.013) expression. In squamous cell carcinoma, pERK expression was significantly related to pMEK (P < 0.001) expression. EGFR or KRAS mutations were significantly associated with TTF-1 expression (P = 0.014) in adenocarcinoma. However, EGFR and KRAS mutation status were not significantly associated with pERK, pMEK, and pmTOR expressions in adenocarcinoma. By univariate analyses, high pERK expression was associated with better 5-year PFS (P = 0.018) in adenocarcinoma and high pMEK expression was associated with worse 5-year PFS (P = 0.043) in squamous cell carcinoma. In multivariate analyses, pERK (P = 0.040) and pMEK (P = 0.048) expression were independent prognostic factors.

Conclusions: There were different patterns of phospho-protein expression in adenocarcinoma and squamous cell carcinoma. pERK expression was a good prognostic factor only in adenocarcinoma and pMEK expression was a poor prognostic factor only in lung squamous cell carcinoma.

Citation Format: Sang-Won Um, Jingxiang Chen, Carrie Lee, Norman E. Sharpless, Patrick J. Roberts, D. Neil Hayes. Differential phosphorylation of signaling targets as a function of RAS mutation status and tumor histology in non-small cell lung cancer. [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 2400. doi:10.1158/1538-7445.AM2013-2400
Abstract 2417: The INHERIT EGFR study: Investigating hereditary risk from T790M.
Authors: Oxnard Geoffrey R. , Root Elizabeth J. , Sable-Hunt Alicia , Rainville Irene , Dahlberg Suzanne E. , Heng Jennifer C. , Jackman David M. , Janne Pasi A. , Garber Judy E.
Published: August 2014

Background: The EGFR T790M mutation, commonly associated with acquired resistance to EGFR kinase inhibitors, has also been described rarely as a germline mutation in association with familial lung cancer. In collaboration with the Addario Lung Cancer Medical Institute (www.ALCMI.net), we initiated a prospective trial to identify patients and families carrying germline EGFR mutations in order to characterize their lung cancer risk.

Preliminary data: In a prior study (GR Oxnard et al, JTO, 2012), it was identified that patients with lung cancers found to harbor EGFR T790M at diagnosis have a 50% chance of carrying an underlying germline T790M mutation. This suggests that by focusing on patients already identified to carry T790M in their cancer, it is possible to enrich for a germline mutation that may be otherwise too rare to study prospectively.

Subject eligibility: The following groups are eligible: (1) Patients with a cancer (lung or other) harboring an EGFR T790M mutation on tumor genotyping; lung cancers that acquired EGFR T790M only following treatment with an EGFR kinase inhibitor are ineligible. (2) First-degree relatives of patients found to carry a germline EGFR mutation. (3) Patients who already are known to carry a germline EGFR mutation on prior testing. Subjects are referred to cohort 1 based upon genotyping results from academic centers, oncology practices, or partnering commercial laboratories (e.g. Response Genetics, Los Angeles, CA).

Trial design: Subjects may present at a participating cancer center or may participate remotely through a study website (www.Dana-Farber.org/T790Mstudy). Eligible patients submit a saliva and/or blood specimen for central testing in a CLIA lab. After counseling, patients carrying germline EGFR mutations have the option of inviting first-degree relatives to participate. Genetic counseling is coordinated at the participating center or offered over the phone. Chest CT scans are collected from germline carriers and analyzed centrally to study nodule prevalence and characteristics. Patients are then clinically followed for 2 years.

Funding: Supported by grants from the Conquer Cancer Foundation of ASCO and the Bonnie J. Addario Lung Cancer Foundation.

Citation Format: Geoffrey R. Oxnard, Elizabeth J. Root, Alicia Sable-Hunt, Irene Rainville, Suzanne E. Dahlberg, Jennifer C. Heng, David M. Jackman, Pasi A. Janne, Judy E. Garber. The INHERIT EGFR study: Investigating hereditary risk from T790M. [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 2417. doi:10.1158/1538-7445.AM2013-2417
Abstract 2421: Smoking, cancer treatment, and design of clinical trials.
Authors: Warren Graham W. , Marshall James R. , Cummings K. Michael
Published: August 2014

Purpose: Smoking has been associated with poor outcomes in cancer patients, but a detailed review of the effects of smoking on cancer treatment outcomes is necessary to effectively implement structured tobacco assessments and cessation into clinical trials design.

Methods: We searched Pubmed citations between 1950 and 2011 with the objective of identifying any reports relating smoking with cancer and non-cancer related outcomes in cancer patients. Our search yielded 34,000 pertinent citations of on smoking and cancer. Over 5000 of these citations appeared to include data on smoking as it might be related to cancer treatment outcomes and of these more than 800 were felt to have sufficient information to examine how smoking affects cancer treatment outcomes. Results were cataloged in tables and cross-references according to cancer disease site.

Results: The vast majority of pertinent reports provide evidence showing that smoking increases the risk of cancer and non-cancer related mortality as well as treatment related adverse events. The adverse effects of smoking and treatment outcomes are observed in all major cancer disease sites (lung, head/neck, breast, gastrointestinal, gynecologic, leukemia/lymphoma, prostate, and non-prostate genitourinary). A limited number of studies examine how smoking cessation affects treatment outcomes and generally these studies show that cessation of tobacco use is associated with fewer adverse treatment outcomes. Measurement of smoking behavior is a problem due to the reliance on self-reported behavior and lack of standardization in classifying smoking status. In most studies a patients’ cigarette use is categorized into ever smoking or current/former/never smoking; however, there is a broad diversity of definitions used making it less clear exactly how smoking affects treatment outcomes. Despite methodologic issues in published reports, results demonstrate a consistent pattern that smoking increases the risk of adverse outcomes across a broad spectrum of cancer disease sites. Conclusions: This review provides evidence that smoking adversely affects cancer related and non-cancer related outcomes in patients. What is less certain is how past and/or current smoking might differentially modify treatment outcomes for patients. Clinical trials must give strong consideration for prior tobacco history, current tobacco use, and efforts to track tobacco cessation during and following cancer treatment.

Citation Format: Graham W. Warren, James R. Marshall, K. Michael Cummings. Smoking, cancer treatment, and design of clinical trials. [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 2421. doi:10.1158/1538-7445.AM2013-2421