bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison Analysis of Single Loop Resonator Based Miniaturized Triple-Band Planar Monopole Antennas
Authors: Wu Kaimin , Huang Yongjun , Wen Ruiming , Li Jian , Wen Guangjun
Published: July 2015

We systemically analyze and discuss a group of miniaturized triple-band planar monopole antennas by integrating with a single loop resonator (SLR) as part of the radiation patch. Two configurations (rectangular and circular shaped radiation patches) are presented and the corresponding SLRs are integrated in the patches to contribute additional operating frequencies. Two feed techniques (coplanar waveguide and microstrip feeds) are performed to couple the electromagnetic waves. The conventional planar monopole antennas without the SLRs are also analyzed comparatively. Numerical and experimental methods are performed to demonstrate and discuss the proposed designs.
Fabrication of a Microtubular La0.6Sr0.4Ti0.2Fe0.8O3−δMembrane by Electrophoretic Deposition for Hydrogen Production
Authors: Lee Kyoung-Jin , Choe Yeong-Ju , Lee Jun-Sung , Hwang Hae-Jin
Published: July 2015

Microtubular type La0.6Sr0.4Ti0.2Fe0.8O3−δ(LSTF) membranes were prepared by electrophoretic deposition (EPD). The oxygen permeation and hydrogen production behavior of the membranes were investigated under various conditions. LSTF green layer was successfully coated onto a carbon rod and, after heat treatment at 1400°C in air, a dense LSTF tubular membrane with a thickness of 250 mm can be obtained. The oxygen permeation and hydrogen production rate were enhanced by CH4in the permeate side, and the hydrogen production rate by water splitting was 0.22 mL/min·cm2at 1000°C. It is believed that hydrogen production via water splitting using these tubular LSTF membranes is possible.
Integrated Production-Distribution Scheduling Problem with Multiple Independent Manufacturers
Authors: Hao Jianhong , Cao Lisi , Jiang Dakui
Published: July 2015

We consider the nonstandard parts supply chain with a public service platform for machinery integration in China. The platform assigns orders placed by a machinery enterprise to multiple independent manufacturers who produce nonstandard parts and makes production schedule and batch delivery schedule for each manufacturer in a coordinate manner. Each manufacturer has only one plant with parallel machines and is located at a location far away from other manufacturers. Orders are first processed at the plants and then directly shipped from the plants to the enterprise in order to be finished before a given deadline. We study the above integrated production-distribution scheduling problem with multiple manufacturers to maximize a weight sum of the profit of each manufacturer under the constraints that all orders are finished before the deadline and the profit of each manufacturer is not negative. According to the optimal condition analysis, we formulate the problem as a mixed integer programming model and use CPLEX to solve it.
Forecasting RMB Exchange Rate Based on a Nonlinear Combination Model of ARFIMA, SVM, and BPNN
Authors: Xie Chi , Mao Zhou , Wang Gang-Jin
Published: July 2015

There are various models to predict financial time series like the RMB exchange rate. In this paper, considering the complex characteristics of RMB exchange rate, we build a nonlinear combination model of the autoregressive fractionally integrated moving average (ARFIMA) model, the support vector machine (SVM) model, and the back-propagation neural network (BPNN) model to forecast the RMB exchange rate. The basic idea of the nonlinear combination model (NCM) is to make the prediction more effective by combining different models’ advantages, and the weight of the combination model is determined by a nonlinear weighted mechanism. The RMB exchange rate against US dollar (RMB/USD) and the RMB exchange rate against Euro (RMB/EUR) are used as the empirical examples to evaluate the performance of NCM. The results show that the prediction performance of the nonlinear combination model is better than the single models and the linear combination models, and the nonlinear combination model is suitable for the prediction of the special time series, such as the RMB exchange rate.
Analysis of Product Complexity considering Disruption Cost in Fast Fashion Supply Chain
Authors: Sardar Shaheen , Lee Young Hae
Published: July 2015

Outsourcing in the textile industry has been playing an important role in the global economy for six decades. Recently, reshoring is an emerging trend due to various complexities involved in supply chain management. As compared with basic textile and apparel products, fast fashion products are complex in their own way. A single assortment contains several new styles, colors, and sizes with unpredictable demand and urgent deadlines. Numerous assortments run simultaneously in the supply chain. For each assortment, the garment manufacturer has to source various types of fabrics and materials from different suppliers and then manufacture the garments to ship within the deadlines. This complexity contributes to supply chain disruption. This paper develops a model to estimate supply chain disruption cost as a function of fast fashion product complexity in the global outsourcing environment. Estimation of disruption cost will help us to increase visibility and eliminate the bottlenecks in supply chain. Model conclusions are used to develop a method to manage the level of product complexity from the global supply chain perspective. Several strategies are proposed to manage the impact of product complexity on supply chain design.
Energy Conditions of Built-In Inflation Models inf(T)Gravitational Theories
Authors: Nashed Gamal G. L.
Published: July 2015

We examine the validity of energy conditions of built-in inflation models inf(T)gravitational theories. For this purpose, we formulate the inequalities of energy conditions by assuming the flat and nonflat Friedmann-Robertson-Walker (FRW) universe. We find the feasible constraints on the constants of integration and evaluate their possible ranges graphically for the consistency of these energy conditions for flat, closed, and open universes. We constrain the constants of integration for flat space-time from the inflation epoch while the closed and open universe constants are constrained from late universe.
Electrocardiographic Predictors of Cardiovascular Mortality
Authors: Mozos Ioana , Caraba Alexandru
Published: July 2015

Cardiovascular diseases are the main causes of mortality. Sudden cardiac death may also appear in athletes, due to underlying congenital or inherited cardiac abnormalities. The electrocardiogram is used in clinical practice and clinical trials, as a valid, reliable, accessible, inexpensive method. The aim of the present paper was to review electrocardiographic (ECG) signs associated with cardiovascular mortality and the mechanisms underlying those associations, providing a brief description of the main studies in this area, and consider their implication for clinical practice in the general population and athletes. The main ECG parameters associated with cardiovascular mortality in the present paper are the P wave (duration, interatrial block, and deep terminal negativity of the P wave in V1), prolonged QT and Tpeak-Tend intervals, QRS duration and fragmentation, bundle branch block, ST segment depression and elevation, T waves (inverted, T wave axes), spatial angles between QRS and T vectors, premature ventricular contractions, and ECG hypertrophy criteria.
Lonicerae Japonicae Flos and Lonicerae Flos: A Systematic Pharmacology Review
Authors: Li Yujie , Cai Weiyan , Weng Xiaogang , Li Qi , Wang Yajie , Chen Ying , Zhang Wei , Yang Qing , Guo Yan , Zhu Xiaoxin , Wang Hainan
Published: July 2015

Lonicerae japonicae flos, a widely used traditional Chinese medicine (TCM), has been used for several thousand years in China.Chinese Pharmacopeiaonce included Lonicerae japonicae flos of Caprifoliaceae family and plants of the same species named Lonicerae flos in general in the same group.Chinese Pharmacopeia(2005 Edition) lists Lonicerae japonicae flos and Lonicerae flos under different categories, although they have the similar history of efficacy. In this study, we research ancient books of TCM, 4 main databases of Chinese academic journals, and MEDLINE/PubMed to verify the origins and effects of Lonicerae japonicae flos and Lonicerae flos in traditional medicine and systematically summarized the research data in light of modern pharmacology and toxicology. Our results show that Lonicerae japonicae flos and Lonicerae flos are similar pharmacologically, but they also differ significantly in certain aspects. A comprehensive systematic review and a standard comparative pharmacological study of Lonicerae japonicae flos and Lonicerae flos as well as other species of Lonicerae flos support their clinical safety and application. Our study provides evidence supporting separate listing of Lonicerae japonicae flos and Lonicerae flos inChinese Pharmacopeiaas well as references for revision of relevant pharmacopeial records dealing with traditional efficacy of Lonicerae japonicae flos and Lonicerae flos.
Time Series Analysis and Forecasting for Wind Speeds Using Support Vector Regression Coupled with Artificial Intelligent Algorithms
Authors: Jiang Ping , Qin Shanshan , Wu Jie , Sun Beibei
Published: July 2015

Wind speed/power has received increasing attention around the earth due to its renewable nature as well as environmental friendliness. With the global installed wind power capacity rapidly increasing, wind industry is growing into a large-scale business. Reliable short-term wind speed forecasts play a practical and crucial role in wind energy conversion systems, such as the dynamic control of wind turbines and power system scheduling. In this paper, an intelligent hybrid model for short-term wind speed prediction is examined; the model is based on cross correlation (CC) analysis and a support vector regression (SVR) model that is coupled with brainstorm optimization (BSO) and cuckoo search (CS) algorithms, which are successfully utilized for parameter determination. The proposed hybrid models were used to forecast short-term wind speeds collected from four wind turbines located on a wind farm in China. The forecasting results demonstrate that the intelligent hybrid models outperform single models for short-term wind speed forecasting, which mainly results from the superiority of BSO and CS for parameter optimization.
Curved Nanotube Structures under Mechanical Loading
Authors: Yazdani Sarvestani Hamidreza , Naghashpour Ali
Published: July 2015

Configuration of carbon nanotube (CNT) has been the subject of research to perform theoretical development for analyzing nanocomposites. A new theoretical solution is developed to study curved nanotube structures subjected to mechanical loadings. A curved nanotube structure is considered. A nonlocal displacement‐based solution is proposed by using a displacement approach of Toroidal Elasticity based on Eringen’s theory of nonlocal continuum mechanics. The governing equations of curved nanotube structures are developed in toroidal coordinate system. The method of successive approximation is used to discretize the displacement‐based governing equations and find the general solution subjected to bending moment. The numerical results show that all displacement components increase with increasing the nonlocal parameter. The present theoretical study highlights the significance of the geometry and nonlocal parameter effects on mechanical behavior of nanotube structures.
Weight Change in Breast Cancer Survivors Compared to Cancer-Free Women: A Prospective Study in Women at Familial Risk of Breast Cancer
Authors: Gross Amy L. , May Betty J. , Axilbund Jennifer E. , Armstrong Deborah K. , Roden Richard B.S. , Visvanathan Kala
Published: July 2015

Background: This study prospectively examines weight gain in breast cancer survivors compared with cancer-free women from a familial risk cohort.

Methods: Absolute and percent weight change over 4 years was compared among 303 breast cancer survivors and 307 cancer-free women matched on age and menopausal status, from the same familial risk cohort. Linear and logistic regression was used to estimate the association between survivor status and weight gain.

Results: Overall, breast cancer survivors gained significantly more weight [β = 3.06 pounds; 95% confidence intervals (CI), 0.94–5.17] than cancer-free women. Significant weight gain was observed in survivors diagnosed less than 5 years prior to baseline (β = 3.81 pounds; 95% CI, 1.22–6.29) and women with estrogen receptor (ER)-negative tumors (β = 7.26 pounds; 95% CI, 2.23–12.30). Furthermore, survivors treated with chemotherapy were 2.1 times more likely to gain at least 11 pounds during follow-up compared with cancer-free women (OR, 2.10; 95% CI, 1.21–3.63). Weight gain was even greater among survivors who took statins while undergoing chemotherapy treatment (Pinteraction = 0.01).

Conclusion: This is the first study to demonstrate that weight gain is an important issue in breast cancer survivors with a familial risk. In the first five years posttreatment, breast cancer survivors gain weight at a faster rate than cancer-free women, particularly after chemotherapy and statin use but not after hormone therapy alone.

Impact: Our findings provide support for the development of weight gain interventions for young breast cancer survivors with a familial risk. Cancer Epidemiol Biomarkers Prev; 24(8); 1262–9. ©2015 AACR.
Deep Sequencing in Conjunction with Expression and Functional Analyses Reveals Activation of FGFR1 in Ewing Sarcoma
Authors: Agelopoulos Konstantin , Richter Günther H.S. , Schmidt Eva , Dirksen Uta , von Heyking Kristina , Moser Benjamin , Klein Hans-Ulrich , Kontny Udo , Dugas Martin , Poos Kathrin , Korsching Eberhard , Buch Thorsten , Weckesser Matthias , Schulze Isabell , Besoke Regina , Witten Anika , Stoll Monika , Köhler Gabriele , Hartmann Wolfgang , Wardelmann Eva , Rossig Claudia , Baumhoer Daniel , Jürgens Heribert , Burdach Stefan , Berdel Wolfgang E. , Müller-Tidow Carsten
Published: July 2015

Purpose: A low mutation rate seems to be a general feature of pediatric cancers, in particular in oncofusion gene-driven tumors. Genetically, Ewing sarcoma is defined by balanced chromosomal EWS/ETS translocations, which give rise to oncogenic chimeric proteins (EWS-ETS). Other contributing somatic mutations involved in disease development have only been observed at low frequency.

Experimental Design: Tumor samples of 116 Ewing sarcoma patients were analyzed here. Whole-genome sequencing was performed on two patients with normal, primary, and relapsed tissue. Whole-exome sequencing was performed on 50 Ewing sarcoma and 22 matched normal tissues. A discovery dataset of 14 of these tumor/normal pairs identified 232 somatic mutations. Recurrent nonsynonymous mutations were validated in the 36 remaining exomes. Transcriptome analysis was performed in a subset of 14 of 50 Ewing sarcomas and DNA copy number gain and expression of FGFR1 in 63 of 116 Ewing sarcomas.

Results: Relapsed tumors consistently showed a 2- to 3-fold increased number of mutations. We identified several recurrently mutated genes at low frequency (ANKRD30A, CCDC19, KIAA0319, KIAA1522, LAMB4, SLFN11, STAG2, TP53, UNC80, ZNF98). An oncogenic fibroblast growth factor receptor 1 (FGFR1) mutation (N546K) was detected, and the FGFR1 locus frequently showed copy number gain (31.7%) in primary tumors. Furthermore, high-level FGFR1 expression was noted as a characteristic feature of Ewing sarcoma. RNA interference of FGFR1 expression in Ewing sarcoma lines blocked proliferation and completely suppressed xenograft tumor growth. FGFR1 tyrosine kinase inhibitor (TKI) therapy in a patient with Ewing sarcoma relapse significantly reduced 18-FDG-PET activity.

Conclusions: FGFR1 may constitute a promising target for novel therapeutic approaches in Ewing sarcoma. Clin Cancer Res; 21(21); 4935–46. ©2015 AACR.
Abstract A05: Identification of RASA1 as a novel melanoma tumor suppressor gene
Authors: Sung Hyeran , Kanchi Krishna L. , Messina Jane , Lee Ji-Hyun , Ding Li , Wilson Richard K. , Weber Jeffrey S. , Kim Minjung
Published: July 2015

Hyperactivation of the Ras-Raf-Mitogen-activated protein kinase (MAPK) pathway via frequent activating mutations in NRAS and BRAF has been commonly observed in melanoma. Novel mutations in other components of this pathway such as MEK1, MEK2, MAP3K5, and MAP3K9 have been reported recently by high-throughput sequencing efforts. In addition, NF1 (neurofibromatosis type 1), one of the Ras GTPase activating proteins (RasGAPs), has been shown to be mutated or suppressed in melanoma. However, importance of other RasGAPs has not been addressed in melanoma.

To obtain a comprehensive view of melanoma genomes, we conducted whole genome sequencing (WGS) of 15 metastatic melanomas and matched normal PBMC genomes from 13 melanoma patients. All melanoma genomes from these 13 patients contained at least one mutation in genes of Ras-Raf-MAPK pathway (MAPK1, MAP3K1, MAP4K2, MAP3K14, NRAS, and BRAF). In addition, we identified two novel, clustered somatic missense mutations (p.Tyr472His and p.Leu481Phe) in RASA1 (RAS p21 protein activator 1, p120RasGAP) that negatively regulates Ras by catalyzing the hydrolysis of active Ras-GTP to inactive Ras-GDP. In this study, we addressed functional roles of RASA1 in melanoma tumorigenesis. Ectopic expression of wild-type RASA1 in human melanoma cell lines SKMEL28 and WM983C (with BRAF V600E) decreased, while RASA1 Y472H mutant enhanced soft agar colony formation, tumor growth, and Ras activity. The RASA1 L481F mutant lost its tumor suppressive activity. The siRNA- or shRNA-mediated knockdown of RASA1 promoted soft agar colony formation, tumor growth, and RAS activation in human melanoma cell lines IGR1 and KML1 (with BRAFV600E). Mechanistically, RASA1 required RasGAP activity to suppress colony formation and showed higher activity toward R-Ras isoform among Ras superfamily of small GTPases. To determine RASA1 level in melanocytic lesions, immunohistochemical analysis of RASA1 on human melanoma tissue microarray (TMA) was performed. RASA1 expression was frequently down-regulated in metastatic melanoma samples (11.4% (4/35) of lymph node metastasis and 3.4% (1/29) of distal metastases) compared to primary melanomas (33.3% (21/63)) and dysplastic nevi (44.1% (15/34)). Thus, these data support that RASA1 is a novel melanoma tumor suppressor that is inactivated by loss of expression or by mutation.

Citation Format: Hyeran Sung, Krishna L. Kanchi, Jane Messina, Ji-Hyun Lee, Li Ding, Richard K. Wilson, Jeffrey S. Weber, Minjung Kim. Identification of RASA1 as a novel melanoma tumor suppressor gene. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr A05.
Abstract A08: CADM1 is a TWIST1 regulated suppressor of melanoma invasion
Authors: Hartsough Edward J. , Weiss Michele B. , Rosenbaum Sheera R. , Aplin Andrew E.
Published: July 2015

Melanoma is the deadliest form of skin cancer; however, with early detection prior to metastatic dissemination, patients generally have a good prognosis. Recently, our lab has implicated the transcription factor TWIST1 in the progression of melanoma towards metastasis. In melanoma cells, we found that RAS-RAF-MEK-ERK (ERK1/2 pathway) signaling increases TWIST1 expression, which promotes invasive properties in the dermal microenvironment, at least in part by enhancing levels of the matrix metalloproteinase, MMP1. Other TWIST1-regulated targets are poorly described. In this study, we compared expression-profiling data from cells expressing shRNA against TWIST1 or cells overexpressing TWIST1 in order to identify TWIST1-regulated genes. KEGG and GO analysis revealed that TWIST1 is responsible for regulating a number of genes involved in cellular adhesion. We found that TWIST1 levels inversely correlate with levels of cell adhesion molecule 1 (CADM1, also known as NECL-2, IGSF4, TSLC1, SynCAM). Chromatin immunoprecipitation (ChIP) studies and promoter assays demonstrated that TWIST1 physically interacts with the CADM1 promoter and this is associated with reduced CADM1 levels. Additionally, CADM1 expression is inversely associated with ERK1/2 signaling and TWIST1 expression. Modulation of cellular CADM1 levels did not affect proliferation rates; however, overexpression of CADM1 augmented cell-cell interaction, cell aggregation, and reduced trans-endothelial invasion. Furthermore, exogenous CADM1 inhibited serum-directed migration and invasion through Matrigel-coated Boyden chambers, whereas knockdown of CADM1 was associated with enhanced migratory and invasive properties. Taken together, these data provide evidence that CADM1 is negatively regulated by TWIST1 and may act as a suppressor of melanoma invasion.

Citation Format: Edward J. Hartsough, Michele B. Weiss, Sheera R. Rosenbaum, Andrew E. Aplin. CADM1 is a TWIST1 regulated suppressor of melanoma invasion. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr A08.
Abstract A25: WWOX phosphorylation at Ser14 enhances melanoma docking and growth in the lung and liver in mice
Authors: Su Wan-Pei , Wang Wan-Jen , Chang Nan-Shan
Published: July 2015

Understanding of the fundamental cell behavior would help develop strategies for cancer stem cell eradication. Metastatic cancer cells tend to have epigenetically inactivated WWOX gene and are highly aggressive. Time-lapse microscopy revealed that they sense the surrounding WWOX-expressing normal or cancer cells, run away from them, and divide rapidly in order to survive. With time of growth and increased space pressure, these cancer cells grow as spheres with the expression of pluripotent stem cell markers such as Oct4, SSEA-4, TRA-1-60, and others. WWOX is re-expressed in the cancer stem cell spheres with S14 phosphorylation (pS14), suggesting that pS14WWOX is a potential cancer stem cell marker. Notably, organs, expressing pS14WWOX, are susceptible for melanoma cell docking and homing in vivo. Suppression of WWOX phosphorylation at Ser14 by micromolar levels of full-length Zfra1-31 or truncated Zfra4-10 prevents melanoma growth in the lung and liver in mice. Zfra stimulates non-T and non-B memory Hyal-2+ CD3- CD19- lymphocytes in the mouse spleens by binding to membrane hyaluronidase Hyal-2 and probably suppressing the TGF-beta/Hyal-2/WWOX/Smad4 signaling. In vitro education of Hyal-2+ CD3- CD19- cells suppresses melanoma growth in vivo.

Citation Format: Wan-Pei Su, Wan-Jen Wang, Nan-Shan Chang. WWOX phosphorylation at Ser14 enhances melanoma docking and growth in the lung and liver in mice. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr A25.
Abstract A33: Loss of tumor suppressors KAI1 and p27 identifies a unique subgroup of primary melanoma patients with poor survival
Authors: Cheng Yabin , Zhang Guohong , Tang Yun , Chen Guangdi , Safaee Gholamreza , Rotte Annand , Martinka Magdalena , McElwee Kevin , Zhou Youwen
Published: July 2015

Objective: Melanoma is a highly heterogeneous disease. A certain subgroup of patients with primary melanomas exhibits a greater potential to develop metastatic disease and thus has worse survival. The present study attempts to identify the molecular features shared by metastatic melanomas and the subgroup of primary melanomas, and use the information to assist clinicians to design personalized therapy for primary melanoma patients.

Patients and methods: Seven previously reported biomarkers, including BRAF, Dicer, Fbw7, KAI1, MMP2, p27 and Tip60, were investigated. A training cohort with 250 melanoma patients (145 primary melanomas and 105 metastatic melanomas) and an independent cohort with 92 primary melanoma patients were used for the study. Logistic regression analysis was used to identify discriminate biomarkers of metastatic melanoma from primary melanoma. Kaplan-Meier survival and multivariate Cox proportional hazard regression analysis were performed to assess the significance of the molecular signature in the prognosis of melanoma.

Results: In the training cohort, we found the loss expression of KAI1 and p27 to be most significant between metastatic and primary melanoma. The primary melanoma patients with loss expression of both KAI1 and p27 had poor 5-year survival in both training cohort (P = 0.002) and independent cohort (P = 0.03). Multivariate Cox regression analysis showed that the KAI1-/p27- signature was an independent factor for disease- specific survival (P = 0.004). More important, compared to KAI1 and p27 as an individual prognostic marker, the KAI1-/p27- signature is more closely associated with melanoma patient survival: the P value is 0.004 for KAI1-/p27-, and 0.044, 0.181 for KAI1 and p27, respectively.

Conclusion: Loss of both KAI1 and p27 defines a subgroup of primary melanoma patients with poor prognosis, and the combination signature is better than individual biomarker. The clinical significance of KAI1 and p27 warrants further assessment in prospective clinical trials.

Citation Format: Yabin Cheng, Guohong Zhang, Yun Tang, Guangdi Chen, Gholamreza Safaee, Annand Rotte, Magdalena Martinka, Kevin McElwee, Youwen Zhou. Loss of tumor suppressors KAI1 and p27 identifies a unique subgroup of primary melanoma patients with poor survival. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr A33.
Abstract B15: Combination chemotherapy in melanoma using EZH2 inhibitor
Authors: Sengupta Deepanwita , Avaritt Nathan L. , Tackett Alan J.
Published: July 2015

Melanoma is the deadliest variety of skin cancer, which according to the American Cancer Society (ACS) will account for around 9710 deaths and diagnosis of around 76,100 new melanoma cases in the current year. According to the ACS, melanoma is also the fastest growing cancer, and hence it is vital to investigate the pathogenesis involved in the advanced stage of melanoma, and design effective therapeutics for its treatment.

Aberrant activity of the histone methyltransferase enhancer of zeste homolog 2 (EZH2) has been reported to be associated with different types of cancer. The EZH2 catalyzes the deposition of the repressive mark histone H3 lysine 27 trimethylation (H3K27me3) on the promoter of the target genes using the catalytic SET domain, thereby epigenetically silencing the expression of the target genes. This is because the H3K27me3 mark results in chromatin compaction which reduces the accessibility of various transcription factors to the gene, and hence adversely affects transcription. Several of such EZH2 target genes have been found to be tumor suppressor genes like RUNX3, DAB2IP, and E-cadherin, and silencing of such tumor suppressor genes is a chief mechanism by which EZH2 leads to cancer progression. Hence derepression of the tumor suppressor genes by inhibition of EZH2 could be an effective treatment strategy in different types of cancer.

There has been no study till date investigating the role of EZH2 in melanoma. Our laboratory is investigating the role of EZH2 in the pathogenesis involved in the advanced stage of melanoma, to enable the design of effective therapeutic strategies for the treatment of melanoma. Drugs targeting epigenetic modifiers are becoming increasingly popular because of the fact that most epigenetic modifications and their effects can be reversed using these drugs. Our lab has strong evidence that metastatic (or advanced) melanoma is associated with upregulation of EZH2 and H3K27me3. We have also observed that EZH2 inhibition inhibits proliferation and invasion, and also induces apoptosis in metastatic melanoma cells. Recently our lab is performing combination chemotherapeutic studies in vitro, where EZH2 inhibitor GSK126 (S-adenosyl-methionine competitive inhibitor) is being used in combination with conventional chemotherapeutics like cisplatin and Bcl-2 inhibitors. Our results strongly suggest that when EZH2 inhibitor is used in combination with cisplatin or Bcl-2 inhibitor in metastatic melanoma cell lines, the inhibition is synergistic. This synergistic effect of GSK126 and cisplatin or Bcl-2 inhibitor has been found to be caspase-mediated.

Citation Format: Deepanwita Sengupta, Nathan L. Avaritt, Alan J. Tackett. Combination chemotherapy in melanoma using EZH2 inhibitor. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr B15.
Abstract B27: Wnt5A-expressing melanoma cells show classical markers of senescence following radiation and therapeutic stress, but retain the ability to metastasize and proliferate at distant sites
Authors: Webster Marie R. , Xu Mai , Kinzler Kathryn , Kaur Amanpreet , Appleton Jessica , O'Connell Michael P. , Marchbank Katie , Valiga Alexander , Dang Vanessa , Perego Michela , Zhang Gao , Slipicevic Ana , Keeney Frederick , Lehrmann Elin , Wood William , Becker Kevin , Kossenkov Andrew V. , Herlyn Meenhard , Murphy Maureen , Weeraratna Ashani T.
Published: July 2015

Senescence is a cytostatic cell fate that can be induced by oncogene activation, replicative stress or therapy-induced stress. Therapy-induced senescence (TIS) of tumor cells can be initiated by anti-cancer compounds or radiation. TIS is generally considered to be irreversible, and as such this process is believed to be a desirable therapeutic outcome. In this study, we show that melanomas characterized by high levels of the signaling molecule Wnt5A respond to irradiation by undergoing growth arrest, and by expressing classical markers of senescence, including positivity for senescence-associated beta-galactosidase (SA-β-gal), senescence associated heterochromatic foci (SAHF), H3K9Me chromatin marks, and PML bodies. Even though these cells are positive for classical markers of senescence, we find that they retain their ability to migrate and invade. Further, these Wnt5A-high cells are able to proliferate following metastasis and form new tumors in the lung, while expressing increased markers of senescence. Silencing Wnt5A reduces expression of these senescence markers and decreases invasiveness. Finally, Wnt5A-high melanoma cells treated with PLX4720, a BRAFV600E targeted therapy, also increase markers of a TIS response, while remaining invasive. The combined data challenge the paradigm that TIS is a desirable therapeutic endpoint. They also point to Wnt5A as a master regulator of this senescence-like stress-induced phenotype in melanoma.

Citation Format: Marie R. Webster, Mai Xu, Kathryn Kinzler, Amanpreet Kaur, Jessica Appleton, Michael P. O'Connell, Katie Marchbank, Alexander Valiga, Vanessa Dang, Michela Perego, Gao Zhang, Ana Slipicevic, Frederick Keeney, Elin Lehrmann, William Wood, III, Kevin Becker, Andrew V. Kossenkov, Meenhard Herlyn, Maureen Murphy, Ashani T. Weeraratna. Wnt5A-expressing melanoma cells show classical markers of senescence following radiation and therapeutic stress, but retain the ability to metastasize and proliferate at distant sites. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr B27.
Abstract B32: Responses of direct in vivo melanoma xenograft cells to targeted therapeutics
Authors: Basken Joel , Fujita Mayumi , Ahn Natalie
Published: July 2015

The purpose of this study was to investigate the mechanism of resistance of a melanoma cell line generated from a direct in vivo xenograft (DIVX) model. Although BRAF and MKK1/2 inhibitors have produced positive clinical responses in melanoma patients with BRAF-V600 mutations, between 25-50% of patients show intrinsic resistance. Deciphering the mechanisms of intrinsic resistance will be critical for developing future treatment strategies. MB947P cells were established from a direct in vivo xenograft of a resected metastatic melanoma and exhibited innate resistance to apoptosis induced by MKK1/2 inhibitor (AZD6244/selumetinib). In contrast, the established metastatic melanoma cell line, WM239A, showed a strong apoptotic response, induced by inhibitor. Microarray expression profiling was carried out on MB947P and WM239A cells in the presence and absence of drug. The results showed that in MB947P cells inhibitor-dependent induction of Bim was minimal. In contrast, Bim was strongly induced by inhibitor in WM239A cells, where siRNA knockdown of Bim demonstrated its requirement for the apoptotic response. This suggests that the lack of apoptotic response in MB947P cells may be caused by a block in Bim induction through an unknown mechanism. Finally, co-culture of MB947P and WM239A cells showed partial inhibition of the apoptotic response in WM239A cells after AZD6244 treatment. This suggests the potential existence of a soluble factor in MB947P cells that confers resistance to apoptosis in WM239A cells. We propose that mechanisms for resistance to apoptosis in direct in vivo xenograft cell lines may ultimately inform mechanisms for resistance in patient tumors.

Citation Format: Joel Basken, Mayumi Fujita, Natalie Ahn. Responses of direct in vivo melanoma xenograft cells to targeted therapeutics. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr B32.
Abstract B33: PKC resistance in uveal melanoma is mediated by activation of AKT and reversed by BYL719, the PI3Kα inhibitor
Authors: Musi Elgilda , Ambrosini Grazia , Schwartz Gary K.
Published: July 2015

Uveal melanoma is the one of most common intraocular malignancies of the adult eye. Recently, Protein Kinase C (PKC) inhibitors have been used, as single agent and in combination, to treat GNAQ/GNA11-mutant uveal melanoma. However, patients may develop resistance to drug therapy. We have studied mechanisms of PKC resistance in uveal melanoma by exposing GNAQ/GNA11 mutant cell lines (92.1, Omm1.3 and Omm1) to increasing concentrations of the PKC inhibitor AEB071 for extended periods of time. Cell survival, as measured by cell proliferation assays, indicates IC50s of > 5μM, ten times higher than the sensitive parental cells. Western blot analysis indicates increased expression of p-MARCKS (the PKC substrate), p-ERK and p-AKT in the resistant cells. When treated with AEB071, there was no evidence of PKC enzyme inhibition and no suppression of p-MARCKS, p-ERK or p-AKT. Though resistant to AEB071, treatment of the resistant cells with BYL719, a PI3Kα inhibitor, shows inhibition of cell growth (IC50s between 1 and 2 μM) with inhibition of p-AKT similar to that of the parental cells. Furthermore, this effect on growth inhibition can be enhanced by combining AEB071 with BYL719 resulting in inhibition of p-ERK, p-S6 and p-AKT despite persistent activation of p-MARCKS. Analysis of p-glycoprotein (MDR) in the PKC resistant cell lines showed no differences when compared to the parental cell lines, suggesting that this effect is not due to differences in drug accumulation. These studies support the combination of AEB071 and BYL719 as a means to maintain sensitization to PKC inhibitor therapy despite the onset of drug resistance. A clinical trial with AEB071 and BYL719 will be initiated shortly.

Citation Format: Elgilda Musi, Grazia Ambrosini, Gary K. Schwartz. PKC resistance in uveal melanoma is mediated by activation of AKT and reversed by BYL719, the PI3Kα inhibitor. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr B33.
Abstract B38: Melanoma patient-derived xenografts accurately models the disease and develop fast enough to guide treatment decisions
Authors: Einarsdottir Berglind O. , Bagge Roger Olofsson , Bhadury Joydeep , Jespersen Henrik , Nilsson Lisa M. , Mattsson Jan , Truvé Katarina , Naredi Peter , Nilsson Ola , Lopez Marcela Davila , Stierner Ulrika , Ny Lars , Nilsson Jonas A.
Published: July 2015

The development of novel therapies against melanoma would benefit from individualized tumor models to ensure the rapid and accurate identification of biomarkers of therapy response. Previous studies have suggested that patient-derived xenografts (PDXes) could be used in biomarker of response discovery. However, the clinical utility of PDXes in guiding real-time treatment decisions have only been reported in anecdotal forms. In the present study we aimed to develop a platform of melanoma patient-derived xenografts (PDXes), to assess whether treatment responses in mice would be applicable as a pre-selection tool for clinical trials. To that end, tumor biopsies from patients with stage III and IV metastatic malignant melanoma were transplanted into immunocompromised mice. Most melanoma biopsies generated serially transplantable PDX models, and their histology, and mutation status resembled their corresponding patient biopsy. RNAseq demonstrated that the expression profiles of the tumors are maintained throughout several passages in mice. The potential treatment for one patient was revealed by an in vitro drug screen and treating PDXes with the MEK inhibitor trametinib. In another patient, the BRAF mutation predicted the response of both the patient and its corresponding PDXes to MAP kinase pathway-targeted therapy. Importantly, in this unselected group of patients, the time from biopsy for generation of PDXes until death was significantly longer than the time required to reach the treatment phase of the PDXes. Thus, we demonstrate that it would be clinically meaningful to use this type of platform for a large proportion of melanoma patients as a pre-selection tool in clinical trials.

Citation Format: Berglind O. Einarsdottir, Roger Olofsson Bagge, Joydeep Bhadury, Henrik Jespersen, Lisa M. Nilsson, Jan Mattsson, Katarina Truvé, Peter Naredi, Ola Nilsson, Marcela Davila Lopez, Ulrika Stierner, Lars Ny, Jonas A. Nilsson. Melanoma patient-derived xenografts accurately models the disease and develop fast enough to guide treatment decisions. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr B38.
Abstract IA18: Adoptive T-cell transfer for metastatic melanoma: What are the relevant antigens?
Authors: Yang James C.
Published: July 2015

The transfer of cultured tumor-reactive T-cells into patients with metastatic melanoma can cause tumor regressions when other immunotherapies have failed. A single transfer of T-cells can mediate complete and durable regressions in some patients. The two main sources of T-cells for transfer have been from the native T-cells infiltrating the tumor (tumor infiltrating lymphocytes; TIL) or from peripheral blood lymphocytes retrovirally engineered with a tumor-reactive T-cell receptor (TCR). In the latter situation, a variety of tumor-associated antigens have been targeted with cloned receptors and although responses are seen, in some cases unacceptable toxicity has occurred due to on-target, off-tumor immune recognition. Although melanoma TIL often contain T-cells that also target the same nominal antigens, their transfer is rarely associated with such undesirable toxicities. In addition, melanoma TIL frequently contain T-cells which recognize the autochthonous tumor, while TIL from other tumors infrequently show this reactivity. Recent data have implicated the presence of abundant tumor-specific mutations in melanoma as the explanation for these various observations. These appear to be significant antigenic targets in therapeutic TIL that cause no collateral damage, and their high frequency as shown by tumor exomic sequencing of melanomas may explain that tumor's high degree of natural immunogenicity. These lessons learned from studying T-cell transfer for melanoma can now be applied to other cancers and a recent example has demonstrated the potential and promise of this approach.

Citation Format: James C. Yang. Adoptive T-cell transfer for metastatic melanoma: What are the relevant antigens? [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr IA18.
Abstract IA21: Skin pigmentation pathways and melanoma risk
Authors: Fisher David E.
Published: July 2015

Melanoma risk is well known to be strongly linked to UV exposure. Yet despite this important linkage, melanoma incidence increases relentlessly, at one of the steepest rates of all human cancers. Prevention approaches that include education, sunscreen, avoidance of tanning beds, and sun-safe behavior messages are excellent in principal, but have not as yet produced sufficient decreases in melanoma incidence. Here we present studies which explore cutaneous responses to ultraviolet radiation exposure. These results indicate the presence of multiple cutaneous responses and pathways that are triggered by UV and which modulate not only skin signaling, but also systemic effects of this cutaneous UV exposure. These pathways include pigmentation or tanning. Other studies will describe a murine “redhair/lightskin” melanoma model of melanoma which identifies roles for pigmentation intermediates in melanoma formation. Finally, the MITF transcription factor plays central roles in both melanocyte and melanoma growth, survival, and differentiation. Studies will be described which characterize several MITF transcriptional targets, some of which may represent novel therapeutic opportunities.

Citation Format: David E. Fisher. Skin pigmentation pathways and melanoma risk. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr IA21.
Abstract IA22: The immunobiology of the melanoma tumor microenvironment and efficacy of immunotherapies
Authors: Gajewski Thomas F.
Published: July 2015

Two major categories of melanoma metastases have been observed based on gene expression profiling and confirmatory assays. One subgroup of patients has an inflamed phenotype that includes expression of chemokines, T cell markers, and other immunoregulatory factors. In contrast, the other major subset lacks this phenotype and appears to display immune “exclusion”. The optimal immunotherapeutic interventions may be distinct in these two global subsets. The T cell-inflamed tumor microenvironment subset shows the highest expression of negative regulatory factors, including PD-L1, IDO, and FoxP3+ Tregs, and evidence for T cell-intrinsic anergy has also emerged aided by a recently defined functional role of EGR2. In addition, the mechanism of induction of these inhibitory mechanisms has been elucidated—PD-L1 and IDO are induced by IFN-γ, and Tregs are largely recruited by the chemokine CCL22, both being produced by activated CD8+ effector T cells. Preclinical experiments have confirmed a critical role for each of these mechanisms in limiting anti-tumor T cell efficacy in vivo, giving candidate treatment strategies for translation back into the clinic. These include anti-PD-1/PD-L1 mAbs, IDO inhibitors, and approaches to deplete CD25+ Tregs and/or reverse anergy. The presence of multiple inhibitory mechanisms in the same tumor microenvironment argues that combination therapies may be advantageous. Preclinical data have indicated synergy between anti-CTLA-4 +/- anti-PD-L1 +/- IDO inhibition. The mechanism of synergy is striking, as it correlates with a marked improvement of IL-2 production and proliferation of tumor-infiltrating CD8+ T cells. Clinical translation of these combination immunotherapies is promising and ongoing. In contrast to the T cell-inflamed melanomas, a new paradigm may be needed to promote de novo inflammation in cases of the non-T cell-infiltrated tumor microenvironment. Natural innate immune sensing of tumors appears to occur via the host STING pathway, type I IFN production, and cross-priming of T cells via CD8α+ DCs. New strategies are being developed to engage or mimic this pathway as a therapeutic endeavor. The molecular mechanisms that mediate the presence or absence of the T cell-inflamed tumor microenvironment in patients are being elucidated and should open up additional new treatment approaches.

Citation Format: Thomas F. Gajewski. The immunobiology of the melanoma tumor microenvironment and efficacy of immunotherapies. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr IA22.
Abstract IA31: Combining targeted therapy and immune checkpoint blockade to improve responses
Authors: Wargo Jennifer A.
Published: July 2015

There have been significant advances in treatment of metastatic melanoma over the past several years through the use of targeted therapy and immune checkpoint blockade. Oncogenic mutations in the BRAF gene occur in over half of melanoma tumors, and treatment with agents targeting this mutation results in objective responses in the majority of patients. However significant limitations exist as responses are not durable, with the majority of patients progressing on therapy within 6-10 months. Another area of significant progress involves the use of immune checkpoint blockade, such as with monoclonal antibodies blocking CTLA-4 and the PD-1 pathway. In contrast, treatment of patients with immune checkpoint inhibitors results in a lower response rate than those observed with targeted therapy, but ones that tend to be more durable. With these advances, we are posed with therapeutic dilemmas with regard to timing and sequence of therapy. Namely, there is significant debate as to whether to begin treatment with targeted therapy versus immune checkpoint blockade upfront, and at which point to change treatment strategy.

There is growing evidence of potential synergy in combining treatment with BRAF-targeted therapy and immune checkpoint blockade, and this concept is being empirically tested in clinical trials. There a strong scientific rationale for combining these therapies, as treatment with BRAF inhibitors is associated with increased immunogenicity of melanoma tumors within 2 weeks of therapy - with increased melanoma antigens and CD8+ T cell infiltrate, and decreased immunosuppressive cytokines and VEGF. BRAF inhibitors may also directly activate T cells through paradoxical activation of the MAPK pathway. However there is a concurrent increase in expression of immunomodulatory molecules (such as PD-L1) within 2 weeks of therapy which is likely related to T cell infiltrate, though also suggest a possible immune mechanism of resistance. Clinical trials and studies in murine models are currently underway to explore synergy between these therapies, and insights gained will be discussed herein. It is becoming increasingly apparent that complexities exist regarding appropriate timing and sequence in these combination strategies to optimize responses and minimize toxicity. Ultimately, ideal approaches using combined BRAF-targeted therapy and immune checkpoint blockade will be built on a deep understanding of the molecular and immune effects of each of these therapies in isolation, as well as in combination.

Citation Format: Jennifer A. Wargo. Combining targeted therapy and immune checkpoint blockade to improve responses. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Melanoma: From Biology to Therapy; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(14 Suppl):Abstract nr IA31.