bluuebunny/crossref_metadata_2025 huggingface dataset:
Fostering Achievement of Low-, Average-, and High-Achievers Students in Biology through Structured Cooperative Learning (STAD Method)
Authors: Yaduvanshi Sangeeta , Singh Sunita
Published: February 2019

Cooperative learning is one among the most innovative and popular strategies of learning for present century students. It is theoretically grounded and extensively researched teaching-learning practice which is believed to foster the achievement of all types of students. Since cooperative learning is in the nascent stage in India and it is not much trendy method of teaching, the investigator carried out the present study. The present study is an experimental investigation that explores the impact of the structured cooperative learning strategy (STAD method) on the achievement of low-achievers, average-achievers, and high-achievers students in biology at the secondary level. The pretest and posttest experimental design was used, and control and experimental groups are equated on the basis of pretest scores. The sample consisted a total of 63 students of ninth class students from Varanasi city. Data were collected and analyzed with t-test, and analysis of covariance (ANCOVA) was performed to test the hypotheses at 0.05 levels of significance with the use of the biology achievement test (BAT). The results revealed that students taught by the cooperative learning strategy perform better on the BAT at three levels of the cognitive domain of knowledge, understanding, and applying, than those taught using the conventional method of instruction. The study revealed that low-achievers, average-achievers, and high-achievers students of the experimental group outperform the control group. So, it can be concluded from this study that the STAD method of structured cooperative learning fosters the achievement of low-, average-, and high-achievers students in Indian context. It was therefore recommended that teachers should be encouraged to use the cooperative instructional strategy to teach biology and other subjects in secondary schools to facilitate learning of higher levels of cognitive domains to meet challenges of the twenty-first century.
Design and Modeling of a Novel Transformable Land/Air Robot
Authors: Wang He , Shi Jiadong , Wang Jianzhong , Wang Hongfeng , Feng Yiming , You Yu
Published: February 2019

This paper describes a novel transformable land/air robot that is capable of terrestrial locomotion and aerial locomotion. What is unusual about the robot is that it can transform between the two modes of locomotion at will through the transformable mechanism, allowing the robot to overcome large obstacles in their mission environment. The wheel mechanism of the robot is shared by both terrestrial and aerial locomotion, instead of simply adding a quadrotor to a wheeled mobile robot. The objective of this paper is to design the robot and establish the kinematic and dynamic models for the transformable process. Herein, we focus on the design of the driving wheels and transformable mechanism. A series of experiments about the energy analysis and the transformation from aerial locomotion mode to terrestrial locomotion mode were performed with the physical prototype; the experiment results confirmed the validity of our design and the theoretical analysis that are helpful to optimize the key parameters in our design. Moreover, our work can provide a reference for the study of the flying car.
Electrode Composite LiFePO4@Carbon: Structure and Electrochemical Performances
Authors: Huynh Le Thanh Nguyen , Nguyen Hoang Hai Au , Tran Thi Thuy Dung , Nguyen Thi Thu Trang , Nguyen Thi My Anh , La Thi Hang , Tran Van Man , Le My Loan Phung
Published: February 2019

This work aimed at preparing the electrode composite LiFePO4@carbon by hydrothermal and the calcination process was conducted at 600, 700, and 800°C. The structure and morphology were determined by X-ray diffraction (XRD), SEM, Raman spectroscopy, X-ray photon spectroscopy (XPS), and thermal analysis. The XRD refinement’s results point out the orthorhombic structure without impurity phase and the high crystalline of synthesized olivines. The results of Raman spectroscopy and XPS confirmed the pure olivine phase as well as the successful carbon coating on the surface of olivines’ powders. Moreover, the calcinated temperature affected the morphology as well as the electrochemical performance of synthesized olivines. The electrochemical measurements were conducted by cyclic voltammetry and galvanostatic cycling test. The diffusion coefficients were calculated from cyclic voltammetry curves and reached 1.09 × 10−12 cm2/s for LFP600, 2.28 × 10−11 cm2/s for LFP700, and 3.27 × 10−12 cm2/s LFP800. The cycling test at rate C/10 exhibited an excellent cyclability with discharge capacity of 145 mAh/g for LFP600, 170 mAh/g for LFP700, and 160 mAh/g for LFP800.
Borneol for Regulating the Permeability of the Blood-Brain Barrier in Experimental Ischemic Stroke: Preclinical Evidence and Possible Mechanism
Authors: Chen Zi-xian , Xu Qing-qing , Shan Chun-shuo , Shi Yi-hua , Wang Yong , Chang Raymond Chuen-Chung , Zheng Guo-qing
Published: February 2019

Borneol, a natural product in the Asteraceae family, is widely used as an upper ushering drug for various brain diseases in many Chinese herbal formulae. The blood-brain barrier (BBB) plays an essential role in maintaining a stable homeostatic environment, while BBB destruction and the increasing BBB permeability are common pathological processes in many serious central nervous system (CNS) diseases, which is especially an essential pathological basis of cerebral ischemic injury. Here, we aimed to conduct a systematic review to assess preclinical evidence of borneol for experimental ischemic stroke as well as investigate in the possible neuroprotective mechanisms, which mainly focused on regulating the permeability of BBB. Seven databases were searched from their inception to July 2018. The studies of borneol for ischemic stroke in animal models were included. RevMan 5.3 was applied for data analysis. Fifteen studies investigated the effects of borneol in experimental ischemic stroke involving 308 animals were ultimately identified. The present study showed that the administration of borneol exerted a significant decrease of BBB permeability during cerebral ischemic injury according to brain Evans blue content and brain water content compared with controls (P<0.01). In addition, borneol could improve neurological function scores (NFS) and cerebral infarction area. Thus, borneol may be a promising neuroprotective agent for cerebral ischemic injury, largely through alleviating the BBB disruption, reducing oxidative reactions, inhibiting the occurrence of inflammation, inhibiting apoptosis, and improving the activity of lactate dehydrogenase (LDH) as well as P-glycoprotein (P-GP) and NO signaling pathway.
Serum Leucine-Richα2-Glycoprotein as a Biomarker for Monitoring Disease Activity in Patients with Systemic Juvenile Idiopathic Arthritis
Authors: Shimizu Masaki , Inoue Natsumi , Mizuta Mao , Nakagishi Yasuo , Yachie Akihiro
Published: February 2019

To investigate whether serum leucine-richα2-glycoprotein (LRG) levels are useful as a marker of disease activity in systemic juvenile idiopathic arthritis (s-JIA), we determined serum LRG levels in fifty-nine s-JIA patients, 15 with other subtypes of JIA, 7 with Kawasaki disease (KD), 7 with influenza A infection (flu), 7 with enterohemorrhagicEscherichia coli(EHEC) infection, and 20 healthy controls (HC). Results were compared with the clinical features of s-JIA and serum cytokine levels including interleukin- (IL-) 6, IL-18, and soluble tumor necrosis factor receptors I and II. Serum LRG levels in active s-JIA were higher compared to those in other subtypes of JIA, EHEC, flu patients, and HC. Serum LRG levels were normalized in the inactive s-JIA phase after treatment. Serum LRG levels were positively correlated with serum C-reactive protein and ferritin levels. Serum LRG levels reflected s-JIA disease activity and thus may be useful for monitoring s-JIA disease activity.
Developing a Novel and Convenient Model for Investigating Sweat Gland Morphogenesis from Epidermal Stem Cells
Authors: Hu Tian , Xu Yongde , Yao Bin , Fu Xiaobing , Huang Sha
Published: February 2019

Sweat glands developed from the embryonic epidermis. To elucidate the underlying mechanisms of morphogenesis, a reliable in vitro test system for bioactive screening must be developed. Here, we described a novel and convenient model by coculturing embryonic tissue and epidermal stem cells (ESCs) using Transwell insert for evaluating the effects of soluble morphogens on sweat gland morphogenesis in vitro. Using this coculture system, morphological alteration, histological features, and specific markers were observed. Initial experiments revealed that ESCs cocultured with embryonic paw pad (EPP) tissue demonstrated glandular structure and cytokeratin 8 (K8) and cytokeratin 18 (K18) positive, while ESCs cocultured with embryonic dorsal skin demonstrated “sea snail” structure and K8, K18 negative. Moreover, bone morphogenetic protein 4 (BMP4) and epidermal growth factor (EGF) concentrations were detected in the medium of the EPP group. BMP receptor inhibitor could effectively block the ESC differentiation to sweat glands, while EGF receptor blocker did not show the effect. Our results showed clear benefits of this novel and convenient model in terms of in vitro-in vivo correlation. It was an appropriate alternative for screening of potential bioactives regulating the sweat gland morphogenesis mechanism.
Wide CPW-Fed Multiband Wearable Monopole Antenna with Extended Grounds for GSM/WLAN/WiMAX Applications
Authors: Mandal Danvir , Pattnaik S. S.
Published: February 2019

A novel wide coplanar waveguide- (CPW-) fed multiband wearable monopole antenna is presented. The multiband operation is achieved by generating slanted monopoles of different lengths from an isosceles triangular patch. The different operating frequencies of the proposed antenna are associated with the lengths of the slanted monopoles, which are determined under quarter wavelength resonance condition. The CPW line is used as a multiband impedance-matching structure. The two grounds are slightly extended for better impedance matching. The proposed antenna is designed to cover the 1800 MHz GSM, 2.4 GHz/5.2 GHz WLAN, and 3.5 GHz WiMAX bands. The measured peak gains and impedance bandwidths are about 4.18/3.83/2.6/2.94 dBi and 410/260/170/520 MHz for the 1550-1960 MHz/2.3-2.56 GHz/3.4-3.57 GHz/5.0-5.52 GHz bands, respectively. The calculated averaged specific absorption rate (SAR) values at all the resonant frequencies are well below the standard limit of 2 W/kg, which ensures its feasibility for wearable applications. The antenna performance under different bending configurations is investigated and the results are presented. The reflection coefficient characteristics of the proposed antenna is also measured for different on-arm conditions and the results are compared. A good agreement between experimental and simulation results validates the proposed design approach.
Effect of the Sampling of a Dataset in the Hyperparameter Optimization Phase over the Efficiency of a Machine Learning Algorithm
Authors: DeCastro-García Noemí , Muñoz Castañeda Ángel Luis , Escudero García David , Carriegos Miguel V.
Published: February 2019

Selecting the best configuration of hyperparameter values for a Machine Learning model yields directly in the performance of the model on the dataset. It is a laborious task that usually requires deep knowledge of the hyperparameter optimizations methods and the Machine Learning algorithms. Although there exist several automatic optimization techniques, these usually take significant resources, increasing the dynamic complexity in order to obtain a great accuracy. Since one of the most critical aspects in this computational consume is the available dataset, among others, in this paper we perform a study of the effect of using different partitions of a dataset in the hyperparameter optimization phase over the efficiency of a Machine Learning algorithm. Nonparametric inference has been used to measure the rate of different behaviors of the accuracy, time, and spatial complexity that are obtained among the partitions and the whole dataset. Also, a level of gain is assigned to each partition allowing us to study patterns and allocate whose samples are more profitable. Since Cybersecurity is a discipline in which the efficiency of Artificial Intelligence techniques is a key aspect in order to extract actionable knowledge, the statistical analyses have been carried out over five Cybersecurity datasets.
Mothers’ Perspectives on the Play of Their Children with Attention Deficit Hyperactivity Disorder
Authors: Jasem Zainab A. , Delport Susan M.
Published: February 2019

Background. Play, which is fundamental to children’s lives, can be utilised to enrich their daily experience. However, the available knowledge regarding the impact of play on children with attention deficit hyperactivity disorder (ADHD) is limited. Therefore, this study is aimed at investigating mothers’ perspectives on their children with ADHD in Kuwait regarding their play preferences, the impact of play on ADHD behaviours, and the strategies used to regulate their children’s ADHD symptoms. Methods. Eight mothers of children aged five to ten years old with ADHD were recruited via purposive sampling. Their perspectives were qualitatively explored through one-week diaries of their children’s routines followed by semistructured interviews. The data were analysed using thematic analysis. Results. Four themes emerged: (1) play characteristics for children with ADHD, (2) play preferences, (3) play settings and their impact on ADHD symptoms, and (4) mothers’ values, understanding, and influence on play. Children were found to spend most of their playtime in indoor settings, largely playing video games. The mothers acknowledged the positive influence on their children of outdoor play and the deleterious effect of video games. However, they undervalued the contribution of play to their children’s development. Their strategies for regulating the types of play that diminish their children’s ADHD behaviours were less well understood. The weather and mothers’ other responsibilities were the main barriers. Conclusions. Understanding the play of children with ADHD revealed the requirement for more opportunities for different types of play, with more consideration of those that positively manage ADHD behaviours.
An Improved EKF-SLAM for Mars Surface Exploration
Authors: Zheng Bo , Zhang Zexu
Published: February 2019

In the traditional EKF-SLAM algorithm, the computational complexity and uncertainty will grow up rapidly with the increase of the feature points and the enlargement of the map coverage. As we know, the computational complexity is proportional to the quadratic of the number of feature points contained in a single filtering process. The approach represented in the paper combines EKF-SLAM with local submaps, which can improve the computational efficiency and reduce the computational complexity. At first, an independent local submap is established for the observed feature points. When the number of feature points contained in the local submap reaches a certain threshold value, the local submap is integrated into the global map. At last, the submap is initialized again. The simulation results show that the approach can reduce the computational complexity effectively and increase the computation speed greatly in the case of maintaining the computational accuracy of the traditional EKF-SLAM algorithm.
Advances in Translational Research and Clinical Care in Pancreatic Cancer: Where Are We Headed?
Authors: Satyananda Vikas , Gupta Rohan , Hari Danielle M. , Yeh James , Chen Kathryn T.
Published: February 2019

While significant advances have been made in the treatment of many different solid tumors, pancreatic cancer remains a glaring exception. Overall 5-year survival rates for pancreatic cancer remain in the single digits. While newer chemotherapy regimens such as FOLFIRINOX and nab-paclitaxel/gemcitabine have demonstrated modest improvement in survival benefit for metastatic disease and have improved the resectability rates of previously borderline or locally advanced tumors, clinically significant improvements from immunotherapy and targeted therapy remain to be demonstrated. Regardless, a wealth of basic science research in pancreatic cancer has been directed at understanding its aggressive biology and its resistance to therapy. We present a brief summary of key areas of laboratory research and its translation to clinical care.
Influence of Water Pressure on the Mechanical Properties of Concrete after Freeze-Thaw Attack under Dynamic Triaxial Compression State
Authors: Wang Ruijun , Li Yan , Li Yang , Xu Fan , Li Xiaotong , Fu Tenghuan
Published: February 2019

This study aims at determining the effect of water pressure on the mechanical properties of concrete subjected to freeze-thaw (F-T) attack under the dynamic triaxial compression state. Two specimens were used: (1) a 100 mm × 100 mm × 400 mm prism for testing the loss of mass and relative dynamic modulus of elasticity (RDME) after F-T cycles and (2) cylinders with a diameter of 100 mm and a height of 200 mm for testing the dynamic mechanical properties of concrete. Strain rates ranged from 10−5·s−1 to 10−3·s−1, and F-T cycles ranged from 0 to 100. Three levels of water pressure (0, 5, and 10 MPa) were applied to concrete. Results showed that as the number of F-T cycles increased, the mass loss rate of the concrete specimen initially decreased and then increased, but the RDME decreased. Under 5 MPa of water pressure and at the same strain rate, the ultimate compressive strength decreased, whereas the peak strain increased with the increase in the number of F-T cycles. This result is contrary to the variation law of ultimate compressive strength and peak strain with the increase in strain rate under the same number of F-T times. With the increase in F-T cycles or water pressure, the strain sensitivity of the dynamic increase factor of ultimate compressive strength and peak strain decreased, respectively. After 100 F-T cycles, the dynamic compressive strength under all water pressure levels tended to increase as the strain rate increased, whereas the peak strain decreased gradually.
Synthesis of Metal/Silica/Titania Composites for the Photocatalytic Removal of Methylene Blue Dye
Authors: Ahmad Sana , Saeed Arfa
Published: February 2019

The present work deals with the synthesis of the metal-doped titanium dioxide/silica composite by using the sol-gel method. The structure, morphology, and composition of the prepared samples were characterized by using Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). The prepared composites were used for the photodegradation of methylene blue dye in sunlight. Different parameters like pH, time, and catalytic concentration were varied to optimize the reaction conditions. Maximum 99.8% degradation was observed with the Ni-doped composite. The results indicate extraordinary efficiency of all metal-doped composites for the removal of the harmful organic pollutant like methylene blue.
Prediction of Fatigue Life of Welded Joints Made of Fine-Grained Martensite-Bainitic S960QL Steel and Determination of Crack Origins
Authors: Ślęzak Tomasz
Published: February 2019

Due to growing requirements connected with the utilization of advanced structures, nowadays the modern design processes are developed. One of the crucial issues considered in these processes is proper design of the joints against fatigue in order to fulfill a stated life of operation. In this study, the method of fatigue life prediction based on the criterion of permissible strain range in the notch root is presented. An engaged simplified model of fatigue life prediction was previously developed for mild and carbon steels. The evaluation made during the research has proven that this method can also be used for S960QL high-strength steel characterized by entirely different properties and structure. A considered theoretical model demonstrates satisfactory correlation with experimental data and safely describes the fatigue life of weldments. Furthermore, the predicted fatigue life of studied steel without welds shows great comparability with experimental data. The limit value of the strain range in the notch root was estimated. Below this value of strain, the fatigue life of welded joints is infinite, theoretically. Finally, the impact of the surface imperfections on the fatigue crack initiation was revealed. For paternal material, the origins of cracking were discovered at the places of nonmetallic scale particles. In welded joints, the fatigue cracks initiated at the whole length of the fusion line.
Novel Third-Generation EGFR Tyrosine Kinase Inhibitors and Strategies to Overcome Therapeutic Resistance in Lung Cancer
Authors: Murtuza Ayesha , Bulbul Ajaz , Shen John Paul , Keshavarzian Parissa , Woodward Brian D. , Lopez-Diaz Fernando J. , Lippman Scott M. , Husain Hatim
Published: February 2019

EGFR-activating mutations are observed in approximately 15% to 20% of patients with non–small cell lung cancer. Tyrosine kinase inhibitors have provided an illustrative example of the successes in targeting oncogene addiction in cancer and the role of tumor-specific adaptations conferring therapeutic resistance. The compound osimertinib is a third-generation tyrosine kinase inhibitor, which was granted full FDA approval in March 2017 based on targeting EGFR T790M resistance. The compound has received additional FDA approval as first-line therapy with improvement in progression-free survival by suppressing the activating mutation and preventing the rise of the dominant resistance clone. Drug development has been breathtaking in this space with other third-generation compounds at various stages of development: rociletinib (CO-1686), olmutinib (HM61713), nazartinib (EGF816), naquotinib (ASP8273), mavelertinib (PF-0647775), and AC0010. However, therapeutic resistance after the administration of third-generation inhibitors is complex and not fully understood, with significant intertumoral and intratumoral heterogeneity. Repeat tissue and plasma analyses on therapy have revealed insights into multiple mechanisms of resistance, including novel second site EGFR mutations, activated bypass pathways such as MET amplification, HER2 amplification, RAS mutations, BRAF mutations, PIK3CA mutations, and novel fusion events. Strategies to understand and predict patterns of mutagenesis are still in their infancy; however, technologies to understand synthetically lethal dependencies and track cancer evolution through therapy are being explored. The expansion of combinatorial therapies is a direction forward targeting minimal residual disease and bypass pathways early based on projected resistance.
Genome-Wide Screening and Functional Analysis Identifies Tumor Suppressor Long Noncoding RNAs Epigenetically Silenced in Hepatocellular Carcinoma
Authors: Xu Feiyue , Li Chi Han , Wong Chi Hin , Chen George G. , Lai Paul Bo San , Shao Shengwen , Chan Stephen L. , Chen Yangchao
Published: February 2019

Long noncoding RNAs (lncRNA) play critical roles in the development of cancer, including hepatocellular carcinoma (HCC). However, the mechanisms underlying their deregulation remain largely unexplored. In this study, we report that two lncRNAs frequently downregulated in HCC function as tumor suppressors and are epigenetically silenced by histone methyltransferase EZH2. lncRNAs TCAM1P-004 and RP11-598D14.1 were inhibited by EZH-mediated trimethylation of H3K27me3 at their promoters. Downregulation of TCAM1P-004 and RP11-598D14.1 was frequently observed in HCC tumors compared with adjacent normal tissues. Both lncRNAs inhibited cell growth, cell survival, and transformation in HCC cells in vitro as well as tumor formation in vivo. Using RNA pull-down and mass spectrometry, we demonstrated that TCAM1P-004 bound IGF2BP1 and HIST1H1C, whereas RP11-598D14.1 bound IGF2BP1 and STAU1. These lncRNA–protein interactions were critical in regulating p53, MAPK, and HIF1α pathways that promoted cell proliferation in HCC. Overexpression of EZH2 was critical in repressing TCAM1P-004 and RP11-598D14.1, and EZH2-TCAM1P-004/RP11-598D14.1–regulated pathways were prevalent in human HCC. Aberrant suppression of TCAM1P-004 and RP11-598D14.1 led to loss of their tumor-suppressive effects by disrupting the interaction with IGF2BP1, HIST1H1C, and STAU1, which in turn promoted HCC development and progression. Collectively, these findings demonstrate the role of TCAMP1P-004 and RP11-598D14.1 in suppressing tumor growth and suggest that EZH2 may serve as a therapeutic target in HCC.

Significance:

EZH2-mediated loss of lncRNAs TCAM1P-004 and RP11-598D14.1 hinders the formation of tumor suppressor lncRNA–protein complexes and subsequently promotes HCC growth.
Abstract A02: Targeting EWS-FLI1 dose-dependent epithelial-to-mesenchymal transition
Authors: Katschnig Anna M. , Kauer Maximilian O. , Schwentner Raphaela , Tomazou Eleni M. , Aryee Dave N.T. , Kovar Heinrich
Published: February 2019

Reversible transitions from a proliferative to a nonproliferative but highly migratory and invasive phenotype, widely known as epithelial-to-mesenchymal transition (EMT), is a prerequisite for wound healing and organ development but also a hallmark of cancer. Recently, phenotypic changes resembling EMT were described to occur in Ewing sarcoma upon modulation of expression of the driver oncogene EWS-FLI1. Evidence was provided for the presence of highly migratory EWS-FLI1-low cells in Ewing sarcoma cell lines as well as in about 20% of primary tumors at low frequency (Franzetti et al., 2017). Importantly, when injected into the tail vein of immunosuppressed mice, EWS-FLI1-low cells were more likely to result in lung metastases than EWS-FLI1-high cells upon re-expression of the oncogene. In our study, we identified a mechanism for the cytoskeletal reprograming underlying EWS-FLI1 dose-dependent phenotypic transitions in Ewing sarcoma. We provide evidence for EWS-FLI1 blocking access of the transcriptional coactivators MRTFB and TAZ to the YAP/TEAD transcription factor complex on enhancers of EWS-FLI1 anti-correlated genes. Upon modulation of EWS-FLI1 expression, TEADs get activated downstream of Rho signaling through recruitment of MRTFB/YAP/TAZ, leading to the expression of key cytoskeletal components and regulators involved in the reorganization of the actin cytoskeleton. Based on these findings, we hypothesized that small-molecule inhibitors of the YAP/TAZ/TEAD pathway should interfere with the migratory phenotype of EWS-FLI1-low cells. In fact, the YAP/TAZ inhibitor verteporfin (Visudyne®), which is clinically used as a photosensitive drug in the treatment of age-related macular degeneration, was able to block Ewing sarcoma cell migration at low nanomolar concentrations without affecting tumor cell proliferation. These results suggest that YAP/TAZ inhibitors may be considered as a promising add-on to conventional combination treatments to prohibit tumor cell dissemination and metastasis.

Citation Format: Anna M. Katschnig, Maximilian O. Kauer, Raphaela Schwentner, Eleni M. Tomazou, Dave N.T. Aryee, Heinrich Kovar. Targeting EWS-FLI1 dose-dependent epithelial-to-mesenchymal transition [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A02.
Abstract A08: TGFβb inhibition improves oncolytic herpes viroimmunotherapy in murine models of rhabdomyosarcoma
Authors: Hutzen Brian , Chen Chun-Yu , Wang Pin-Yi , Sprague Les , Hayley Swain M. , Love Julia , Conner Joe , Boon Louis , Cripe Timothy P.
Published: February 2019

Oncolytic viruses are an emerging class of cancer therapeutics that couple cytotoxicity with the induction of an antitumor immune response. Host-virus interactions are complex and modulated by a tumor microenvironment whose immunosuppressive activities can limit the effectiveness of cancer immunotherapies. In an effort to improve this aspect of oncolytic virotherapy, we combined the oncolytic herpes virus HSV1716 with the TGFβR1 inhibitor A8301 to treat syngeneic models of murine rhabdomyosarcoma. Mice that received HSV1716 or A8301 alone showed little to no benefit in efficacy/survival over controls. Conversely, mice given combination therapy exhibited tumor stabilization throughout the treatment regimen, which was reflected in significantly prolonged survival times including some complete responses. In vitro cell viability and virus replication assays showed that the rhabdomyosarcoma cell lines were generally insensitive to HSV1716 and A8301. Likewise, in vivo virus replication assays showed that HSV1716 titers moderately decreased in the presence of A8301. The enhanced efficacy instead appears to be dependent on the generation of an improved antitumor T-cell response as determined by its loss in athymic nude mice and following in vivo depletion of either CD4+ or CD8+ cells. These data suggest that TGFβ inhibition can augment the immunotherapeutic efficacy of oncolytic herpes virotherapy.

Citation Format: Brian Hutzen, Chun-Yu Chen, Pin-Yi Wang, Les Sprague, Swain M. Hayley, Julia Love, Joe Conner, Louis Boon, Timothy P. Cripe. TGFβb inhibition improves oncolytic herpes viroimmunotherapy in murine models of rhabdomyosarcoma [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A08.
Abstract A22: Identification of gene expression differences between primary pediatric tumors and their PDX models
Authors: Pfeil Jacob J. , Lee Alex G. , Sayles Leanne C. , Kephart Ellen T. , Beale Holly C. , Sanders Lauren M. , Morozova Olena , Salama Sofie R. , Sweet-Cordero Alejandro , Haussler David
Published: February 2019

Molecularly targeted therapies inhibit specific cancer pathways and have fewer harmful side effects than cytotoxic chemotherapies; however, the development of targeted therapies for childhood cancers has lagged behind that of adult cancers. Although adult and childhood cancers have different molecular mechanisms, similar druggable cancer pathways may be activated. We propose using gene expression analysis to identify opportunities to reposition FDA-approved targeted therapies for childhood cancer patients. We developed an N-of-1 gene expression approach that leverages a compendium of ~11,000 cancer gene expression profiles. To evaluate the utility of this approach for identifying cancer driver pathways, we analyzed gene expression data for 9 patient-derived xenograft (PDX) tumors. PDXs were tested for sensitivity to selected targeted therapies. Our analysis correctly predicted drug sensitivity in 8 of the 9 PDXs based on overexpression of specific cancer pathways. We also compared gene expression between PDX and matched primary tumor samples (n=7). Using TumorMap analysis, we found that PDX tumors cluster with their matched primary tumor and other samples with a similar cancer diagnosis. We also ran differential expression analysis and found 260 overexpressed genes and 630 underexpressed genes in PDX tumors. Underexpressed genes were associated with immune functions, extracellular matrix proteins, and growth factor signaling. Overexpressed genes were involved in TNF-α; signaling. This analysis confirmed expected changes in gene expression while also identifying TNF-α; signaling as a potential biologic artifact of PDX tumors. These features may influence the results of PDX experiments and should be accounted for when using this model. Overall, the PDX model recapitulates gene expression features of pediatric cancer and is a powerful tool for evaluating novel precision medicine approaches.

Citation Format: Jacob J. Pfeil, Alex G. Lee, Leanne C. Sayles, Ellen T. Kephart, Holly C. Beale, Lauren M. Sanders, Olena Morozova, Sofie R. Salama, Alejandro Sweet-Cordero, David Haussler. Identification of gene expression differences between primary pediatric tumors and their PDX models [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A22.
Abstract A39: Characterizing the epigenetic effects of the histone 3.3 G34W mutation in giant cell tumors of bone
Authors: Deshmukh Shriya , Khazaei Sima , Marchione Dylan , Harutyunyan Ashot , Garcia Benjamin , Jabado Nada
Published: February 2019

Introduction: Pediatric glioblastomas (pGBM) are malignant brain tumors associated with a dismal prognosis. A subset of pGBMs carry mutations of either the Lysine 27 or Glycine 34 (G34) amino acid residues of histone 3 variant genes. The same G34 residue is also mutated in 85-95% of giant cell tumors of bone (GCTs), albeit to Tryptophan (G34W) in GCTs rather than to Arginine or Valine (G34R/V) as in pGBMs. The G34 mutation is predicted to impede access of histone methyltransferases like SetD2 to the nearby Lysine 36 residue, thereby altering the epigenome and transcriptome.

Methods: To elucidate the tumorigenic effect of G34 mutations, we used the gene-editing technology CRISPR/Cas9 to correct the G34W mutation to wild-type in 2 GCT cell lines. We then investigated CRISPR-edited cell lines using functional assays, proteomic, epigenomic, and transcriptomic analyses.

Results: Correction of the G34W mutation to wild-type in CRISPR-edited GCT cells results in phenotypic and functional changes suggestive of reduced tumorigenicity. By mass spectrometry, G34W-mutant GCT cell lines display decreased level of Lysine 36 trimethylation (H3K36me3) on the mutant G34-peptide, similar to G34-mutated pGBM cell lines. However, unlike pGBMs, GCTs display increased levels of Lysine 36 dimethylation (H3K36me2) on the mutant G34W-peptide.

Ongoing Experiments and Analyses: We are currently comparing the level and distribution of multiple histone marks by performing chromatin immunoprecipitation followed by next-generation sequencing (ChIP-Seq) of CRISPR-edited wild-type clones relative to the parent G34W-mutant GCT line. We are also assessing differential gene expression by RNA-Seq and characterizing the methylation signature by 850K DNA methylation array of G34W-mutant GCTs and edited clones.

Conclusion: The G34W-mutation clearly has an impact on tumorigenic potential, as evidenced by in vitro functional assays. The G34W-mutant peptide of GCT cell lines features a distinct profile of post-translational histone modifications compared to G34R- or G34V-mutant peptides of pGBM cell lines. Further investigation could elucidate the epigenetic mechanism(s) through which the G34W mutation confers its tumorigenic properties.

Citation Format: Shriya Deshmukh, Sima Khazaei, Dylan Marchione, Ashot Harutyunyan, Benjamin Garcia, Nada Jabado. Characterizing the epigenetic effects of the histone 3.3 G34W mutation in giant cell tumors of bone [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A39.
Abstract A41: Role of UHRF1 in tumorigenesis and metastasis of osteosarcoma
Authors: Wu Stephanie C. , Benavente Claudia A.
Published: February 2019

Osteosarcoma is the eighth most common form of childhood cancer, with very limited advances in treatment over the past three decades. Osteosarcoma is a highly metastatic cancer, with 15-20% of osteosarcoma patients diagnosed after the cancer has already metastasized, causing the survival rate to be little over 50%. Common sites of metastasis include lung and liver. Loss of the retinoblastoma protein (RB) is associated with poor prognosis in osteosarcoma, as defined by shorter life span and heavier tumor burden. Ubiquitin Like with PHD and Ring Finger Domains 1 (UHRF1) has been identified as a critical downstream effector of the RB/E2F pathway that is overexpressed in various cancers. We have confirmed using human mesenchymal stem cells (MSCs) and osteosarcoma cell lines that UHRF1 is directly regulated by the RB/E2F pathway in the bone lineage, and found that the UHRF1 gene is upregulated and its protein overexpressed in osteosarcoma. Using gain/loss-of-function assays to study the role of UHRF1 in osteosarcoma, we observed that UHRF1 is involved in promoting cell migration. To study the role of UHRF1 in osteosarcoma tumor development, we generated Uhrf1 conditional knockout mice. In this system, we use an Osterix-driven Cre recombinase (Osx-cre) to facilitate preosteoblast-specific loss of various combinations of genes. Osx-cre Tp53; Rb1 double knockout mice give rise to osteosarcoma and have a median survival of 26.7 weeks. Additional loss of Uhrf1 in Osx-cre Tp53; Rb1 double knockout mice (Osx-cre Tp53; Rb1; Uhrf1 triple knockout mice) significantly delays the age of tumor onset to >47 weeks. Furthermore, triple knockout mice have a decreased frequency and number of lung and liver metastasis. In conclusion, we have strong evidences both in vitro and in vivo that UHRF1 is a crucial component in aiding osteosarcoma progression, and have identified its novel role in tumor metastasis. The high metastatic rate and low 5-year survival rate following metastasis of osteosarcoma make UHRF1 a promising target for therapeutics. Further studies on the mechanism by which UHRF1 overexpression contributes to osteosarcomagenesis, including tumor growth and dissemination, will strongly aid the process of drug development for anticancer treatment.

Citation Format: Stephanie C. Wu, Claudia A. Benavente. Role of UHRF1 in tumorigenesis and metastasis of osteosarcoma [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A41.
Abstract A45: Tumor suppressor miR-584-5p regulates MYC and sensitizes MYC-amplified medulloblastoma to vincristine and ionizing radiation
Authors: Abdelfattah Nourhan , Rajamanickam Subapriya , Timilsina Santosh , Subbarayalu Panneerdoss , Onyeagucha Benjamin , Rao Manjeet
Published: February 2019

Despite recent improvements in the overall survival of medulloblastoma (MB), only a modest percentage of patients survive high-risk disease. Furthermore, the toxicity associated with current therapies inflicts debilitating lifelong side effects on surviving MB patients. Using an unbiased microRNA screen, we identified miR-584-5p as a potent tumor suppressor that uniquely sensitizes MB to both ionizing radiation (IR) and vincristine (VCR), which are commonly used in combination to treat MB. Our studies revealed that miR-584-5p inhibited MB stem cell proliferation and consequently growth in vivo. Notably, we discovered that miR-584-5p imparts its tumor-suppressor and therapy-sensitizing effects by inhibiting histone deacetylase 1 (HDAC1), eukaryotic translation initiation factor 4e family member 3 (eIF4E3), and C-MYC. We demonstrated that miR-584-5p overexpression or the knockdown of its target genes including eIF4E3 and HDAC1 resulted in G2-M arrest, DNA damage, spindle defects, and apoptosis in MB cells. Meta-analysis of normal brain and MB tumor samples showed that while miR-584-5p is highly expressed in normal brain and cerebellum, it is highly downregulated across all MB subtypes. miR-584-5p targets, on the other hand, are highly overexpressed in MB tumors compared to normal cerebellum. Taken together, this study highlights a previously unidentified role for miR-584-5p, eIF4E3, and HDAC1 in regulating microtubule dynamics and DNA repair pathways. In addition, our study identifies novel targets that can affect the therapeutic efficacy of VCR and IR in MB and sets the stage for a new paradigm of treating medulloblastoma using microRNA-based therapeutics.

Citation Format: Nourhan Abdelfattah, Subapriya Rajamanickam, Santosh Timilsina, Panneerdoss Subbarayalu, Benjamin Onyeagucha, Manjeet Rao. Tumor suppressor miR-584-5p regulates MYC and sensitizes MYC-amplified medulloblastoma to vincristine and ionizing radiation [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A45.
Abstract A47: Inhibition of histone methyltransferase with novel epipolythiodioxopiperazines (ETP) alkaloids in pediatric high-grade glioma
Authors: Dombrowski Stephen , Abou-Antoun Thamara , Mansour Anthony , Arar Rewa , Movassaghi Mohammad , Murphy Erin , Banerjee Shuvojit , Li Xiao-Nan , Houghton Peter , Bigner Darell , Carcaboso Angel Montero , Rich Jeremy , Recinos Violette
Published: February 2019

Pediatric high-grade glioma (HGG), which ranks among the most lethal of cancers among children, contain two recurrent mutations within the histone H3 gene H3F3A (K27M and G34R/V). It is not known whether these mutations directly alter specific epigenetic regulatory mechanisms that may be involved in tumor proliferation and growth, or contribute to their therapeutic resistance and rapid recurrence. Preliminary findings from our lab have identified that Chaetocin (CH), a pan histone methyltransferase (HMT) inhibitor, and two newly synthesized epipolythiodioxopiperazines (ETP) alkaloids that are structurally and functionally distinct HMT inhibitors can effectively kill pediatric HGG (N=7), including three that contain the H3 K27M mutation. In addition, CH and ETPs can also inhibit tumor sphere formation and cell migration at low concentrations (<0.5uM) through in vitro studies. In H3 K27M tumors, these HMTi were shown to reverse hypomethylation of K27me3 transiently (<48 hours). We also found these HMTis to decrease tumor stem cell expression (Sox2, Oct4), and increase tumor-suppressor activity (PTEN, p53) for three pediatric HGG up to 72 hours. These results suggest ETP effectiveness that acts through direct HMT inhibition may also influence other tumorigenic pathways, which may in part contribute to their aggressive nature. Results from this investigation have advanced our understanding of epigenetic regulation of solid brain tumors, and identified drugs that may specifically target pediatric H3 K27M tumor subtypes and could ultimately improve overall survival in pediatric patients with high-grade glioma.

Citation Format: Stephen Dombrowski, Thamara Abou-Antoun, Anthony Mansour, Rewa Arar, Mohammad Movassaghi, Erin Murphy, Shuvojit Banerjee, Xiao-Nan Li, Peter Houghton, Darell Bigner, Angel Montero Carcaboso, Jeremy Rich, Violette Recinos. Inhibition of histone methyltransferase with novel epipolythiodioxopiperazines (ETP) alkaloids in pediatric high-grade glioma [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr A47.
Abstract B26: Novel lncRNA regulates PAX3/FOXO1 expression and is essential for RMS tumorigenesis
Authors: Chen Xiang , Herrera-Herrera Mireya , McEvoy Justina
Published: February 2019

Rhabdomyosarcoma (RMS) is a pediatric cancer of the developing skeletal muscle, and the mechanisms that drive this tumor are not completely understood. Despite the genetic alterations discovered in this disease, therapeutics targeting these mutations have not improved patient survival over the past 25 years. Since more than 60% of patients with metastatic disease do not survive, this creates an urgency to identify new vulnerabilities to exploit for therapeutic development. To do this we performed a comprehensive evaluation of the epigenomic landscape across 16 RMS orthotopic patient-derived xenograft tumors. We identified 41 super enhancers unique to RMS tumors. Within these, we discovered 9 novel lncRNAs of which 3 have potential cis-regulatory activity. We hypothesize that these lncRNAs are required for RMS tumorigenesis. Indeed, from preliminary knockdown analysis of one of these lncRNAs, lnc19_31, we observed rapid cell death in RMS cell lines and concomitant upregulation of apoptotic genes based on RNA-seq, such as TP53. Remarkably, we also observed more than 70 cancer consensus genes that were up- or downregulated, including genes that are frequently mutated in RMS patients such as BCOR and FGFR4. Within these we were excited to discover the loss of PAX3/FOX1 translocation, as this is a major driver of the alveolar subtype and often associated with poor prognosis. Taken together, we hypothesize that lncRNA deregulation is required for expression of key oncogenes and tumor suppressor and essential for RMS cell survival and tumorigenesis. These data will shed insight into a new underlying mechanism that drives this disease. These exciting findings present potential tumor vulnerabilities and, thus, new opportunities for future therapeutic discoveries that would otherwise be beyond our reach.

Citation Format: Xiang Chen, Mireya Herrera-Herrera, Justina McEvoy. Novel lncRNA regulates PAX3/FOXO1 expression and is essential for RMS tumorigenesis [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr B26.
Abstract B35: Reactivation of p53 signaling in hepatoblastoma with a stapled peptide dual inhibitor of MDM2 and MDM4
Authors: Woodfield Sarah E. , Patel Roma H. , Ibarra Aryana M. , Chen Zhenghu , Vasudevan Sanjeev A.
Published: February 2019

Introduction: Hepatoblastoma is the most common pediatric liver malignancy. Despite aggressive therapeutic regimens, long-term survival of patients with high-risk, metastatic disease is less than 50%. Most cases of hepatoblastoma do not carry mutations in the TP53 tumor suppressor gene; therefore, we hypothesized that targeting the two major regulators of p53, MDM2 and MDM4, with the stapled peptide dual inhibitor, ATSP-7041, to reactivate p53 signaling would be an effective therapeutic strategy for hepatoblastoma.

Methods: Levels of MDM2 and MDM4 gene expression were examined in HB cell lines and in a cohort of 18 primary HB patient samples from stage III (n=12) and IV (n=6) patients in comparison to expression in four adjacent uninvolved liver samples with qPCR experiments. The effects of the dual MDM2/MDM4 inhibitor ATSP-7041 (Aileron Therapeutics) on the same cell lines were analyzed with MTT assays for cytotoxicity and qPCR experiments to look at changes in mRNA expression of p53 targets Bax, Puma, CDKN1A, and MDM2.

Results: In 16 of 18 samples, MDM4 expression was higher than average in the uninvolved liver samples; in five samples expression was at least 5-fold higher and in four samples expression was at least 10-fold higher. Of note, all four samples with at least 10-fold higher expression were from stage IV patients. In comparison, MDM2 expression was only elevated in one of the 18 patient samples. All three hepatoblastoma cell lines showed expression of both MDM2 and MDM4. Treatment with ATSP-7041 showed cytotoxic (5.7-8.6 μM) effects in all cell lines tested. All cell lines also showed upregulation of p53 targets Bax (1.4- to 2.4-fold), Puma (2.4- to 3.2-fold), CDKN1A (2.4- to 7.2-fold), and MDM2 (6.1- to 8.3-fold) with treatment.

Conclusions: Dual inhibition of MDM2 and MDM4 shows efficacy in preclinical models of hepatoblastoma by upregulating p53 tumor-suppressor signaling in TP53 wild-type cells and is a promising therapeutic strategy for high-risk patients with wild-type TP53.

Citation Format: Sarah E. Woodfield, Roma H. Patel, Aryana M. Ibarra, Zhenghu Chen, Sanjeev A. Vasudevan. Reactivation of p53 signaling in hepatoblastoma with a stapled peptide dual inhibitor of MDM2 and MDM4 [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr B35.