bluuebunny/crossref_metadata_2025 huggingface dataset:
Research on Side Lobe Suppression of Time-Modulated Sparse Linear Array Based on Particle Swarm Optimization
Authors: Liang Lei , Sun Jie , Li Hailin , Liu Jialing , Jiang Yachao , Zhou Jianjiang
Published: August 2019

An efficient pattern synthesis approach is proposed for the synthesis of a time-modulated sparse linear array (TMSLA) in this paper. Due to the introduction of time modulation, the low/ultralow side lobe level can be obtained with a low amplitude dynamic range ratio. Besides, it helps reduce the difficulty of antenna feeding system effectively. Based on particle swarm optimization (PSO) and convex (CVX) optimization, this paper proposes a hybrid optimization method to suppress the grating lobes of the sparse arrays, peak side lobe level (PSLL), and peak sideband level (PSBL). Firstly, the paper utilizes the CVX optimization as a local optimization algorithm to optimize the elements’ switch-on duration time, which reduces the side lobe of the array. Secondly, with the PSBL as the objective function, the paper adopts the PSO as a global optimization algorithm to optimize the elements’ positions and switch-on time instant, which helps reduce the loss of sideband power caused by time modulation. With respect to the time modulation model, variable aperture sizes (VAS) and more flexible pulse-shifting (PS) schemes are used in this paper. Owing to the introduction of time modulation and CVX optimization, the proposed method is much more feasible and efficient than conventional approaches. Furthermore, it has better array pattern synthesis performance. Numerical examples of the TMSLA and comparisons with the reference are presented to demonstrate the effectiveness of the proposed method.
Identification of Potential Biomarkers and Survival Analysis for Head and Neck Squamous Cell Carcinoma Using Bioinformatics Strategy: A Study Based on TCGA and GEO Datasets
Authors: Shen Yujie , Liu Jinhui , Zhang Liqing , Dong Shikun , Zhang Jiacheng , Liu Yaqin , Zhou Han , Dong Weida
Published: August 2019

The mechanism and gene markers of head and neck squamous cell carcinoma (HNSCC), a common malignant tumor, have not yet been identified. The aim of this study was to identify the key genes and pathways associated with HNSCC and to further analyze its molecular mechanism and prognostic significance. In this study, the expression profile chip data GSE6631 from Gene Expression Omnibus (GEO) included paired HNSCC tumor and normal samples from 22 patients; the RNAseq tertiary dataset of HNSCC and corresponding clinical information from The Cancer Genome Atlas (TCGA) included biological information of 12 normal head and neck tissues and 111 HNSCC sample tissues. Differentially expressed genes (DEGs) were screened by R software, and the pathway enrichment analysis of DEGs was performed by DAVID, String, and Sytoscape software programs. Combining the GEO and the TCGA databases, we used bioinformatics technology to screen out 50 DEGs in HNSCC and enrich the biological functions and key pathways of HNSCC. Then we performed Gene Ontology (GO) enrichment analysis, the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis, protein-protein interaction (PPI) analysis, and survival analysis on these DEGs. Using CMap, we identified candidate small molecules that might reverse HNSCC gene expression. Finally, four most important small molecules that could provide more reliable biomarkers for early diagnosis and individualized control of HNSCC were identified.
Anticipating Stock Market of the Renowned Companies: A Knowledge Graph Approach
Authors: Liu Yang , Zeng Qingguo , Ordieres Meré Joaquín , Yang Huanrui
Published: August 2019

An increasing number of the renowned company’s investors are turning attention to stock prediction in the search for new efficient ways of hypothesizing about markets through the application of behavioral finance. Accordingly, research on stock prediction is becoming a popular direction in academia and industry. In this study, the goal is to establish a model for predicting stock price movement through knowledge graph from the financial news of the renowned companies. In contrast to traditional methods of stock prediction, our approach considers the effects of event tuple characteristics on stocks on the basis of knowledge graph and deep learning. The proposed model and other feature selection models were used to perform feature extraction on the websites of Thomson Reuters and Cable News Network. Numerous experiments were conducted to derive evidence of the effectiveness of knowledge graph embedding for classification tasks in stock prediction. A comparison of the average accuracy with which the same feature combinations were extracted over six stocks indicated that the proposed method achieves better performance than that exhibited by an approach that uses only stock data, a bag‐of‐words method, and convolutional neural network. Our work highlights the usefulness of knowledge graph in implementing business activities and helping practitioners and managers make business decisions.
Biomechanical Effects of Different Auxiliary-Aligner Designs for the Extrusion of an Upper Central Incisor: A Finite Element Analysis
Authors: Savignano R. , Valentino R. , Razionale A. V. , Michelotti A. , Barone S. , D’Antò V.
Published: August 2019

Aim. To evaluate the biomechanical effects of four different auxiliary-aligner combinations for the extrusion of a maxillary central incisor and to define the most effective design through finite element analysis (FEA). Materials and Methods. A full maxillary arch (14 teeth) was modelled by combining two different imaging techniques: cone beam computed tomography and surface-structured light scan. The appliance and auxiliary element geometries were created by exploiting computer-aided design (CAD) procedures. The reconstructed digital models were imported within the finite element solver (Ansys® 17). For the extrusion movement, the authors compared the aligner without an attachment with three auxiliary-aligner designs: a rectangular palatal attachment, a rectangular buccal attachment, and an ellipsoid buccal attachment. The resulting force-moment (MF) system delivered by the aligner to the target tooth and the tooth displacement were calculated for each scenario. Results. The maximum tooth displacement along the z-axis (0.07 mm) was obtained with the rectangular palatal attachment, while the minimum (0.02 mm) was obtained without any attachments. With the ellipsoid attachment, the highest undesired moments Mx and My were found. The rectangular palatal attachment showed the highest Fz (2.0 N) with the lowest undesired forces (Fx = 0.4 N; Fy = −0.2 N). Conclusions. FEA demonstrated that the rectangular palatal attachment can improve the effectiveness of the appliance for the extrusion of an upper central incisor.
Mutant IDH Sensitizes Gliomas to Endoplasmic Reticulum Stress and Triggers Apoptosis via miR-183-Mediated Inhibition of Semaphorin 3E
Authors: Zhang Ying , Pusch Stefan , Innes James , Sidlauskas Kastytis , Ellis Matthew , Lau Joanne , El-Hassan Tedani , Aley Natasha , Launchbury Francesca , Richard-Loendt Angela , deBoer Jasper , Chen Sheng , Wang Lei , von Deimling Andreas , Li Ningning , Brandner Sebastian
Published: August 2019

Human astrocytomas and oligodendrogliomas are defined by mutations of the metabolic enzymes isocitrate dehydrogenase (IDH) 1 or 2, resulting in the production of the abnormal metabolite D-2 hydroxyglutarate. Here, we studied the effect of mutant IDH on cell proliferation and apoptosis in a glioma mouse model. Tumors were generated by inactivating Pten and p53 in forebrain progenitors and compared with tumors additionally expressing the Idh1 R132H mutation. Idh-mutant cells proliferated less in vitro and mice with Idh-mutant tumors survived significantly longer compared with Idh-wildtype mice. Comparison of miRNA and RNA expression profiles of Idh-wildtype and Idh-mutant cells and tumors revealed miR-183 was significantly upregulated in IDH-mutant cells. Idh-mutant cells were more sensitive to endoplasmic reticulum (ER) stress, resulting in increased apoptosis and thus reduced cell proliferation and survival. This was mediated by the interaction of miR-183 with the 5′ untranslated region of semaphorin 3E, downregulating its function as an apoptosis suppressor. In conclusion, we show that mutant Idh1 delays tumorigenesis and sensitizes tumor cells to ER stress and apoptosis. This may open opportunities for drug treatments targeting the miR-183–semaphorin axis.

Significance:

The pathologic metabolite 2-hydroxyglutarate, generated by IDH-mutant astrocytomas, sensitizes tumor cells to ER stress and delays tumorigenesis.
Abstract 1002: An association between Annexin A1 (ANXA1) and loss of progesterone receptor expression in estrogen receptor-positive breast cancer
Authors: Ahn Sung Gwe , Bae Soong June , Cha Chihwan , Yoon Chang Ik , Park So Eun , Jeong Joon
Published: August 2019

Purpose: There is a strong body of evidence that low expression of progesterone receptor has a negative prognostic value in estrogen receptor (ER)-positive breast cancer. To discover key factors associated with PR-loss in ER-positive cancer, we conducted cell line experiments and investigated clinical samples.

Methods: We generated PR knock-down MCF7 and T47D cell lines using lentiviral transfection with PR shRNA. Gene expression microarray analyses were performed using Illumina Chip. Annexin A1 (ANXA1) was evaluated for tissue microarray (TMA) using HER2-negative human breast carcinoma.

Results: When PR-intact and PR-knockdown cells were compared using gene-expression profiles, ANXA1 was commonly identified as a molecule associated PR-knockdown in both MCF7 and T47D cell lines. Further, the mRNA expression of ANXA1 was elevated by PR-knockdown in both cell lines. In HER2-negative patients (n=370), ANXA1-positive tumors were associated with a lack of PR expression (P<0.001). The patients with ANXA1-positivity had a poor survival outcome in terms of overall survival and metastasis-free survival.

Conclusions: Our findings inform that ANXA1 may act a role in tumor progression of PR-lacking breast cancer.

Citation Format: Sung Gwe Ahn, Soong June Bae, Chihwan Cha, Chang Ik Yoon, So Eun Park, Joon Jeong. An association between Annexin A1 (ANXA1) and loss of progesterone receptor expression in estrogen receptor-positive breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1002.
Abstract 1013: Role of mitochondria and glutamine metabolism in therapy resistance: Combined targeting of estrogen receptor alpha and exportin 1 in endocrine resistant breast cancers
Authors: Cotul Eylem Kulkoyluoglu , Smith Brandi Patrice , Chang Hua , Landesman Yosef , Duong Kevin , Erdogan Zeynep Madak
Published: August 2019

Majority of breast cancer specific deaths in women with ERα (+) tumor occur due to metastases that are resistant to endocrine therapy. There is a critical need for novel therapeutic approaches to prevent or delay recurrence of ERα (+) tumors. The objective of this study was to elucidate the role of mitochondrial pathways that are activated in the presence of single therapies and undermine therapy effectiveness in ER (+) breast cancers. In our previous studies, we identified Exportin 1 (XPO1), a nuclear export protein, as an important player in endocrine resistance progression. Selinexor (SXR), an XPO1 antagonist, has been evaluated in multiple later stage clinical trials in patients with relapsed and /or refractory hematological and solid tumor malignancies. Using a combination of transcriptomics, kinase arrays, metabolomics and metabolic flux experiments, we identified glutamine metabolism pathways to be rewired during endocrine resistance. In limited media conditions mimicking nutrient deprived tumor microenvironment, endocrine resistant cells were more dependent on mitochondria for energy production. Their glucose and fatty acid dependency decreased in the presence of SXR and cells were more dependent on glutamine. The effect of glutamine was dependent on conversion of the glutamine to glutamate and mitochondrial complex 1 activity. In order to examine metabolites that might result in the observed phenotype we performed GC/MS whole metabolite profiling and identified aminoacid metabolism pathways to be upregulated when cells were treated with SXR. We demonstrated that combined targeting of XPO1 and ERα rewires metabolic pathways and shuts down both glycolytic and mitochondrial pathways that would eventually lead to autophagy. Remodeling metabolic pathways to regenerate new vulnerabilities in endocrine-resistant breast tumors is novel, and given the need for better strategies for improving therapy response of relapsed ERα (+) tumors, our findings show great promise for uncovering the role ERα-XPO1 crosstalk plays in reducing cancer recurrences.

Citation Format: Eylem Kulkoyluoglu Cotul, Brandi Patrice Smith, Hua Chang, Yosef Landesman, Kevin Duong, Zeynep Madak Erdogan. Role of mitochondria and glutamine metabolism in therapy resistance: Combined targeting of estrogen receptor alpha and exportin 1 in endocrine resistant breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1013.
Abstract 1016: The androgen receptor splice variant AR-V7 associates with the glucocorticoid receptor following androgen blockade
Authors: Bueter Eric , Hosfield David , McAuley Erin , Selman Phillip , Greene Geoffrey , Conzen Suzanne D. , Szmulewitz Russell Z.
Published: August 2019

Expression of the constitutively active androgen receptor splice variant AR-V7 is associated with resistance to anti-androgen therapies and worse clinical outcomes in prostate cancer. Dimerization is a requirement for AR-V7 transcriptional activity, but whether that occurs through homodimerization or heterodimerization remains poorly understood. Because NR3C-family nuclear hormone receptors rely on a highly conserved D-box motif within the DNA-binding domain to mediate dimerization, we hypothesized that AR-V7 can heterodimerize with other NR3C-family nuclear hormone receptors, with a particular interest in the glucocorticoid receptor (GR). Like AR-V7, GR is upregulated in prostate cancer following androgen blockade, regulates canonical AR target genes, and is associated with resistance to anti-androgen therapies. Using NanoBRET technology in HEK293 cells, we found that AR-V7 interacts with liganded NR3C-family nuclear hormone receptors, including the full-length androgen receptor, the progesterone receptor, the mineralocorticoid receptor, and, most notably, GR. Using co-immunoprecipitation and proximity ligation assays, we demonstrate that AR-V7 associates with GR following androgen blockade in both the VCaP cell line and the CWR-22RV1 cell line, two models of prostate cancer that express endogenous AR-V7 and GR. Preliminary ChIP-Seq experiments in these cell lines suggest AR-V7 and GR chromatin occupancy at the same genomic sites in a ligand-dependent manner. Taken altogether, this data suggests AR-V7 can utilize GR as an alternative binding partner to initiate heterodimerization and transcriptional activation.

Citation Format: Eric Bueter, David Hosfield, Erin McAuley, Phillip Selman, Geoffrey Greene, Suzanne D. Conzen, Russell Z. Szmulewitz. The androgen receptor splice variant AR-V7 associates with the glucocorticoid receptor following androgen blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1016.
Abstract 1018: GnRH antagonists have direct inhibitory effects on castration-resistant prostate cancer via intracrine androgen and AR-V7 expression
Authors: Cucchiara Vito , Yang Joy C. , Liu Chengfei , Adomat Hans , Guns Emma , Gleave Martin E. , Gao Allen C. , Evans Christopher P.
Published: August 2019

Hormone therapy is currently the mainstay in the management of locally advanced and metastatic prostate cancer (PCa). Degarelix (Firmagon, Deg), a GnRH receptor antagonist demonstrated some advantages over the LHRH agonist Leuprolide (Leup) by avoiding “testosterone flare” and lower FSH levels. We compared the effect of Deg and Leup on prostate cancer (PC) cell progression in vitro and in vivo. GnRHR2 was readily detectable in PC cells. AR transcriptional activity reported by PSA-Luc assay was modulated by both Leup and Deg. In LNCaP and C4-2B MDVR cells, 20µM Deg significantly reduced the cell viability (p<0.01). GnRH-antagonist (alone or in combination with AA or Enza) counteracted the transactivation activity of AR by reducing AR-FL and AR variants at the protein level. In C4-2BMDVR cells, Deg reduced AR-V7 protein expression by 26 to 40% alone or combined with AA and Enza compared to control. Leup, however, enhanced variant expression. Deg reduced AR-variant levels from 17 to 41% in monotherapy or combinations compared to control. In mice, Leup slightly suppressed tumor growth compared to controls (p>0.05). However, tumors in Deg-treated group were 1.5-fold smaller than Leup-treated group but 1.7-fold bigger than surgical castration-group. Ki67 IHC staining confirmed the difference in tumor proliferation among groups. Measurements of intratumoral steroids by LC-MS from tumors, serum samples or cell pellets confirmed that Deg decreased the levels of testosterone and steroidogenesis pathway intermediates, comparable to surgical castration; while Leup had no inhibitory effect. Our results suggest a selective mechanism of action of Deg against AR-variants. The present study provides additional molecular insights regarding the mechanism of Deg compared to GnRH agonist therapy which may have clinical implications.

Citation Format: Vito Cucchiara, Joy C. Yang, Chengfei Liu, Hans Adomat, Emma Guns, Martin E. Gleave, Allen C. Gao, Christopher P. Evans. GnRH antagonists have direct inhibitory effects on castration-resistant prostate cancer via intracrine androgen and AR-V7 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1018.
Abstract 1032: Establishing an in vitro model of lipopolysaccharide-induced tumorospheres formation of prostate epithelial cells for drug screening
Authors: Jiang Jiajia , Lian Xueqi , Tang Sijie , Li Xiaohua
Published: August 2019

Tumorospheres forming assay not only was utilized to enrich cancer stem cell from bulk cancer cells, also was widely used to analyze the ability of clonogenic growth and self-renewal capability of cancer stem cells. Meanwhile, chronic inflammation-induced epithelial transformation has been documented in a broad range of cells. However, the effect of chronic inflammatory agent on tumorosphere formation has never been tested. In current study, the classic pro-inflammatory agent lipopolysaccharide was utilized to establish an in vitro model through inducing prostate tumoroshpere formation. Then, trichostatin A, a panel inhibitor of histone deacetylases, was tested in this model for its anti-tumor effect. The results showed that treatment with lipopolysaccharide stimulated prostate cancer DU145 and PC3 cells undergoing migration and invasion with decreased the ratio of CK8 and CK18 versus vimentin. It also induced tumorospheres formation in suspension culture with increased a panel of tumor stem cell markers transcription (e.g. CD44, α-intergrin, nestin etc) and promoted phosphorylation of Stat3 (p-Stat3) and pro-oncogene c-Myc expression in vitro. Treatment with trichostatin A showed to restrain the above lipopolysaccharide-induced EMT and tumorosphere formation with decreasing the level of p-Stat3, p-Erk, p-c-jun, c-Myc, CD44 and SOX-2. Finally, blocking Stat3 signaling pathway by treatment with trichostatin A and/or small molecule compound Stattic, an inhibitor of p-Stat3, abrogated LPS-induced cancer stem-like tumorosphere formation. Taken together, our data demonstrated that lipopolysaccharide induced cancer stem-like tumorospheres formation successfully, which could be served as a simple and easy handling in vitro model for anti-tumor drug screening.

Citation Format: Jiajia Jiang, Xueqi Lian, Sijie Tang, Xiaohua Li. Establishing an in vitro model of lipopolysaccharide-induced tumorospheres formation of prostate epithelial cells for drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1032.
Abstract 1038: Metformin inhibits epithelial-mesenchymal transition in human pancreatic cancer cell lines
Authors: Yoshida Juichiro , Ishikawa Takeshi , Endo Yuki , Matsumura Shinya , Ohta Takayuki , Mizushima Katsura , Hirai Yasuko , Okayama Tetsuya , Sakamoto Naoyuki , Inoue Ken , Katada Kazuhiro , Kamada Kazuhiro , Uchiyama Kazuhiko , Takagi Tomohisa , Naito Yuji , Itoh Yoshito
Published: August 2019

Background: Epithelial-mesenchymal transition (EMT) is considered to be a crucial event in the development of cancer metastasis. Metformin is an antidiabetic drug used in treatment of type 2 diabetes. Recently, increasing evidence has indicated that metformin can inhibit tumor growth, however, effect of metformin on EMT remains still well unknown.

Methods: We investigated whether metformin inhibits EMT in the human pancreatic cancer cell lines (PANC-1, BXPC-3, and MIAPaCa-2). Pancreatic cancer cells were stimulated TGF-β1 (10ng/ml) with or without metformin(10mM). After 48h, we evaluated for EMT-related factors using western blot analyses, immunocytochemistry and RT-PCR. Wound healing assay was performed to determine cell migration.

Results: The cells stimulated TGF-β1 acquired elongated and fusiform morphology, and metformin inhibited the TGF-β1-induced morphological changes in PANC-1 and BXPC-3 cells. Wound healing assay of PANC-1 and MIAPaCa-2 cells showed that significant wound healing was seen after 24h in cells treated with TGF-β1 compared with the control, and metformin inhibited wound closure. E-cadherin expression weakened after exposure to TGF-β1, but it was reversed by metformin in these cell lines. Metformin treatment reduced the expression of mesenchymal markers (Vimentin, N-cadherin and αSMA) induced by TGF-β1 in these cell lines. Exposure of cells to TGF-β1 resulted in the phosphorylation of mTOR, and exposure of cells to metformin resulted in the phosphorylation of AMPK and blocked the phosphorylation of mTOR in BXPC-3 cells. Furthermore, cell viability was no significantly change between exposing metformin or not.

Discussion: Our results indicate that metformin inhibits TGF-β1 induced EMT in human pancreatic cancer cells. Moreover, our results suggest that metformin induces the phosphorylation of AMPK and inhibits TGF-β1 induced mTOR activation in BXPC-3 cells.

Citation Format: Juichiro Yoshida, Takeshi Ishikawa, Yuki Endo, Shinya Matsumura, Takayuki Ohta, Katsura Mizushima, Yasuko Hirai, Tetsuya Okayama, Naoyuki Sakamoto, Ken Inoue, Kazuhiro Katada, Kazuhiro Kamada, Kazuhiko Uchiyama, Tomohisa Takagi, Yuji Naito, Yoshito Itoh. Metformin inhibits epithelial-mesenchymal transition in human pancreatic cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1038.
Abstract 1046: A breast cancer MET reporter cell line model for drug discovery and development
Authors: Enuameh Metewo S. , Shu Weiguo , Newman Robert , Kumari Sangeeta
Published: August 2019

World-wide, cancer metastasis continues to be the leading cause of death in cancer (DIZON et al. 2016). Although epithelial-to-mesenchymal transition (EMT) and mesenchymal-to-epithelial transition (MET) have been implicated in the incidence of cancer metastasis and drug resistance, their impact in cancer progression and patient survival is not fully understood (NIETO et al. 2016). This is partly due to the lack of suitable in vitro models. Thus, to facilitate the utility of the EMT concept in therapeutic development, we have utilized some of the basic biology of EMT/MET, to create a novel advanced in vitro model for use in both basic research and discovery of new anti-EMT drugs.

In breast cancer, vimentin (VIM) intermediate filament (IF) proteins are generally upregulated during EMT and down-regulated during MET (GILLES et al. 1999; THIERY and SLEEMAN 2006; RICHARDSON et al. 2012; LAMOUILLE et al. 2014). Here we employed CRISPR/Cas9 gene editing to generate a VIM RFP (red fluorescent protein) reporter in the MDA-MB-231 breast adenocarcinoma cell line. The VIM RFP C-terminal fusion gene at the endogenous VIM locus, enables end-point or real-time tracking of the MET status as cells transition from epithelial to mesenchymal phenotype under distinct conditions. We have validated the cell line at the nucleic acid (genomic and mRNA) and protein levels as well as in cell-based assays. Bio-functional evaluation of the MDA-MB-231 VIM RFP cell line shows sensitivity to metastatic breast cancer drugs axitinib (tyrosine kinase inhibitor) and U0126 (MEK1/2 inhibitor) via the inhibition of the inherent signaling pathways which impact EMT. These effects provide the basis for the use of this cell line in high throughput screening (HTS) applications including the discovery of new anti-EMT drugs for metastatic breast cancer. Furthermore, the MDA-MB-231 VIM RFP reporter cell line is also a convenient and sensitive model for studying the mechanisms of metastasis and for basic translational science research.

Citation Format: Metewo S. Enuameh, Weiguo Shu, Robert Newman, Sangeeta Kumari. A breast cancer MET reporter cell line model for drug discovery and development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1046.
Abstract 1049: Targeting YAP/TAZ pathway inhibits Ewing sarcoma metastasis
Authors: Bierbaumer Lisa , Katschnig Anna M. , Radic-Sarikas Branka , Petro Jeffrey R. , Mühlbacher Karin , Aryee Dave N. , Pötsch Anna R. , Högler Sandra , Kenner Lukas , Uren Aykut , Kovar Heinrich
Published: August 2019

The term epithelial to mesenchymal transition (EMT) and its reverse (MET) describe the dynamic and reversible metamorphosis of cells from a highly organized to a loose migratory phenotype through cytoskeletal reorganization causing cellular plasticity. EMT/MET are inherent to normal embryonal development and wound healing, but cancer cells hijack the underlying mechanisms to enable oscillations between proliferation, invasion and migration that cause metastasis, the major killer of cancer patients. Recently, fluctuations of the oncoprotein EWS-FLI1 were identified to drive EMT/MET in Ewing sarcoma. We previously demonstrated that, in presence of EWS-FLI1, transcriptional co-activators MRTFB and TAZ are largely blocked from associating with TEAD and its target genes keeping tumor cells in a poorly migratory, highly proliferative state. In contrast, under EWS-FLI-low conditions, MRTFB and TAZ associate with YAP/TEAD complexes on chromatin resulting in cytoskeletal target gene activation and phenotypic transition to a highly migratory and low proliferative state. We therefore hypothesized that pharmacologic inhibition of YAP/TAZ/TEAD protein interaction should prevent Ewing sarcoma cells from EMT and consequently interfere with their metastatic potential. Verteporfin is a small molecule safely used in the treatment of age-related macular degeneration. Independent of its photosensitizing activity exploited in ophthalmology, it is a YAP/TAZ pathway -blocking compound. In vitro treatment of EWS-FLI1-low Ewing sarcoma cells with verteporfin resulted in decreased YAP/TAZ/TEAD complex formation in the nanomolar range, reversal of the de-repression of a EWS-FLI1 controlled EMT transcriptional signature, and inhibition of tumor cell migration in a Boyden chamber assay. Upon orthotopic implantation of TC71 Ewing sarcoma cells into the tibial crest of SCID beige mice, intra-peritoneal treatment of mice with 25mg verteporfin/kg/day before and after amputation of the affected limb led to a drastic decrease of lung metastases without affecting primary tumor growth and without obvious general toxicity. Therefore, YAP/TAZ pathway blockade holds promise as a potential metastasis-preventive strategy in the treatment of Ewing sarcoma patients with primary localized disease.

Citation Format: Lisa Bierbaumer, Anna M. Katschnig, Branka Radic-Sarikas, Jeffrey R. Petro, Karin Mühlbacher, Dave N. Aryee, Anna R. Pötsch, Sandra Högler, Lukas Kenner, Aykut Uren, Heinrich Kovar. Targeting YAP/TAZ pathway inhibits Ewing sarcoma metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1049.
Abstract 1050: Characterization of human cancer cell line xenografts in humanized mice
Authors: Stevens Christina , Maddage Christopher , Lasky Kerri , Rios-Doria Jonathan , Koblish Holly
Published: August 2019

Studying immune-oncology (IO) agents that target the human immune system in mice have posed challenges for oncology researchers. Since the commercial introduction of humanized mice, antitumor efficacy studies can now be performed with human cancer cells within mice bearing the components of a human immune system. However, development and characterization of these models is necessary to understand which model may be best for different agents. To this end, we characterized A375, A549, Caki-1, H1299, H1975, HCC827, HCT116, KU19-19, MDA-MB-231, and RKO human cancer cell lines in CD34+ humanized NSG mice for tumor growth rate and immune cell profiling in tumor and spleen compartments. We found that CD4+ T cell composition in both the spleen and tumor varied among the models, with A375, Caki-1, and RKO expressing higher levels of CD4+ T cells compared to the other models. Many of the models had moderate levels of CD8+ T cells in the tumor. Spleen analysis revealed higher NK cell populations in mice bearing Caki-1 tumors, while A549 tumor samples had a higher percentage of NK cells relative to the other models. Surprisingly, many of the tumor models showed an abundance of myeloid cells in the spleen and tumor compartments, although these were mostly comprised of B cells and dendritic cells and lower levels of macrophages and MDSCs. The expression level and percentage of cells expressing PD-1, and PD-L1 in the models were also determined. Furthermore, the antitumor response of select models to PD-1 and PD-L1 antibodies will be presented. In summary, these data demonstrate that there are tumor-intrinsic factors that influence the immune cell repertoire within tumors and spleen. These data may also aid in selection of tumor models to test antitumor activity of novel IO agents.

Citation Format: Christina Stevens, Christopher Maddage, Kerri Lasky, Jonathan Rios-Doria, Holly Koblish. Characterization of human cancer cell line xenografts in humanized mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1050.
Abstract 1051: Dianhydrogalactitol (VAL-083) reduces glioblastoma tumor growth upon bevacizumab-induced hypoxia, in vivo
Authors: Golebiewska Anna , Oudin Anais , Steino Anne , Niclou Simone P. , Bacha Jeffrey , Brown Dennis M.
Published: August 2019

Background: Standard-of-care for glioblastoma (GBM) includes surgery, radiation and temozolomide (TMZ). Nearly all tumors recur and 5-year survival is less than 3%. Unmethylated promoter status for O6-methylguanine-DNA-methyltransferase (MGMT) is a validated biomarker for TMZ-resistance. Second-line treatment with bevacizumab has not only failed to improve survival, but has also been shown to induce intratumor hypoxia, which is implicated in increased chemoresistance. VAL-083 is a bi-functional DNA-targeting agent that readily crosses the blood-brain barrier and accumulates in brain tumor tissue. VAL-083 induces DNA double-strand breaks at N7-guanine and cancer cell-death in GBM cancer stem cells (CSCs) and non-CSCs, independent of MGMT. We have previously shown that bevacizumab treatment upregulates expression of glucose transporters GLUT-1/GLUT-3 on GBM cells. We hypothesized that, based on its unique monosaccharide backbone structure, VAL-083 may benefit from bevacizumab-induced GLUT transporter upregulation leading to enhanced VAL-083 uptake and anti-tumor activity.

Methods: To investigate the in vivo anti-tumor effect of VAL-083+bevacizumab, we used a orthotopic patient-derived xenograft GBM model. All mice carried MGMT-unmethylated, temozolomide-resistant recurrent T16 GBM tumors as detected by MRI 35 days post-implantation. Mice were grouped into control, bevacizumab, VAL-083, and VAL-083+bevacizmab. Tumor progression was measured by MRI on days 49 and 56, and tumor growth rate was calculated for the entire study (day 35 vs. 56) and for the last 7 days (day 49 vs. 56).

Results: Tumors were significantly smaller in VAL-083-treated mice both compared to control (-83%, p<0.001) and compared to bevacizumab-treated (-75%, p<0.001) mice. Additionally, analysis of tumor growth in-time showed significantly reduced tumor growth rate for VAL-083+bevacizumab compared to VAL-083 alone (p<0.01).

Conclusions: These results show strong in vivo anti-tumor efficacy of VAL-083 against MGMT-unmethylated, TMZ-resistant recurrent GBM. This effect was further augmented in combination with bevacizumab, providing rationale of clinical investigation of VAL-083 in combination with bevacizumab in the treatment of GBM.

Citation Format: Anna Golebiewska, Anais Oudin, Anne Steino, Simone P. Niclou, Jeffrey Bacha, Dennis M. Brown. Dianhydrogalactitol (VAL-083) reduces glioblastoma tumor growth upon bevacizumab-induced hypoxia, in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1051.
Abstract 1053: A patient-derived xenograft model demonstarates the effectiveness of combined oxaliplatin and 5-fluorouracil for advanced small bowel adenocarcinoma
Authors: Yamano Tomoki , Song Jihyung , Kimura Kei , Babaya Akihito , Hamanaka Michiko , Kataoka Kozo , Beppu Naohito , Noda Masafumi , Ikeda Masataka , Tomita Naohiro
Published: August 2019

Background: Combination oxaliplatin and capecitabine therapy is recommended to treat advanced small bowel adenocarcinoma (SBA). However, SBA is rare, and limited preclinical studies and randomized clinical trials have been conducted.

Methods: A patient-derived xenograft (PDX) was established from an advanced SBA patient with liver metastasis and peritoneal metastasis. Growth inhibition by oxaliplatin, irinotecan, 5-fluorouracil (5-FU), and 5-FU in combination with oxaliplatin, irinotecan, or paclitaxel was assessed using the PDX. Drug sensitivity against oxaliplatin, irinotecan, 5-FU, and paclitaxel was assessed using SBA cells from the PDX. The patient’s clinicopathological and genetic features were compared with the PDX to evaluate it as a preclinical model.

Results: 5-FU, but not oxaliplatin and irinotecan showed statistically significant growth inhibition compared to control group (P = 0.0024). Among the combination treatment with 5-FU, oxaliplatin showed the highest tumor growth inhibition rate of 0.84 or 1.1, which was 0.46 for irinotecan and 0.49 for paclitaxel, respectively. IC50 of SBA cells was significantly lower against oxaliplatin and higher against paclitaxel compared with colon cancer cell line HT29. These results corresponded to the clinical response to FOLFOX (folinic acid+5-FU+oxaliplatin) and FOLFIRI (folinic acid+5-FU+irinotecan). PDX pathological and genetic features resembled those of the patient.

Conclusions: We established a preclinical model comprising the PDX from SBA and SBA cells from the PDX. This model showed that the combination of oxaliplatin and 5-FU was a

reasonable standard therapy for advanced SBA. A preclinical model comprising PDX and tumor cells from PDX seems to be useful for rare tumors including SBA.

Citation Format: Tomoki Yamano, Jihyung Song, Kei Kimura, Akihito Babaya, Michiko Hamanaka, Kozo Kataoka, Naohito Beppu, Masafumi Noda, Masataka Ikeda, Naohiro Tomita. A patient-derived xenograft model demonstarates the effectiveness of combined oxaliplatin and 5-fluorouracil for advanced small bowel adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1053.
Abstract 1062: Development and characterization of patient-derived xenograft models for acute myeloid leukemia
Authors: Ulrich Michelle , Anderson Martha , Thurman Robert , Li Fu
Published: August 2019

Drug development for acute myeloid leukemia (AML) remains challenging, in part, due to the lack of preclinical models that reflect the biology of AML patients. For example, while the majority of AML patients express both CD33 and CD123, only a small fraction of AML cell lines express CD123. Similarly, most AML cell lines have a differentiated phenotype, which may bias the evaluation towards certain types of AML. To address these challenges, we developed a dozen patient derived xenograft (PDX) models to aid preclinical development of AML drugs. Bone marrow samples from 32 AML patients were implanted intravenously into NSG mice. Thirteen of them led to successful expansion of human CD45+ cells in the bone marrow, as demonstrated by flow cytometry. Twelve of these established disease in a secondary passage. Using multicolor flow cytometry analysis, we found that expression of CD33 and CD123 were preserved from AML patients and engrafted disease in NSG mice. Moreover, RNA-sequencing analysis demonstrated that gene expression largely remained consistent between the first two passages. Detailed genetic analysis demonstrated the preservation AML-relevant mutations, such as FLT3-ITD mutations and IDH2 mutations in the PDX model. In summary, we have successfully established AML PDX models that reflect the antigen expression and molecular genetics of AML patients. These models may facilitate the future development of antibody-based therapeutics.

Citation Format: Michelle Ulrich, Martha Anderson, Robert Thurman, Fu Li. Development and characterization of patient-derived xenograft models for acute myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1062.
Abstract 1090: β-Hydroxy-β-Methylbutyrate supplementation preserves muscle mass and reduces tumor growth in obese mice
Authors: Coleman Michael F. , Liu Kristyn A. , Tang Xiaohu , Fabela Salvador , Lashinger Laura M. , Cui Zhengrong , Hursting Stephen D.
Published: August 2019

Background: Pancreatic ductal adenocarcinoma (PDAC) remains very challenging to treat with mean 5-year survival of approx. 6%, leading to PDAC’s status as the 4th most deadly cancer in the US. Diet induced obesity (DIO) has been shown to promote both increased incidence and growth of PDAC. Simultaneously DIO promotes muscle loss and the loss of immune surveillance in tumors. Cachexia, a chronic catabolic process in which muscle mass is lost, is a common feature of PDAC. We have previously shown that while leucine driven mTOR activation protects muscle mass in tumor bearing mice it also promotes tumor growth. Hence, we sought to test the potential of a leucine metabolite, β-hydroxy-β-methyl-butyrate, to protect against cachexia in a PDAC model while antagonizing DIO mediated tumor growth. We further sought to determine if HMB treatment would alter gemcitabine response in tumors from obese animals.

Methods: C57BL/6 mice consuming either control or high fat diet, bearing Panc02 tumors, were treated with HMB alone or in combination with gemcitabine. Muscle size was determined using Feret’s diameter measured from H&E stained cross sections. Immunohistochemistry staining was performed for Ki-67 and CD3. Tumor transcriptomic analysis was preformed using Affymetrics microarray. In vitro characterization HMB modulated signaling in C2C12 myotubes, Panc02 cells, and bone marrow derived macrophages treated with pro-inflammatory signaling molecules.

Results: a) HMB significantly increases muscle fiber size and mTOR signaling; b) HMB reduces DIO increased tumor volume; c) HMB significantly synergizes with gemcitabine to suppress PDAC growth in obese mice; and d) HMB reverts DIO mediate immune suppression and promotes tumor immune surveillance.

Discussion: Antagonism of DIO promoted PDAC tumor growth, by HMB, and promotion of immune surveillance may offer significant synergy with standard therapy or, immune targeted therapies in PDAC.

Citation Format: Michael F. Coleman, Kristyn A. Liu, Xiaohu Tang, Salvador Fabela, Laura M. Lashinger, Zhengrong Cui, Stephen D. Hursting. β-Hydroxy-β-Methylbutyrate supplementation preserves muscle mass and reduces tumor growth in obese mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1090.
Abstract 1096: Response of pancreatic ductal adenocarcinoma to gemcitabine using human organotypic slice cultures
Authors: Chang Jae Hyuck , Lim Chae Yoon , Lee Weon Sun , Lee Guk Jin
Published: August 2019

Background/Objective: The oragnotypic tumor slice culture model resembles the in vivo situation of tumor. It may serve as a proper ex vivo platform for response to anti-tumor therapies. We aimed to evaluate the response of pancreatic ductal adenocarcinoma (PDA) cells and helper, cytotoxic, and regulatory T cells to gemcitabine using human PDA slice culture model.

Methods: PDA tissues were obtained from patients who were operated on for pancreatic resection. The tumors were sliced by a vibratome, and the tumor slices were then cultured until 5 days with 0.1 μM, 1 μM, and 10 μM gemcitabine for 1 or 2 days. Tumor responses were evaluated by number of tumor cells and proportion of Ki-67 or caspase-3 positively expressed cells in the tumor. CD4+, CD8+, and FOXP3+ cells were analyzed by immunohistochemistry staining.

Results: Gemcitabine treatment to PDA slices exerted proportional cytotoxic effects in terms of tumor cell number, and Ki-67 and cleaved caspase-3 expression. Mean Ki-67 positive cells in PDA cells tended to decrease according to increase of gemcitabine concentration (case 1, 48 hr treatment; 65.7% in control vs. 27.9% in 0.1 μM, p=0.025). Mean caspase-3 positive cells in PDA cells tended to increase according to increase of gemcitabine concentration (case 1, 48 hr treatment; 8.6% in control vs. 18.6% in 0.1 μM, p=0.042). Mean PDA cells per x200 tended to decrease according to increase of gemcitabine concentration (case 2, 48 hr treatment; 78.3 in 1.0 μM vs. 27.3 in 10 μM, p=0.042). Mean CD4+, CD8+, and FOXP3+ cells/total cells tended to decrease according to increase of gemcitabine concentration; in case 3 (24hr treatment), mean CD4+ cells/total cells decreased as 2.6%, 1.6%, 1.5%, and 0.5%, mean CD8+ cells/total cells decreased as 2.6%, 1.5%, 1.4%, and 1.1%, and mean FOXP3+ cells/total cells were 0.2%, 0.3%, 0.2%, and 0.1% in control, 0.1 μM, 1 μM, and 10 μM gemcitabine, respectively. However, mean CD8+/FOXP3+ cell ratio did not significantly decrease, and even increase from 4.4 in 0.1 μM to 8.6 in 1 μM (p=0.039).

Conclusions: Tumoricidal effect of gemcitabine could be demonstrated using organotypic slice culture model. T cells in the tumor decreased according to increase of gemcitabine concentration; however CD8+/FOXP3+ cell ratio relatively maintained.

Citation Format: Jae Hyuck Chang, Chae Yoon Lim, Weon Sun Lee, Guk Jin Lee. Response of pancreatic ductal adenocarcinoma to gemcitabine using human organotypic slice cultures [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1096.
Abstract 1098: Enhanced AKT-dependent metabolic fitness and antitumor effect of T lymphocytes in lung cancer model
Authors: Kim Sung Eum , Pyo Kyoung-Ho , Lim Sang Bin , Yun Mi Ran , Kim Jae Hwan , Lee Ji Min , Jo Ha Ni , Xin Chun-Feng , Cho Jae Seok , Cho Byoung Chul
Published: August 2019

Introduction: PI3K signaling plays a pivotal role in balancing of immune responses. This study is to elucidate the role of PI3K-AKT signaling in regulating T cell metabolic fitness.

Methods: PI3K/AKT inhibitors including BKM120 were used to evaluate the contribution of PI3K-AKT signaling on the metabolic fitness of T cell. Glucose metabolism and T cell activity were measured by flow cytometry, IHC, RT-PCR. Oxygen consumption rate (OCR) and extra cellular acidification rate (ECAR) were examined by seahorse. In addition, anti-tumor effects of T cells were evaluated by adaptive transfer in LLC2 tumor bearing mouse.

Results: Compare to vehicle, BKM120-treated antigen-specific CD8+ T cells co-cultured with LLC2 cancer cells maintained cytotoxic effects under hypoglycemic and hypoxic conditions. Treatment of BKM120 significantly reduced tumor growth, compared to vehicle in syngeneic models (LLC2, KLN205), whereas it did not affect tumor growth in nude mice, which suggests role of activation of antitumor immunity, augmented by BKM120. Compared to vehicle, BKM120 significantly induced higher infiltration of CD8+ T cells and reduction of Foxp3+ regulatory T cell and CD11b+/Gr-1+ myeloid derived suppressor cells. Intriguingly, increased glucose uptake through overexpression of glucose transporter (GLUT1) was observed upon BKM120 treatment. Glut1 overexpression occurred through the release of feedback inhibition of mTORC2 followed by rebound AKT activation. In addition, glycolysis-related genes (Glut1, Hk, Pgam, Pdk and Pgm) and OCR/ECAR were significantly augmented in BKM120/selective PI3K-delta inhibitor (Idelalisib)-treated group. The enhanced glycolysis via PI3K signaling inhibition was significantly more prominent by Idelalisib, compared to BKM120 or selective PI3K-alpha inhibitor (BYL-719). The antigen-specific T cells pretreated with BKM120 ex vivo exhibited substantially more potent antitumor effects, compared to untreated T cells, in LLC2 tumor-bearing nude mice.

Conclusion: PI3K inhibitor, specifically selective PI3K-delta inhibitor, promotes AKT-dependent metabolic fitness of T cells which sustain anti-cancer effects under nutrient/oxygen-deprived tumor microenvironment. These data support a rationale for combination of selective PI3K-delta inhibitor with checkpoint inhibitors anti-PD1 for patient with Lung cancer.

Citation Format: Sung Eum Kim, Kyoung-Ho Pyo, Sang Bin Lim, Mi Ran Yun, Jae Hwan Kim, Ji Min Lee, Ha Ni Jo, Chun-Feng Xin, Jae Seok Cho, Byoung Chul Cho. Enhanced AKT-dependent metabolic fitness and antitumor effect of T lymphocytes in lung cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1098.