bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 854: Combination effect of low dose Cytarabine and pan-RAF inhibitor LY3009120 in AML cells with RAS mutations
Authors: PARK Jihyun , Yoon Sung-Soo , Koh Youngil 
Published: August 2019

Abstract
The alterations of RAS oncogenes were known to be involved in various cancers. In acute myeloid leukemia (AML), there are 6% of alterations in KRAS and 13% in NRAS. The RAS/RAF/MEK/ERK cascade has roles in promoting hematopoietic cell growth, inhibiting apoptosis, and is related to chemotherapy resistance. Fortunately, the recent advance in pharmacology suggests the advent of potentially efficacious RAS inhibitors. However, current RAS inhibitor development is focused on solid tumors. We, therefore, tried to identify if potential RAS targeting molecule would be effective in hematologic malignancies. Here we focused on the combination of LY3009120, a pan-RAF inhibitor that targets RAF with widely used antimetabolite agent cytarabine on AML cell line.
We used AML cell lines, HL-60, NB-4 which have NRAS/KRAS mutation and KG-1 as RAS wild-type control cell line. In the cytotoxicity test, we first performed a single effect of cytarabine and LY3009120 in the AML cell lines using cytotoxicity assay. Based on the single treat experiment, cytarabine and LY3009120 were simultaneously treated on three AML cell lines at the indicated concentration. Above all, there was a strong synergistic effect on combination treat of cytarabine containing LY3009120 in HL-60 (56.8%, p=0.0001) and NB-4 (66%, p&lt;0.0001) compared with KG-1 cell line (8%).
In order to examine the changes in RAF/MAPK pathways, we demonstrated western blotting using antibodies to RAS, RAF, MAPK signaling molecules. In HL-60, the phosphorylation of MEK was slightly increased by the single treat of cytarabine or LY3009120. However, the combination treatment of cytarabine and LY3009120 decreased the phosphorylation of MEK. And more, in NB-4, the phosphorylation of ERK (T202/Y204) was not affected by single treatment while the combination treatment decreased ERK phosphorylation at low concentration of LY3009120. Furthermore, we investigated the underlying mechanisms of the combined effect, PARP protein cleavage indicating cell death was examined. PARP cleavage was observed in HL-60 and NB-4 after treatment, whereas KG-1 did not show any change in PARP after treatment.
In conclusion, we show the synergistic effect between cytarabine and LY3009120 on AML cell lines which have the mutation on RAS gene. Even at the low concentration of cytarabine, combining LY3009120 significantly increases the anti-cancer effect. With our data, we suggest that novel therapeutic strategies for AML patients with RAS mutation.
Citation Format: Jihyun PARK, Sung-Soo Yoon, Youngil Koh. Combination effect of low dose Cytarabine and pan-RAF inhibitor LY3009120 in AML cells with RAS mutations [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 854.
Abstract 855: Analysis of a novel variant of HER2 detected by clinical sequencing in cancer of unknown primary
Authors: Harada Yohei , Kashiwada Tomomi , Sato Akemi , Abe Tomonori , nakashima Chiho , Sueoka Eisaburo , Kimura Shinya , Sueoka-Aragane Naoko 
Published: August 2019

Abstract
A patient with cancer of unknown primary underwent an NGS-based multiplex gene assays (OncoPrime). As a result, a novel variant of HER2 (E401G) which is a missense alteration located in the extracellular domain of the HER2 protein was detected. This alteration has not been reported (COSMIC, Nov 2018) and its effect on protein function is unknown. Sanger sequencing using genomic DNA from peripheral blood sample showed that the variant was a somatic variant. HER2 E401G variant was confirmed by Sanger sequencing using genomic DNA extracted from the formalin-fixed paraffin-enbedded tumor tissues. In spite of somatic variant, the variant allele frequency was relatively high (55.8%). This result suggested the amplification of HER2 gene, FISH analysis confirmed the copy number gain of HER2 gene (21 copies). The prediction tools in silico (SIFT, PolyPhen-2 and PROVEAN) suggested pathogenicity of the variant, so we are now performing the functional analysis of the mutant HER2 protein. Many variants of unknown significance have been identified through clinical sequencing, establishing a database of integrated information of variant is essential to implement precision medicine of oncology. This study was approved by the Ethics Committee and the patient provided written informed consent for the use of genomic and clinical data for research purposes.
Citation Format: Yohei Harada, Tomomi Kashiwada, Akemi Sato, Tomonori Abe, Chiho nakashima, Eisaburo Sueoka, Shinya Kimura, Naoko Sueoka-Aragane. Analysis of a novel variant of HER2 detected by clinical sequencing in cancer of unknown primary [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 855.
Abstract 857: Identification and functional analysis of FGFR2 binding proteins in diffuse-type gastric carcinoma
Authors: Shirakihara Takuya , Sakai Ryuichi 
Published: August 2019

Abstract
Diffuse-type gastric carcinoma (DGC) is highly invasive subtype of gastric adenocarcinomas and frequently exhibit scattered peritoneal metastasis. Previous studies have shown that genes of receptor tyrosine kinases (RTKs) such as FGFR2 or Met are amplified in DGC cell lines with high frequency. Hence, these cells exhibit oncogene addiction to sustained activity of these RTKs for maintenance of malignant phenotype. In order to gain novel insight in the downstream signaling pathway of DGC-specific RTKs, phosphoproteomic analysis was performed. Phosphotyrosine-containing proteins associated with RTKs were purified through two sequential rounds of immunoprecipitation from lysates of RTK-amplified DGC cell lines. Here we will show the initial functional data of transferrin receptor (TFRC), one of the identified proteins by MS analysis. In the DCG cells with FGFR2 amplification, TFRC proteins were found to bind FGFR2 and were phosphorylated at Tyr20. Moreover, shRNA knockdown of TFRC as well as inhibitor of FGFR2 revealed effective impairment in cell proliferation. These results suggest that TFRC has essential roles in malignancy of FGFR2-dependent DGC.
Citation Format: Takuya Shirakihara, Ryuichi Sakai. Identification and functional analysis of FGFR2 binding proteins in diffuse-type gastric carcinoma [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 857.
Abstract 858: Oncogenic EGFR mutations as genomic biomarkers for cetuximab and panitumumab response in colorectal adenocarcinoma
Authors: Kim Nayoung , Cho Jeonghee 
Published: August 2019

Abstract
Somatic mutations of Epidermal Growth Factor Receptor (EGFR) occur in ~3% of colorectal cancer patients. Here, through systematic functional screening of 21 recurrent EGFR mutations selected from public datasets, we show that 11 colon cancer-derived EGFR mutants; G63R, E114K, R165Q, R222C, S492R, P596L, K708R, E709K, G719S, G724S and L858R, are oncogenic, and able to transform cells in a ligand-independent manner. We demonstrate that cellular transformation by these mutants requires receptor dimerization. Importantly, the EGF-induced and constitutive oncogenic potential of these EGFR mutants are inhibited by cetuximab or panitumumab in vivo and in vitro.
Taken together, we propose that a subset of EGFR mutations can serve as genomic predictors for response to anti-EGFR antibodies and that metastatic CRC patients with such mutations may benefit from these drugs as part of the first-line therapy.
Citation Format: Nayoung Kim, Jeonghee Cho. Oncogenic EGFR mutations as genomic biomarkers for cetuximab and panitumumab response in colorectal 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 858.
Abstract 860: Solution structure of a dGMP fill-in G-quadruplex forms in PDGFR-b promoter
Authors: Wang Kaibo , Dickerhoff Jonathan , Wu Guanhui , Lin Clement , Yang Danzhou 
Published: August 2019

Abstract
Platelet-derived growth factor receptor beta (PDGFR-β) is an important cell-surface-receptor tyrosine kinase implicated in PDGF signaling pathways. The PDGFR-β signaling pathway has been demonstrated as an essential component in control of the cellular growth, proliferation, survival, motility and differentiation. Overexpression of PDGFR-β is associated with tumor growth, angiogenesis, and migration, making PDGFR-β an attractive target for anticancer therapeutics. In the PDGFR-β gene promoter, a highly GC-rich proximal promoter region -165 to -139 nt upstream of the transcriptional start site is crucial for basal promoter activity, which can form G-quadruplexes (G4s). These structures can be stabilized by the G4-interactive molecules to inhibit the PDGFR-β transcriptional activity. Our previous structure study showed that the G4 structures formed in the PDGFR-β gene promoter always contain a broken G-strand. The broken-strand structure suggests a possibility of the formation of a G-vacancy-bearing G-quadruplex (GVBQ) structure, which can be complemented and stabilized by guanine and guanine derivatives. Elevated levels of guanine and guanine derivatives/metabolites are found in tumor cells, suggesting that the GVBQs may play important regulatory roles in tumor cell pathological processes, such as PDGFR-β transcriptional regulation. Using NMR, CD, EMSA and DMS footprinting experiments, herein, we contributed the first atomic resolution structure of a dGMP fill-in G4 formed in PDGFR-β gene promoter in potassium solution. Surprisingly, our results showed that solution dGMP preferentially inserts from the 5' end tetrad instead of the G-22 position of the previously determined structure, and maybe in dynamic equilibrium with our previously reported G-quadruplex. The dGMP, like other intramolecular guanines, that forms four Hoogsteen hydrogen bonds with adjacent guanines and stabilizes by electronic coordination with potassium cation as well as the π-π stacking interaction with the middle tetrad guanine. Moreover, a series of guanine derivatives were investigated to fill in this GVBQ, and found that only these derivatives that keep the ability to form Hoogsteen hydrogen bonds in G-tetrad can serve as the substrate, providing insights into designing new guanine derivatives anticancer drugs.
Citation Format: Kaibo Wang, Jonathan Dickerhoff, Guanhui Wu, Clement Lin, Danzhou Yang. Solution structure of a dGMP fill-in G-quadruplex forms in PDGFR-b promoter [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 860.
Abstract 861: Precise excision of an oncogenic allele of tumor by Adenovirus-based CRISPR/Cas9 system induces tumor regression
Authors: Koo Taeyoung , Yoon A-Rum , Bae Sangsu , Kim Jin-Soo , Yun Chae-Ok 
Published: August 2019

Abstract
Approximately 15% of non-small cell lung cancer patients possesses several mutations in the epidermal growth factor receptor (EGFR) gene, which plays a critical role in tumor progression. To this end, we demonstrate that an adenovirus-mediated co-delivery of Cas9 and a guide RNA targeting oncogenic EGFR mutation commonly found in lung cancer can selectively remove oncogenic allele while being inactive against wild-type EGFR allele. An EGFR mutant harboring a single-nucleotide missense mutation (CTG to CGG) was targeted as this mutation leads to generation of protospacer-adjacent motif sequence recognized by the Cas9 protein of S. pyogenes. The CRISPR/Cas9-mediated excision of mutant EGFR allele with high indel rate led to potent lung cancer cell killing. Importantly, intratumoral administration of Ad-based CRISPR/Cas9 system led to potent tumor growth inhibition and complete tumor regressions in number of mice harboring EGFR mutant xenograft tumors. Collectively, our findings show that precise and controlled excision of oncogenic mutation by Ad-mediated expression of CRISPR/Cas9 system offers a powerful genomic surgical strategy to excise oncogenic mutations to treat cancers.
Note: This abstract was not presented at the meeting.
Citation Format: Taeyoung Koo, A-Rum Yoon, Sangsu Bae, Jin-Soo Kim, Chae-Ok Yun. Precise excision of an oncogenic allele of tumor by Adenovirus-based CRISPR/Cas9 system induces tumor regression [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 861.
Abstract 869: Molecular characterization of a pediatric high-grade glioma (pHGG) mouse model harboring the H3.3 G34R mutation
Authors: Whalen Matthew J. , Garcia-Fabiani Maria B. , Núñez Felipe J. , Lowenstein Pedro R. , Castro Maria G. 
Published: August 2019

Abstract
Pediatric high-grade glioma (pHGG) accounts for 8-12% of central nervous system tumors and has a 90% mortality rate. Consequently, novel therapies are needed to advance treatment options and improve survival rates. A means to do this is through designing a mouse model that accurately mirrors pHGG in humans to further study this type of pHGG and design treatment options. We focused on the analysis of pHGG harboring mutations on histone H3.3 at glycine 34 (H3.3 G34R) in combination with ATRX and TP53 mutations. We generated a mouse model for pHGG using the sleeping-beauty transposase system and the delivery of plasmids into the sub-ventricular zone of neonatal mice. This results in the transposon-mediated integration of the genetic lesions into the brain cells, thereby allowing the generation of tumors with the desired molecular alterations. Two groups of mice have been developed; the control group (WT H3.3 group) bears tumors generated with the NRASV12 overexpression and two shRNA that target the Tp53 and Atrx genes. The H3.3 G34R group of mice harbors tumors generated with NRASV12 expression, sh-TP53, sh-ATRX, and H3.3 G34R expression. RNA-sequencing was carried out to analyze the transcriptome signature associated with the H3.3 G34R tumor phenotype. Several genes related to the activation of the immune response were differentially upregulated in the H3.3 G34R tumors, such as Ciita, CD74, and Irfl, amongst others. Some of these deregulated genes were validated by immunohistochemistry. Understanding the tumor transcriptome and characterizing the tumor’s immune microenvironment are crucial for the development of gene and immune therapies designed to target H3.3 G34R pHGG.
Citation Format: Matthew J. Whalen, Maria B. Garcia-Fabiani, Felipe J. Núñez, Pedro R. Lowenstein, Maria G. Castro. Molecular characterization of a pediatric high-grade glioma (pHGG) mouse model harboring the H3.3 G34R mutation [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 869.
Abstract 872: Genetic models reveal that the novel VGLL2-NCOA2 fusion oncogene leverages embryonic programs for sarcomagenesis
Authors: Kendall Genevieve C. , Watson Sarah , Xu Lin , LaVigne Collette , Murchison Whitney , Rakheja Dinesh , Tirode Franck , Delattre Olivier , Amatruda James 
Published: August 2019

Abstract
Rhabdomyosarcoma (RMS) is an aggressive pediatric cancer characterized by a misregulation of skeletal muscle developmental pathways. To date, identified oncogenic drivers predominantly include RAS mutations or chromosomal translocations and gene fusions between PAX3 or PAX7 and FOXO1. RNAseq analysis of sarcomas with non-canonical gene fusions has identified new potential genetic drivers of tumorigenesis that have not been rigorously functionally validated for their transformation capacity and biological activity. One new fusion is a chromosomal translocation and inversion between chromosomes 6 and 8, which acts to juxtapose two transcriptional co-activators, VGLL2 and NCOA2. This VGLL2-NCOA2 fusion was identified in congenital rhabdomyosarcoma clinical cohorts by us and others, and characterizes aggressive RMS that express MYOD and MYOG histological markers. However, evidence of VGLL2-NCOA2 transformation capacity has not been verified, hindering insights into its functional contributions to tumorigenesis. Here, we interrogate the function ofVGLL2-NCOA2 using complementary genomic patient data and zebrafish model systems. We utilized the Tol2 transposon system to express mosaic human VGLL2-NCOA2 during early development, and found that VGLL2-NCOA2 is sufficient for tumorigenesis, and results in aggressive tumors with high penetrance by 75 days of age in zebrafish. Further, the histology of zebrafish tumors resembles the human disease, and tumors express markers indicative of RMS such as myog and desma. A cross-species RNAseq of patient and zebrafish VGLL2-NCOA2 RMS tumors highlights a significant enrichment and overlap between gene expression signatures. Finally, mapping the gene expression signatures of VGLL2-NCOA2 zebrafish RMS tumors along the spectrum of zebrafish embryogenesis indicates a clustering with developmental stages corresponding to early somitogenesis, highlighting their arrested developmental nature. Hence, we have generated the first animal model of human VGLL2-NCOA2 tumorigenesis, and have applied this model to understand the biology and identify potential therapeutic targets for this newly identified disease.
Citation Format: Genevieve C. Kendall, Sarah Watson, Lin Xu, Collette LaVigne, Whitney Murchison, Dinesh Rakheja, Franck Tirode, Olivier Delattre, James Amatruda. Genetic models reveal that the novel VGLL2-NCOA2 fusion oncogene leverages embryonic programs for sarcomagenesis [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 872.
Abstract 875: Targeting Nrf2 driven glutathione de novo synthesis as a novel strategy to suppresses IDH1-mutated glioma
Authors: Liu Yang , Yu Di , Cai Sabrina , Yang Chunzhang 
Published: August 2019

Abstract
Background: Isocitrate dehydrogenase (IDH) mutation frequently occurs in low grade glioma and secondary glioblastoma, characterizing by the abnormally catalyzation of alpha-ketoglutarate (α-KG) to D-2-hydroxyglutarate (D-2-HG), which induces CpG island methylator phenotype, as well as metabolic reprogramming and high reactive oxygen species (ROS) generation. Our previous data showed that nuclear factor-erythroid 2-related factor 2 (Nrf2) plays a crucial role in maintaining redox homeostasis of IDH1-mutated glioma cells, while the detail mechanism remains elusive. Our recent data showed that glutathione de novo synthesis is governed by Nrf2, which makes great contribution to ROS scavenging in IDH1-mutated tumors and could serve as a potential therapeutic target.
Methods: In this study, we explored the important role of glutathione de novo synthetic pathway in maintaining ROS homeostasis by small RNA interference of key enzymes GCLC, GCLM and SLC7A11. We also investigated the correlation between Nrf2 and glutathione de novo synthetic pathway by Chromatin Immunoprecipitation. We further investigated the therapeutic value of Nrf2 inhibitor, Brusatol, in IDH1-mutated cancer both in vitro and in vivo.
Results: Our data showed that IDH1 mutation cells exhibit high level steady-state of ROS generation and detoxification. Prolonged Nrf2 activity enhanced transcriptional activity of key enzymes in glutathione de novo synthesis, such as GCLC, GCLM and SLC7A11. Targeting Nrf2 using Brusatol suppressed glutathione synthesis in IDH1-mutated cells, resulted in cytotoxic effect and suppressed xenograft growth in vivo.
Conclusions: IDH1-mutated glioma highly depends on Nrf2 activity to maintain ROS homeostasis. Enhanced glutathione de novo synthesis is an important mechanism that governed by Nrf2 and plays critical role in ROS scavenging. Targeting ROS scavenging might be a novel approach to treat gliomas with IDH1 mutation.
Citation Format: Yang Liu, Di Yu, Sabrina Cai, Chunzhang Yang. Targeting Nrf2 driven glutathione de novo synthesis as a novel strategy to suppresses IDH1-mutated glioma [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 875.
Abstract 88: Stable luciferase expressing cell lines for in vivo xenograft and syngeneic tumor model bioluminescence imaging
Authors: Foulke John , Chen Luping , Miagkov Alexei , Turner-Gillies Elizabeth , Volpe Lysa-Anne , Tian Fang 
Published: August 2019

Abstract
Human and mouse cancer cell lines are used in xenograft and syngeneic models, respectively, for the in vivo study of tumor formation and development, metastases, measuring tumor burden in whole animals and monitoring response to therapeutic treatment. Whole animal in vivo imaging has been widely applied by researchers due to the ease of operation in visualizing in vivo biological events, eliminating the requirement for animal subject sacrifice, allowing for continual monitoring/imaging of a single individual animal and reducing the amount of inter-animal variation. Luciferase reporters provide a relatively simple, robust, and highly sensitive means to measure biological processes and to assess drug efficacy in animal models through in vivo bioluminescence imaging. Here we report on the generation of a large panel of cell lines which express high levels of luciferase and -have broad applications for in vitro and in vivo studies. The panel includes human and mouse cell lines, which represent various common cancer and tissue types, such as breast, colon, lung, pancreas, prostate, and skin. After introduction of a Lenti-LUC2 luciferase reporter into the parental cell lines, single cell cloning was performed to isolate stable clones with high luciferase expression. We quantified both the relative and absolute in vitro bioluminescence signals within the cells and analyzed the signal-to-noise ratio. In addition, the cell lines were characterized and authenticated using cell morphology, growth kinetics, and STR analysis. In addition, luciferase labeled human cell lines were administered into immune-deficient mice either by subcutaneous or orthotropic injection to establish xenograft models. Luciferase labeled mouse cell lines were tested in the associated breed of mice to evaluate tumor growth in the syngeneic mouse model. The bioluminescence live imaging was performed using the Xenogen IVIS imaging system and the in vivo bioluminescent radiance was analyzed. Although growth can be evaluated by both traditional caliper measurement and bioluminescence in subcutaneous mouse models, only live bioluminescent imaging enables continuous monitoring tumor metastasis or analysis of tumor growth of the intracranial brain tumor models. In summary, luciferase expressing cell lines are valuable tools for elucidating mechanisms involved in tumorigenesis, studying tumors in vivo, and screening anti-cancer compounds for drug discovery and development.
Citation Format: John Foulke, Luping Chen, Alexei Miagkov, Elizabeth Turner-Gillies, Lysa-Anne Volpe, Fang Tian. Stable luciferase expressing cell lines for in vivo xenograft and syngeneic tumor model bioluminescence imaging [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 88.