bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4378: Discovery and characterization of two novel drivers of hepatocellular carcinoma
Authors: Feddersen Charlotte R. , Dupuy Adam J. 
Published: July 2016

Abstract
Liver cancer is globally the sixth most commonly diagnosed and the third most deadly cancer. Hepatocellular carcinoma (HCC) is the most common form of the disease and a difficult cancer to treat due to inadequately understood pathways involved in tumorigenesis and progression. To obtain a better understanding of key HCC driver mutations, we performed a forward genetic screen using Sleeping Beauty transposon mutagenesis in a damaged liver microenvironment. Briefly, T2Onc3; Rosa26-SB11lsl/lsl;Alb-Cre mice were treated with carbon tetrachloride (CCl4) during ongoing transposition. We then sequenced the resulting liver tumors using linker mediated PCR (LM-PCR) and Illumina sequencing. We analyzed the sequenced data using gene-centric common insertion site (gCIS) analysis that allowed us to determine common insertion sites across tumors. These common insertion sites are referred to as candidate driver mutations.
This screen demonstrated the role of Glioma associated protein-2 (Gli2) and Fidgetin (Fign) as candidate driver mutations in a significant subset of HCC tumors (20% and 10% respectively). Gli2 is a key member of the Hedgehog signaling pathway, acting as a transcription factor for genes promoting cell proliferation. Fign is a relatively uncharacterized protein in the AAA-ATPase family with the ability to sever microtubules in vitro. We validated Gli2 as a driver of HCC by generating a hepatocyte specific Gli2-overexpression mouse model using hydrodynamic tail vein injection of Gli2 cDNA into FAH-/- mice. Mice underwent rapid tumorigenesis, regardless of CCl4 treatment, demonstrating that Gli2 is a strong oncogene. Additionally we have demonstrated that overexpression of Fign drives cell proliferation and promotes stabilization of the primary cilium in hepatocytes, a novel finding that provides insight into the mechanism of Fign-driven HCC.
The discovery of two novel HCC candidate driver mutations, validation of Gli2 as a bona fide HCC oncogene, and mechanistic insight into these commonly mutated genes in HCC will allow for future research into understanding HCC tumorigenesis and potential therapeutic options.
Citation Format: Charlotte R. Feddersen, Adam J. Dupuy. Discovery and characterization of two novel drivers of hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4378.
Abstract 4381: High-throughput in vivo systems biology using implantable microdevices
Authors: Jonas Oliver 
Published: July 2016

Abstract
We have developed an implantable device that can perform drug sensitivity testing of a large number of anti-cancer drugs inside the living tumor. This device contains reservoirs which release microdoses of single agents or drug combinations into distinct spatial regions of the tumor in local concentrations that are representative of concentrations achieved during systemic drug treatment. The efficacy of each of up to 30 individual drugs or combinations can be assessed from a single specimen without systemic drug exposure through minimally invasive biopsy. This assay takes into consideration physiologic effects that contribute to drug response by allowing drugs to interact with the living tumor in its native microenvironment.
Using this technology, we have been able to assess phenotypic drug response for up to 100 therapies in a single mouse, probing all clinically relevant single and combination treatments related to the estrogen receptor, CDK4/6 and PI3K pathways in ER+ PDX mouse models. We also examined a range of targeted and cytotoxic drug therapies in parallel before, during and after systemic drug treatments to investigate emerging drug resistance at the phenotypic level in BRAF-mutant melanoma models. The results demonstrate new patterns of escape and adaptive signaling mechanisms, and may offer clues for effective combination therapy.
Citation Format: Oliver Jonas. High-throughput in vivo systems biology using implantable microdevices. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4381.
Abstract 440: Quantification of somatic chromosomal rearrangements in circulating cell-free DNA from ovarian cancers
Authors: Harris Faye R. , Kovtun Irina , Smadbeck James , Multinu Francesco , Jatoi Aminah , Kalli Kimberly R. , Murphy Stephen J. , Halling Geoffrey C. , Kosari Farhad , Johnson Sarah H. , Mariani Andrea , Vasmatzis George 
Published: July 2016

Abstract
Circulating tumor DNA (ctDNA) provides great potential for the non-invasive clinical assessment of tumor burden. We developed a monitoring protocol using next generation mate-pair sequencing to identify chromosomal rearrangement signatures in primary tumors to follow ctDNA in blood. Primary tumors of 10 patients with ovarian cancer were sequenced, and individualized tumor-specific panels were designed to detect the junctions in ctDNA. A novel algorithm was developed and applied to data generated by quantitative PCR to calculate the percent of ctDNA. Selected junctions, some with potential as therapeutic targets, were detected in pre-surgically drawn blood in eight out of ten patients. Of these eight, three demonstrated the continued presence of circulating tumor DNA post-surgery, and five had undetectable levels, consistent with their documented presence or absence of disease. The detection of somatic junctions derived from ctDNA could be an effective way to monitor ovarian cancer patients for relapse and therapy response.
Citation Format: Faye R. Harris, Irina Kovtun, James Smadbeck, Francesco Multinu, Aminah Jatoi, Kimberly R. Kalli, Stephen J. Murphy, Geoffrey C. Halling, Farhad Kosari, Sarah H. Johnson, Andrea Mariani, George Vasmatzis. Quantification of somatic chromosomal rearrangements in circulating cell-free DNA from ovarian cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 440.
Abstract 4403: SMYD2-mediated lysine methylation regulates nuclear transport of beta-catenin in human cancer cells
Authors: Deng Xiaolan , Nakamura Yusuke , Hamamoto Ryuji 
Published: July 2016

Abstract
The canonical Wnt-signal pathway is a major signaling system in cancer cells as well as stem/progenitor cells. It is well-known that binding of Wnt molecules to cell surface receptors, including a Frizzled (Fz) protein and LDL receptor-related protein 6, initiates a transduction cascade leading to stabilization of the transcription coactivator β-catenin, which then enters the nucleus to form a transcriptional complex with T cell factor (TCF) or lymphoid enhancer factor (LEF) to activate the expression of TCF/LEF downstream genes, such as cyclin D1 (CCND1) and c-Myc. However, the molecular mechanisms that regulate nuclear localization of β-catenin and its transcriptional activation are not fully understood. Here we demonstrate that the protein lysine methyltransferase SMYD2 methylates β-catenin at lysine 133, which is located in an amino-terminal domain (NTD), and regulates nuclear transport of β-catenin. After treatment with SMYD2-specific siRNAs in hepatocellular carcinoma SNU475 and SNU449 cells, we fractionated cell components and conducted western blot analysis and found that nuclear localization of β-catenin was significantly decreased after knockdown of SMYD2; we also confirmed this result by immunocytochemistry. In addition, we prepared a lysine 133-substituted β-catenin mutant expression vector and transfected it into 293T cells with a SMYD2 expression vector. We observed that protein amount of lysine 133-substituted β-catenin in the nucleus is significantly lower than that of wild-type β-catenin by western-blot and immunocytochemical analyses. These results indicate that SMYD2-mediated methylation plays a critical role in the nuclear transport of β-catenin in cancer cells. This is a novel mechanism of the canonical Wnt-signal pathway regulated by lysine methylation.
Citation Format: Xiaolan Deng, Yusuke Nakamura, Ryuji Hamamoto. SMYD2-mediated lysine methylation regulates nuclear transport of beta-catenin in human cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4403.
Abstract 4407: Inhibition of STAT3 in primary trastuzumab-resistant HER2-positive cells leads to decrease cell viability
Authors: Helu Xousaen M. , Fernandez Alfida , De Leon Daisy D. 
Published: July 2016

Abstract
Breast Cancer is the second leading cause of cancer related death in women. Approximately, 20%-30% of breast cancer patients overexpress Human Epidermal Growth Factor Receptor 2 (HER2). HER2 Is a Tyrosine Kinase receptor involved in proliferation and survival and its overexpression makes it a good target for patients with HER2 positive breast cancer. Trastuzumab (TRA) is a recombinant monoclonal antibody that binds HER2 receptor and inhibits its signaling. Unfortunately, clinical studies have demonstrated that 30% of patients that overexpress HER2 do not respond to Trastuzumab treatment. Therefore, understanding the molecular mechanisms underlying Trastuzumab resistance is essential to develop new therapies to treat these breast cancer patients. Previous studies in our lab demonstrated that a decrease in cell growth of HER2 overexpressing cells occurred when IGF2 and HER2 were inhibited. Nevertheless, these cells overcame the growth inhibition and the resulting proliferative growth coincided with an increase in STAT3 activation. STAT3 is a transcription factor that promotes cell proliferation and can regulate apoptosis in cancer cells. These findings lead us to hypothesize that blocking STAT3 together with IGF2 and HER2 will inhibit proliferation and cell viability of Trastuzumab resistant breast cancer cells. We treated Trastuzumab-resistant HER2 positive JIMT1 cells with HER2, IGF2, and/or STAT3 inhibitors. After 24 hours of treatment, there was a marked difference in cell morphology observed between the treatment groups. Cell viability was measured using WST-1, a reagent that measures mitochondrial activity. A significant decrease in cell viability was observed when cells were treated for 24 hours with a commercially available STAT3 inhibitor (Sttatic). There was no difference between the treatment groups when STAT3 was inhibited by Stattic or when STAT3 was inhibited in combination with IGF2 and/or HER2 inhibitors. These findings suggest that in Trastuzumab-resistant HER2 positive JIMT1 cells, STAT3 activation overcomes the inhibition of the HER2 and IGF2 proliferation pathways resulting in cell death prevention. Our study demonstrate that Trastuzumab-resistant HER2 positive JIMT1 cells can activate STAT3 as an alternative pathway to evade apoptosis and promote chemoresistance. Our data also illustrate the challenge of treating cancer based on single therapies since tumors have many alternative pathways to promote growth.
Citation Format: Xousaen M. Helu, Alfida Fernandez, Daisy D. De Leon. Inhibition of STAT3 in primary trastuzumab-resistant HER2-positive cells leads to decrease cell viability. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4407.
Abstract 441: Association of interferon inducible genes with tumor immune microenvironment and chemotherapy resistance in high-grade serous epithelial ovarian cancer
Authors: Au Katrina K. , Meunier Liliane , Le Page Cécile , Graham Charles H. , Craig Andrew WB , Childs Timothy , Francis Julie-Ann , Squire Jeremy , Mes-Masson Anne-Marie , Koti Madhuri 
Published: July 2016

Abstract
Chemotherapy resistance is a major hurdle in high-grade serous epithelial ovarian cancer (HGSC) management. We previously reported differential expression of interferon inducible genes in pre-treatment tumours from chemoresistant and sensitive HGSC tumors. STAT1 expression was evaluated as a prognostic and predictive biomarker via immunohistochemistry in Phase I (n = 183) and Phase II (n = 550) biomarker validation studies conducted on HGSC tumours accrued from the Terry Fox Research Institute- Canadian Ovarian Experimental Unified Resource (TFRI-COEUR). Tumor STAT1 expression levels significantly correlated with the density of tumor infiltrating CD8+ T lymphocytes in both Phase I and Phase II cohorts. STAT1 expression significantly associated with progression free survival and response to chemotherapy in both Phase I and Phase II validation studies. Significant positive correlation between STAT1 expression levels and intratumoral CD8+ T cell density was observed. Intratumoral CD8+ T cell infiltration did not associate with progression free survival or response to chemotherapy in both cohorts. Interestingly, the prognostic relevance of CD8+ T cell was enhanced in combination with STAT1 in both cohorts. These findings provide evidence that STAT1 as an independent biomarker and a combination of CD8+ T cell infiltration with STAT1 could be novel immune-based prognostic and predictive biomarkers in HGSC. Findings from the current study will aid in patient stratification for novel immunomodulatory therapies for the management of chemotherapy resistance in HGSC.
Citation Format: Katrina K. Au, Liliane Meunier, Cécile Le Page, Charles H. Graham, Andrew WB Craig, Timothy Childs, Julie-Ann Francis, Jeremy Squire, Anne-Marie Mes-Masson, Madhuri Koti. Association of interferon inducible genes with tumor immune microenvironment and chemotherapy resistance in high-grade serous epithelial ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 441.
Abstract 4420: Heparan sulfate proteoglycans mediate SW13 oncogenic potential and response to granulin
Authors: Davis McKale , Hull Elizabeth 
Published: July 2016

Abstract
The SW13 adrenal adenocarcinoma cell line exists as two epigenetically and phenotypically distinct subtypes. One is a highly proliferative epithelial subtype lacking the expression of key tumor suppressor proteins, Brm and Brg1, the intermediate filament protein vimentin, and the cell cycle check point protein CD44 (SW13-), while the other is a slow growing, mesenchymal-like subtype which expresses all of these proteins (SW13+). Evidence strongly suggests that subtype determination is epigenetic in nature, as HDAC inhibitors can induce an SW13- to SW13+ subtype switch, but the cellular and environmental factors that regulate cellular phenotype have yet to be characterized. We performed a comprehensive microarray analysis and identified heparan sulfate proteoglycan (HSPG) binding as a primary pathway affected between these two subtypes. HSPGs are glycoproteins present in the extracellular matrix that can have tumor-promoting or tumor-suppressing activities depending on the number and sulfation status of attached sugar chains known as glycosaminoglycans (GAGs). Quantitative RT-PCR was then used to confirm the increased expression of the HSPGs, serglycin and glypican-3, as well as the HSPG-binding growth factors granulin and fibroblast growth factors-1 and -2 in SW13+ cells. Furthermore, HDAC inhibition was also shown to increase the expression of these genes in a time-dependent manner. As granulin over-production is associated with cancer progression in a number of human cancers, we were interested in examining the role of this HSPG binding growth factor further. A pro-granulin ELISA showed that SW13+ cells secrete significantly more granulin than SW13- cells. Intriguingly however, treatment with exogenous granulin increases growth rates in the already highly proliferative SW13- subtype, but had no effect on SW13+ growth. We hypothesized that the pleiotropic response of SW13 cells to growth factor treatment may be the result of HSPG post-translational modifications, so we assessed total GAG sulfation was by dimethylmethylene blue assay, and indeed the SW13+ cells were found to have significantly higher levels of total GAG sulfation than SW13- cells. Thus, transcriptional and post-translational modifications of HSPGs may contribute to SW13 phenotype control, and ultimately may affect tumorigenesis.
Citation Format: McKale Davis, Elizabeth Hull. Heparan sulfate proteoglycans mediate SW13 oncogenic potential and response to granulin. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4420.
Abstract 4421: Molecular features of hepatocellular carcinoma arising from non-alcoholic fatty liver
Authors: Takeda Makoto , Sakaguchi Takanori , Kiuchi Ryouta , Hiraide Takanori , Shibasaki Yasushi , Morita Yoshihumi , Kikuchi Hirotoshi , Setou Mitsutoshi , Konno Hiroyuki 
Published: July 2016

Abstract
(Background)
The incidence of hepatocellular carcinoma (HCC) arising from non-alcoholic fatty liver (NAFL) has been recently increasing according to increase of patients having metabolic syndrome. HCC, which shows steatosis, cellular ballooning, Mallory-Denk bodies, inflammation and fibrosis in cancer tissue, are recognized as steatohepatitic HCC (SH-HCC). This HCC phenotype is more frequently found in NAFL patients than in viral-hepatitic patients.
(Purpose)
The purpose of this study is to elucidate molecular features of SH-HCC with NAFL background.
(Methods)
To profile SH-HCC-related genes, mRNA expression patterns in SH-HCCs were compared with those in viral hepatitis HCCs by RNA array. RNA expression of the identified SH-HCC related gene was analyzed in three human liver carcinoma cell lines (HepG2, Huh7, and HLE) in normal conditioned medium. By mimicking the lipid-rich environment, these cell lines were also cultured in the presence of saturated (palmitic or stearic acid, recognized as endoplasmic reticulum stressors) or unsaturated fatty acids (oleic acid: OA). To address the role of SH-HCC-related gene, cell proliferation and apoptosis were assessed in the presence or absence of fatty acids, by knock-down of the objective gene using siRNA.
(Results)
(1) Caveolin (cav)-1 and -2 were identified as SH-HCC related genes (1.63 and 1.58 fold expression of cav-1 and -2, respectively) by RNA microarray analysis.
(2) HLE and Huh7 showed high and low expression of both cav-1 and -2 mRNA, whereas HepG2 did not express either on qRT-PCR.
(3) In the presence of OA, HepG2 and Huh7 did not well proliferate, whereas HLE cell proliferation increased with concentration of OA and culture duration.
(4) Knock-down of cav-1 and -2 canceled OA-induced increase of cell proliferation. In the same condition, expression of cleaved caspase 3 was increased, suggesting the promotion of apoptosis.
(Consideration)
Cav-1 and -2, the important protein for endocytosis and cellular signaling, are highly expressed in SH-HCC. These proteins would increase cell proliferation and inhibit apoptosis in the presence of unsaturated fatty acids, presumably by using these fatty acids as energy source or by caveolae-related anti-apoptotic process.
Citation Format: Makoto Takeda, Takanori Sakaguchi, Ryouta Kiuchi, Takanori Hiraide, Yasushi Shibasaki, Yoshihumi Morita, Hirotoshi Kikuchi, Mitsutoshi Setou, Hiroyuki Konno. Molecular features of hepatocellular carcinoma arising from non-alcoholic fatty liver. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4421.
Abstract 4437: DNA methylation analysis in cisplatin resistant ovarian cancer cells and recurrent ovarian cancer patient samples
Authors: Granados Michaela L. , Hudson Laurie G. , Luo Li , Samudio-Ruiz Sabrina L. 
Published: July 2016

Abstract
Acquisition of platinum resistance following first line platinum therapy is commonly observed in ovarian cancer patients and is a major obstacle to clinical effectiveness. Currently there are no options available to prevent platinum resistance; however, studies show that demethylating agents may resensitize patients to platinum therapy thereby demonstrating DNA methylation as a critical contributor to the development of platinum resistance. Using a cellular model of platinum resistance we have previously shown that resistant cells have increased DNA methyltransferase (DNMT) activity as well as increased global DNA methylation. These observations were dependent on cisplatin induced activation of the Epidermal Growth Factor Receptor (EGFR) which we previously identified as a novel a regulator of DNMT activity and DNA methylation. Inhibition of the EGFR conferred during a platinum resistance paradigm decreased DNMT activity, decreased global methylation and attenuated resistance in our platinum resistant model. We hypothesized that further genome wide analysis of DNA methylation in resistant, parental and EGFR inhibited cell lines at specific CG sites would reveal targets of methylation important for the development of platinum resistance. Using the Illumina 450K methylation array, we found few significant alterations in DNA methylation for given genes between control and cisplatin resistant (CPR) cells. However, additional DNA methylation analysis with the 450K methylation array of patient tumor samples at initial diagnosis as well as after relapse, while not significant, showed similar trends in the data for certain genes namely CSRNP3, LOC400904, LYPD6, MTMR9L, NIPAL4 and RBP7. We are still evaluating the effects of EGFR inhibition in vitro on DNA methylation patterns. We suggest that while the in vitro model of platinum resistance corresponds to patient samples in some aspects, ovarian cancer cells display a great deal of heterogeneity in DNA methylation as described by others and a much larger sample size is necessary to draw concrete conclusions. Current studies are evaluating gene expression in control and resistant cells to validate whether alterations in DNA methylation coincide with changes in expression.
Citation Format: Michaela L. Granados, Laurie G. Hudson, Li Luo, Sabrina L. Samudio-Ruiz. DNA methylation analysis in cisplatin resistant ovarian cancer cells and recurrent ovarian cancer patient samples. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4437.