bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2764: Detection of early gastric cancer with Sox17 DNA methylation analysis of gastric washes
Authors: Watanabe Yoshiyuki , Oikawa Ritsuko , Yamamoto Hiroyuki , Itoh Fumio 
Published: July 2016

Abstract
Aim: Although minimal invasive treatment is widely accepted for early stage of gastric cancer (EGC), we still do not have any appropriate risk markers to detect residual neoplasia and some potential of recurrence. We previously reported that aberrant DNA methylation is an early and frequent process in gastric carcinogenesis and could be useful for detection of gastric neoplasia (Gastroenterology 2009, Tumor Biology 2011). Our goal is to verify the potency of our three selected candidate genes (Sox17, MINT25, miR34) as a treatment marker for EGC. Method: We conducted a multicenter, prospective study using selected three candidate genes (Sox17, MINT25 and miR34) by Methylated CpG Island Amplification Microarray (MACAM) based analysis and analyzed the DNA methylation levels before and after endoscopic resection (ER) for EGC using gastric washes. We statistically analyzed sensitivity, specificity and AUC with DNA methylation levels of before / after ER and information of recurrent cases in 2 years. Results: 352 EGC cases were registered and endoscopically removed. 182 cases could followed endoscopically for 2 years (51.7%). Sensitivity, specificity and AUC were as follows. Sox17 (70.0%, 48.1%, 0.646), MINT25 (70.0%, 35.2% 0.555), miR34 (65.0%, 49.4%, 0.578). Conclusion: We have developed a new methodology for gastric cancer detection by DNA methylation in gastric washes. It is easy and useful for early detection of recurrence after endoscopic treatment in GCa patients.
Citation Format: Yoshiyuki Watanabe, Ritsuko Oikawa, Hiroyuki Yamamoto, Fumio Itoh. Detection of early gastric cancer with Sox17 DNA methylation analysis of gastric washes. [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 2764.
Abstract 2770: DERL3 hypermethylation alters rhabdomyosarcoma cell metabolism
Authors: Llinas Pere , Lopez Paula , Esteller Manel 
Published: July 2016

Abstract
Physiological cellular activity requires a precise control of the proteome. However, cancer cells have a distorted protein profile. It is likely that intrinsic defects in protein homeostasis participate in human tumorigenesis, such as alterations in de novo protein synthesis, or defects in protein degradation. In this last case, the ubiquitin (Ub) proteasome system (UPS) targets a variety of proteins, including functional proteins that are no longer needed. Without appropriate protein homeostasis maintained by the UPS, healthy cells can undergo malignant transformation, and this observation has been therapeutically exploited by the development of proteasome inhibitors as anticancer agents. Many of the cytoplasm UPS-degraded proteins are retrotranslocated from the endoplasmic reticulum (ER). In this regard, ER possesses a quality control mechanism, termed the ER-associated degradation mechanism (ERAD), that is induced in response to ER stress by a transcriptional program, known as the unfolded protein response (UPR), which leads to the accelerated degradation of unfolded proteins. Within the ERAD pathway, the Derlin proteins play a critical role. It has been proposed that Derlins form an export channel in the membrane of the ER through which the ERAD substrates pass to reach the proteasome.
A previous study in our lab has shown that DERL3 silencing by DNA methylation deregulate SLC2A1 (GLUT1) degradation, promoting the Warburg effect. SLC2A1 is a key glucose receptor up-regulated in most solid tumors, allowing cancer cells to get the glucose they need for tumor progression.
In this study we use an embryonic rhabdomyosarcoma cell line as a model to find out the role of DERL3 in these tumors, as the percentage of promoter hypermethylation is these tumors is approximately 60%. Rhabdomyosarcoma (RMS) is one of the leading causes of cancer related deaths among children, being the histological variant of embryonic rhabdomyosarcoma the most common soft tissue sarcoma of childhood and adolescence, with 350 cases per year in USA.
The restoration of DERL3 expression in vitro promotes GLUT1 degradation and GLUT4 up-regulation, becoming cells sensitive to insulin. Moreover, it has been shown that DERL3 confers sensitivity against an ERAD related drug, Eeyarestatin I, opening a new approach for RMS treatment. All together, these initial findings highlight the role of DERL3 in RMS metabolism.
Citation Format: Pere Llinas, Paula Lopez, Manel Esteller. DERL3 hypermethylation alters rhabdomyosarcoma cell metabolism. [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 2770.
Abstract 2771: Epigenetic regulation of the cocaine and amphetamine regulated transcript (CART) in cancer
Authors: Connor Kate , Das Sudipto , Moran Bruce , Mulrane Laoighse , Gallagher William M. , O’Connor Darran 
Published: July 2016

Abstract
The cocaine and amphetamine regulated transcript (CART) is well documented in the central nervous system for its role in appetite regulation and reward pathways. In recent studies CART has been proven to be a poor prognostic marker in lymph node-negative, estrogen receptor (ER)-positive breast cancer patients, and has also been shown to be associated with worse survival in small bowel carcinoid tumours. The regulation of CART in these different cancer types however remains largely unknown. The presence of a large CpG island in the putative promoter region and the lack of expression of CART in many cancer cell lines suggest a possible epigenetic mechanism of suppression.
A panel of various cancer cell lines (breast, colorectal, endometrial and ovarian) were assessed for endogenous CART expression via western blot and RT-PCR. Cells were then treated with forskolin and leptin; agents which have been previously shown to induce CART expression in the CNS. Cells were also treated with CART conditioned media, which has been previously shown to be the only factor capable of inducing CART expression. Treatment was carried out with varying doses of two DNA methyltransferase inhibitors, 5-Aza-2′-deoxycytidine and RG108, followed by examination of CART expression post treatment. Bisulfite sequencing of the promoter region of CART was performed in all cell lines and a small selection of formalin fixed paraffin embedded (FFPE) tissue samples. Whole methylome sequencing was carried out to confirm this promoter region of CART and study the global effects of treatment with epigenetic modifiers and CART peptide.
Assessment of a collection of cell lines via western blot has shown CART expression to be negative across the panel. Moreover, treatment with forskolin and leptin did not result in re-expression of CART. Treatment with 5-Aza-2′-deoxycytidine has however resulted in increased CART mRNA levels, and subsequent bisulfite sequencing of the promoter region of CART has revealed dense methylation at the CpG island. Treatment with CART conditioned media may also induce promoter demethylation and subsequent re-expression of CART. Collectively, this data suggests the potential involvement of DNA methylation in the regulation of CART.
Citation Format: Kate Connor, Sudipto Das, Bruce Moran, Laoighse Mulrane, William M. Gallagher, Darran O’Connor. Epigenetic regulation of the cocaine and amphetamine regulated transcript (CART) in 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 2771.
Abstract 2784: Mitochondrial genomic backgrounds affect nuclear DNA methylation and gene expression
Authors: Vivian Carolyn J. , Brinker Amanda E. , Gooden Gerald C. , Legendre Christophe , Turpin Samuel , Koestler Devin C. , Salhia Bodour , Welch Danny R. 
Published: July 2016

Abstract
Increasing evidence shows mitochondrial DNA (mtDNA) genetics contribute to complex diseases like cancer, but the mechanisms responsible are mostly unknown. A new genetically engineered mouse model was created that contains nuclear DNA from one mouse strain and mtDNA from different mouse strain(s), designated Mitochondrial Nuclear Exchange (MNX). We have demonstrated that specific mtDNA-nDNA combinations could modify metastasis efficiency. The underlying mechanism is hypothesized to be due to mtDNA-driven changes in nDNA expression, but how mtDNA regulates the coordinated gene expression is not yet known. Since DNA methylation is an important epigenetic modification that occurs in all vertebrate genomes and retrograde and anterograde cross-talk exists between the nuclear and mitochondrial genomes, we hypothesized that the nuclear epigenome, specifically methylation, changes as a result of mitochondrial haplotype. To test this hypothesis, we performed Meth-Seq using the Agilent Mouse SureSelectXT, RNA-Seq using Illumina HiSeq 2500®, and Affymetrix GeneChip® Mouse Transcriptome Array. Initial studies were performed using four male mouse brains (8 wk) that were pooled from different litters and cages of wild-type and MNX mice. Significant and selective differential DNA methylation and gene expression patterns were observed and indicate that there are some global, some subtle, but mostly reproducible, differences between these strains. Pathway analysis show clustering of changes involving cell adhesion, ion channels, cell surface receptors, metabolism and molecules involved in transport. Together these observations provide insights into how mtDNA could be altering epigenetic regulation and thereby contribute to cancer pathogenesis. Support: Susan G. Komen for the Cure (SAC11037), Natl Fndn Cancer Res, Steiner Family Fund for Metastasis Research, Kansas Bioscience Authority CA134981, P30-CA168524
Citation Format: Carolyn J. Vivian, Amanda E. Brinker, Gerald C. Gooden, Christophe Legendre, Samuel Turpin, Devin C. Koestler, Bodour Salhia, Danny R. Welch. Mitochondrial genomic backgrounds affect nuclear DNA methylation and gene expression. [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 2784.
Abstract 2799: Exosomes from hypoxic pancreatic cancer cells confer resistance to subsequent hypoxia insult
Authors: Patton Mary C. , Zubair Haseeb , Patel Girijesh K. , Singh Seema , Singh Ajay P. 
Published: July 2016

Abstract
Pancreatic cancer (PC) remains one of the leading causes of cancer-associated deaths in the US due to its highly aggressive nature and lack of effective treatments. Pancreatic tumors are characterized by a dense, desmoplastic reaction and poor vasculature leading to reduced oxygen levels within the tumor microenvironment. It is believed that this encounter of pancreatic tumor cells to intratumoral hypoxic condition is a major cause of their persistent survival under a variety of stresses, increased aggressiveness and therapeutic-resistance. However, underlying molecular mechanisms responsible for PC's adaptation to hypoxia (along with enhanced growth and aggressiveness) remain elusive. In this study, we examined the role of exosomes, which are nanoscale biological vesicles with a lipid bilayer membrane, in imparting survival benefit to PC cells. For this, we first examined the effect of conditioned media (CM) from PC cells cultured under hypoxic (0.1% O2, H-CM) or normoxic (21% O2, N-CM) conditions on the growth of fresh PC cells cultured under hypoxic (0.1% O2) conditions. We found that H-CM conferred significant survival benefit to cancer cells under hypoxia as compared to N-CM. Thereafter, we isolated exosomes from N-CM (N-exo) or H-CM (H-exo) and used them to treat pancreatic cancer cells under hypoxic culture conditions. We observed that H-exo imparted a significantly higher survival advantage to cancer cells under hypoxia as compared to N-exo. Interestingly, protein quantification indicated the presence of higher levels of exosomes from cells cultured under hypoxic conditions as compared to those grown in normoxic environment. Western blot analysis with anti-CD9 and anti-CD63 antibodies confirmed enrichment of exosomes. Moreover, Dynamic Light Scattering-based size distribution suggested that H-exo were smaller in size compared to N-exo. Further studies are underway to identify the molecular determinants in exosomes responsible for inducing growth-promoting effects under hypoxia. Overall, our data provides the first evidence that hypoxic PC cells secrete exosomes that enhance survivability under hypoxic conditions.
Citation Format: Mary C. Patton, Haseeb Zubair, Girijesh K. Patel, Seema Singh, Ajay P. Singh. Exosomes from hypoxic pancreatic cancer cells confer resistance to subsequent hypoxia insult. [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 2799.
Abstract 2802: Exogenous pyruvate supports oxygen-independent tumor cell proliferation by serving as an oxygen surrogate to maintain homeostasis of NAD+/NADH
Authors: Yin Chengqian , He Dan , Chen Shuyang , Sang Nianli 
Published: July 2016

Abstract
Hypoxia is usually associated with solid tumors and ischemic lesions. Because oxygen is the common final electron acceptor in a variety of metabolic reactions, hypoxia leads to cell injury and death in normal tissues but promotes tumor progression and metastasis. Hypoxic cells use pyruvate from glycolysis as electron acceptor to support ATP production; anaerobic glycolysis, in which the glycolytic product pyruvate is converted to lactate, has been well established as an adaptive energy strategy for hypoxic cells. In tumor cells, pyruvate may also be used as a substrate to support an active biosynthesis, as well as to maintain the homeostasis of NAD+/NAD. However, whether exogenous pyruvate promotes hypoxic tumor cell proliferation by serving as an electron acceptor or a biosynthetic substrate remains unclear. By using both hypoxia and ρ0 cell with defective mitochondrial electron transfer chain, we show that exogenous pyruvate sustains the proliferation of tumor cells that cannot use oxygen as the electron acceptor. Pyruvate-derived metabolites including acetyl-coA, α-ketoglutarate, succinate, alanine, and aspartate were not able to substitute pyruvate to support oxygen-independent proliferation. Knockdown of metabolic enzymes including pyruvate carboxylase, pyruvate dehydrogenase and citrate synthetase showed no negative effects on pyruvate-facilitated hypoxic cell proliferation, whereas knockdown of lactate dehydrogenase significantly inhibited proliferation, indicating that the crucial role of exogenous pyruvate is to serve as the electron acceptor other than a substrate for proliferating biosynthesis. Consisting with this notion, we show that exogenous pyruvate significantly increased lactate generation and increased the ratio of NAD+/NADH. We also show that in our model cells, hypoxia did not cause accumulation of reactive oxygen species, suggesting that pyruvate-facilitated proliferation of hypoxic cells is not achieved by alleviating oxidative stress. Finally, we show that well-oxygenated cells release pyruvate, providing an in vivo source of pyruvate exogenous to the hypoxic cells. Taken together, our data support a novel model that well-oxygenated cells release pyruvate, which is utilized by hypoxic cells as an electron acceptor to facilitate proliferation. This model provides a potential new therapeutic target to suppress tumor progression.
Citation Format: Chengqian Yin, Dan He, Shuyang Chen, Nianli Sang. Exogenous pyruvate supports oxygen-independent tumor cell proliferation by serving as an oxygen surrogate to maintain homeostasis of NAD+/NADH. [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 2802.
Abstract 2809: ROS signaling by NADPH oxidase 5 (Nox5) modulates proliferation of human melanoma UACC-257 cells and prolongs growth in the absence of serum or growth factors
Authors: Antony Smitha , Wu Yongzhong , Haines Diana C. , Jiang Guojian , Meitzler Jennifer L. , Juhasz Agnes , Lu Jiamo , Roy Krishnendu K. , Doroshow James H. 
Published: July 2016

Abstract
Reactive oxygen species (ROS) produced by the NADPH oxidase (Noxs) homologs participate in signaling cascades regulating proliferation. Recently, we have reported substantial overexpression of Nox5 in several human cancers including those of prostate, ovary and melanomas. Despite being upregulated in many human cancers and implicated in promoting cell proliferation, the molecular mechanisms modulated by Nox5 are poorly understood. In this study, the functional significance of Nox5 was assessed in human UACC-257 and WM-852 melanoma cells by generating Nox5-overexpressing and knockdown cells. Stable overexpression of Nox5 in UACC-257 cells resulted in enhanced cell growth, increased BrdU positive cells and decreased protein tyrosine phosphatase activity. Additionally, these cells had increased normoxic Hif-1α expression and decreased p27Kip1 expression. Conversely, knockdown of endogenous Nox5 in UACC-257 cells resulted in decreased cell growth, decreased BrdU positive cells, decreased Hif-1α expression and increased p27Kip1 expression. Likewise, in human WM-852 melanoma cells, transient overexpression of Nox5 decreased p27Kip1 expression and knockdown of endogenous Nox5 resulted in increased p27Kip1 and gamma-H2AX expression with decreased cell growth. Cadherin switch, loss of E-cadherin expression and upregulation of N-cadherin was observed in UACC-257 Nox5 overexpressing cells indicative of an invasive phenotype; conversely, an upregulation of E-cadherin expression in Nox5 knockdown cells was noted. Cell invasion assay through matrigel-coated membranes also demonstrated enhanced invasion by Nox5 overexpressing cells. Additionally, 3D cultures of Nox5 overexpressing UACC-257 cells exhibit an amoeboid morphology compared to that of the Nox5 knockdown cells that were mesenchymal, suggestive of an amoeboid - mesenchymal (AMT) transition. Strikingly, UACC-257 cells that overexpress Nox5 were able to proliferate in serum-free conditions for over a month. In summary, our findings suggest that ROS signaling by Nox5 in human melanoma could modulate multiple signaling networks that regulate Hif-1α and p27Kip1 expression, contributing in part, to cell proliferation and the ability to grow in the absence of serum or growth factors.
Citation Format: Smitha Antony, Yongzhong Wu, Diana C. Haines, Guojian Jiang, Jennifer L. Meitzler, Agnes Juhasz, Jiamo Lu, Krishnendu K. Roy, James H. Doroshow. ROS signaling by NADPH oxidase 5 (Nox5) modulates proliferation of human melanoma UACC-257 cells and prolongs growth in the absence of serum or growth factors. [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 2809.
Abstract 2818: An unbiased tumor cell panel profiling method to identify drug-drug interactions reveals synergy between the CDK4 and CDK 6 inhibitor abemaciclib and the Raf dimer and pan-Raf inhibitor LY3009120 in Ras mutant cancers
Authors: Gong Xueqian , Wu Wenjuan , Chio Li-Chun , Pratt Susan , King Constance , Webster Yue , Lallena Maria Jose , Boehnke Karsten , Torres Raquel , Iversen Philip , Reinhard Christoph , Chen Shih-Hsun , Bechmann Richard , Peng Sheng-Bin , Buchanan Sean 
Published: July 2016

Abstract
Drug sensitivity profiling across genomically characterized panels of tumor cells can identify the molecular determinants of drug response. By testing compound combinations in an unbiased format, the same methodology can be used to identify the genomic context of drug-drug synergy. Based on this principle, we developed an unbiased combination screening protocol to identify synergistic interactions with LY3009120, a novel Raf dimer inhibitor that inhibits all three Raf isoforms (Peng et al. 2015, Cancer Cell 28:384-98). Inhibitors of the Ras-MAPK pathway have proven very effective in the treatment of BRAF-mutant melanoma but are, in general, only partially effective in the treatment of BRAF-mutant colorectal cancer and Ras mutant cancers. LY3009120 combined with various compounds was screened across panels of genomically characterized tumor cells. These screens identified a strong synergy with abemaciclib, an inhibitor of cyclin dependent kinases 4 and 6 (CDK4 and CDK6). Statistical analysis of effects in over 500 cancer cell lines showed that mutations in BRAF or Ras family genes were strongly associated with sensitivity to this combination. Strong synergy was observed in skin, colorectal, lung and pancreatic cancers with Ras/Raf mutations, but was also observed in various cancer cells wild type for Ras pathway genes. This included tumor types sensitive to single agent abemaciclib, such as mantle cell lymphoma, ER+ breast cancers, certain leukemias, squamous non-small cell lung cancer, and/or lung cancer with receptor tyrosine kinase activation. In vitro and in vivo analyses of the effects of the combination treatment on signaling pathways in KRAS mutant cancers led to potential mechanistic explanations for the differing efficacy of the combination, which manifests as regression of tumor xenografts in rodent models.
Citation Format: Xueqian Gong, Wenjuan Wu, Li-Chun Chio, Susan Pratt, Constance King, Yue Webster, Maria Jose Lallena, Karsten Boehnke, Raquel Torres, Philip Iversen, Christoph Reinhard, Shih-Hsun Chen, Richard Bechmann, Sheng-Bin Peng, Sean Buchanan. An unbiased tumor cell panel profiling method to identify drug-drug interactions reveals synergy between the CDK4 and CDK 6 inhibitor abemaciclib and the Raf dimer and pan-Raf inhibitor LY3009120 in Ras mutant 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 2818.