bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5053: Preliminary results from the Reproducibility Project: Cancer Biology - a replication of 50 high-impact cancer cell biology papers from 2010-2012
Authors: William Gunn , Iorns Elizabeth , Silva Elizabeth , Nosek Brian
Published: May 2015

The lack of reproducibility of preclinical research is a significant and growing problem which slows basic research and leads to fruitless clinical trials. An increasing number of reports have found discrepancies in published preclinical studies across scientific disciplines. For example:

* Amgen found that 47 of 53 “landmark” oncology publications could not be reproduced.

* Bayer found that their internal results contradicted academic publications in 43 of 67 oncology and cardiovascular projects.

* Dr. John Ioannidis and his colleagues found that of 432 publications purporting sex differences in hypertension, multiple sclerosis, or lung cancer, only one data set was reproducible.

These studies, and the many others that report similar results, highlight a significant problem in the development of new therapies to treat disease. With increasing reports of discrepancies in preclinical publications, pharmaceutical companies are being forced to re-evaluate their reliance on academic research. In fact, Bayer recently decided to halt nearly two-thirds of target-validation projects.In this session, we'll report the work we have done to address this issue. We will bring together researchers and representatives from funders and publishers to discuss the issue of reproducibility. We will share funder and publisher initiatives to promote reproducibility and also discuss research practices that lead to more reproducible research such as using proper statistical tests, reporting all experimental data, experimental blinding, and identification and validation of research reagents. We'll also share preliminary results from an Arnold Foundation-funded study to replicate the 50 most high impact cancer biology studies from 2010-1012.

Reproducibility ResearchAmgen47 of 53 “landmark” oncology publications could not be reproducedBayerinternal results contradicted academic publications in 43 of 67 oncology and cardiovascular projectsDr. John Ioannidis and colleagues432 publications purporting sex differences in hypertension, multiple sclerosis, or lung cancer, only one data set was reproducible

Note: This abstract was not presented at the meeting.

Citation Format: Gunn William, Elizabeth Iorns, Elizabeth Silva, Brian Nosek. Preliminary results from the Reproducibility Project: Cancer Biology - a replication of 50 high-impact cancer cell biology papers from 2010-2012. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5053. doi:10.1158/1538-7445.AM2014-5053
Abstract 5054: Smoking prevention intervention with school classes at a university hospital by thoracic surgeon und pulmonologist
Authors: Tomaszek Sandra , Schuurmans Macé M. , Schneiter Didier , Weder Walter , Hillinger Sven
Published: May 2015

Purpose: Smoking prevention in schoolchildren with the aim to inform and prevent smoking initiation has been widely studied and has shown variable results. Interventions provided by physicians in a hospital setting have been rarely reported. Here we show the feasibility and gain of knowledge of our smoking prevention project in a hospital setting.

Methods: Interventions performed from November 2009 - February 2013 were evaluated. Overall 470 children participated in our preventive intervention. A 7-item questionnaire was provided to the school classes (Grades 6 to 10) before and after a two-hour smoking prevention intervention consisting of anatomical models, oral presentations, videos, patient interviews and hands-on lung function tests. The goal was to show the anatomical and physiological basics as well as age-based information about the harms of smoking. During the intervention the children have been motivated to be actively involved. Class selection has been performed for groups of children in a highly vulnerable phase of age before smoking initiation.

Results: The baseline questionnaire was completed by 457 children, the one after intervention by 426. The knowledge about which organs are affected by smoking increased from 6.8-99.6% to 65.5-99.5% (p<0.01). While only 59.5% knew that only a minority of people is able to quit smoking successfully, 94.1% answered the question correctly after intervention (p<0.001). Prior to the intervention only 78.1% believed that minor tobacco consumption is not damaging which increased to 89.4% after the teaching session (p<0.05). Smoking hookah was believed to be less harmful than cigarettes by 30.2% of children decreasing to 7.7% after the intervention (p<0.001).

Conclusion: Information on health effects provided by lung specialists in the hospital leads to a statistically significant increase in knowledge as assessed by a short questionnaire. The intervention is feasible and well received. This kind of interventions might help to prevent schoolchildren from smoking in a highly vulnerable phase of age.

Citation Format: Sandra Tomaszek, Macé M. Schuurmans, Didier Schneiter, Walter Weder, Sven Hillinger. Smoking prevention intervention with school classes at a university hospital by thoracic surgeon und pulmonologist. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5054. doi:10.1158/1538-7445.AM2014-5054
Abstract 5057: The role of health locus of control and self-efficacy beliefs in sunscreen use: Evidence from 2012 Health Information National Trends Survey
Authors: He Xiaofei
Published: May 2015

Sun exposure has been identified as a cause of skin cancer (Armstrong & Kricker, 2001). Extensive empirical evidence shows that sunscreen use was associated with lower skin cancer incidence (Green et al., 1999, 2011). However, only 32% U.S. adults used sunscreen regularly (NCI, 2010); the percentage was even lower (21.7%) among adolescents (Eaton et al., 2012). To reduce skin cancer burden, sunscreen use promotion should be enhanced.

Locus of control, the extent to which people believe they have control over their lives with external and internal dimensions (Rotter, 1954), is a significant predictor of behavioral changes (Bundek et al., 1993). Internal locus of control, i.e., beliefs that reinforcements are dependent upon one's own behavior, is positively related with behavioral changes, e.g., weight loss (O'Bryan, 1972) and health information seeking (Wallston & Maides, 1976), etc. Self-efficacy, the extent to which one believes to be capable of performing the recommended behavior (Bandura, 1977), is a robust predictor for behavioral changes including smoking cessation (Norman, 1994) and AIDS prevention (Yzer, 1998), etc.

However, little research focuses on the role of locus of control and self-efficacy in promoting sunscreen use by using nationally representative cross-sectional data. The present study bridges this gap by examining the following two hypotheses:

H1: People with high external locus of control were less likely to use sunscreen.

H2: People with high health self-efficacy were more likely to use sunscreen.

Results show that people high in external locus of control were less likely to use sunscreen (B=-0.160, p=0.001) and people high in health self-efficacy were more likely to use sunscreen (B= 0.331, p=0.046), both of which were statistically significant. Males, Pacific Islanders, homemakers, and non-immigrants were more likely to use sunscreen. The findings attest to the necessity of enhancing people's internal locus of control and self-efficacy with the purpose of promoting sunscreen use. Public health efforts could be allocated based on the socio-demographic findings.

Citation Format: Xiaofei He. The role of health locus of control and self-efficacy beliefs in sunscreen use: Evidence from 2012 Health Information National Trends Survey. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5057. doi:10.1158/1538-7445.AM2014-5057
Abstract 5076: Rapid assessment of AML genes for mutation detection and copy number variation
Authors: Andersen Mark , Rhodes Kate , Roman Steve , VanLoy Cristina , Broomer Adam , Allen Michael , Topacio Denise , Liu Guoying , Hyland Fiona
Published: May 2015

Targeted sequencing using the Ion AmpliSeq™ Library kit combined with the Ion PGM™ sequencing instrument is a fast and effective method to identify genetic variants in cancer samples. A new targeted primer panel for amplification of genes involved in Acute Myeloid Leukemia (AML) has been developed by Life Technologies. The panel covers 19 genes characterized using over 300 specific primer pairs in two highly multiplexed PCRs. To demonstrate the coverage efficiency, we evaluated libraries prepared from whole blood and isolated genomic DNA. When libraries from 4 individuals were run on a single Ion 318™ Chip, the average coverage depth was >3000x, with >97% of the target bases covered >500X. Additionally, >90% of reads were on-target. The panel was tested on control samples and analyzed using the Ion Reporter™ Software, and expected variants, including copy number variations (CNVs) were detected with high sensitivity and specificity.

Citation Format: Mark Andersen, Kate Rhodes, Steve Roman, Cristina VanLoy, Adam Broomer, Michael Allen, Denise Topacio, Guoying Liu, Fiona Hyland. Rapid assessment of AML genes for mutation detection and copy number variation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5076. doi:10.1158/1538-7445.AM2014-5076
Abstract 5087: Peli 1 targets BubR1 for ubiquitinational degradation and induces aneuploidy transformation
Authors: Kim Suhyeon , Park Hye-Young , Song Ha Rim , Go Heounjeong , Cho Hyeseong , Chung Doo Hyun , Lee Chang-Woo
Published: May 2015

Disruption of the BubR1-mediated function negatively impacts mitotic checkpoint control, increases chromosomal instability, and is associated with infertility, life span and cancer. Therefore, BubR1 plays an important role in preventing abnormal mitotic cells with chromosomal aneuploidy from achieving neoplastic aneuploidy, however it remains unclear how the signalling activates or inactivates the BubR1 function during the neoplastic transformation. Here, we report that Peli 1, a new family of signal-responsive E3 ubiquitin ligase, directly interacts with BubR1. This interaction promotes the ubiquitin-mediated degradation, and prevents the kinetochore association of BubR1. In addition, cells expressing Peli 1 led to severe mitotic defects and displayed severe chromosomal aneuploidy in vitro and in vivo. Together, these results suggest that Peli 1 expression acts as a potent causative signalling against chromosome integrity by mediating the ubiquitinational degradation of BubR1, and thus contributes to the development of neoplastic chromosome aneuploidy.

Citation Format: Suhyeon Kim, Hye-Young Park, Ha Rim Song, Heounjeong Go, Hyeseong Cho, Doo Hyun Chung, Chang-Woo Lee. Peli 1 targets BubR1 for ubiquitinational degradation and induces aneuploidy transformation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5087. doi:10.1158/1538-7445.AM2014-5087
Abstract 5090: Estrogen receptor-alpha promotes multi-centrosomes by inhibition of BRCA1-Rad51 binding
Authors: Jung Youn-Sang , Chun Ho-Young , Woo Tae-Gyun , Park Bum-Joon
Published: May 2015

Von Hippel Lindau gene (VHL) is frequently deleted in renal cell carcinoma and known to be negative regulator of Hypoxia inducible factor-1 (HIF-1). However, its tumor suppressive functions has not been fully explained by HIF-1 suppression, because loss of pVHL is appeared in initial stage of RCC tumorigenesis and induces the defect in cytokinesis or centrosome instability. Concerning this, we previously revealed that estrogen receptor-alpha is primary target of E3 ligase function of pVHL. In this study, we demonstrate that ER-α is responsible for mitotic defect and drug resistance in VHL-deficient cells. Activated ER-α can interact with BRCA1 or Rad51 and disrupt the BRCA1-Rad51-mediated centrosome maintaining. Thus, VHL-deficient cells, like as BRCA1-deficient cells, possessed multi-centrosome and multi-nucleated phenotypes. In addition, multi-centrosomal features rendered the resistance to Taxol. Since it is mediated by ER-α in nucleus, blocking of ER-α translocation by specific inhibitor, Faslodex, can ameliorate multi-centrosomes and induce Taxol sensitivity in RCC and BRCA1-deficient breast cancers. These results suggest that pVHL is critical for proper cell division and suppression of ER-α in RCC and BRCA1-deficient breast cancer is one of useful strategy for improving the drug sensitivity.

Citation Format: Youn-Sang Jung, Ho-Young Chun, Tae-Gyun Woo, Bum-Joon Park. Estrogen receptor-alpha promotes multi-centrosomes by inhibition of BRCA1-Rad51 binding. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5090. doi:10.1158/1538-7445.AM2014-5090
Abstract 5104: Identification of cancer-specific COPI inhibitors and their associated apoptotic cell death pathways
Authors: Oliver David J. , Chaparala Anusha , Okafor Emeka , Camacho Carlos J. , Doemling Alexander S. , Wieland Felix T. , Roninson Igor B. , Shtutman Michael
Published: May 2015

The COPI complex is a heptameric complex involved in multiple aspects of intravesicular transport. Recently, Shtutman et al. identified the ζ1 subunit of the COPI complex as a potential target for cancer therapy1. The ζ subunit of COPI is coded for by two genes, COPZ1 and COPZ2. In normal tissue, both COPZ genes are expressed and the protein products are functional isoforms within the COPI complex. In numerous different cancers, COPZ2 is silenced and the cells must depend on the ζ1 isoform for COPI function. When ζ1 is inhibited by siRNAs, cancer cells undergo apoptotic cell death while normal cells, which retain the ζ2 isoform, survive. Thus, the ζ1 subunit of the COPI complex is a potential target for cancer therapy. Here we present our work to develop and test small molecule inhibitors of ζ1-containing COPI complexes, as well as the identification of the apoptotic cell death pathways activated upon ζ1 depletion with siRNAs. We have identified a small molecule which disrupts the formation of COPI vesicles in biochemical and immunofluorescence studies, and also kills cells in a ζ1-specific manner. In addition we have shown that the unfolded protein response (UPR) is activated by ζ1 depletion but that it is not the primary activator of apoptotic cell death in cancer cells. The identification of small molecules which can inhibit ζ1-containing COPI will be a valuable tool for cancer therapy as well as the study of COPI biology. In addition, elucidation of the mechanism of activation and execution of apoptosis upon COPI depletion will provide insight into methods for combination therapy.

Citation Format: David J. Oliver, Anusha Chaparala, Emeka Okafor, Carlos J. Camacho, Alexander S. Doemling, Felix T. Wieland, Igor B. Roninson, Michael Shtutman. Identification of cancer-specific COPI inhibitors and their associated apoptotic cell death pathways. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5104. doi:10.1158/1538-7445.AM2014-5104
Abstract 5107: The role of PERIOD2 for radioprotection against ionizing radiation in mice bone marrow
Authors: Alexandrou Aris , Menaa Cheikh , Liu Chris , Pai Steven , Xiao Kai , Fan Ming , Juma Shuaib , Fu Loning , Murphy William J. , Li Jian Jian
Published: May 2015

Background: Circadian deregulation is associated with the development of numerous diseases, including cancer, diabetes, and several neurological disorders. A recent study shows that the onset of carcinogenicity is specifically linked to the time of exposures (day or night) to ultraviolet light and ionizing radiation in mice. This effect is believed to be mediated through radiation-induced DNA damage and deregulation of the WNT/β-Catenin signaling pathway, hence affecting cell survival and proliferation.

Results: The current studies aim to elucidate the contribution of a core circadian gene, Period2 (Per2), in the radioprotective response of mice bone marrow. Epithelial cells and mice bone marrow express PER2 in contrast to cancer cells. Bone marrow isolated from Per2 knockout mice exhibit radioresistence to high doses of ionizing radiation (HDIR; 2Gy) compared to wild-type mice controls suggesting that Per2 expression was responsible for radiation-induced cell death. This finding was translated by increased granulocyte-macrophage (GM-CFUs) clonogenic survival and reduced cell apoptosis. Mechanistic analysis showed that Per2 knockout mice bone marrow expressed higher levels of procaspase3 and less expression of active caspase3 after HDIR treatment compared to wild-type mice bone marrow and sham irradiated controls. Furthermore, Per2 knockout mice treated with 12Gy total body radiation (TBIR) showed an 80% survival verses 30% survival in wild-type mice. Whole genome-wide sequencing detected signal transduction pathways reflecting enhanced DNA damage repair capacity in knockout mice bone marrow, hence defining the role of PER2 in the radioresistant phenotype in vivo.

Conclusions: This data is the first evidence supporting the critical role of PER2 in protecting mice hematopoietic stem cells (HSCs) from HDIR via enhancement of genes involved in DNA repair signaling networks.

Citation Format: Aris Alexandrou, Cheikh Menaa, Chris Liu, Steven Pai, Kai Xiao, Ming Fan, Shuaib Juma, Loning Fu, William J. Murphy, Jian Jian Li. The role of PERIOD2 for radioprotection against ionizing radiation in mice bone marrow. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5107. doi:10.1158/1538-7445.AM2014-5107
Abstract 5114: The chemomodulatory effects of resveratrol and didox on herceptin cytotoxicity in breast cancer cell lines
Authors: Ellatif Ghada A. Abd , Tadros Marianne Georges , Naim Ashraf Bahey Eldin Abd El , Khalifa Amany Ibrahim , Alabd Ahmad M.
Published: May 2015

Herceptin is considered an essential treatment option for double negative breast cancer. Resveratrol and didox are known chemopreventive agents with potential anticancer properties. The aim of the current study is to investigate the influence of resveratrol and didox on the cytotoxicity profile of herceptin in her-2 receptor positive and her-2 receptor negative breast cancer cell lines. The IC50's of herceptin in T47D and MCF-7 were 0.126±0.01 ng/ml and 26.71±1.85 ng/ml respectively. Equitoxic combination of herceptin with resveratrol or didox in T47D significantly reduced the IC50 to 0.05±0.01 and 0.029±0.006 ng/ml, respectively and similar results were obtained in MCF-7. The gene expression of Bcl-xl was markedly decreased in T47D cells following treatment with herceptin/resveratrol compared to herceptin alone. Immunocytochemical staining of her-2 receptor in T47D cells showed a significant reduction after treatment with herceptin/resveratrol combination compared to herceptin alone. On the other hand, combination of herceptin and didox had no significant effect on her-2 receptor expression. Cell cycle analysis showed an arrest at G2/M phase for both cell lines following treatment with herceptin alone and in either combinations regimen. In conclusion the combination of herceptin with resveratrol and didox improved the cytotoxic profile of herceptin in T47D (her-2 receptor positive cell line) and to a lesser extent in MCF-7 (her-2 receptor negative cell line).

Citation Format: Ghada A. Abd Ellatif, Marianne Georges Tadros, Ashraf Bahey Eldin Abd El Naim, Amany Ibrahim Khalifa, Ahmad M. Alabd. The chemomodulatory effects of resveratrol and didox on herceptin cytotoxicity in breast cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5114. doi:10.1158/1538-7445.AM2014-5114
Abstract 5115: The Bcl-2 Homology Domain-3 Mimetic, Obatoclax, chemosensitizes muscle invasive bladder cancer cell lines to cisplatin treatment
Authors: Steele Thomas , Pham Dan , Phan Kathy , Ghosh Paramita , White Ralph deVere , Vinall Ruth L.
Published: May 2015

The primary objective of this study was to determine whether inhibition of anti-apoptotic Bcl-2 family protein activity causes decreased clonogenicity and/or increased apoptosis of muscle invasive bladder cancer (MI-BC) cell lines when used in combination with cisplatin, the standard of care treatment for MI-BC, and/or pre-miR-34a, a miRNA that targets several components of pathways know to promote tumor progression and chemoresistance. Obatoclax, a Bcl-2 homology domain-3 mimetic that has been demonstrated to prevent Bcl-2, Bcl-xL, and Mcl-1 from interacting with and inhibiting Bax and Bak, was used for this study. The impact of Obatoclax, in combination with cisplatin and/or pre-miR-34a, on clonogenic potential and apoptosis was assessed in three MI-BC cell lines (T24, TCCSuP, 5637) using clonogenic assay and annexin V flow cytometry respectively. The ability of Obatoclax to prevent interaction between Bcl-2, Bcl-xL, and Mcl-1, with Bax and Bak was assessed via immunoprecipitation and western blot. The impact of treatments on apoptosis/survival pathways was assessed via western blot. Obatoclax mediated a dose-dependent increase in apoptosis in all three MI-BC cell lines. Obatoclax was able to decrease binding of Bak to Bcl-xL, and cause decreased expression of Bcl-2 and Bcl-xL. Unexpectedly, Obatoclax also mediated a dose-dependent decrease in cdk4, cdk6, and cyclin D1. Combining Obatoclax with cisplatin or/and pre-miR-34a caused further inhibition of clonogenic potential of all three cell lines compared to treatment with single agents. Of the combination treatments, Obatoclax combined with cisplatin mediated the biggest increase in apoptosis levels. Treatment with Obatoclax, cisplatin, and pre-miR-34a actually decreased apoptosis levels. Western blot revealed that treatment with Obatoclax was unable to overcome the effect of pre-miR-34a on causing increased Bcl-2 and Bcl-xL expression. Our data demonstrate that Obatoclax chemosensitizes MI-BC cell lines to cisplatin and indicate that in vivo testing of this combination treatment is warranted. The data do not support further testing of pre-miR-34a in this setting. With the current 5-year survival rate for MI-BC being only 35%, clearly new and improved treatment options are needed for this disease.

Citation Format: Thomas Steele, Dan Pham, Kathy Phan, Paramita Ghosh, Ralph deVere White, Ruth L. Vinall. The Bcl-2 Homology Domain-3 Mimetic, Obatoclax, chemosensitizes muscle invasive bladder cancer cell lines to cisplatin treatment. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5115. doi:10.1158/1538-7445.AM2014-5115
Abstract 5130: Functional role of the crosstalk between arginine methylation and phosphorylation of hnRNPK during DNA damage
Authors: Yang Jen-Hao
Published: May 2015

HnRNPK is an RNA/DNA binding protein participating in chromatin remolding, RNA processing and DNA damage response. In addition, elevation of hnRNPK has been associated with tumor development and progression. Moreover, hnRNPK is known as co-transcription factor of p53 to regulate cell cycle arrest and apoptosis upon DNA damage. A variety of post-translational modifications of hnRNPK has been identified and shown to regulate its function, including phosphorylation, ubiquitination, sumoylation and methylation. However, the functional significance of hnRNPK arginine methylation remains unclear. In this study, we characterize the function of two essential arginine methylation of hnRNPK and demonstrate that they are able to inhibit the nearby PKCδ phosphorylation. Since PKCδ is a pro-apoptotic kinase upon DNA damage, we further investigate whether these essential arginine methylations of hnRNPK participate in the regulation of cell apoptosis.

Citation Format: Jen-Hao Yang. Functional role of the crosstalk between arginine methylation and phosphorylation of hnRNPK during DNA damage. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5130. doi:10.1158/1538-7445.AM2014-5130
Abstract 5141: Inhibition of HDAC1 leads to subtype switching in the SW13 adrenal adenocarcinoma line
Authors: Lam Jessica M. , Daggett Juliane , Leyva Kathryn J. , Hull Elizabeth E.
Published: May 2015

HDAC (histone deacetylase) enzymes are highly expressed in cancer cells and HDAC inhibitors have shown promise in clinical trials for certain patients in multiple types of cancer. In this context, the ability of the SW13 human adrenal adenocarcinoma line to switch between two well-established subtypes in response to HDAC inhibition is of interest. However, the previously characterized inhibitor (trichostatin A) is a toxic, broad-spectrum inhibitor affecting both class I and class II HDACs which have roles in the nucleus and cytosol, respectively. Multiple HDAC inhibitors are screened for their ability induce switching between the SW13 subtypes to identify which HDAC enzyme is necessary for controlling SW13 subtype switching. Results suggest that inhibition of class I HDAC1 increases the rate of conversion between SW13- to SW13+ cells. The effect of HDAC1 inhibition may be enhanced by HDAC3 inhibition as MS-275, an inhibitor specific for HDAC1 but with some specificity for HDAC3, is most effective, particularly when compared to the efficacy of depudecin, an HDAC1 specific inhibitor. After conversion to SW13+, there is increased acetylation on histone H3 but not histone H4 when compared to levels in SW13-, suggesting that the role of acetylation on the H3 tail plays an essential role in the subtype switch. SW13- cells do not express Brg1 or Brm, the mutually exclusive ATPase subunits of the SWI/SNF complex, and do not have an active SWI/SNF complex. Altered activity of this complex and the increased histone H3 acetylation may contribute to the extensive changes in gene expression observed between SW13+ and SW13- subtypes.

Citation Format: Jessica M. Lam, Juliane Daggett, Kathryn J. Leyva, Elizabeth E. Hull. Inhibition of HDAC1 leads to subtype switching in the SW13 adrenal adenocarcinoma line. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5141. doi:10.1158/1538-7445.AM2014-5141
Abstract 5145: KDM2B expression regulates ribosome biogenesis and cancer cell growth in a p53-dependent manner
Authors: Penzo Marianna , Casoli Lucia , Sicuro Laura , Galibiati Alice , Pollutri Daniela , Govoni Marzia , Ceccarelli Claudio , Santini Donatella , Taffurelli Mario , Treré Davide , Montanaro Lorenzo
Published: May 2015

Ribosome biogenesis is a cellular process requiring a fine-tuned control, as its deregulation is linked to cancer progression. Indeed, tumors characterized by an intense ribosome biogenesis often display a more aggressive biological and clinical behavior. Ribosomal RNA (rRNA) synthesis is controlled at several levels, the higher one being the epigenetic regulation of chromatin portions containing rRNA genes. KDM2B (Lysine (K)-specific demethylase 2B) is a histone demethylase modulating the accessibility of rRNA genes, thereby repressing their transcription. In particular KDM2B is known to specifically act on trimethylated K4 of histone H3 (H3K4) associated with rDNA. In this study we aimed to define the contribution of KDM2B expression to the biological features of breast cancer, a tumor type whose clinical behavior is known to be related to the rate of ribosome biogenesis. We show that, in primary breast carcinomas, lower KDM2B expression correlates with an increased size of specifically stained nucleolar organizer regions, a morphological parameter directly related to the rate of ribosome biogenesis, and with a poorer prognosis. In vitro, in breast cancer-derived cell lines, siRNA mediated KDM2B knock-down (KD) induced an increase in H3K4 trimethylation and in rRNA transcription. However, this is mirrored by an augmented cell proliferation only in p53 compromised cells, while p53 competent cells undergo cellular senescence and death. The latter effect, appears to be mediated by a p38-mediated phosphorylation of p53, inducing the expression of p15Ink4b and p21Waf1, and is not linked to p53 stabilization. In the long term, stable KDM2B KD in p53-compromised breast cancer cells induced a series of additional biological features including their augmented cellular clonogenic potential and invasive capability. Altogether our data indicate that abnormal JHDM1B-mediated epigenetic activation of rDNA genes elicits a p53-mediated growth arrest, but may promote cancer cell growth when p53 is lacking.

Citation Format: Marianna Penzo, Lucia Casoli, Laura Sicuro, Alice Galibiati, Daniela Pollutri, Marzia Govoni, Claudio Ceccarelli, Donatella Santini, Mario Taffurelli, Davide Treré, Lorenzo Montanaro. KDM2B expression regulates ribosome biogenesis and cancer cell growth in a p53-dependent manner. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5145. doi:10.1158/1538-7445.AM2014-5145
Abstract 5147: Deregulation of G9a and its functional roles in liver cancer
Authors: Wei Lai , Tsang Felice Ho-Ching , Lau Jeffery Chi-Fai , Au Sandy Leung-Kuen , Lee Joyce Man-Fong , Wong Carmen Chak-Lui , Ng Irene Oi-Lin , Wong Chun-Ming
Published: May 2015

Liver cancer, primarily hepatocellular carcinoma (HCC), is the fifth most common cancers and the third leading cause of cancer mortality worldwide. In recent years, deregulated epigenetic mechanisms were found important in a wide range of human carcinogenesis, including HCC. The epigenetic changes elicited by various environmental factors and lifestyles (such as hepatitis B virus (HBV) and hepatitis C virus (HCV) infection, chronic alcohol intake) were suspected to promote HCC development. However, the underlying mechanism and specific gene targets are still unknown. Our previous study suggested that deregulation of epigenetic regulators was a common event in human HCC. Among 90 epigenetic modifiers analyzed in human HCC, Euchromatic histone-lysine N-methyltransferase 2 (EHMT2/G9a) was the most significantly up-regulated one. G9a is a SET domain containing histone methyltransferase specific for histone H3 lysine 9 mono- and di-methylation. Previous work had revealed that G9a is important for embryonic development and adipocyte differentiation. However, little is known about functional roles of G9a in human HCC. We hypothesize that deregulation of G9a may led to aberrant epigenetic changes and contribute to initiation and progression of human HCC. In this study, we found that G9a mRNA expression in primary HCC was 3 folder than their corresponding non-tumor liver and 10 fold higher than normal livers. The up-regulation of G9a was detected in 63% (24/38) of primary HCC samples and was significantly associated with the presence of venous invasion (P = 0.006). The frequent G9a up-regulation in HCC was attributed to gene amplification and microRNA deregulation. We showed that 53% of HCC samples have 3 or more copies of G9a gene. In addition, we identified that miR-1 was a post-transcriptional regulator of G9a. Ectopic expression of miR-1 suppressed G9a 3′-untranslated-region-coupled luciferase activity. Functionally, knockdown of G9a significantly suppressed HCC cell proliferation, migration and colony formation. We also demonstrated that UNC0638, as a G9a specific inhibitor, could effectively inhibit HCC cell growth and cause cell morphology changes. By utilizing nude mice model, we showed that depletion of G9a drastically inhibited in vivo tumorigenicity. Our finding suggested that up-regulation of G9a contributed to the liver carcinogenesis and could be a therapeutic target for HCC treatment.

Citation Format: Lai Wei, Felice Ho-Ching Tsang, Jeffery Chi-Fai Lau, Sandy Leung-Kuen Au, Joyce Man-Fong Lee, Carmen Chak-Lui Wong, Irene Oi-Lin Ng, Chun-Ming Wong. Deregulation of G9a and its functional roles in liver cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5147. doi:10.1158/1538-7445.AM2014-5147
Abstract 5150: JMJD3 suppresses progression of colorectal carcinoma by regulating cell cycle and anti-apoptosis
Authors: Tokunaga Ryuma , Nakagawa Shigeki , Sakamoto Yasuo , Karashima Ryuichi , Ida Satoshi , Imamura Yu , Ishimoto Takatsugu , Iwagami Shiro , Baba Yoshifumi , Miyamoto Yuji , Yoshida Naoya , Baba Hideo
Published: May 2015

Background: JMJD3 has been known to play important roles in transcriptional regulation and cell proliferation by demethylating histone3 lysin27 (H3K27) in various cancers. However, the mechanism underlying JMJD3-mediated transcriptional regulation and the relation with prognosis were unclear in colorectal carcinoma. Here, we investigated the expression of JMJD3 in clinical samples and the function of JMJD3 in colorectal carcinoma cell lines.

Methods: Quantitative real-time PCR were performed to check the expression of JMJD3 mRNA using the carcinoma and normal colorectal tissue in 14 colorectal carcinoma patients who underwent a curative surgery.

In vitro analysis, we investigated the expression of JMJD3 in colorectal carcinoma cell lines by Western Blotting, and we knocked down the expression of JMJD3 by using small interfering RNA in Colo201 and colo320. The influence of JMJD3 on cell growth, apoptosis and cell cycle was assessed by growth assay, apoptosis assay and cell cycle analysis. We checked the changes of cell cycle related genes by real time PCR.

Results: In clinical samples, the expression of JMJD3 in carcinoma tissue was significantly lower than in normal tissue (P=0.02). In vitro analysis, we knocked down the expression of JMJD3 by siRNA targeting JMJD3 and checked the reduction on mRNA and protein level. Knockdown of JMJD3 significantly increased cell proliferation (P=0.02), reduced apoptosis (P=0.003), and increased G2/M phase population (P = 0.05) which was detected by FACS. Moreover, knockdown of JMJD3 significantly decreased the expression of p15INK4B in quantitative real-time PCR.

Conclusion: We showed the expression of JMJD3 in carcinoma tissue was lower than in normal tissue and demonstrated that the down-regulation of JMJD3 resulted in the increase of cell proliferation, acceleration of cell cycle and anti-apoptosis in vitro analysis. These results suggest that JMJD3 is a tumor suppressor gene for colorectal carcinoma.

Note: This abstract was not presented at the meeting.

Citation Format: Ryuma Tokunaga, Shigeki Nakagawa, Yasuo Sakamoto, Ryuichi Karashima, Satoshi Ida, Yu Imamura, Takatsugu Ishimoto, Shiro Iwagami, Yoshifumi Baba, Yuji Miyamoto, Naoya Yoshida, Hideo Baba. JMJD3 suppresses progression of colorectal carcinoma by regulating cell cycle and anti-apoptosis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5150. doi:10.1158/1538-7445.AM2014-5150
Abstract 5154: Targeting the histone demethylase KDM4 subfamily as a potential therapeutic strategy in breast cancer
Authors: Holowatyj Andreana , Ye Qin , Zhang Lihong , Wu Jack , Yang Zeng-Quan
Published: May 2015

Epigenetic alterations, including histone modifications, play fundamental roles in breast cancer initiation and progression. We originally identified and cloned the GASC1 (gene amplified in squamous cell carcinoma 1, also known as KDM4C) gene from an amplified region at 9p24 in esophageal cancer cells; and recently demonstrated that KDM4C/GASC1 is amplified and over-expressed in breast cancer, particularly in the aggressive basal subtype. The KDM4C/GASC1 protein belongs to the KDM4 family of histone demethylases, and although KDM4 family members have a high degree of homology, they may play different roles in various types of breast cancer. The goal of this study is to analyze genomic anomalies and expression levels of KDM4 demethylases in breast cancer, and elucidate the fundamental role and mechanism of their dysregulation in promoting breast tumorigenesis.

We conducted a large-scale meta-analysis of KDM4 demethylase expression across multiple available gene expression studies in breast cancer. Next, we examined KDM4 expression in a panel of non-tumorigenic and cancerous breast epithelial cell lines using quantitative RT-PCR and Western blot assays. We also assessed global methylation (H3K4, H3K9, H3K27 and H3K36) levels by Western blot in a panel of breast cancer cell lines. Finally, we tested a novel KDM4 inhibitor in breast cancer.

We found that the KDM4 members show different expression patterns in subtypes of breast cancer. GASC1/KDM4C expression is high in estrogen receptor (ER)-negative, basal type breast cancers. In contrast, KDM4B expression is significantly higher in ER-positive luminal-type breast cancers. Expression levels of homologs KDM4A and D are not significantly different between ER-+/- breast cancers. Our findings suggest that H3 global methylation levels vary among different breast cancer cell lines. Furthermore, we demonstrated that inhibition of KDM4 with a novel small molecule inhibitor increased H3K9 methylation levels and slowed KDM4-overexpressed breast cancer cell growth in vitro.

In summary, our data indicate that the KDM4 histone demethylase family may contribute to the dysregulation of histone methylation status differently in breast cancer subtypes. Moreover, breast cancer cell lines with defined histone methylation levels will provide a useful model for investigating biological and functional roles of KDM4 histone demethylases, and for developing novel anticancer epi-drugs in breast cancer.

Citation Format: Andreana Holowatyj, Qin Ye, Lihong Zhang, Jack Wu, Zeng-Quan Yang. Targeting the histone demethylase KDM4 subfamily as a potential therapeutic strategy in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5154. doi:10.1158/1538-7445.AM2014-5154
Abstract 5172: Whole exome mutation landscape of 70 commonly used colorectal cancer cell lines
Authors: Sieber Oliver , Mouradov Dmitri , Sloggett Clare , Jorissen Robert , Love Chris , Li Shan , Burgess Antony , Arango Diego , Strausberg Robert , Buchanan Daniel , Wormald Samuel , O'Connor Liam , Wilding Jenny , Bicknell David , Tomlinson Ian , Bodmer Walter , Mariadason John
Published: May 2015

Human colorectal cancer (CRC) cell lines are an important model system for studying the biology of this malignancy as well as therapeutic and biomarker discovery. However, data on the degree to which established CRC cell lines reflect the somatic diversity of primary cancers at the genome level are limited. Using whole exome sequencing and SNP microarray analysis we show that the profile of mutations and DNA copy-number alterations in 70 widely-used CRC cell lines closely resembles that of primary colorectal tumors published by The Cancer Genome Atlas Network. We demonstrate presence of at least two hypermutation phenotypes, consistent with defective DNA mismatch repair and DNA polymerase epsilon (POLE) proof-reading deficiency, and similar mutation signatures in the WNT, MAPK, PI3K, TGF-beta and TP53 pathways. Paired cell lines derived from the same tumor are found to exhibit substantial mutation and DNA copy-number differences, with in silico simulations suggesting that these largely reflect pre-existing tumor cell heterogeneity, but these differences do not obscure known driver genes. Our genome level data indicate that CRC cell lines are representative models of the main molecular subtypes of primary tumors, and will facilitate informed selection of molecularly-defined models for preclinical investigations.

Citation Format: Oliver Sieber, Dmitri Mouradov, Clare Sloggett, Robert Jorissen, Chris Love, Shan Li, Antony Burgess, Diego Arango, Robert Strausberg, Daniel Buchanan, Samuel Wormald, Liam O'Connor, Jenny Wilding, David Bicknell, Ian Tomlinson, Walter Bodmer, John Mariadason. Whole exome mutation landscape of 70 commonly used colorectal cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5172. doi:10.1158/1538-7445.AM2014-5172
Abstract 518: Identification of differentially expressed microRNAs in human hepatocellular adenoma associated with type I glycogen storage disease: a potential utility as biomarkers
Authors: Li Ling-Hui , Chiu Li-Ya , Kishnani Priya S. , Chuang Tzu-Po , Tang Cheng-Yang , Liu Cheng-Yuan , Bali Deeksha , Koeberl Dwight , Austin Stephanie , Boyette Keri , Weinstein David A. , Murphy Elaine , Yao Adam , Chen Yuan-Tsong
Published: May 2015

Background: It is known that malignant transformation to hepatocellular carcinoma (HCC) occurs at a higher frequency in hepatocellular adenoma (HCA) from type I glycogen storage disease (GSD I) compared to HCA from other etiologies. In this study, we aimed to identify differentially expressed miRNAs in GSD Ia HCA as candidates that could serve as putative biomarkers for detection of GSD Ia HCA and/or risk assessment of malignant transformation.

Methods: Utilizing massively parallel sequencing, the miRNA profiling was performed for paired adenomas and normal liver tissues from seven GSD Ia patients. Differentially expressed miRNAs were validated in liver tumor tissues, HCC cell lines and serum using quantitative RT-PCR.

Results: miR-34a, miR-34a*, miR-224, miR-224*, miR-424, miR-452 and miR-455-5p were found to be commonly deregulated in GSD Ia HCA, general population HCA, and HCC cell lines at compatible levels. In comparison with GSD Ia HCA, the upregulation of miR-130b and downregulation of miR-199a-5p, miR-199b-5p, and miR-214 were more significant in HCC cell lines. Furthermore, serum level of miR-130b in GSD Ia patients with HCA was moderately higher than that in either GSD Ia patients without HCA or healthy individuals.

Conclusion: We make the first observation of the distinct miRNA deregulation in HCA associated with GSD Ia. We also provide evidence that miR-130b could serve as a circulating biomarker for detection of GSD Ia HCA. This work provides prominent candidate miRNAs worth evaluating as biomarkers for monitoring the development and progress of liver tumors in GSD Ia patients in the future.

Citation Format: Ling-Hui Li, Li-Ya Chiu, Priya S. Kishnani, Tzu-Po Chuang, Cheng-Yang Tang, Cheng-Yuan Liu, Deeksha Bali, Dwight Koeberl, Stephanie Austin, Keri Boyette, David A. Weinstein, Elaine Murphy, Adam Yao, Yuan-Tsong Chen. Identification of differentially expressed microRNAs in human hepatocellular adenoma associated with type I glycogen storage disease: a potential utility as biomarkers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 518. doi:10.1158/1538-7445.AM2014-518
Abstract 5185: Redefining the somatic landscape of pancreatic adenocarcinoma
Authors: Brown Andrew M.K. , Notta Faiyaz , Lemire Mathieu , Lungu Ilinca , Denroche Robert E. , Yung Christina , Geras Kristen , Stein Lincoln , Bartlett John M. , Hudson Thomas J. , Roehrl Michael H.A. , Gallinger Steven , McPherson John D.
Published: May 2015

Efforts to catalogue genomic abnormalities in pancreatic ductal adenocarcinoma (PDAC) have been challenging due to low tumor cellularity which requires deep sequencing to obtain adequate coverage of tumor DNA. To make whole genome sequencing (WGS) feasible with PDAC tumors, we have developed protocols for sequencing enriched tumor material by laser capture microdissection (LCM) from frozen samples and by flow sorting when fresh tissue is available. The enriched samples have a median tumor cellularity of 88% compared to a median cellularity of 12% in the bulk samples. Furthermore, the enrichment methods may also provide a source of normal or reference material, non-tumor cells in the case of LCM and CD45+ blood cells in the case of flow sorting, and eliminate the need for matched blood samples to call somatic variation. In conjunction with low quantity DNA input protocols for library construction, high quality next generation sequencing libraries are readily produced without sacrificing library complexity that enables somatic mutation, copy-number and structural variant detection at modest sequencing depths (∼50x). These methods are applicable to all tumor types where significant stromal contamination hinders somatic mutation detection. In addition to redefining the rates of known somatically altered genes in PDAC, such as CDKN2A and SMAD4, the high cellularity data set has also allowed us to evaluate PDAC tumor heterogeneity with the ability to detect sub-clonal variation that is not masked by non-tumor DNA. Moreover, in 50% of the samples sequenced to date, we observe evidence of punctuated evolution suggesting a catastrophic event during mutagenesis, which may be associated with the clinically observed rapid progression of the disease. The result of enhanced somatic mutation detection is a more comprehensive picture of the PDAC genomic landscape.

Citation Format: Andrew M.K. Brown, Faiyaz Notta, Mathieu Lemire, Ilinca Lungu, Robert E. Denroche, Christina Yung, Kristen Geras, Lincoln Stein, John M. Bartlett, Thomas J. Hudson, Michael H.A. Roehrl, Steven Gallinger, John D. McPherson. Redefining the somatic landscape of pancreatic adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5185. doi:10.1158/1538-7445.AM2014-5185