bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-181: Tolfenamic acid suppresses intestinal tumorigenesis through inhibiting cyclin D1 translation.
Authors: Zhang Xiaobo , Lee Seong-Ho , Li Qingwang , Baek Seung J.
Published: August 2014

A nonsteroidal anti-inflammatory drug tolfenamic acid (TA) has been shown to suppress cancer cell growth and tumorigenesis in different cancer models. However, the underlying mechanism by which TA exerts its anti-tumorigenic activity is not well understood. Here, we have shown that TA markedly reduced the number of polyps and tumor load in APCmin/+ mice, accompanied with the significant decrease in cyclin D1 expression. Furthermore, we have demonstrated that TA down-regulates cyclin D1 expression in different cancer cells in vitro, coupled by decreasing phosphorylated retinoblastoma protein and increasing cell cycle arrest. Subsequent observation suggests that TA promotes endoplasmic reticulum (ER) stress, resulting in activation of the unfolded protein response (UPR) signaling pathway including phosphorylation of eukaryotic translation initiation factor 2α (eIF2α), which mediates the repression of cyclin D1 translation. Silence of pancreatic ER eIF2α kinase (PERK) restores cyclin D1 expression in the presence of TA. In conclusion, TA inhibits colorectal cancer cell growth in vitro and blocks colorectal tumorigenesis in a mouse model, at least in part, mediated by inhibiting cyclin D1 translation via ER stress.

Support: The University of Tennessee Center of Excellence in Livestock Diseases and Human Health grant (SJB), NIH grant RO1CA108975 (SJB), American Cancer Society grant (RSG-11-133-01-CCE) (S-H Lee), and a grant from the Ministry of Agriculture of the People's Republic of China (2011ZX08008-003) (QL).

Citation Format: Xiaobo Zhang, Seong-Ho Lee, Qingwang Li, Seung J. Baek. Tolfenamic acid suppresses intestinal tumorigenesis through inhibiting cyclin D1 translation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-181. doi:10.1158/1538-7445.AM2013-LB-181
Abstract LB-188: Isoflavone pharmacokinetics and metabolism after consumption of soy and soy-almond bread in men with asymptomatic prostate cancer.
Authors: Ahn-Jarvis Jennifer H. , Clinton Steven K. , Grainger Elizabeth M. , Riedl Kenneth M. , Schwartz Steven J. , Vodovotz Yael
Published: August 2014

Background: Soy phytochemicals, particularly isoflavones, are hypothesized to have anti-prostate cancer activity. Soy isoflavones are present predominantly as glucosides but when combined with β-glucosidase activity in almond meal, a greater conversion to aglycones has been observed. We hypothesized that soy bread will serve as an excellent source of soy polyphenols and that an aglycone-rich soy-almond bread (SAB) compared to a glycoside-rich soy bread (SB) will demonstrate greater bioavailability of soy isoflavones.

Methods: SB and SAB were fed in a 20 week randomized phase II cross-over trial involving 32 men with asymptomatic metastatic prostate cancer and escalating prostate specific antigen (PSA). Isoflavones were quantitated by HPLC in soy breads and isoflavone metabolites in 24h urine. HPLC-tandem mass spectrometry evaluated isoflavone pharmacokinetics in plasma. Biomarkers were determined by chemiluminescence-based assays.

Results: The aglycone composition in the SB and SAB was 18% and 78%, respectively. Compliance was 95±14%, toxicity rare and limited to grade I GI complaints (NIH_NCI criteria). Urinary isoflavone patterns over 24hours revealed that isoflavonoid metabolites were significantly greater with soy bread consumption. Plasma pharmacokinetics following a test meal showed that daidzein (p=0.028) and genistein (p=0.010) absorption was significantly faster from the aglycone-rich SAB. Interestingly, PSA doubling time was prolonged compared to pre-enrollment, in 60% of men (n=30) with soy intervention.

Conclusions: Bread is an excellent vehicle for the delivery of soy phytochemicals with outstanding compliance. Modification of soy bread with a source of β-glucosidase may enhance the initial absorption rates of isoflavones. Food products, such as SAB, with consistent and stable composition of phytochemicals can be developed that are appropriate for long-term chemoprevention studies in humans or as an adjunct to therapy.

Citation Format: Jennifer H. Ahn-Jarvis, Steven K. Clinton, Elizabeth M. Grainger, Kenneth M. Riedl, Steven J. Schwartz, Yael Vodovotz. Isoflavone pharmacokinetics and metabolism after consumption of soy and soy-almond bread in men with asymptomatic prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-188. doi:10.1158/1538-7445.AM2013-LB-188
Abstract LB-206: Target validation of C4.4a (LYPD3) as a target for antibody-drug conjugates for the treatment of squamous cell carcinoma of different origin.
Authors: Leder Gabriele , Cao Yong-Jiang , Linden Lars , Golfier Sven , von Ahsen Oliver , Schneider Claudia , Willuda Joerg , McLean Kirk , Pohlenz Hans-Dieter , Kreft Bertolt , Ziegelbauer Karl
Published: August 2014

C4.4a (LYPD3) is a GPI-anchored cell surface protein with unknown physiological function, although it has been speculated that it might play a role in tumor invasion and metastasis formation. In addition, over-expression of C4.4a correlates with a malignant phenotype and poor prognosis in several cancer indications.

As a result of comprehensive gene expression profiling, we report that the C4.4a transcript is being significantly up-regulated in human non-small cell lung cancer (NSCLC) as well as in other tumors compared to the respective normal tissue. This finding was confirmed in IHC studies showing strong C4.4a protein expression on the surface of tumor cells particularly in lung squamous cell carcinoma. Interestingly, a high incidence of C4.4a protein expression was found in a number of additional cancers such as stomach, bladder, colorectal, ovarian, prostate, hepatocellular as well as SCC of the head and neck. In marked contrast to tumors, C4.4a expression in normal tissues is restricted to esophagus, cervix and the outer epithelial layer of skin (stratum spinosum and stratum granulosum). Employing FACS and IHC analyses, several model systems were identified expressing C4.4a in vitro and in vivo. Subsequently, Mab-Zap assays were used to demonstrate efficient internalization of C4.4a in endogenously expressing cells as well as in cells transfected to over-express C4.4a.

In summary, the observed strong over-expression of C4.4a in tumors, its very limited expression in normal tissue as well as its ability to rapidly internalize make it a promising target for the generation of antibody-drug conjugates for the treatment of several cancer indications, in particular lung squamous cell carcinoma.

Citation Format: Gabriele Leder, Yong-Jiang Cao, Lars Linden, Sven Golfier, Oliver von Ahsen, Claudia Schneider, Joerg Willuda, Kirk McLean, Hans-Dieter Pohlenz, Bertolt Kreft, Karl Ziegelbauer. Target validation of C4.4a (LYPD3) as a target for antibody-drug conjugates for the treatment of squamous cell carcinoma of different origin. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-206. doi:10.1158/1538-7445.AM2013-LB-206
Abstract LB-218: Role of ERBB family negative regulator Mig6 in estimating erlotinib responsiveness.
Authors: Izumchenko Eugene , Chang Xiofei , Sidransky David
Published: August 2014

The sensitivity of some tumors to anti-epidermal growth factor receptor EGFR tyrosine kinase inhibitors (TKIs) can be explained by the presence of EGFR tyrosine kinase (TK) domain mutations. However, such mutations are rare and the clinical benefit of a subset of patients with wild-type EGFR may be underestimated due to the lack of appropriate predictive markers to stratify them from larger number of patients who do not response.

In this study we sought to elucidate mechanisms of resistance to EGFR-targeted therapies in tumors that harbor no TK sensitizing mutations in order to identify markers capable of guiding the decision to incorporate these drugs into chemotherapeutic regimens. Here we show that mitogen-inducible gene 6 (Mig6), a negative regulator of EGFR, was selectively upregulated by the PI3K-AKT pathway during the evolution of resistance to the EGFR tyrosine kinase inhibitor erlotinib, resulting in decreased EGFR phosporylation. Moreover, we found that high Mig6/EGFR ratio was highly correlated with erlotinib sensitivity, AKT activity, loss of PTEN and epithelial-mesenchymal transition (EMT) status in the panel of 20 lung and head and neck cancer cell lines.

Since depletion of Mig6 in erlotinib resistant cell lines didn't render significant increase in drug sensitivity and did not affect AKT activity, we tested whether in resistant cells, activation of kinases other then ERBB family can sustain high level of AKT activation thus suspending erlotinib sensitivity in Mig6-depleted cells. Surprisingly, we found that despite significant deactivation of EGFR, HER2 and HER3 kinases in erlotinib resistant cell lines, activation of PDGFR, FGFR, VEGFR, IGFR or FAK kinases was upregulated in these cells. Here we show that combined treatment with erlotinib and regorafinib (a novel multi-target inhibitor for VEGFR, PDGFR, FGFR, Kit, RET and Raf-1) markedly enhances erlotinib-induced growth inhibition in erlotinib-resistant cells.

Collectively, our data highlights the importance of Mig6 in estimating erlotinib responsiveness. Furthermore, our data suggests that by inhibiting EGFR activity in sensitive neoplastic cells, elevated expression of Mig6 triggers acquisition of alternative growth factor pathways that can sustain high level of AKT activity, thus further promoting erlotinib resistance.

Citation Format: Eugene Izumchenko, Xiofei Chang, David Sidransky. Role of ERBB family negative regulator Mig6 in estimating erlotinib responsiveness. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-218. doi:10.1158/1538-7445.AM2013-LB-218
Abstract LB-229: Whole genome sequencing reveals genetic landscape of hepatocellular carcinoma.
Authors: Mao Mao , Zheng Hancheng , Kan Zhengyan , Xu Jiangchun , Liu Xiao , Li Shuyu , Barber Thomas , Gong Zhuolin , Gao Huan , Hao Ke , Willard Melinda , Hauptschein Robert , Rejto Paul , Fernandez Julio , Wang Guan , Zhang Qinghui , Wang Bo , Chen Ronghua , Wang Jian , Lee Nikki , Zhou Wei , Lin Zhao , Peng Zhiyu , Yi Kang , Chen Shengpei , Li Lin , Fan Xiaomei , Yang Jie , Ye Rui , Ju Jia , Wang Kai , Estrella Heather , Deng Shibing , Wei Ping , Qiu Ming , Wulur Isabella , Liu Jiangang , Ehsani Mariam , Zhang Chunsheng , Loboda Andrey , Sung Wing Kin , Aggarwal Amit , Poon Ronnie , Fan Sheung Tat , Wang Jun , Hardwick James , Reinhard Christoph , Dai Hongyue , Li Yingrui , Luk John
Published: August 2014

Hepatocellular carcinoma (HCC) is one of the most deadly cancers worldwide and has no effective treatment, yet the molecular basis of hepatocarcinogenesis remains largely unknown. Here we report findings from a whole genome sequencing (WGS) study of 88 matched HCC tumour/normal pairs, 81 of which are HBV positive, seeking to identify genetically altered genes and pathways implicated in HBV-associated HCC. We find β-catenin to be the most frequently mutated oncogene (15.9%) and TP53 the most frequently mutated tumour suppressor (35.2%). The Wnt/β-catenin pathway, altered in 62.5% of cases, is likely to act as the major oncogenic driver in HCC. TP53 alterations appear to cause increased levels of genomic arrangement and chromosomal instability. We identified chromothripsis in 5 HCC genomes (5.7%) recurrently affecting chromosomal arms 1q and 8q. We also identified recurrent HBV integration events at the known and putative cancer-related genes such as TERT, MLL4 and CCNE1, which showed upregulated gene expression in tumour versus normal tissue. The frequently altered genes and pathways in HCC reflect classical cancer hallmarks. This study identified several prevalent and actionable mutations that provide a path towards therapeutic intervention of the disease.

Citation Format: Mao Mao, Hancheng Zheng, Zhengyan Kan, Jiangchun Xu, Xiao Liu, Shuyu Li, Thomas Barber, Zhuolin Gong, Huan Gao, Ke Hao, Melinda Willard, Robert Hauptschein, Paul Rejto, Julio Fernandez, Guan Wang, Qinghui Zhang, Bo Wang, Ronghua Chen, Jian Wang, Nikki Lee, Wei Zhou, Zhao Lin, Zhiyu Peng, Kang Yi, Shengpei Chen, Lin Li, Xiaomei Fan, Jie Yang, Rui Ye, Jia Ju, Kai Wang, Heather Estrella, Shibing Deng, Ping Wei, Ming Qiu, Isabella Wulur, Jiangang Liu, Mariam Ehsani, Chunsheng Zhang, Andrey Loboda, Wing Kin Sung, Amit Aggarwal, Ronnie Poon, Sheung Tat Fan, Jun Wang, James Hardwick, Christoph Reinhard, Hongyue Dai, Yingrui Li, John Luk. Whole genome sequencing reveals genetic landscape of hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-229. doi:10.1158/1538-7445.AM2013-LB-229
Abstract LB-23: Meta-analysis of genome-wide association studies identifies multiple loci associated with chronic lymphocytic leukemia.
Authors: Berndt Sonja I. , Skibola Christine F. , Joseph Vijai , Camp Nicola J. , Nieters Alexandra , Wang Zhaoming , Cozen Wendy , Monnereau Alain , Kelly Rachel S. , de Sanjose Silvia , Linet Martha S. , Riboli Elio , Vineis Paolo , Hjalgrim Henrik , Cerhan James R. , Chanock Stephen J. , Rothman Nathaniel , Slager Susan L.
Published: August 2014

Despite limited discovery stages (<1,125 cases), genome-wide association studies (GWAS) have successfully identified 13 loci associated with risk of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL); however, these loci only explain a portion of the heritability. To identify additional genetic loci associated with CLL susceptibility, we conducted the largest meta-analysis of GWAS, to date, including 3,100 CLL cases and 7,667 controls of European ancestry from 25 independent studies genotyped on four different SNP microarrays. Association testing was conducted assuming a log-additive genetic model, adjusting for age, sex, and significant principal components. Meta-analyses were performed using the fixed effects inverse variance method based on the beta estimates and standard errors from each study. Promising loci were taken forward for replication in three additional studies totaling 788 cases and 4,872 controls. In the meta-analysis of both the discovery and replication studies, we conclusively identified ten independent SNPs in nine novel loci that reached genome-wide significance (P<5 x 10−8): 10q23.31 (FAS; P=1.22x10−14), 18q21.33 (BCL2; P=2.66x10−12 and P=7.76x10−11), 11p15.5 (C11orf21; P=2.15x10−10), 4q25 (LEF1; P=4.24x10−10), 2q33.1 (CASP8; P=2.50x10−9), 9p21.3 (CDKN2B-AS1; P=1.27x10−8), 18q21.32 (PMAIP1; P=2.51x10−8), 15q15.1 (BMF; P=2.71x10−10), and 2p22.2 (QPCT; P=1.68x10−8) as well as an independent signal at an established locus (2q13, ACOXL, P=2.08x10−18). We also found evidence for two additional promising loci that reached marginal genome-wide significance (P<2.0x10−7) at 8q22.3 (ODF1; P=5.40x10−8) and 5p15.33 (TERT; P=1.92x10−7). Although further studies are required, the proximity of five of the novel loci to genes involved in apoptosis suggests a possible unifying underlying biological mechanism. Together with the previously established loci, these genetic loci explain ~17% of the familial risk of CLL.

Citation Format: Sonja I. Berndt, Christine F. Skibola, Vijai Joseph, Nicola J. Camp, Alexandra Nieters, Zhaoming Wang, Wendy Cozen, Alain Monnereau, Rachel S. Kelly, Silvia de Sanjose, Martha S. Linet, Elio Riboli, Paolo Vineis, Henrik Hjalgrim, James R. Cerhan, Stephen J. Chanock, Nathaniel Rothman, Susan L. Slager. Meta-analysis of genome-wide association studies identifies multiple loci associated with chronic lymphocytic leukemia. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-23. doi:10.1158/1538-7445.AM2013-LB-23
Abstract LB-232: Tumor clonality detection using next generation sequencing data.
Authors: Dees Nathan D. , Miller Christopher A. , White Brian S. , Schierding William , Vij Ravi , Tomasson Michael H. , Welch John S. , Graubert Timothy A. , Walter Matthew J. , Ley Timothy J. , DiPersio John F. , Mardis Elaine R. , Wilson Richard K. , Ding Li
Published: August 2014

A tumor's somatic variants, regardless of their significance in disease etiology, serve as genetic markers that capture the evolutionary history of clones and thus may be used for identifying them. We have previously identified steps along this evolutionary trajectory by discovering a broad range of variants, including exonic, intronic, and intergenic mutations, using whole genome sequencing (WGS) and by obtaining deep read counts of those variants using capture-based, targeted re-sequencing. Most of these variants, in most of the tumors analyzed, form clusters. This has revealed that most tumors are multi-clonal in composition with cellular sub-populations having distinct genomes harboring diverse somatic variants. Understanding the clonal diversity and composition of a tumor sample may have implications for clinical treatment, while the clonal evolution across tumor and relapse samples provides insight into disease progression.

Here we describe our approach for inferring clones using standard single nucleotide variant (SNV) and copy number alteration (CNA) data obtained from next-generation sequencing of a normal sample and one or more diseased (e.g., tumor or relapse) samples. Variants are clustered according to their deep-read-count-derived frequencies using a variational Bayesian approach to Beta mixture modeling, through which outliers and the number of clusters are identified automatically. The method's probabilistic interpretation provides a quantifiable measure of confidence in a resulting cluster, for example, through a standard error of the mean. We show that amplifications and deletions may induce artifactual clones, thus highlighting the importance of accounting for copy number events in interpreting SNV data for inferring clones. We demonstrate the performance of our approach using published acute myeloid leukemia tumor and relapse pairs as well as unpublished multiple myeloma WGS data sets. However, the approach is applicable to solid tumors as well and has been used to cluster six-dimensional variant data from a breast tumor and five relapses originating from the same patient.

Citation Format: Nathan D. Dees, Christopher A. Miller, Brian S. White, William Schierding, Ravi Vij, Michael H. Tomasson, John S. Welch, Timothy A. Graubert, Matthew J. Walter, Timothy J. Ley, John F. DiPersio, Elaine R. Mardis, Richard K. Wilson, Li Ding. Tumor clonality detection using next generation sequencing data. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-232. doi:10.1158/1538-7445.AM2013-LB-232
Abstract LB-235: Functional SNP in MICA binding to SP1 is associated with HCV-induced hepatocellular carcinoma.
Authors: Lo Paulisally H. , Tanikawa Chizu , Nakamura Yusuke , Matsuda Koichi
Published: August 2014

Hepatitis C virus (HCV) infection is the major cause of hepatocellular carcinoma (HCC) in Japan. To understand the genetic basis of HCV related diseases, identification of the susceptibility gene involving HCV-induced HCC is important. We previously identified the association of SNP rs2596542 in the 5′ flanking region of the MHC class I polypeptide-related sequence A (MICA) gene with the risk of HCV-induced HCC. In the current study, we performed detailed functional analysis of 12 candidate SNPs in the promoter region. A SNP rs2596538 located at 2.8 kb upstream of the MICA gene affected the binding of a nuclear protein(s) to the genomic segment including this SNP. By electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay, we identified that transcription factor Specificity Protein 1 (SP1) can bind to the protective G allele, but not to the risk A allele. In addition, reporter construct containing the G allele exhibited higher transcriptional activity than that containing the A allele. SNP rs2596538 exhibited the stronger association with HCV-induced HCC (P = 1.82 x10−5 and OR = 1.34) than the marker SNP rs2596542. We also found significantly higher serum level of soluble MICA (sMICA) in HCV-induced HCC patients carrying the G allele than those carrying the A allele (P = 0.00616). In summary, we have identified a functional SNP that is associated with expression of MICA and risk for HCV-induced HCC.

Citation Format: Paulisally H. Lo, Chizu Tanikawa, Yusuke Nakamura, Koichi Matsuda. Functional SNP in MICA binding to SP1 is associated with HCV-induced hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-235. doi:10.1158/1538-7445.AM2013-LB-235
Abstract LB-244: MiR-34a/c-dependent PDGFR-α/β reduction inhibits tumorigenesis and sensitizes TRAIL-induced apoptosis in lung cancer.
Authors: Jeon Young-Jun , Nuovo Gerard J. , Midddleton Justin , Joshi Pooja , Nazaryan Natalya , Di Leva Gianpiero , Romano Giulia , Crawford Melissa , Nana-Sinkam Patrick , Garofalo Michela , Croce Carlo M.
Published: August 2014

Lung cancer is the most common cause of cancer death worldwide. Despite years of research, the prognosis for patients with lung cancer remains dismal. The most frequent type, non-small cell lung cancer (NSCLC) (85%), shows an overall five year survival of 15%. Isoforms of platelet-derived growth factor receptor (PDGFR) and its ligand, PDGF, constitute a family of receptors and ligands involved in proliferative and prosurvival signaling within cells. MicroRNAs (miRNAs), a class of ~ 22 nt endogenous RNAs, are important regulators of gene expression and have been implicated in the regulation of critical processes that are deregulated in cancer cells, as proliferation differentiation and apoptosis. Alterations in miRNA expression in cancer have been documented in numerous studies and suggest that miRNAs critically contribute to the cancer cell phenotype. In mammalians, the miR-34 family comprises three processed miRNAs that are encoded by two different genes: miR-34a is encoded by its own transcript, whereas miR-34b and miR-34c share a common primary transcript. In this study we found that miR-34a/c and not miR-34b, suppress the metastatic capacity of NSCLC by targeting platelet-derived growth factor receptor alpha and beta (PDGFR-α and PDGFR-β). We also observed an inverse relation between PDGFR-α/β mRNAs and miR-34a/c expression in lung tumor compared to normal lung samples. Finally, miR34-a/c-dependent PDGFR-α/β suppression or knocking-down of PDGFR-α/β by siRNAs sensitized NSCLC cells to TNF-related apoptosis inducing ligand (TRAIL)-induced apoptosis. Taken together our findings could be helpful for future anti-tumor therapy of lung cancer.

Citation Format: Young-Jun Jeon, Gerard J. Nuovo, Justin Midddleton, Pooja Joshi, Natalya Nazaryan, Gianpiero Di Leva, Giulia Romano, Melissa Crawford, Patrick Nana-Sinkam, Michela Garofalo, Carlo M. Croce. MiR-34a/c-dependent PDGFR-α/β reduction inhibits tumorigenesis and sensitizes TRAIL-induced apoptosis in lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-244. doi:10.1158/1538-7445.AM2013-LB-244
Abstract LB-260: CCR5 antagonists block basal breast cancer and prostate cancer metastasis in vivo.
Authors: Pestell Richard G.
Published: August 2014

The identification of new therapeutic targets and treatments to reduce tumor metastasis homing requires alternative interrogation approaches. Interrogation of microarray analysis of 2,254 human breast cancers demonstrated increased expression of CCL5 and its receptor CCR5, but not CCR3, in the basal and HER-2 genetic subtypes of breast cancer. Genome wide interrogation of pathways activated in patient normal breast vs. tumor identified upregulation of a CCR5 signaling module. Human breast cancer cell lines expressed CCR5 by flow cytometry, and displayed a functional response to CCL5 by calcium mobilization assays and invasion assays. Using isogenic oncogene transformed breast and prostate cancer cell lines, we show oncogene transformation induces CCR5 expression and that the subpopulation of cells that express functional CCR5 display increased invasiveness. CCR5 promoted metastasis homing in vivo. The CCR5 antagonists Maraviroc and Vicriviroc, developed to block CCR5 HIV co-receptor function, reduced in vitro invasion of basal breast cancer and prostate cancer cell lines without affecting cell proliferation or viability. In a series of preclinical mouse models, Maraviroc decreased breast pulmonary metastasis. The isogenic prostate cancer cell lines metastasized to bones in immune-competent mice representing an ideal model for testing anti-metastasis therapies. Maraviroc and Vicriviroc reduced prostate cancer metastasis to brain, bones and lungs. Our findings provide evidence for a key role of CCL5/CCR5 in the metastasis of basal breast cancer and prostate cancer cell lines, and suggest that CCR5 antagonists may be used as an adjuvant therapy to reduce the risk of metastasis in patients with the basal breast cancer subtype and prostate cancer.

Citation Format: Richard G. Pestell. CCR5 antagonists block basal breast cancer and prostate cancer metastasis in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-260. doi:10.1158/1538-7445.AM2013-LB-260
Abstract LB-330: Therapeutic activity of a combination of immunostimulatory monoclonal antibodies (anti-B7-H1, CD137 and OX40) on a c-myc-driven spontaneous transgenic model of hepatocellular carcinoma.
Authors: Morales-Kastresana Aizea , Sanmamed Miguel F. , Rodriguez Inmaculada , Palazon Asis , Martinez-Forero Ivan , Labiano Sara , Hervas-Stubbs Sandra , Sangro Bruno , Ochoa Carmen , Azpilikueta Arantza , Bolaños Elixabet , Gütgemann Ines , Melero Ignacio
Published: August 2014

Several immunostimulatory monoclonal antibodies (ISmAbs) that derepress and unleash antitumor immune responses are showing efficacy in cancer clinical trials. MAbs anti-B7-H1 (PD-L1) block a critical inhibitory pathway for T cells, while antibodies anti-CD137 and OX40 provide intense T cell costimulation. Documenting efficacy of immunotherapies on spontaneous tumors arising in oncogene transgenic mice is considered more predictive than experiments on transplanted tumors. A combination of these ISmAbs has been tested on a transgenic mouse model of spontaneous primary liver cancer in which c-myc drives transformation and the tumor cells express ovalbumin as a surrogate transgenic antigen. The induced tumor lymphocyte infiltrates and immune mechanisms of action were studied. The triple combination of mAbs clearly extended survival of mice bearing hepatocellular carcinomas (HCC) and synergized with adoptive T cell therapy with activated TCR-transgenic T cells that recognize OVA. Mice undergoing therapy showed a clear increase in the tumor tissue infiltration by activated and blastic CD8 T lymphocytes expressing the ISmAb-targeted receptors. The triple combination of ISmAbs did not result in enhanced OVA-specific CTL activity but other antigens in cell lines derived from such HCC were recognized by the elicited tumor infiltrating T lymphocytes. Indeed adoptive transfer of OT-1 cells to tumor bearing mice resulted in their tolerization, unless the triple mAb therapy was instigated. Our results emphasize the role of combinational immunotherapy approaches including ISmAbs to impact on agressive and highly T cell-tolerizing hepatocellular carcinomas.

Citation Format: Aizea Morales-Kastresana, Miguel F. Sanmamed, Inmaculada Rodriguez, Asis Palazon, Ivan Martinez-Forero, Sara Labiano, Sandra Hervas-Stubbs, Bruno Sangro, Carmen Ochoa, Arantza Azpilikueta, Elixabet Bolaños, Ines Gütgemann, Ignacio Melero. Therapeutic activity of a combination of immunostimulatory monoclonal antibodies (anti-B7-H1, CD137 and OX40) on a c-myc-driven spontaneous transgenic model of hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-330. doi:10.1158/1538-7445.AM2013-LB-330
Abstract LB-339: Increase in tumor-associated macrophages (TAMs) after antiangiogenic therapy is associated with poor survival in recurrent glioblastoma (GBM) patients.
Authors: Lu-Emerson Christine , Snuderl Matija , Kirkpatrick Nathaniel D. , Goveia Jermaine , Taylor Jennie , Davidson Christian , Huang Yuhui , Riedemann Lars , Ivy S. Percy , Duda G. Dan , Ancukiewicz3 Marek , Plotkin Scott R. , Chi Andrew , Gerstner Elizabeth R. , Eichler April F. , Dietrich Jorg , Stemmer-Rachamimov Anat O. , Batchelor Tracy T. , Jain Rakesh K.
Published: August 2014

Antiangiogenic therapy is associated with increased radiographic responses in glioblastomas (GBMs), but tumors invariably recur. Because tumor-associated macrophages (TAMs) have been shown to mediate escape from antiangiogenic therapy in preclinical models, we examined the role of TAMs in recurrent glioblastoma (rGBM) patients. We compared autopsy brain specimens from 20 rGBM patients who received antiangiogenic treatment and chemoradiation (AAT+) to 8 patients who received chemotherapy and/or radiotherapy without antiangiogenic therapy, or no treatment (AAT-). TAMs were morphologically and phenotypically analyzed using flow cytometry and immunohistochemistry (IHC) for CD68, CD14, CD163, and CD11b expression. Flow cytometry showed an increase in TAMs in the AAT+ patients. IHC analysis demonstrated an increase in CD68+ TAMs in the tumor bulk (p<0.01) and infiltrative areas (p=0.02) in AAT+ patients. We also observed an increase in CD11b+ cells in the tumor bulk (p<0.01) and an increase in CD163+ TAMs in infiltrative tumor (p=0.02). Of note, an increased number of CD11b+ cells in bulk and infiltrative tumor (p=0.05 and p=0.05, respectively) correlated with poor overall survival in patients who first received antiangiogenic therapy at recurrence. In summary, rGBMs showed an increased infiltration in myeloid populations in the tumor bulk and in the infiltrative regions after antiangiogenic therapy. Higher numbers of CD11b+ cells correlated with poor survival in rGBM patients. These data suggest that TAMs may participate in escape from antiangiogenic therapy and may represent a potential biomarker of resistance and a potential therapeutic target in rGBM.

Citation Format: Christine Lu-Emerson, Matija Snuderl, Nathaniel D. Kirkpatrick, Jermaine Goveia, Jennie Taylor, Christian Davidson, Yuhui Huang, Lars Riedemann, S. Percy Ivy, G. Dan Duda, Marek Ancukiewicz3, Scott R. Plotkin, Andrew Chi, Elizabeth R. Gerstner, April F. Eichler, Jorg Dietrich, Anat O. Stemmer-Rachamimov, Tracy T. Batchelor, Rakesh K. Jain. Increase in tumor-associated macrophages (TAMs) after antiangiogenic therapy is associated with poor survival in recurrent glioblastoma (GBM) patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-339. doi:10.1158/1538-7445.AM2013-LB-339
Abstract LB-345: Hypoxia-induced reprogramming cause normal glia to mimic tumor-associated glial cells.
Authors: Leiss Lina , Manini Ivana , Calderon Marta , Enger Per Øyvind
Published: August 2014

Cancer-associated fibroblasts (CAFs) have been shown to foster tumor growth in several solid cancers. In the brain, the stroma is mainly composed of glial cells such as astrocytes and oligodendrocytes whose role in brain tumor-stroma interactions are poorly characterized. We have previously identified a unique gene expression profile of tumor-associated glial cells (TAGs) by fluorescence activated cell sorting (FACS) of glial cells from GFP/Nod Scid mice engrafted with human glioblastoma biopsies. Little is known about the mechanisms mediating these expression changes, but both direct tumor cell-glial cell contact and environmental stress such as hypoxia are external cues that may induce these changes. Thus, we investigated how hypoxia impacted on the phenotype of glial cells from normal mouse brains. Briefly, mouse brains were enzymatically digested to obtain single cell suspensions, and then glial cells were isolated by FACS with simultaneous removal of immune cells (CD11b+) and endothelial cells (CD31+). These cells were then seeded at a density of 50,000 cells/well in a 48-well plate and incubated under either normoxic (21% oxygen) or hypoxic (0.5% oxygen) conditions for seven days. Notably, glial cells upregulated Sox2, HIF-2a, Nestin and Vimentin in hypoxia compared to normoxic conditions. In addition glial cells also upregulated angiogenic factors such as VEGF, Angiopoietin 2 and FGF2. Furthermore, cell cycle analysis revealed a significantly higher fraction of cycling cells in hypoxia than in normoxia. To investigate how glial cells and tumor cells interact, we made co-cultures of glial cells isolated from DsRed+ mice and GFP-transfected human glioma cells. Fluorescence imaging of the co-cultures show that these two cell types establish cell contacts in vitro, and that the glial cells alter their morphology in co-cultures compared to monocultures of glial cells alone. These changes strongly resembled the changes observed in tumor-associated glial cells, suggesting that hypoxia is a major contributor to the phenotype of tumor-associated glial cells. Furthermore direct physical contact between tumor cells and glial cells may lead to phenotypical changes in the glial cells. Ongoing work aims at further characterizing the TAG population to reveal its cell composition and address the role of various subpopulations of glial host cells.

Citation Format: Lina Leiss, Ivana Manini, Marta Calderon, Per Øyvind Enger. Hypoxia-induced reprogramming cause normal glia to mimic tumor-associated glial cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-345. doi:10.1158/1538-7445.AM2013-LB-345
Abstract LB-39: Sphingosine kinase activity is not required for tumor cell viability.
Authors: Wesche Holger , Brown Matthew L. , Carlson Timothy J. , Coxon Angela , Frank Brendon , Gustin Darin J. , Jeffries Shawn , Li Shyun , Li Yihong , Morgenstern Kurt , Morgenstern Kurt , Plant Matthew , Rex Karen , Schmidt Joanna , Shen Shanling , Walker Nigel , Wickramasinghe Dineli , Xu Guifen
Published: August 2014

Sphingosine kinase activity is not required for tumor cell viability

Holger Wesche, Matthew L. Brown, Timothy J. Carlson, Angela Coxon, Brendon Frank, Darin J. Gustin, Shawn Jeffries, Shyun Li, Yihong Li, Kurt Morgenstern, Matthew Plant, Karen Rex, Joanna Schmidt, Shanling Shen, Nigel Walker, Dineli Wickramasinghe, Mariwil Wong, Guifen Xu

Contribution from the Departments of Oncology Research, Medicinal Chemistry, Molecular Structure and Characterization and Pharmacokinetics and Drug Metabolism, Amgen Inc., South San Francisco, 1120 Veterans Blvd., South San Francisco, Ca, 94080.

Sphingosine kinases (SPHKs) are enzymes that phosphorylate the lipid sphingosine, leading to the formation of sphingosine-1-phosphate (S1P). In addition to the well established role of extracellular S1P as a mitogen and potent chemoattractant, SPHK activity has been postulated to be an important intracellular regulator of apoptosis. According to the proposed rheostat theory, SPHK activity shifts the intracellular balance from the pro-apoptotic sphingolipids ceramide and sphingosine to the mitogenic S1P, thereby determining the susceptibility of a cell to apoptotic stress. Despite numerous publications with supporting evidence, a clear experimental confirmation of the impact of this mechanism on tumor cell viability in vitro and in vivo has been hampered by the lack of suitable tool reagents. Utilizing a structure based design approach, we developed potent and specific SPHK1/2 inhibitors. These compounds completely inhibited intracellular S1P production in human cells and attenuated vascular permeability in mice, but did not lead to reduced tumor cell growth in vitro or in vivo. These results show that the SPHK rheostat does not play a major role in tumor cell viability, and that SPHK inhibition may not offer an advantage over S1P neutralization in the treatment of cancer.

Citation Format: Holger Wesche, Matthew L. Brown, Timothy J. Carlson, Angela Coxon, Brendon Frank, Darin J. Gustin, Shawn Jeffries, Shyun Li, Yihong Li, Kurt Morgenstern, Kurt Morgenstern, Matthew Plant, Karen Rex, Joanna Schmidt, Shanling Shen, Nigel Walker, Dineli Wickramasinghe, Guifen Xu. Sphingosine kinase activity is not required for tumor cell viability. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-39. doi:10.1158/1538-7445.AM2013-LB-39
Abstract LB-45: Multiple melanoma suppressors are downregulated by loss of human chromosome 10.
Authors: Kwong Lawrence N. , Chin Lynda
Published: August 2014

Increasingly high-resolution genomic studies have made it clear that recurrent focal deletions and amplifications in cancer can selectively target specific oncogenes and tumor suppressors. In melanoma, MITF, MDM2, BRAF, CDKN2A, and PTEN are some of the more well-validated oncogenes and tumor suppressors that are targeted by such focal copy number-changing aberrations. However, there are also many recurrent large losses and gains that affect up to whole chromosome arms. One hypothesis is that, in targeting single genes such as PTEN, large chromosomal losses may inadvertently hit other loci. However, gathering evidence is pointing towards a possible functional role for these other collateral loci in affecting tumor progression. Here, we ask whether recurrent losses in melanoma may broadly target multiple tumor suppressors acting at different steps in tumor progression. We demonstrate via in vivo and in vitro shRNA screens that multiple genes on human chromosome 10, other than PTEN, affect both tumor growth and invasion.

Citation Format: Lawrence N. Kwong, Lynda Chin. Multiple melanoma suppressors are downregulated by loss of human chromosome 10. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-45. doi:10.1158/1538-7445.AM2013-LB-45
Abstract LB-61: Vemurafenib promotes RAS wild-type tumor formation in a mouse model of HPV-driven cutaneous squamous cell carcinoma.
Authors: Holderfield Matthew , Lorenzana Edward , Weisburd Ben , Lomovasky Lisa , Ghoddusi Majid , Daniel Dylan , Pryer Nancy , McCormick Frank , Stuart Darrin
Published: August 2014

A subset of melanoma patients (18-30%) treated with Vemurafenib (and other RAF inhibitors) develop spontaneous cutaneous squamous-cell carcinoma (cSCC) and/or keratoacanthoma (KA). Preclinical studies demonstrate that RAF inhibitors paradoxically activate the MAPK pathway and stimulate growth of RAS mutated cells. Based on these observations it has been proposed that drug induced MAPK activation in pre-cancerous tissues may account for the high incidence of cSCC and KA in treated patients. Because human cSCC is frequently positive for human papillomavirus (HPV) DNA, we hypothesized that RAF inhibitors promote tumorigenesis in HPV infected epidermis. To investigate this possibility, we used a transgenic murine model (K14-HPV16) of cSCC, driven by basal keratinocyte-specific expression (K14 promoter) of HPV type 16 early genes including E6 and E7, which inactivates p53 and pRB tumor suppressors. Exposure to Vemurafenib elevated MAPK markers in epidermis and increased cSCC incidence from 22% to 70%. Exome sequencing revealed that 100% of cSCCs from untreated mice harbored activating RAS mutations, yet 55% of cSCCs from Vemurafenib treated mice were RAS wild-type. Concomitant treatment with a MEK inhibitor reduced cSCC frequency, and MEK inhibitor alone was sufficient to completely regress established cSCCs. Together, these results suggest that cSCC is dependent upon MAPK activation induced either by a RAS oncogene or alternatively by exposure to RAF inhibitors in HPV infected keratinocytes.

Citation Format: Matthew Holderfield, Edward Lorenzana, Ben Weisburd, Lisa Lomovasky, Majid Ghoddusi, Dylan Daniel, Nancy Pryer, Frank McCormick, Darrin Stuart. Vemurafenib promotes RAS wild-type tumor formation in a mouse model of HPV-driven cutaneous squamous cell carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-61. doi:10.1158/1538-7445.AM2013-LB-61
Abstract LB-69: Novel WBC gene expression assay for prostate cancer detection.
Authors: Kassis Amin I. , Wu Qi , Joy Maliackal Poulo , Balagurumoorthy Pichumani , Loda Massimo , Evan Carolyn , Petrozziello Gillian , Kantoff Philip W. , Adelstein S. James
Published: August 2014

In recent studies, we showed that several oncogenes and cancer-associated genes and proteins are differentially expressed in phagocytic WBCs of tumor-bearing mice (colon, lung, melanoma, and prostate), compared to non-phagocytic WBCs from the same mice as well as to phagocytic WBCs obtained from non-tumor-bearing mice. Prompted by the translational potential of these findings, we carried out whole genome microarray analysis of total RNA samples isolated from peripheral blood phagocytic (macrophages and neutrophils) and nonphagocytic (T cells) WBCs of prostate cancer patients (n = 58) and blood donors (n = 36). Comparison of the genomic expression profiles of macrophages vs. T cells or neutrophils vs. T cells of these patients to those identified in blood donors revealed the presence of gene signatures that are unique to the patient population. These differentially expressed genes were used to generate receiver operating characteristic curves. The area under the curve was 1.000 for macrophages and 0.980 for neutrophils, indicating the high accuracy of the assay. While the PSA cutoff of 4 ng/mL failed to identify 14 of the 58 patients, the assay reported herein correctly identified 57 out of the 58 patients and all 36 controls (sensitivity = 0.983; specificity = 1.000) when macrophages were used; with neutrophils, there were 7 false negatives (sensitivity = 0.879; Specificity = 1.000). These promising results suggest that the approach lays the foundation for a next generation diagnostic blood test for prostate-cancer.

Citation Format: Amin I. Kassis, Qi Wu, Maliackal Poulo Joy, Pichumani Balagurumoorthy, Massimo Loda, Carolyn Evan, Gillian Petrozziello, Philip W. Kantoff, S. James Adelstein. Novel WBC gene expression assay for prostate cancer detection. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-69. doi:10.1158/1538-7445.AM2013-LB-69

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract LB-71: Carcinogenic significance of integrated hepatitis b viral factors and its genotype in hepatocarcinogenesis.
Authors: Banerjee Soma , Datta Somenath , Roychoudhury Shrabasti , Dasgupta Debanjali , Ghosh Amit , Roy Gaurav , Datta Simanti , Chowdhury Abhijit
Published: August 2014

Integration of Hepatitis B Virus (HBV) is one of the major causes underlying Hepatocellular carcinoma (HCC) development. Recent studies revealed that altered expression of multiple cellular genes due to multiple random integration of viral factors lead to development of HCC. The present study aims to identify the carcinogenic significance of integrated HBV genome and its genotype in HCC development.

Host-Viral Junctions were determined in HCC (T) /adjacent tissues (NT) and cirrhotic (-HCC) tissues by sequencing of Alu PCR product followed by sequence blast. Host gene expression was checked by real time PCR. Competition between two coexisting genotypes was studied by replication assay using genotype specific PCR from media and viral core particle isolated from genotype specific plasmid transfected HepG2 cell line. Similarly, each viral genotype mediated DNA damage (γH2aX foci by confocal microscopy), ROS generation (DCFDA quantitation by Facs analysis and ER stress marker GRP78 luciferase assay), homologous recombination efficiency were also compared.

HBV integration was observed in 66.7% of T (8/12), 33.3% (4/12) NT and 75% (6/8) cirrhotic tissue samples in 18, 4 and 8 different locations respectively. These viral integration analysis within or close to several host genes, such as genes involved in differentiation, signaling, stress response, cell cycle, telomere regulations (8/30, 26.7%) etc and most of them showed altered expression. Interestingly, C-terminal truncated HBX which lost its growth suppressive domain were observed more in T (5/8, 62.5%) than NT (2/4, 50%) tissue samples. Most importantly, upon genotype analysis of each five clones in serum and tissue showed genotype C preferentially integrated in coexistence of both C and D genotype as “free virus” in tissue but D genotype observed in circulation. Genotype specific PCR from HBV/C & D cotransfected media showed that in presence of HBV/C, D replicates better than C but HBV/C with high recombination frequency generates more DNA damage and facilitate viral integration.

Thus random integration of HBV in host chromosome causes alterations in oncogenic gene expression and these alterations presumably lead to clonal selection of hepatocyte that acquires a growth advantage and proceeds towards HCC. Genotype of HBV is an important determining factor for HBV integration and liver inury.

Citation Format: Soma Banerjee, Somenath Datta, Shrabasti Roychoudhury, Debanjali Dasgupta, Amit Ghosh, Gaurav Roy, Simanti Datta, Abhijit Chowdhury. Carcinogenic significance of integrated hepatitis b viral factors and its genotype in hepatocarcinogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-71. doi:10.1158/1538-7445.AM2013-LB-71