bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 710: Telomerase-specific oncolytic adenovirus OBP-301 kills quiescent peritonially disseminated cancer cells.
Authors: Yano Shuya , Tazawa Hiroshi , Kishimoto Hiroyuki , Fujiwara Toshiyoshi , Hoffman Robert M.
Published: August 2014

MKN45 human gastric cancer cells can develop peritonitis carninomatosa where the cancer cells form aggregated structures. The cells are mostly in G0/G1 phase, as imaged by fluorescent ubiquitination-based cell cycle indicator (FUCCI). However, most cytotoxic agents target only proliferating cancer cells. The telomerase-specific adenovirus, OBP-301 was injected into the peritoneal cavity 7 days after inoculation of FUCCI-expressing MKN45. Control mice had approximately 5 ml bloody ascites with big tumor masses in the peritoneal cavity after 60 days. Cisplatin-treated mice also had approximately 5 ml bloody ascites with a big tumor masses in the peritoneal cavity. Paclitaxel-treated mice also had big tumor masses. There were many tumor spheres in the bloody ascites, and the tumors in the peritoneal cavity consisted of cancer cells mostly in G0/G1 phase. In contrast, OBP-301 treated mice had no ascites and only small tumor nodules consisting of cancer cells in S/G2 phases in the peritoneal cavity. These data suggest that OBP-301 is effective against peritonially disseminated cancer cells which are mostly in G0/G1.

Citation Format: Shuya Yano, Hiroshi Tazawa, Hiroyuki Kishimoto, Toshiyoshi Fujiwara, Robert M. Hoffman. Telomerase-specific oncolytic adenovirus OBP-301 kills quiescent peritonially disseminated cancer 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 710. doi:10.1158/1538-7445.AM2013-710
Abstract 713: Three-dimensional in vivo blood flow mapping in tumors using optical frequency domain imaging.
Authors: Blauvelt David , Mohan Nishant , Bouma Brett , Jain Rakesh , Padera Timothy , Vakoc Benjamin
Published: August 2014

Angiogenesis in the tumor microenvironment is a chaotic process that results in a dysfunctional vascular network. Unlike normal blood vessels, tumor vessels are dense, tortuous, and leaky. The disorganized nature of the tumor vascular network often leads to unequal perfusion in different parts of the tumor. While some vessels see high blood flow rates, others can be virtually stagnant. This heterogeneity in blood flow can affect oxygen distribution, and, as a result, the efficacy of radiation and chemotherapy. Furthermore, knowing how blood flow varies as a tumor develops is critical for our comprehension of tumor angiogenesis. Finally, our understanding of how anti-angiogenic and chemotherapeutic treatments affect tumor vessels depends on our ability to monitor blood flow. Unfortunately, existing strategies for measuring blood flow are limited by low spatial resolution, long imaging times, or an inability to measure flow over whole tumor volumes. Here, we use optical frequency domain imaging (OFDI) to measure blood flow in individual vessels over an entire tumor volume growing in a mouse. Previously, we have used OFDI to obtain high-resolution images of tumor vasculature. Our new technique is based on an association between the speed of moving blood and the dynamics of optical speckle patterns that are intrinsically generated in OFDI. By employing novel methods to measure these speckle patterns and then relate them to an underlying flow velocity, we are able to generate high-resolution three-dimensional maps of blood flow within tumors. Our results expand the potential of OFDI to characterize the tumor microvasculature by adding wide-field blood flow measurements to its demonstrated ability to characterize microvascular morphology, tumor anatomy, tissue viability, and lymphatic vessel morphology.

Citation Format: David Blauvelt, Nishant Mohan, Brett Bouma, Rakesh Jain, Timothy Padera, Benjamin Vakoc. Three-dimensional in vivo blood flow mapping in tumors using optical frequency domain imaging. [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 713. doi:10.1158/1538-7445.AM2013-713
Abstract 717: Detection of premalignancy of esophageal mucosa by multispectral imaging of autofluorescence from protoporphyrin IX.
Authors: Horiuchi Toshikatsu , Morimoto Yuji , Taniguchi Hiroaki , Umetsu Araya , Ogura Shun-ichiro , Iwaya Keiichi , Shinomiya Nariyoshi
Published: August 2014

Background: We have recently found that chemically induced esophageal carcinoma emitted a strong fluorescence originating from protoporphyrin IX (PpIX). The aim of this study was to detect early changes in the esophageal mucosa for clarifying the mechanism of carcinogenesis. For this purpose, we established a novel method for detection of premalignancy based on multispectral fluorescence imaging (MFI) of autofluorescence (AF) from tissue. Methods: We made an N-nitroso-methylbutylamine (NMBA)-induced rat esophageal tumor model (15 mg/L in daily drinking water) and extracted the esophagus. The mucosal side was illuminated by an excitation blue light (405 nm) and the emitted light was captured on an EM-CCD detector (512 x 512 pixels), of which each pixel collected a spectrum ranging from 450 nm to 800 nm. The extracted tissues were further analyzed to measure porphyrins by high-performance lipid chromatography (HPLC). Results: When the mucosa was illuminated by the excitation light, AF with a peak at 630 nm was observed in preneoplastic lesions and HPLC revealed that the origin of AF was PpIX. MFI based on the spectral difference clearly visualized preneoplastic lesions: the spectral pattern of the AF from preneoplastic lesions was different from that from normal mucosa, and the spectral difference became greater with increase in the duration of NMBA administration and the concentration of PpIX also increased with the time course. Conclusion: The results showed that MFI of tissue AF has a potential for cancer screening.

Citation Format: Toshikatsu Horiuchi, Yuji Morimoto, Hiroaki Taniguchi, Araya Umetsu, Shun-ichiro Ogura, Keiichi Iwaya, Nariyoshi Shinomiya. Detection of premalignancy of esophageal mucosa by multispectral imaging of autofluorescence from protoporphyrin IX. [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 717. doi:10.1158/1538-7445.AM2013-717
Abstract 728: Fluorescence-guided surgery for human-patient colon cancer tumorgrafts using a fluorophore-conjugated anti-CEA antibody.
Authors: Hiroshima Yukihiko , Maawy Ali , Zhang Yong , Chishima Takashi , Tanaka Kuniya , Bouvet Michael , Endo Itaru , Hoffman Robert M.
Published: August 2014

One of the most important prognostic factors for overall survival in colorectal cancer patients is the complete resection of tumor. We have previously demonstrated the effective fluorescence-guided surgery of primary and metastatic cancer with the use of fluorescent probes in mouse models. In this study, we determined the effectiveness of fluorescence-guided surgery using a fluorophore-conjugated anti-CEA antibody for human-patient colon cancer tumorgrafts in nude mouse models. Colon-cancer-patient tumor specimens were initially established subcutaneously in NOD/SCID mice immediately after surgery. The patient tumors were then harvested from NOD/SCID mice and passed orthotopically in nude mice. Eight weeks after orthotopic implantation of human colon cancer tumorgraft, a monoclonal anti-CEA antibody conjugated with Alexa 488 was delivered to tumor-bearing mice as a single intravenous dose 24 hours before laparotomy. Using fluorescence imaging, the primary tumor was clearly visible at laparotomy. The tumor was completely resected under fluorescence navigation. Histologic evaluation of the resected specimen revealed there were no cancer cells in the margin. The cancer cells were strongly stained by anti-CEA antibody conjugated with Alexa488. Fluorescence-guided surgery using a fluorophore-conjugated anti-CEA antibody should improve intraoperative staging and efficacy of resection for CEA-positive colorectal cancer.

Citation Format: Yukihiko Hiroshima, Ali Maawy, Yong Zhang, Takashi Chishima, Kuniya Tanaka, Michael Bouvet, Itaru Endo, Robert M. Hoffman. Fluorescence-guided surgery for human-patient colon cancer tumorgrafts using a fluorophore-conjugated anti-CEA antibody. [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 728. doi:10.1158/1538-7445.AM2013-728
Abstract 729: Chimeric antibodies to tumor antigen CEA improve detection of human colon cancer for fluorescence-guided surgery.
Authors: Metildi Cristina A. , Kaushal Sharmeela , Luiken George A. , Hoffman Robert M. , Bouvet Michael
Published: August 2014

Mouse models of human colon cancer were established with fragments of a CEA-expressing patient colon tumor. Mice received tail vein injections of a human-mouse chimeric anti-CEA-Alexa-488 antibody or mouse anti-CEA-Alexa-488 antibody 24 hours prior to fluorescence-guided surgery (FGS). Pre- and post-operative images were obtained to assess for completeness of resection. Mice were subsequently followed for 6 months post-operatively to assess for tumor recurrence and overall survival. At termination, all tumor lesions were harvested and evaluated histologically. Furthermore, the chimeric antibody was tested on frozen tumor and normal tissue arrays comparing it to the mouse antibody. The chimeric antibody demonstrated improved sensitivity and specificity in labeling the CEA-expressing tumor compared to the mouse antibody. The conjugation efficiency of the fluorophore to the chimeric antibody was 2-fold higher than that of the mouse antibody. On frozen tumor tissue arrays, the signal intensity of the chimeric antibody was significantly brighter compared to the mouse antibody indicating improved binding to tumor tissue. The chimeric antibody demonstrated improved specificity as shown by the lower signal intensity in normal human tissue samples compared to the mouse antibody indicating decreased binding of the chimeric antibody. The animals injected with the chimeric antibody had more effective FGS than the animals injected with the mouse antibody indicating improved visualization of tumor margins by the chimeric antibody. The humanized chimeric antibody has greater potential for clinical application for FGS than a mouse antibody.

Citation Format: Cristina A. Metildi, Sharmeela Kaushal, George A. Luiken, Robert M. Hoffman, Michael Bouvet. Chimeric antibodies to tumor antigen CEA improve detection of human colon cancer for fluorescence-guided surgery. [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 729. doi:10.1158/1538-7445.AM2013-729
Abstract 760: Fem1b attenuates the transcriptional activity of Gli1 by promoting the ubiquitin mediated degradation of Gli1.
Authors: Gilder Andrew S. , Lu Deyin , Maher Joseph F.
Published: August 2014

Gli1 is a member of the Gli family of zinc finger transcription factors that functions in stem cell biology, and as an oncogene in malignant transformation. The Hedgehog signaling pathway in flies and mammals shares homology to the sex determination pathway of C. elegans in which the Gli-like transcription factor TRA-1 is regulated by the ubiquitin ligase adapter FEM-1. Here we provide evidence that the mammalian FEM-1 homolog Fem1b regulates Gli1 transcriptional activity. Consistent with this, we demonstrate an interaction between Fem1b and Gli1, and show that Fem1b promotes the ubiquitylation and proteasome-mediated degradation of Gli1. Fem1b mediated ubiquitylation of Gli1 was dependent on the VHL box domain of Fem1b, shown to interact with Elongin B/C-Cul2-box protein ubiquitin ligases. Fem1b reduced Gli1 transcriptional activity in a reporter assay, and this reduction was also dependent on the VHL box of Fem1b. Given that Gli1 up-regulation promotes tumorigenesis in glioblastoma, medulloblastoma, basal cell carcinoma, and a variety of epithelial malignancies, we hypothesize that Fem1b antagonizes the oncogenic capacity of Gli1 by targeting Gli1 for ubiquitin-mediated destruction in the proteasome.

Citation Format: Andrew S. Gilder, Deyin Lu, Joseph F. Maher. Fem1b attenuates the transcriptional activity of Gli1 by promoting the ubiquitin mediated degradation of Gli1. [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 760. doi:10.1158/1538-7445.AM2013-760
Abstract 762: Genome-wide mapping of binding sites for the DEK oncoprotein by ChIP-seq.
Authors: Sanden Carl , Järvstråt Linnea , Ullmark Tove , Nilsson Björn , Gullberg Urban
Published: August 2014

The DEK oncogene is overexpressed in a wide range of cancers, including acute myeloid leukemia (AML). While DEK was initially described as a protein binding to certain topological chromatin structures, recent studies implicate DEK in the epigenetic and transcriptional regulation of specific genes. To assess the global gene binding of DEK, we performed chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq). We demonstrate that DEK does not bind evenly throughout the genome, but rather accumulates around transcription start sites, with binding to promoters and initial parts of the coding sequences. Furthermore, gene set enrichment analysis revealed that DEK preferentially binds genes associated with epigenetic regulation of gene expression. Our results represent the first genome-wide characterization of DEK binding, and further establish DEK as a transcriptional regulator with a specific set of target genes.

Citation Format: Carl Sanden, Linnea Järvstråt, Tove Ullmark, Björn Nilsson, Urban Gullberg. Genome-wide mapping of binding sites for the DEK oncoprotein by ChIP-seq. [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 762. doi:10.1158/1538-7445.AM2013-762
Abstract 765: DDX31 regulates p53 tumor suppressive activity in renal cell carcinomas.
Authors: Katagiri Toyomasa , Fukawa Tomoya , Matsuo Taisuke , Kanayama Hiro-omi
Published: August 2014

Renal cell carcinoma (RCC) is the most common cancer of the kidney, and up to 30% of patients with RCC present with metastatic disease, yet its oncogenic origins are poorly understood. At an early stage, RCC can be cured by surgical resection, which is the most effective treatment for localized RCC tumors .Moreover, cytokine therapies, such as interleukin-2 or IFN-a, are widely used as a first-line treatment for metastatic disease. Therefore, it is highly important to develop a new molecular target agent(s) against RCC. However, these treatments can be associated with severe toxicity. Therefore, it is highly important to develop a new molecular target agent(s) against RCC. To identify therapeutic targets for cancer and understand the detailed molecular mechanism of carcinogenesis, we analyzed the expression profiling of clear cell RCC (ccRCC), which is a major histologic type of RCC, and identified DEAD (Asp-Glu-Ala-Asp) box polypeptide 31 (DDX31), a novel member of the DEAD box protein family, which is frequently up-regulated in the vast majority of human RCCs. Immunohistochemistry analysis indicated that DDX31 positivity was an independent prognostic factor for RCC patients. RNAi-mediated knockdown of DDX31 significantly suppressed RCC cell growth. Concordantly, expressing exogenous DDX31 promoted the growth of HEK293 cells. We also found that endogenous DDX31 interacted and colocalized with nucleophosmin (NPM1), involved in ribosome biogenesis and regulation of p53–HDM2 pathway, in nucleoli. Knockdown of DDX31 or NPM1 decreased pre-ribosome RNA biogenesis. Interestingly, in DDX31-knockdown cells, NPM1 was translocated from nucleoli to the nucleoplasm or cytoplasm, and then bound to HDM2, which prevented HDM2 from interacting with the p53 protein, resulting in p53 stabilization by inhibiting p53 ubiquitination. Our findings suggest that the DDX31–NPM1 complex plays critical roles in cell proliferation of RCC.

Citation Format: Toyomasa Katagiri, Tomoya Fukawa, Taisuke Matsuo, Hiro-omi Kanayama. DDX31 regulates p53 tumor suppressive activity in renal cell carcinomas. [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 765. doi:10.1158/1538-7445.AM2013-765
Abstract 778: Suppression of lung adenocarcinoma progression by NKX2-1 is dependent on p53 Status.
Authors: Lee Huei , Chen Po-Ming , Wu Tzu-Chin , Cheng Yawen , Chen Chih-Yi
Published: August 2014

This article investigates the mechanistic aspects of NKX2-1 as a tumour suppressor and its oncogenic properties in lung adenocarcinoma. We show that NKX2-1 is a transcriptional factor that directly regulates p53 transcription. In p53 wild-type cells, NKX2-1 interacts with SP1 to down-regulate IKKβ transcription. In p53 mutated cells, NKX2-1 elevates the mutant p53/NF-Y complex to up-regulate IKKβ transcription. NKX2-1-mediated IKKβ expression is responsible for the efficacy of soft agar growth, invasion, and xenograft lung tumour nodule formation. Moreover, tumour aggressiveness attenuated by NKX2-1-mediated IKKβ is predominately through the shuffling of Foxo3a between the cytosol and nucleus of lung cancer cells to modulate E-cadherin expression. A consistent association of NKX2-1 with IKKβ and E-cadherin expression was observed in tumours from lung adenocarcinoma patients. P53 mutated tumours with high NKX2-1 expression have poorer overall survival (OS) and relapse-free survival (RFS) rates than those with low-NKX2-1 expression. Conversely, p53 wild-type tumours with high-NKX2-1 tumours have more favourable OS and RFS rates than those with low-NKX2-1 expression. As such, we demonstrate that aberrant transcription of IKKβ underlines the ability of NKX2-1 to act as a tumour suppressor or to display oncogenic properties in lung adenocarcinoma.

Citation Format: Huei Lee, Po-Ming Chen, Tzu-Chin Wu, Yawen Cheng, Chih-Yi Chen. Suppression of lung adenocarcinoma progression by NKX2-1 is dependent on p53 Status. [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 778. doi:10.1158/1538-7445.AM2013-778
Abstract 797: Targeted next generation sequencing of FFPE lung cancer tissue.
Authors: Endris Volker , Stenzinger Albrecht , Penzel Roland , Warth Arne , Schirmacher Peter , Weichert Wilko
Published: August 2014

In view of a personalized medicine, the screening of somatic mutations in an individual tumor that predict therapeutic outcome has become an increasing need in the clinical and molecular diagnostics. In most laboratories, Sanger sequencing has become an accepted standard, but due to the limited screening throughput and the increasing demand for targeted sequencing, the application of next-generation sequencing in the molecular diagnostic has turned into focus. Whole-genome or whole-exome sequencing approaches provide a comprehensive view of an individual tumor mutation load, but the current high costs and the excess of information without a direct clinical implication limit the routine use of this technology. In this study, we evaluated a targeted resequencing approach that focuses on somatic hotspot cancer mutations based on semiconductor sequencing. We especially took attention on the reliability of the method concerning sample quality, mutation type and reproducibility. We used DNA extracted from formalin-fixed, paraffin embedded (FFPE) tumor tissues (biopsy and resection) of different tumor degree and overall quality that were previously screened for the presence of EGFR mutations (exon 18-21) by Sanger sequencing. 190 amplicons covering hotspot mutations in 46 genes were generated in a multiplex PCR reaction (AmpliSeqTM), barcoded and 8 samples were run on a single IonTorrent 318 chip. We were able to successfully sequence all samples with a mean coverage rate of 670.000 reads (AQ20:560.000) and an average read depth of 2947 AQ20 reads / amplicon. In all but one sample, variant calling identified the EGFR mutation, missing only one low level 9bp insertion in exon 20. Moreover, we identified additional 43 mutations in 17 genes and uncovered three previously unknown EGFR amplifications. Taken together, amplicon-based semiconductor sequencing is a powerful and cost-effective method working with low-quality DNA material, enabling routine diagnostic next-generation sequencing.

Citation Format: Volker Endris, Albrecht Stenzinger, Roland Penzel, Arne Warth, Peter Schirmacher, Wilko Weichert. Targeted next generation sequencing of FFPE lung cancer tissue. [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 797. doi:10.1158/1538-7445.AM2013-797
Abstract 809: High-resolution genomic profiling reveals gains of CCND1 and EGFR associated with disease progression in Taiwanese oral cavity squamous cell carcinomas.
Authors: Chien Huei-Tzu , Huang Shiang-Fu , Chen I-How , Liao Chun-Ta , Wang Hung-Ming , Cheng Sou-De , Hsieh Ling-Ling
Published: August 2014

The genetic mechanisms underlying Taiwanese oral cavity squamous cell carcinoma (OSCC) are still largely unknown. We used high-resolution single nucleotide polymorphism microarrays (Affymetrix 500K) to detect copy number alterations (CNAs) in 26 OSCC tumor tissues. Genomic copy number was determined by calculating the intensity ratio between the OSCC and a set of 270 normal DNAs from HapMap project. Segmented copy numbers for each tumor were inferred with the hidden markov model (HMM) using the Partek GS software. Genomic Identification of Significant Targets in Cancer (GISTIC) analysis (version 2.0) was used to identify significant CNAs using a residual Q-value cutoff of 0.25. A total of 25 significant regions (2 focal amplifications and 23 deletions) were identified. The well known oncogene EGFR and CCND1 are resided in these two focal amplification regions (7p11.2 and 11q13.3), respectively. Several candidate cancer related genes such as FHIT (3p14.2), FAT (4q35.1), SH3GL2 (9p22.2) and ARHGAP21 (10p12.1) are known to locate in the identified 23 deletion regions. Furthermore, we found that 7p11.2 amplification and 4q35.2 deletion was mutually exclusive by ‘inter-target relationships’ (ITR) analysis using dimension reduction permutation (DRP) methods. Copy number gains of EGFR and CCND1 were also confirmed along with fluorescence in situ hybridization (FISH) and TaqMan CN assay (Hs01818912_cn and Hs00655630_cn), respectively. We found that the concordance rate was 70.59% for EGFR and 75.00% for CCND1. To further explore the clinical value of copy number gains of EGFR and CCND1 in OSCC, 182 OSCC cases were analyzed. Preliminary results indicated that gains of CCND1 were associated with tumor differentiation, lymph node metastasis and lymph node extracapsular spread (LNECS); while amplification of EGFR gene was significantly associated with tumor stage and lymph node metastasis but not LNECS. Furthermore, gains of CCND1 were significantly associated with poor disease-free and overall survival. On the other hand, amplification of EGFR was not associated with either disease-free or overall survival. Taken together, we have identified that gains of CCND1 and EGFR gene were significantly associated with disease progression in Taiwanese oral cavity squamous cell carcinomas.

Citation Format: Huei-Tzu Chien, Shiang-Fu Huang, I-How Chen, Chun-Ta Liao, Hung-Ming Wang, Sou-De Cheng, Ling-Ling Hsieh. High-resolution genomic profiling reveals gains of CCND1 and EGFR associated with disease progression in Taiwanese oral cavity squamous cell carcinomas. [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 809. doi:10.1158/1538-7445.AM2013-809
Abstract 810: Integrative genomic and transcriptomic analysis in idenfitication of biomarkers of chemoresistance in serous epithelial ovarian cancer.
Authors: Koti Madhuri , Gooding Robert J. , Nuin Paulo , Haslehurst Alexandria , Crane Colleen , Weberpals Johanne , Childs Timothy , Bryson Peter , Dharsee Moyez , Evans Kenneth , Feilotter Harriet E. , Park Paul C. , Squire Jeremy A.
Published: August 2014

Resistance to platinum-based chemotherapy remains a major impediment in the treatment of serous epithelial ovarian cancer. The objective of this study was to use gene expression and copy number profiling to delineate major deregulated pathways and biomarker networks associated with the development of intrinsic chemotherapy resistance with exposure to standard first-line therapy for ovarian cancer. The study cohort comprised 28 high grade serous ovarian cancer patients divided into two groups based on their varying sensitivity to first-line chemotherapy using progression free survival (PFS) as a surrogate of response. Twelve patient tumors demonstrating relative resistance to platinum based chemotherapy corresponding to shorter PFS (less than 6 months) were compared to 16 tumors from platinum-sensitive patients (PFS more than 18months). Molecular profiling was performed using Affymetrix high-resolution microarray platforms to permit global comparisons of gene expression levels and copy number profiles between tumors from the resistant group with the sensitive group. Microarray data analysis revealed a set of 227 discriminating genes of which expression levels may be influencing differential chemotherapy response between the two groups. Pathway analysis of these genes showed the,PI3K,NFkB and IGF1 networks as some of the significant networks distinguishing the chemotherapy resistant with the sensitive group. Copy number analysis performed using Nexus copy number version 6.1 revealed differences in the chromosomal regions, 4q31.22, 5q13.2, 9p24.3, 2p23.2, 16q21, 6q14.1, 7p22.3, 12p13 and Xq.

Integrative copy number and gene expression profiling will delineate the drivers of chemotherapy resistance in patients undergoing standard platinum-based treatment of ovarian cancer. Future studies to validate these markers are necessary to apply this knowledge to biomarker-based clinical trials.

Citation Format: Madhuri Koti, Robert J. Gooding, Paulo Nuin, Alexandria Haslehurst, Colleen Crane, Johanne Weberpals, Timothy Childs, Peter Bryson, Moyez Dharsee, Kenneth Evans, Harriet E. Feilotter, Paul C. Park, Jeremy A. Squire. Integrative genomic and transcriptomic analysis in idenfitication of biomarkers of chemoresistance in serous epithelial ovarian 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 810. doi:10.1158/1538-7445.AM2013-810
Abstract 833: S100A4 interacts with p53 in the nucleus promoting its degradation.
Authors: Orre Lukas , Panizza Elena , Kaminskyy Vitaliy , Gräslund Torbjörn , Zhivotovsky Boris , Lehtiö Janne
Published: August 2014

Background.

S100A4 is a calcium binding protein which has no known enzymatic activity, but it exerts its function by binding and regulating other molecules, among which have been described Myosin and p53. S100A4 has been shown to be upregulated in several types of cancers, correlating with poor prognosis and metastasis.

In a previous study (Orre et al., 2007), S100A4 has been shown to be involved the cellular response to ionizing irradiation in a p53 dependent manner; our aim here was therefore to investigate the cellular interaction partners of S100A4 in response to p53 and then, based on this data, to further elucidate the cellular function of S100A4.

Observations.

Nutlin-3A is a compound that specifically stabilizes p53 through inhibiting the interaction between p53 and mdm2. We could show an interaction between S100A4 and p53 via immunoprecipitation in the NSCLC cell line A549 (which has a high expression of S100A4 and a wild-type p53), and that this interaction is increased after Nutlin-3A treatment. The interaction has been independently confirmed via Proximity Ligation Assay, showing as well that the protein complexes are localized in the nucleus.

Since this interaction increased after inhibition of the mdm2-p53 interaction, we hypothesized that S100A4 could be involved in regulation of p53 degradation; therefore we generated two cell lines from A549, infected with S100A4 shRNA (shS100A4) or an Empty Vector shRNA (shEV) as a control. Knockdown of S100A4 resulted in stabilization of p53 and transactivation of p53 transcriptional targets; it also resulted in higher sensitivity to cisplatin-mediated apoptotic cell death, measured as a drop in mitochondrial membrane potential, caspase-3 activation and AV/PI assay.

We confirmed this finding by an independent method using transient S100A4 down regulation via siRNA; this resulted in an increase in p53 level and of p21 and MDM2 level in A549 adenocarcinoma cells. S100A4 siRNA resulted also in p53-dependent growth arrest, that was reversed after co-transfecting p53 siRNA.

Conclusions.

Our data show that S100A4 interacts with p53 increasing its degradation, resulting in higher cell viability and lower apoptosis after cisplatin treatment. A possible correlation in patients between S100A4 overexpression and cisplatin resistance could be used to drive treatment selection.

Citation Format: Lukas Orre, Elena Panizza, Vitaliy Kaminskyy, Torbjörn Gräslund, Boris Zhivotovsky, Janne Lehtiö. S100A4 interacts with p53 in the nucleus promoting its degradation. [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 833. doi:10.1158/1538-7445.AM2013-833
Abstract 84: The role of redox state in human breast cancer progression.
Authors: Jorgenson Tonia C. , Zhong Weixiong , Oberley Terry D.
Published: August 2014

We are investigating the role of reduction-oxidation (redox) state in human breast cancer progression. Physiologic levels of oxidants and antioxidants within subcellular compartments regulate and coordinate normal cell functions. Cells in a state of redox imbalance undergo adaptation and/or altered behavior. We propose to investigate the thioredoxin 1 (TRX1) redox couple in human breast cancer progression. This molecule is known to interact with several molecules involved in breast carcinogenesis, including redox-sensitive transcription factors and estrogen receptor (ER) alpha. We hypothesize human breast cancer tissues are in a state of redox imbalance favoring a more reducing environment, and post-translational redox modifications involving TRX1, thioredoxin reductase 1 (TRXR1), and thioredoxin interacting protein (TXNIP) are involved. Preliminary data demonstrated redox imbalance in comparisons of breast cancers to surrounding tissues. TRX1 protein levels were 1.5-fold increased, while TRXR1 and TXNIP were 10-fold and 2.4-fold decreased, respectively. The redox state of TRX1 demonstrated a 4.7-fold increase in the reduced form versus the oxidized, and TRX1 activity was 2.1-fold increased. Our future plans involve analyzing redox-regulated protein subcellular distribution patterns of TRX1, TRXR1, TXNIP, and ER alpha utilizing immunohistochemistry in human breast cancer tissue microarrays varying in histological grade; determine protein levels of TRX1, TRXR1, and TXNIP, redox state of TRX1, and activities of TRX1 and TRXR1 in human breast cancer cell lines relative to normal primary epithelial cells in cell culture; and investigating the effects of modulating the extra- and intracellular redox state on cell invasion in hormone dependent/independent breast cancer cell lines.

Citation Format: Tonia C. Jorgenson, Weixiong Zhong, Terry D. Oberley. The role of redox state in human breast cancer progression. [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 84. doi:10.1158/1538-7445.AM2013-84
Abstract 844: Peroxisome proliferator-activated receptor γ agonists abrogate tumor invasion and promote TRAIL-induced apoptosis in malignant glioma.
Authors: Choi Kyungsun , Chong Kyuha , Lee Jungwhoi , Choi Hojun , Choi Chulhee
Published: August 2014

Glioblastoma multiforme is the most common brain tumors and have a worse prognosis, so it is needed to develop effective treatments. Peroxisome proliferator-activated receptor gamma (PPAR-γ) agonists, including 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and synthetic ligands, troglitazone and rosiglitazone, are known to sensitize tumor cells to apoptosis via various molecular mechanisms. In this study, we investigated the therapeutic potentials of combined treatment with PPAR-γ agonists and TRAIL in TRAIL-resistant astrocytoma cells. TRAIL alone and each PPAR-γ agonist exhibited no cytotoxic effects; while the pre-incubation with PPAR-γ agonists enhanced dramatically TRAIL-induced cell death, which was abrogated by a broad spectrum caspase inhibitor, z-VAD-fmk. And also, the combined treatment with PPAR-γ agonists and TRAIL increased the enzymatic activity of caspase-3. These results indicate that PPAR-γ agonists might promote TRAIL-mediated apoptosis by modulating the caspase activity. In addition, the expression of anti-apoptotic proteins including FLIP, Bcl-2, Bcl-xL and XIAP was suppressed by the combined treatment. Besides the cytotoxic effect, treatment with PPAR-γ agonists significantly inhibited glioma invasion in vitro and in vivo. These results collectively suggest that PPAR-γ agonists may provide a promising strategy to treat malignant glioma.

Citation Format: Kyungsun Choi, Kyuha Chong, Jungwhoi Lee, Hojun Choi, Chulhee Choi. Peroxisome proliferator-activated receptor γ agonists abrogate tumor invasion and promote TRAIL-induced apoptosis in malignant glioma. [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 844. doi:10.1158/1538-7445.AM2013-844
Abstract 850: Sirt1-mediated suppression of cell death in breast cancer.
Authors: Mortenson Jeffrey B. , Weerasekara Vajira K. , Andersen Josh
Published: August 2014

Several lines of evidence suggest that protein lysine acetylation pathways are deregulated in cancer (1). Moreover, deacetylase inhibitors are emerging as important anti-tumor therapeutics, suggesting that the forced reprogramming of protein-lysine acetylation is toxic to tumor cells. In this study we show that Sirt1, an NAD+-dependent Sirtuin deacetylase that promotes cancer cell survival, is aberrantly mislocalized to the cytoplasm of breast tumor cells. Moreover, the depletion of cytosolic Sirt1 by siRNA sensitizes breast tumor cells to paclitaxel-induced death. Previously, we developed a biotin-switch proteomics approach to identify cytosolic Sirt1 substrates (2). This approach yielded a variety of substrates with roles in metabolism, survival, and oxidative stress signaling. Our current work focuses on three of the proteins identified as Sirt1 substrates: SOD1, DJ-1, and 14-3-3z. SOD1 and DJ-1 both suppress oxidative stress-induced death, and high levels of 14-3-3z expression suppress chemotherapy-induced apoptosis and correlate with negative patient outcomes in breast cancer. Our preliminary results suggest that acetylation of DJ-1 and SOD1 suppress their anti-oxidant functions, while acetylation of 14-3-3z disrupts its binding to pro-survival proteins. Taken together, our data support a model in which cytosolic Sirt1 activates multiple pathways that work together to promote tumor cell survival.

Citation Format: Jeffrey B. Mortenson, Vajira K. Weerasekara, Josh Andersen. Sirt1-mediated suppression of cell death in breast 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 850. doi:10.1158/1538-7445.AM2013-850
Abstract 859: miR-638, a novel tumor suppressor for ER-negative breast cancer.
Authors: Peng Jin , Fu Yebo , Brem Rachel F. , Teal Christine B. , Fu Sidney W.
Published: August 2014

MicroRNAs (miRNAs) are endogenous non-coding RNAs of 19∼25 nt in length, which regulate many protein-coding genes. miRNA dysregulation is observed in a variety of cancers, including breast cancer. miRNAs have emerged as an important set of biomarkers. miRNA profiling studies have led to the identification of miRNAs that are aberrantly expressed in human breast cancer, with miR-10b, miR-125b and miR-145 being down-regulated and miR-21, and 155 being up-regulated. In our previous miRNA expression profiling studies using microdissected FFPE tissues, we identified 8 clusters of miRNAs, which discrete the normal and breast lesions such as ADH, DCIS and IDC. miR-638 is one of the differentially expressed miRNAs during breast cancer progression. We first sought to determine the expression of miR-638 in different breast cancer cell lines such as MCF-7, Hs578T, T47D and MDA-MB-231 using qRT-PCR, which shows a very low expression compared to normal tissues. This led us to hypothesize that miR-638 may function as a tumor suppressor during breast cancer development. To determine the role of miR-638 in both ER+ and ER- breast cancer cells, we transfected miR-638 mimic oligos and scrambled oligo mocks into MCF-7 (ER+), T47D (ER+), MDA-MB-231 (ER-) and Hs578T (ER-) cells. After 48 hours, we found that the proliferation rate was significantly down-regulated in ER- cells but not in ER+ cells by MTT assay. Our preliminary matrigel analysis shows that miR-638 overexpressing MDA-MB-231 cells had 3∼4 fold invasiveness decrease compared to the control. Further, our TargetScan analysis revealed that BRCA1 is one of the direct targets of miR-638 among the 30 conservative target genes. We found that overexpression of miR-638 resulted in an upregulation of BRCA1 mRNA expressions in ER- cell lines, MDA-MB-231 and Hs578T but not in ER+ cell lines, T47D and MCF-7. Taken together, these findings showed that miR-638 may function as a tumor suppressor in ER- breast cancer, possibly in part by upregulating BRCA1 tumor suppressor gene. Further functional analysis is underway to decipher the exact role of miR-638 in ER- breast cancer, which could become a therapeutic target.

Citation Format: Jin Peng, Yebo Fu, Rachel F. Brem, Christine B. Teal, Sidney W. Fu. miR-638, a novel tumor suppressor for ER-negative breast 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 859. doi:10.1158/1538-7445.AM2013-859
Abstract 866: Cetuximab treatment reduces VEGF expression and targeting of radiolabeled bevacizumab in breast cancer xenografts.
Authors: Heskamp Sandra , Boerman Otto C. , Molkenboer-Kuenen Janneke D.M. , Oyen Wim J.G. , van der Graaf Winette T.A. , van Laarhoven Hanneke W.M.
Published: August 2014

Introduction: Bevacizumab (anti-VEGF) and cetuximab (anti- EGFR) are approved for the treatment of cancer. However, in advanced colorectal cancer, the combination of bevacizumab and cetuximab did not improve survival (Tol et al. NEJM 2009). Up to now, the reason for the lack of activity of the combination therapy remains unclear. Previously, we have shown that bevacizumab reduces the vascular density of xenografts and consequently limits the delivery of cetuximab to tumors. Thereby, it can potentially limit the therapeutic efficacy of cetuximab. Alternatively, cetuximab treatment could alter VEGF expression, limiting tumor targeting by bevacizumab. The aim of this study was to determine the effect of cetuximab treatment on VEGF expression and targeting of bevacizumab to the tumor.

Material and methods: Mice with subcutaneously implanted SUM149 xenografts, which express EGFR and VEGF, were treated intraperitoneally with 1 mg/kg cetuximab (twice a week). Before start of treatment and after 7, 14, and 21 days, the uptake of radiolabeled bevacizumab in the tumor was measured by immunoSPECT/CT. Three days prior to scanning, mice received 15 MBq of 111In-bevacizumab. After scanning, tumors were dissected for ex vivo biodistribution and immunohistochemical analysis of VEGF and CD34 expression.

Results: After cetuximab treatment, tumor targeting of bevacizumab was significantly reduced on immunoSPECT/CT compared to untreated tumors. Tumor uptake measured ex vivo was 28.9 ± 4.0 %ID/g for untreated tumors, compared to 18.2 ± 2.8 %ID/g for tumors treated 21 days with cetuximab (p = 0.009). Immunohistochemical analysis showed that the vascular density was unaltered while VEGF expression decreased during cetuximab treatment.

Conclusion: Cetuximab treatment reduced VEGF expression and targeting of radiolabeled bevacizumab to the tumor. This could at least partly explain why the combination of bevacizumab and cetuximab does not result in improved therapeutic efficacy.

Citation Format: Sandra Heskamp, Otto C. Boerman, Janneke D.M. Molkenboer-Kuenen, Wim J.G. Oyen, Winette T.A. van der Graaf, Hanneke W.M. van Laarhoven. Cetuximab treatment reduces VEGF expression and targeting of radiolabeled bevacizumab in breast cancer xenografts. [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 866. doi:10.1158/1538-7445.AM2013-866
Abstract 872: Novel potent and orally active STAT3 inhibitors for cancer therapies.
Authors: Yang Zhengduo , Chen Haijun , Chu Lili , Zhang Yusong , Ding Chunyong , Liu Huiling , Zhou Jia , Shen Qiang
Published: August 2014

Signal transducers and activators of transcription (STATs) are proteins which have dual roles as cytoplasmic signaling proteins and transcription factors that activate sets of genes including those contribute to tumorigenesis. Among the STATs family members, STAT3 has been shown to play an important role in breast carcinogenesis and is often overexpressed in primary breast tumors. Increased STAT3 transcriptional activity is reported to correlate with ER-negative phenotype in breast cancer cell lines and in primary human invasive ductal breast carcinomas. Therefore, STAT3 is a promising target for breast cancer therapies. Current strategies of inhibiting STAT3 activity did not sufficiently block STAT3 activation in cancer cells due to their limited potency, poor solubility or bioavailability. Despite several STAT3 inhibitors have been advanced into early clinical trials, there is an urgent need to develop orally active, potent STAT3 inhibitors as cancer therapeutics. We have designed, synthesized and screened a series of novel STAT3 inhibitors utilizing fragment-based drug design strategy. A number of small molecules with novel scaffolds have been discovered with low-micromolar to nanomolar potency in inhibiting the proliferation of various human breast cancer and pancreatic cancer cells. The identified compounds HJC0152, HJC0123, HJC0125, and HJC0416 significantly suppress STAT3 promoter activity and STAT3 phosphorylation (pY705), reduce total STAT3 protein expression, induce cell growth arrest, and early apoptosis in ER-negative breast cancer cell line MDA-MB-231. These novel compounds have demonstrated a better efficacy in suppressing the growth of MDA-MB-231 breast xenograft tumors via intraperitoneal and oral administrative routes, with remarkably improved aqueous solubility and less toxicity than the lead compounds. In conclusion, we have developed a new class of orally active STAT3 inhibitors with superior drug-like properties, for further evaluation in cancer therapeutic and preventive settings. This work was supported by The University of Texas MD Anderson Cancer Center DFI Funding Program.

Citation Format: Zhengduo Yang, Haijun Chen, Lili Chu, Yusong Zhang, Chunyong Ding, Huiling Liu, Jia Zhou, Qiang Shen. Novel potent and orally active STAT3 inhibitors for cancer therapies. [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 872. doi:10.1158/1538-7445.AM2013-872
Abstract 885: Human organic cation transporter 1 (hOCT1) as a mediator of bendamustine cytotoxic action.
Authors: Arimany-Nardi Cristina , Montraveta Arnau , Lee-Vergés Eriong , Koepsell Hermann , Colomer Dolors , Pastor-Anglada Marçal
Published: August 2014

Bendamustine hydrochloride is a cytotoxic agent that encompasses structural features of alkylating and nucleoside-derived drugs. This drug is currently used in the treatment of several haemotological malignancies; among them Chronic Lymphocytic Leukemia (CLL). CLL is the most common leukemia in Western countries and is characterized by the accumulation of monoclonal CD5+ B cells in the blood, bone marrow, lymph nodes, spleen and other organs. The pathways facilitating the uptake of bendamustine hydrochloride into target cells still remain poorly understood. Here we have addressed this issue by monitoring the interaction of the drug with selected plasma membrane transporter candidates, which might be relevant to bendamustine pharmacokinetics and pharmacodynamics. We have used a panel of HEK293 cells stably expressing selected drug transporter proteins to prove that bendamustine is able to interact with human Organic Cation Transporter 1 (hOCT1) with a Ki of 140+/-10 μM, as deduced from cis-inhibition studies using tritiated 1-methyl-4-phenylpyridinium (MPP+) as hOCT model substrate. Genetic variants known to modify substrate/transporter interactions, some of them with high allelic frequency in humans, were also tested and, in particular, hOCT1M420Del showed a significant decrease in its affinity for the drug (Ki value 234+/-24 μM, p<0.05 vs the wild type transporter). The whole panel of hOCT1 and hOCT1 allelic variants were also tested for bendamustine-induced cytotoxicity and data consistent with bendamustine being a hOCT1 substrate were obtained. This was based upon the finding that hOCT1 expression triggered increased sensitivity to the drug and variants known to decrease drug-transporter affinity interactions were also shown to induce resistance. hOCT1 is expressed in CLL cells and genotyping of a cohort of 125 CLL patients is now allowing the analysis of the relationships between particular genetic variants and ex vivo bendamustine-induced cytotoxicity, thus opening the possibility that hOCT1 genotyping might be a useful tool in optimizing bendamustine-based therapies.

Citation Format: Cristina Arimany-Nardi, Arnau Montraveta, Eriong Lee-Vergés, Hermann Koepsell, Dolors Colomer, Marçal Pastor-Anglada. Human organic cation transporter 1 (hOCT1) as a mediator of bendamustine cytotoxic action. [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 885. doi:10.1158/1538-7445.AM2013-885