bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3200: Spliced variant of T-cell intercellular antigen 1 (TIA-1) in colorectal cancer.
Authors: Hamdollah-Zadeh Maryam , Ye Xi , Domingo Enric , Kerr David J. , Ladomery Mike R. , Harpar Steve J. , Bates Dave O.
Published: August 2014

Alternative splicing regulates many cancer-associated genes. Splice isoforms that are found in tumors could have diagnostic value and may provide potential drug targets. We identified an endogenous splice variant of TIA-1 (T-cell intercellular antigen-1), short TIA-1 (shTIA-1), expressed in invasive colon cancer cells, which was not expressed in non-invasive adenoma cell lines. In this study we investigated the frequency of shTIA-1 expression in human normal and cancer tissue and if it correlates with advanced-stage of colon cancer.

RNA was extracted from 25μm thick scrolls of formalin fixed paraffin embedded sections of 40 colorectal carcinoma (20 Duke's, 20 Duke's C stage) and normal samples. RT-PCR was carried out to detect full length (flTIA-1) and shTIA-1. Immunohistochemical CD31 staining of microvessels in the same samples was used to assess microvessel density (MVD) and correlate the shTIA isoform expression and stages of the tumors. The results showed that flTIA-1 was less frequently found in tumour than in normal tissues (p=0.002). shTIA-1 was more highly expressed than flTIA-1 and in significantly higher portion of tumors than normal samples (p=0.0106). There was a significantly lower proportion of Duke's B than Duke's C grade tumours with detectable shTIA-1 expression (p=0.018). CD31 staining showed samples with shTIA-1 associated with increased MVD. We did not find any association between stages and MVD, as there was no statistically significant difference in MVD between Duke's B and Duke's C. However tumours had higher MVD than normal tissues from the same patients. These results indicate that a splice variant of TIA-1 is expressed in human colorectal carcinoma and is associated with increased angiogenesis.

Citation Format: Maryam Hamdollah-Zadeh, Xi Ye, Enric Domingo, David J. Kerr, Mike R. Ladomery, Steve J. Harpar, Dave O. Bates. Spliced variant of T-cell intercellular antigen 1 (TIA-1) in colorectal 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 3200. doi:10.1158/1538-7445.AM2013-3200
Abstract 3206: Multistage vectored siRNA targeting ataxia-telangiectasia mutated for breast cancer therapy.
Authors: Xu Rong , Huang Yi , Mai Jun Hua , Zhang Guo Dong , Guo Xiao Jing , Xia Xiao Jun , Koay Eugene J , Liu Xue Wu , Ferrari Mauro , Shen Hai Fa
Published: August 2014

The ataxia-telangiectasia mutated (ATM) protein plays a central role in DNA damage response and cell cycle checkpoints, and may be a promising target for cancer therapy if normal tissue toxicity could be avoided. Our strategy to target ATM for breast cancer therapy involves the use of liposomal-encapsulated, gene-specific ATM small interfering RNA (siRNA) delivered with a well-characterized porous silicon-based multistage vector (MSV) delivery system (MSV/ATM). Here we have shown that biweekly treatment of MSV/ATM suppressed ATM expression in tumor tissues, and consequently inhibited growth of MDA-MB-231 orthotopic tumor in nude mice. At the therapeutic dosage, neither free liposomal ATM siRNA nor MSV/ATM triggered acute immune response in BALB/c mice, including changes in serum cytokines, chemokines or colony-stimulating factors. Weekly treatments of mice with free liposomal ATM siRNA or MSV/ATM for 4 weeks did not cause significant changes in body weight, hematology, blood biochemistry, or major organ histology. These results indicate that MSV/ATM is biocompatible and efficacious in inhibiting tumor growth, and that further preclinical evaluation is warranted for the development of MSV/ATM as a potential therapeutic agent.

Citation Format: Rong Xu, Yi Huang, Jun Hua Mai, Guo Dong Zhang, Xiao Jing Guo, Xiao Jun Xia, Eugene J Koay, Xue Wu Liu, Mauro Ferrari, Hai Fa Shen. Multistage vectored siRNA targeting ataxia-telangiectasia mutated for breast cancer therapy. [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 3206. doi:10.1158/1538-7445.AM2013-3206
Abstract 3212: Capillary electrophoresis for upstream and downstream biopharmaceutical development.
Authors: Nguyen Uyen , Ramirez Francisco , Mack Scott , Darling Susan , Proctor John , Boge Annegret
Published: August 2014

In order to bring an antibody or other biologic drug to market, each step of the process needs to be carefully monitored. Charge heterogeneity and apparent MW via SDS-PAGE are two of the most commonly assessed parameters. Ideally, similar methods should be used from the beginning of development (low expression, complex matrix) to late in production and QC (high concentration, pure material).Capillary techniques such as iCE and CE-SDS are currently heavily used in downstream product development but pose challenges for small scale screening and characterization during cell culture and formulation development. Here we present the Simple Western that combines in one novel instrument, Peggy, capillary electrophoresis with an immunoassay to provide highly reproducible and fully automated analysis of monoclonal antibodies. This sensitive technology measures either size or charge in complex samples and provides critical charge heterogeneity, size, and product titer information without the need for sample purification. Data will be presented demonstrating the application of the Simple Western technique and the iCE technology for the analysis of monoclonal antibodies against VEGF and the ability of the two techniques to provide consistency of data across the whole range of product development. In addition we show examples for the unique capability of this technology to assess affinity information for these anti VEGF antibodies to different charge isoforms of VEGF.

Citation Format: Uyen Nguyen, Francisco Ramirez, Scott Mack, Susan Darling, John Proctor, Annegret Boge. Capillary electrophoresis for upstream and downstream biopharmaceutical development. [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 3212. doi:10.1158/1538-7445.AM2013-3212
Abstract 3215: Detecting low abundance therapeutic targets for ovarian cancer using engineered gold nanoparticles.
Authors: Giri Karuna , Arvizo Rochelle
Published: August 2014

Gold nanoparticles (AuNPs) have been widely studied for use in disease therapeutics as targeting and imaging agents, drug delivery vehicles and as self-therapeutics. When AuNPs interact with the biological milieu they form a corona layer, which is predominantly composed of proteins. The outer “soft” layer is dynamic and here proteins are free to exchange over time. The inner “hard” layer on the other hand consists of proteins firmly bound to the NP surface. The Vromans effect predicts that given a limited surface area, low affinity, high abundance proteins, that first attach to the surface, are over time replaced by high affinity, low abundance proteins. Our aim was to enrich low abundance proteins on the AuNP surface to probe for differentially expressed proteins that are not detected by traditional methods of protein identification. We studied the binding of ovarian cell lysates (cancerous and non cancerous) to 10 nm sized positively and negatively charged AuNPs. The formation of corona around the AuNPs was characterized by dynamic light scattering and zeta potential measurements. The composition of the protein corona was identified using mass spectrometry. Proteins were also identified from the cell lysate pool to serve as a control to assess protein enrichment on NP surface. Using this approach, Hepatoma Derived Growth Factor (HDGF) was identified as a low abundance protein that was detected by mass spectrometry in cancer cells only after enrichment on the surface of positively charged AuNPs. HDGF is overexpressed in multiple ovarian cancer cells lines and knock down using siRNA shows decrease in cell proliferation, G2/M arrest and apoptosis. This suggests an important role of HDGF in the tumorigenicity of ovarian cancer, which could serve as an important therapeutic target for the disease.

Citation Format: Karuna Giri, Rochelle Arvizo. Detecting low abundance therapeutic targets for ovarian cancer using engineered gold nanoparticles. [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 3215. doi:10.1158/1538-7445.AM2013-3215
Abstract 3220: A microfluidic, multi-analyte, high sensitivity, immunoassay platform for rapid biomarker detection.
Authors: Pande Rajiv , Leamon John H. , Putnam Marty , Pitta M , Svancara D , Tu J
Published: August 2014

Traditionally, protein analytes or biomarkers have been measured individually in ELISAs. However, it is becoming increasingly clear that multiple markers are associated with multivariate diseases like cancer. Despite demand, the adoption of multi-analyte biomarker tests in clinical research has been severely limited for many reasons including technical concerns regarding assay reproducibility, time and the labor-intensive nature of assay panel development, and reported non-correlation with conventional ELISA data. To address this issue, we have developed a novel fully automated desktop immunoassay platform that enables simultaneous interrogation of four analytes in eight individual samples on a single disposable microfluidic cartridge in under an hour. We have devised a unique approach to immobilize antibodies that results in assays with high sensitivities (femtomolar) and broad dynamic ranges (>4 logs). Twenty microliters of patient sample are split into parallel channels containing individual immunoassays. By segregating each assay to a discrete location, the multi-analyte assay mimics single-analyte ELISAs, thus enabling each assay to operate under its optimal conditions. This methodology combines the specificity of a traditional ELISA immunoassay, where the sample is assayed by a singular pair of capture and detect antibodies eliminating cross-reactivity and other negative interactions from other antibody pairs while offering the benefits of a multiplexed antigen analysis. We describe the system's design, speed, sensitivity and reproducibility using data generated with a custom-built 4-analyte pro-inflammatory panel cartridge (IL-1β, IL-6, IL-10 and TNFα) on clinical samples. We demonstrate that our assay system is ideal for quantitative analysis of soluble protein markers in clinical research and in clinical trials.

Citation Format: Rajiv Pande, John H. Leamon, Marty Putnam, M Pitta, D Svancara, J Tu. A microfluidic, multi-analyte, high sensitivity, immunoassay platform for rapid biomarker 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 3220. doi:10.1158/1538-7445.AM2013-3220
Abstract 3222: Broad spectrum metabolite profiling reveals metabolic finger and footprinting which correlate to gene expression signatures of triple-negative breast cancer (TNBC).
Authors: Beatty Alexander , Fink Lauren , Strigun Alexander , Rennefahrt Ulrike , Schmitz Oliver , Schiewe Hajo , Moeller Niels , Noppinger Patricia R. , Reszka Regina , Peterson Jeffrey R.
Published: August 2014

TNBC constitutes ∼20% of all breast cancers and disproportionately affects younger women and African-American women. This genomically heterogeneous disease is defined by the lack of expression of estrogen and progesterone receptors and the lack of amplification of HER2/neu. TNBC is resistant to standard breast cancer therapies. Due to the lack of a predominant genetic lesion associated with the disease, identification of metabolic fingerprints within TNBC is a critical first step for identification of molecular drivers and development of targeted therapies. Metabolite profiling (metabolomics) of a wide range of metabolites can be used to globally profile the metabolic state of cancer cells. Nine widely used TNBC-derived cell lines already characterized by gene expression signatures were analyzed by metabolomics. Two different approaches were combined: (i) untargeted, broad profiling including LC-MS/MS and GC-MS, and (ii) targeted UPLC-MS/MS Energy platform technology which covers polar key metabolites mainly reflecting energy status and nucleotide biosynthesis. Finally, 239 high quality metabolites were statistically evaluated in cell lysates and 128 metabolites in spent media samples collected from all cell lines. Using innovative bioinformatics and data mining tools we identified metabolic alterations in energy utilization, lipid synthesis, and other pathways of importance to highly proliferative cells that differ significantly from an untransformed control cell line (MCF-10A). Major metabolic changes combining results of metabolic fingerprinting and footprinting and their biological relevance will be discussed.

Citation Format: Alexander Beatty, Lauren Fink, Alexander Strigun, Ulrike Rennefahrt, Oliver Schmitz, Hajo Schiewe, Niels Moeller, Patricia R. Noppinger, Regina Reszka, Jeffrey R. Peterson. Broad spectrum metabolite profiling reveals metabolic finger and footprinting which correlate to gene expression signatures of triple-negative breast cancer (TNBC). [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 3222. doi:10.1158/1538-7445.AM2013-3222
Abstract 3224: Metabolite profiling as tool for the identification of differentiating and prognostic markers of prostate carcinoma.
Authors: Kristiansen Glen , Reszka Regina , Kamlage Beate , Bethan Bianca , Lein Michael , Stephan Carsten , Jung Klaus
Published: August 2014

Background and objective: Metabolomic research offers a deeper insight into biochemical changes in cancer metabolism and is a promising tool for identifying novel biomarkers. We aimed to evaluate the diagnostic and prognostic potential of metabolites in prostate cancer (PCa) tissue after radical prostatectomy.

Material and methods: 107 matched-paired tissue samples collected after radical prostatectomy were subjected to the MxPTM Broad Profiling by gas chromatography- mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (Patent WO 2010/139711 A1: "Means and methods for diagnosing prostate carcinomas"). Aminoadipic acid, cerebronic acid, gluconic acid, glycerophosphoethanolamine, 2- hydroxybehenic acid, isopentenyl pyrophosphate, maltotriose, 7-methylguanine, and tricosanoic acid were related to clinicopathological variables like prostate volume, tumor stage, Gleason score, preoperative prostate-specific antigen (PSA), and disease recurrence in the follow-up. Non-parametric statistical tests, receiver- operating characteristics (ROC) and univariate and multivariate analyses (Kaplan- Meier curve; Cox regression) were performed.

Results: All metabolites showed higher concentrations in malignant than in non- malignant samples except for gluconic acid and maltotriose, which had lower levels in tumors. ROC analyses showed a clear differentiation for all metabolites with a maximal area under the curve of 0.86 for tricosanoic acid. However, the metabolites were not related to tumor stage and Gleason grade. "Aminoadipic acid, gluconic acid, and maltotriose levels were associated with tumor recurrence (Kaplan-Meier analysis) and were, together with tumor stage and Gleason score, a successful metabolite combination in the multivariate Cox regression model for the prediction of tumor recurrence."

Conclusions: This exemplary study performed with selected metabolites from a global metabolic profiling investigation proves that metabolites in prostate carcinoma tissue can be used, in combination with traditional pathological and histomorphological parameters, as promising diagnostic and prognostic tools.

Citation Format: Glen Kristiansen, Regina Reszka, Beate Kamlage, Bianca Bethan, Michael Lein, Carsten Stephan, Klaus Jung. Metabolite profiling as tool for the identification of differentiating and prognostic markers of prostate 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 3224. doi:10.1158/1538-7445.AM2013-3224
Abstract 3237: Combination therapy of paclitaxel and EW-7197, a novel ALK5 inhibitor prolonged the life span of breast tumor bearing mice.
Authors: Kim Sol J. , Son Jee Y. , Kim Min J. , Park Sang A. , Lee Seon J. , Park So Y. , Kim Seung W. , Kim Dae-Kee , Nam Jeong S. , Sheen Yhun Y.
Published: August 2014

TGF-β has multifunctional effects on carcinogenesis. In the early stage tumor, TGF-β acts as a tumor suppressor but in later stage of tumor, TGF-β promotes tumor invasiveness and metastasis. TGF-βRI kinase has been the target of anti-metastatic drugs, so we have synthesized new ALK5 inhibitor (EW-7197) and investigated anti-metastatic effects of EW-7197. The treatment of the EW-7197 also inhibited the luciferase activity in the lung of 4T1-Luc cell-orthotopic implanted mouse compared to that of untreated animal. The levels of MMP2 and MMP9 were decreased in the lung and breast tumor of the EW-7197 treated mice. To determine the inhibitory effect of EW-7197 on TGF-β1-induced transcriptional activation, we performed luciferase reporter gene assay in breast cancer cells. EW-7197 effectively blocked TGF-β1-induced luciferase activity in the cells in a dose-dependent manner. TGFß signaling, the phosphorylation of smad2/3 was decreased by EW-7197. TGFß-stimulated mesenchymal markers of epithelial mesenchymal transition (EMT) were inhibited by ALK5 inhibitor concomitant treatment. And also, EW-7197 showed higher inhibition on cell motility and cell invasion based on wound healing assay or invasion assay. Further, we investigated combination effect of EW-7197 and paclitaxel. To investigate anti-metastatic effect we did experiment on BALB/c-nu/nu MB231-luc cells xenografted mice by treating EW-7197 and paclitaxel. Both EW-7197 alone group and EW-7197 and paclitaxel combination group showed inhibition of metastasis and the expression of lung metastatic markers were inhibited. We experiment on BALB/c-nu/nu MB231 cells xenografted mice for survival analysis. EW-7197 and paclitaxel combination therapy lengthen the life span of mice. In summary, EW-7197 could be effective therapeutic choice for breast cancer metastasis. Also EW-7197 and paclitaxel combination therapy could be more effective than paclitaxel alone therapy for anti-metastatic effect and survival.

Citation Format: Sol J. Kim, Jee Y. Son, Min J. Kim, Sang A. Park, Seon J. Lee, So Y. Park, Seung W. Kim, Dae-Kee Kim, Jeong S. Nam, Yhun Y. Sheen. Combination therapy of paclitaxel and EW-7197, a novel ALK5 inhibitor prolonged the life span of breast tumor bearing mice. [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 3237. doi:10.1158/1538-7445.AM2013-3237
Abstract 3243: DIM inhibits growth of human gastric cancer through modulation of the hippo signaling pathway.
Authors: Li Xiu Juan , Lee Hwan Hee , Kim Soo Mi
Published: August 2014

3, 3-diindolylmethane (DIM) has been reported that it has anti-proliferative effects in many cancer cells. However, the biological functions of DIM in human gastric cancer cells are unknown. Genetic and biological studies have confirmed the importance of the novel hippo tumor suppressor pathway in regulating cell proliferation, apoptosis, organ size, and tumorigenesis in mammals. Thus, the purpose of this study was to investigate the cytotoxic effects of DIM in human gastric cancer cells and to elucidate whether DIM induces cell death by activating the hippo signaling pathway. Four human gastric cancer cell lines (SNU1, SNU16, SNU484, and SNU638) were used to test the response of DIM. MTT, cell cycle and western blot analyses were conducted. DIM significantly inhibited the proliferation of human gastric cancer cells in a dose-dependent manner. The percentage of G1 phase cells increased 24h after being treated by DIM. DIM reduced CDK2, CDK4, CDK6, CyclinD1 protein levels and increased p53 protein levels. DIM also induced cleaved poly (ADP-ribose) polymerase, cleaved-caspase-9 levels, and diminished pro-caspase-3 protein expression levels. Additionally, DIM increased P-LATS1, Mob1, P-Mob1, P-Yap protein levels and reduced Yap protein levels. Moreover, DIM inhibited the growth of human esophageal tumors in an orthotopic mouse model. These results suggest that DIM can sensitize gastric cancers by modulating the hippo signaling pathway linked to tumorigenesis.

Citation Format: Xiu Juan Li, Hwan Hee Lee, Soo Mi Kim. DIM inhibits growth of human gastric cancer through modulation of the hippo signaling pathway. [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 3243. doi:10.1158/1538-7445.AM2013-3243

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 3255: MLN0128, an investigational selective TORC1/2 kinase inhibitor, demonstrates potent antitumor activity in preclinical models of soft tissue sarcoma.
Authors: Patwardhan Parag , Deraje Shyamprasad Vasudeva , Tap William D. , De Stanchina Elisa , Schwartz Gary K.
Published: August 2014

Sarcomas, cancers of mesodermal origin, represent a heterogenous group of malignancies with limited treatment options currently available. The mammalian target of rapamycin (mTOR), a serine-threonine protein kinase, consists of two distinct multi-protein complexes, TORC1 and TORC2, which regulate protein synthesis, cell proliferation, and metabolism. Rapamycin, a TORC1 inhibitor, and its analogs (rapalogs) have been disappointing in sarcoma clinical studies but inhibitors of TORC1/2 kinase have the potential of resulting in better outcomes. MLN0128, an investigational potent small molecule TORC1/2 inhibitor, has shown promising results in preclinical studies of breast and other cancer types. We tested the in vitro anti-proliferative activity of MLN0128 in a panel of sarcoma cell lines including malignant peripheral nerve sheath tumor, synovial, Ewing's and osteosarcoma. Our results indicate that when compared to rapamycin, MLN0128 is highly efficient in blocking proliferative activity of all the sarcoma cell lines tested at nanomolar concentrations irrespective of the tumor subtype. We also carried out western blot analysis to test the efficacy of MLN0128 in blocking TORC1/2 signaling pathway. Inhibition of TORC1 pathway by rapamycin in sarcomas has been shown to release negative feedback inhibition of receptor tyrosine kinases like IGF1-R and activate AKT (p-AKT). Our data suggest that MLN0128, unlike rapamycin, effectively blocks both the phosphorylation of S6 and 4EBP1, the downstream substrates of TORC1, and specifically inhibits AKT phosphorylation at Ser473, the downstream substrate of TORC2. This superior in vitro activity of MLN0128 may be attributed to its apparent improved TORC1 inhibition. Taken together, our data supports further clinical research of MLN0128 in sarcoma. Targeting TORC1/2 pathways using MLN0128 either alone or in combination with inhibitors of receptor tyrosine kinases could provide a new therapeutic approach in this disease.

Citation Format: Parag Patwardhan, Shyamprasad Vasudeva Deraje, William D. Tap, Elisa De Stanchina, Gary K. Schwartz. MLN0128, an investigational selective TORC1/2 kinase inhibitor, demonstrates potent antitumor activity in preclinical models of soft tissue sarcoma. [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 3255. doi:10.1158/1538-7445.AM2013-3255
Abstract 3263: Evaluation of the activity of rigosertib, an inhibitor of PI3K and MAPK pathways, in a preclinical model of colorectal cancer.
Authors: Purkey Alicia , Song Eun-Kee , Quackenbush Kevin , Bagby Stacey , Arcaroli John J. , Jimeno Antonio , Messersmith Wells
Published: August 2014

Background: Rigosertib is an inhibitor of the PI3K and MAPK kinase pathways and is currently being evaluated in clinical trials for pancreatic cancer and hematologic diseases. The PI3K/AKT and MAPK (RAS/RAF/MEK/ERK) signaling pathways have been identified to play a central role in the development and progression of CRC by modulating many different downstream targets that are essential for enhancing cellular survival and proliferation. Mutations in the PI3K (PIK3CA) and MAPK (RAS) pathways have been identified and their importance is well-established in oncogenesis and as drivers of tumor growth. The objective of this study is to determine the anti-proliferative and anti-tumor effects of rigosertib in a preclinical model of colorectal cancer (CRC).

Experimental Design: We used colorectal cancer cell lines and a patient-derived explant model to investigate the growth inhibitory activity of rigosertib. We treated 25 CRC cell lines with rigosertib for 72 hours and measured the effect on proliferation by a sulforhodamine B (SRB) assay. We treated 5 patient-derived CRC explants with rigosertib (250 mg/kg/day-daily) for 28 days to determine anti-tumor activity in vivo. A tumor growth inhibition inhibition (TGI) ≥ 50% was considered sensitive.

Results: Twenty-four CRC cell lines were sensitive to rigosertib with an IC50 > 1μM. The cell lines MIP-101, Lovo, and Caco-2 were the most sensitive reaching an IC50 at 0.0625 μM. The Colo678 cell line was resistant to rigosertib with an IC50 > 1μM. There was no association between mutational status in PIK3CA or RAS and sensitivity to rigosertib. Zero out of 5 CRC explants showed sensitivity to rigosertib.

Conclusion: Rigosertib is a potent growth inhibitor in vitro of the majority of CRC cell lines tested with an IC50 < 1μM. Our upcoming studies will test the inhibitory effects of rigosertib on an additional 15 CRC explants in an attempt to develop biomarkers of sensitivity or resistance.

Acknowledgements: The work was supported by a grant from Onconova Therapeutics® Inc.

Citation Format: Alicia Purkey, Eun-Kee Song, Kevin Quackenbush, Stacey Bagby, John J. Arcaroli, Antonio Jimeno, Wells Messersmith. Evaluation of the activity of rigosertib, an inhibitor of PI3K and MAPK pathways, in a preclinical model of colorectal 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 3263. doi:10.1158/1538-7445.AM2013-3263
Abstract 3270: Improved response of mitogen activated protein kinase kinase 1/2-dependent tumors using combination drug therapy.
Authors: Andersen Nicholas John , Boguslawski Elissa , Froman Roe , Duesbery Nicholas
Published: August 2014

Mitogen activated protein kinase kinase 1/2 (MEK1/2) inhibitors have failed to demonstrate a sustained clinical benefit for the treatment of melanoma or other MEK1/2 dependent cancers. We hypothesize combined treatment with inhibitors of MEK1/2 plus another cancer pathway would provide a more durable tumor response for MEK1/2 driven cancers. To test this we examined in vitro synergy between MEK1/2 inhibitor PD0325901 and inhibitors to eleven common cancer pathways using MEK1/2-driven melanoma cell lines and canine hemangiosarcoma cell isolates. Combination indices (CI) were calculated following the methods of Chou and Talalay (Chou and Talalay, 1984). Of the combinations tested, tyrosine kinase receptor inhibitors sorafenib and dasatinib, and the dual PI3 kinase and mTOR inhibitor rapamycin showed synergy (CI<0.7) with PD0325901. To validate these results we subsequently tested synergy for PD0325901 plus sorafenib in vivo using mouse xenograft models for melanoma and canine angiosarcoma. We conclude dual inhibition of MEK1/2 and receptor tyrosine kinases or mTOR signaling are useful strategies to treat MEK1/2 dependent cancers.

Citation Format: Nicholas John Andersen, Elissa Boguslawski, Roe Froman, Nicholas Duesbery. Improved response of mitogen activated protein kinase kinase 1/2-dependent tumors using combination drug therapy. [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 3270. doi:10.1158/1538-7445.AM2013-3270
Abstract 3273: TAK1 inhibition sensitizes neuroblastoma cells to chemotherapy.
Authors: Fan Yihui
Published: August 2014

Treatment failure in high risk neuroblastoma is largely due to development of chemoresistance. NF-κB activation is one of the resistance mechanisms for cancer cells to escape from chemotherapy-induced cell-death. TAK1 is an essential component in genotoxic stresses-induced NF-κB activation; however, the role of TAK1 in the development of chemoresistance in neuroblastoma remains unknown. Using a panel of neuroblastoma cell lines, we found that TAK1 inhibitor 5Z-7-oxozeaenol significantly augmented the cytotoxic effects of doxorubicin (Dox) and etoposide (VP-16) on neuroblastoma cell lines. TAK1 inhibition also enhanced the inhibitory effect of Dox and VP-16 on anchorage-independent growth. Treatment of neuroblastoma cells with 5Z-7-oxozeaenol blocked Dox-induced NF-κB activation and enhanced Dox-induced apoptosis. Moreover, 5Z-7-oxozeaenol could overcome the established chemoresistance in LA-N-6 neuroblastoma cells. Using an orthotopic neuroblastoma mouse model, we found that 5Z-7-oxozeaenol significantly enhanced chemotherapeutic efficacy in vivo. Together, we provide a proof-of-concept that TAK1 inhibition significantly enhances the sensitivity of neuroblastoma cells to chemotherapy-induced cell-death and can serve as an effective adjunct to current chemotherapeutic regimens for high risk disease. Since the requirement of TAK1 kinase activity in genotoxic stresses-induced NF-κB activation is not limited by cell type, this suggests that TAK1 inhibition may greatly enhance the chemotherapeutic efficacy in other cancers.

Keywords: Neuroblastoma; TAK1 inhibitor; 5Z-7-oxozeaenol; chemotherapy

Citation Format: Yihui Fan. TAK1 inhibition sensitizes neuroblastoma cells to chemotherapy. [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 3273. doi:10.1158/1538-7445.AM2013-3273

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 3279: Enhanced in vivo inhibition of neuropeptide-mediated castration resistant prostate cancer progression by combination therapy with enzalutamide and saracatinib.
Authors: Yang Joy C. , Ngyuen Hao G. , Gao Allen C. , Evans Christopher P.
Published: August 2014

Introduction: It has been shown that overexpression of the neuropeptides gastrin-releasing peptide (GRP) in LNCaP cells renders the cells castration-resistant through both androgen receptor (AR) activation and intracrine androgen biosynthesis. Aberrant activation of AR by GRP is mediated through Src kinase activation, which may be inhibited by the specific SFK inhibitor saracatinib. Inhibition of cognate AR activation requires anti-androgen. We hypothesize that treatment of this kind of CRPC demands the combination of saracatinib and new generation anti-androgen enzalutamide (MDV-3100).

Materials and methods: LNCaP GRP-Pro cells grown in CS medium were treated with control, saracatinib (2 μM), enzalutamide (10 μM) or the combination. Cell proliferation over 7 days was monitored by MTT assays. Cell cycle distributions over 72 hours of treatment were analyzed by flow cytometry. Cells were collected to prepare nuclear and cytosolic lysates respectively and examined by Western Blotting analysis for AR, pSrc, pAkt, LC-3 and β-actin. Expression of full-length AR and variants were compared by RT-qPCR. Orthotopic injection of GRP-Pro cells into castrated SCID male mice was used as the animal model for drug treatment studies.

Results: Enzalutamide or saracatinib alone only slightly inhibited CR cell growth but the combination brought it down to 27% of the control. Apoptosis was also enhanced with the combination. Enzalutamide only inhibited AR nuclear translocation in regular serum-kept GRP-Pro cells, while saracatinib inhibited the translocation in CS conditions. In the orthotopic model, neither treatments with saracatinib (25 mg/kg/daily) or enzalutamide (10 mg/kg/daily) alone showed any effect on tumor take, tumor weight or serum PSA levels. However, the combination treatment drastically reduced the tumor weights (0.66±0.13 g for controls vs. 0.24±0.07 g for the combo) and serum PSA levels (120.79±27.87 ng/ml vs. 19.19±7.45 ng/ml).

Conclusions: For GRP mediated CRPC growth, neither saracatinib or enzalutamide alone is sufficient to inhibit AR translocation and tumor progression. Combination of the two significantly improved the treatment for the CRPC orthotopic tumor model.

Citation Format: Joy C. Yang, Hao G. Ngyuen, Allen C. Gao, Christopher P. Evans. Enhanced in vivo inhibition of neuropeptide-mediated castration resistant prostate cancer progression by combination therapy with enzalutamide and saracatinib. [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 3279. doi:10.1158/1538-7445.AM2013-3279
Abstract 3284: An in vitro synergism between Ganoderma lucidum and Bacillus Calmette Guérin on premalignant transitional cell carcinoma.
Authors: Yuen John W. , Gohel Mayur-Danny I. , Ng Chi-Fai
Published: August 2014

Introduction: The major challenge for superficial transitional cell carcinoma (TCC) of the urinary bladder is recurrence. In the past three decades, Bacillus Calmette Guérin (BCG) has been used as an effective adjuvant therapy through the eradication of residual adverse cells following the surgical ablation of TCC tumors. Despite BCG has recently suspended for manufacture and shortage in supply, it provides moderate efficacy by reducing 30-40% of the recurrence cases but has high side effects toll of allergic cystitis. A medicinal mushroom - Ganoderma lucidum (GL) is known for its un-disputed scientific evidences which supporting the therapeutic roles on different cancers including TCC. In our previous reports, the triterpene-enriched ethanol extract of GL (GLe) was found to have similar immunological activities with BCG, in addition to apoptosis and telomerase inhibition. Therefore, BCG and GLe synergism was investigated. Brief Methodologies: The drug-drug interactions between BCG and GLe were assessed in an established carcinogen-sensitive human uroepithelial cell (HUC-PC) line, using two treatment schedules, i.e. co-treatment and pre-treatment with GLe. Outcome measures were cytoxicity and cytokines secretion using automated Vi-CELL cell viability analyzer (Beckman Coulter, USA) and FlowCytomix method (Bender MedSystems, Austria) with flow cytometry, respectively. Treated cells and culture media were harvested for measurements at 24 hours and 5 days upon the completion of the treatment schedules. Results and Discussion: In comparison with BCG, GLe was shown to exhibit immediate cytotoxic effects on the HUC-PC cells following incubation, whereas both drugs were individually active in stimulating the production of interleukin (IL)-6, IL-8 and monocyte chemotactic protin-1 (MCP-1). In both treatment schedules, complete cytostasis by maintaining the baseline cell seeding numbers was demonstrated in 24 hours, and cell eradication processes were progressively executed in the next 5 days in dose-dependent manners. By isobologram analysis, the interaction between BCG and GLe on cytotoxicity was shown to be synergistic. However, the pre-treatment of GLe has also resulted in suppression of all cytokines without influencing its cytotoxic effect. Conclusion: The results suggested that pre-treatment of GLe before BCG incubation could be effective to enhance the efficacy in adverse cells eradication, and potentially useful for counteracting the inflammation-associated side effects.

Citation Format: John W. Yuen, Mayur-Danny I. Gohel, Chi-Fai Ng. An in vitro synergism between Ganoderma lucidum and Bacillus Calmette Guérin on premalignant transitional 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 3284. doi:10.1158/1538-7445.AM2013-3284
Abstract 3288: Isolation of anti-neoplastic drug candidates against metastatic breast, lungs and ovarian carcinoma cells.
Authors: Ozes Osman N.
Published: August 2014

Cancer is a leading cause of death in man and women worl-wide. In recent years, wold-wide increase of obesity and diabetes, consumption of food additives and pollution are affecting quality of life and the incidense of all types of cancers are on the rise. Although new treatment regimens are also introduced into clinic war against cancer is still not won.

Treatment of cancer with synthetic chemoterapics come with a unfavorable side effects which often ends therapy. Instead of investing in synthetic compounds, OZES BIOPHARMA is engaged in discovery of anti-neoplastic drug candidates from endemic plants of TURKEY. We have been extracting and analyzing organic compounds from variety of plants consumed as a food. Our search in past two years yielded very enchoraging results and we have discovered 6 novel compounds named OZS 1-6. Our compounds show significant levels of anti-tumor and anti-metastatic activities against human metastatic and non-metastatic breast cancer cells(MDA-231,MCF7), lung cancer cells (A549),

Ovarian cancer cells (A2780) and its cisplatin resistant clone. Currently, we are idientifying the pathways inhibited or activated by our compounds.

Citation Format: Osman N. Ozes. Isolation of anti-neoplastic drug candidates against metastatic breast, lungs and ovarian carcinoma 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 3288. doi:10.1158/1538-7445.AM2013-3288
Abstract 3292: The potential chemomodulatory effects of didox and resveratrol on doxorubicin in colon cancer cells.
Authors: Khaleel Sahar A. , Al-Abd Ahmed M. , Ali Azza A. , Abdel-Naim Ashraf B.
Published: August 2014

Resveratrol and didox are polyhydroxy phenols that possess potential anticancer and chemosensitizing effects. The efficacy of doxorubicin in the treatment of colorectal cancer is limited by multi-drug resistance. Herein, we have assessed the chemomodulatory effect of resveratrol and didox on doxorubicin in colorectal cancer cell lines (HCT116 and HT29). Furthermore, we have investigated the molecular aspects of this interaction. Resveratrol was found to possess anti-cancer properties; however with a relatively low potency in HCT116 (IC50 = 17.5 μM) and HT29 (IC50 = 187 μM). IC50 values of Didox in HCT 116 and HT29 was 105 μM and 501 μM, respectively. In terms of Doxorubicin, the IC50-values were 0.96 μM and 0.88 μM in HCT116 and HT29, respectively. Resveratrol combination with doxorubicin significantly reduced its IC50-values to 0.59 μM in HCT116, and to 0.49 μM in HT29. Moreover, Didox combination significantly increased the potency of doxorubicin showing IC50 shift to 0.4 μM in HCT116, and to 0.29 μM in HT29. The combination between doxorubicin and resveratrol/didox in HCT116 and HT29 was found to be additive (combination index ranged from 0.8-1.2). The pro-apoptotic genes, p53 and bax, were markedly elevated after combination of doxorubicin with resveratrol or didox in both HCT116. On the other hand, the anti apoptotic genes, bcl2 and bclx, were downregulated. Caspase-3 concentration revealed marked increase due to combination of didox with doxorubicin in both cell lines. On the other hand, resveratrol combination with doxorubicin showed marginal elevation of caspase-3 concentration in HT-29 cells while marked elevation in HCT116 cells. In conclusion, didox and resveratrol potentiated the efficacy of doxorubicin in colorectal cancer cell lines.

Citation Format: Sahar A. Khaleel, Ahmed M. Al-Abd, Azza A. Ali, Ashraf B. Abdel-Naim. The potential chemomodulatory effects of didox and resveratrol on doxorubicin in colon 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 3292. doi:10.1158/1538-7445.AM2013-3292
Abstract 3298: Neotargeting HER2 negative cancer cells with Trastuzumab-based photoimmunotherapy by viral transduction of HER2-extracellular domain.
Authors: Ishida Michihiro , Kagawa Shunsuke , Shimoyama Kyoko , Watamabe Shinichiro , Noma Kazuhiro , Tazawa Hiroshi , Hashimoto Yuuri , Matsuoka Junji , Kobayashi Hisataka , Fujiwara Toshiyoshi
Published: August 2014

The prognosis of HER2 positive breast cancer has been improved by Trastuzumab which features high specificity and limited side effects. Numerous randomized trials have confirmed the efficacy and safety of Trastuzumab in HER2-overexpressing cancers. However, many patients who initially respond to Trastuzumab ultimately develop disease progression. Strategies to overcome Trastuzumab-refractory cancer are sought, and drug-conjugation is one of candidates. Trastuzumab-IR700 which is Trastuzumab conjugated with a near-infrared (NIR) photosensitizer, phthalocyanine dye, IR700, has recently been developed as a new type of molecular-target photoimmunotherapy(PIT). This PIT can induce target-selective necrotic cell death only at the place which have Trastuzumab's target part and where exposure of NIR light. Disappointingly, HER2-targeting therapy is applicable to only Her2-expressig tumors which consist of approximately 20-25% of primary breast cancers. In previous study, we developed the adenoviral vector to express HER2-extracellular domain (HER2-ECD), Ad/HER-ECD, and confirmed that it efficiently induced HER2 expression for HER2 negative and HER2-downregulated human cancer cells. So, we hypothesized that Ad/HER-ECD expand the range of tumor entities suitable for Trastuzumab-based PIT. By using Ad/HER-ECD and Trastuzumab-IR700 with PIT we revealed that the direct cell membrane destruction are induced at HER2 negative cancer cells, similarly at HER2 positive cancer cells, in vitro. These results suggest that combination of virally-transduction of target antigen and an antibody-based PIT would expand and potentiate target therapy even for target-negative or attenuated cancer cells.

Citation Format: Michihiro Ishida, Shunsuke Kagawa, Kyoko Shimoyama, Shinichiro Watamabe, Kazuhiro Noma, Hiroshi Tazawa, Yuuri Hashimoto, Junji Matsuoka, Hisataka Kobayashi, Toshiyoshi Fujiwara. Neotargeting HER2 negative cancer cells with Trastuzumab-based photoimmunotherapy by viral transduction of HER2-extracellular domain. [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 3298. doi:10.1158/1538-7445.AM2013-3298
Abstract 3299: Salmonella commensal subspecies: a new model for the treatment of human cancer.
Authors: Ly Vy Khan , Viloria Edward , Saxena Priyanka , Sokhom Bossakara , McClelland Michael , Arrach Nabil
Published: August 2014

Salmonella can colonize and survive in both hypoxic and oxygenated regions of solid tumors. Avirulent Salmonella mutants have been investigated as anticancer agents in human clinical trials. The strains used, derived from Salmonella enterica subspecies I (sspI) strains, proved to be highly toxic. Salmonella non-subspecies I (nsspI) strains are known as commensals of cold-blooded species and rarely cause disease in humans. Therefore, nsspI strains could present a safer alternative for the treatment of human cancer. Here, we explore the potential of nsspI strains for use as an anticancer therapy. Strains representing the five nsspI subspecies were tested for virulence and tumor targeting in mice, and for their ability to invade and kill human breast, prostate, and colorectal cancer cell lines. We identified nsspI strains that were successful at tumor targeting and capable of invading and killing a variety of cancer cell lines in vitro while being incapable of causing disease in mice. The nsspI strains were engineered to express two key therapeutic enzymes: cytosine deaminase and uracil phosphoribosyltransferase. After invading cancer cells in culture, intracellular bacteria from all five nsspI classes caused significant cytotoxicity in the presence of 5-FC. We conclude that nsspI strains could serve as potential improved anticancer vectors, to be used in combination with a therapeutic gene, for the treatment of primary or metastatic tumors. Due to their safety profile, nsspI strains present an excellent alternative to conventional Salmonella sspI strains for the treatment of human cancer.

Citation Format: Vy Khan Ly, Edward Viloria, Priyanka Saxena, Bossakara Sokhom, Michael McClelland, Nabil Arrach. Salmonella commensal subspecies: a new model for the treatment of human 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 3299. doi:10.1158/1538-7445.AM2013-3299

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.