bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5499: Kinase inhibitor selectivity profiling utilizing a versatile TR-FRET kinase binding assay platform
Authors: Lebakken Connie S. , Koepnick Elizabeth , Bruinsma Rica , Riddle Steve 
Published: February 2011

Abstract
There is great potential for the utilization of selective kinase inhibitors in the treatment of many clinical indications, especially in oncology and autoimmune conditions. Kinase inhibitor selectivity profiling has become an important tool during lead development to understand a compound's selectivity, potential off-target effects and possible new indications for the compound. We have utilized the LanthaScreen® Eu Kinase Binding Assay platform to develop a simple, robust means for profiling compounds across the kinome utilizing pre-plated reagents and controls. The assay is based on competitive displacement of a tracer (a fluorophore conjugated to a kinase inhibitor scaffold) from specific kinases of interest. Binding of the tracer to a kinase is detected by addition of a europium-labeled anti-tag antibody. Binding of the tracer and antibody to a kinase results in a high degree of FRET, whereas displacement of the tracer with a kinase inhibitor results in a loss of FRET. This platform is useful to explore activation-state binding selectivity, time-dependent binding, and binding to kinases for which substrates have not been identified. The assay detects both ATP-competitive as well as non-competitive, allosteric kinase inhibitors. We will present data demonstrating the utility of these reagents for kinase profiling efforts with a variety of kinase inhibitors.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5499.
Abstract 55: Glucose metabolism influences migration of high-grade glioma in vitro by modification of TGF-beta2
Authors: Leukel Petra , Seliger Corinna , Vollmann Arabel , Doerfelt Anett , Bogdahn Ulrich , Hau Peter 
Published: February 2011

Abstract
Introduction: Lactate dehydrogenase type A (LDH-A) is a key metabolic enzyme catalyzing pyruvate into lactate and is excessively expressed in several human tumors. Transforming growth factor-beta2 (TGF-beta2) is a key regulator of invasion in high-grade gliomas, partially remodeling the extracellular matrix (ECM) and inducing proteinases. Thrombospondin1 (THBS-1) is an extracellular protein important for activation and processing of TGF-beta2. A microarray of LDH-A knocked-down glioma cell RNA showed down regulation of THBS-1 and TGF-beta2. In this study, we tested the hypothesis that LDH-A influences TGF-beta2 activation by up regulation of THBS-1 leading to an enhanced migration of high-grade glioma in vitro.
Methods: We performed LDH-A knock-down by transient transfection of glioma cells with small interfering RNA directed against LDH-A (siLDH-A). Expression levels of TGF-beta2 and THBS-1 in siLDH-A transfected cells were investigated using microarrays, RT-PCR, Western Blot and ELISA. Migration of transfected cells was investigated by Boyden Chamber and scratch assays.
Results: siLDH-A suppresses TGF-beta2 in high-grade glioma and decreases the expression of THBS-1 on the RNA and protein level. THBS-1 leads to an increased level of activated TGF-beta2 in supernatants of siLDH-A treated cells. In migration assays, siLDH-A leads to a decreased migration of high-grade glioma cells.
Discussion: We demonstrate, for the first time, that knockdown of LDH-A can decrease the RNA and protein level of TSP-1 and consecutively the processing of TGF-beta2. Additionally, knockdown of LDH-A decreases the RNA level of TGF-beta2. Both results may contribute to an enhanced level of TGF-beta2 and increased migration, given that LDH-A is expressed. An increased expression of LDH-A has been found in aerobic glycolysis, a mechanism well known from several human cancers. Recent paper showed that LDH-A is able to bind RNA. Thus we suppose LDH-A could influence the level of TGF-beta2 RNA by RNA stabilization. Together with our recent results that show that TGF-beta enhances migration in high-grade gliomas, we demonstrate a new panel of interactions between lactate metabolism and TGF-beta2 that might be crucial for glioma migration and possibly invasion.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 55.
Abstract 5500: Biochromatography for the screening of new drugs for the estrogen related receptors
Authors: Sanghvi Mitesh , Moaddel Ruin , Frazier Chester , Gandhari Mukesh , Wainer Irving W. 
Published: February 2011

Abstract
Estrogen related receptors, ERRα, ERRβ, and ERRγ, form a subfamily of orphan nuclear receptors that have significant amino acid homology with the estrogen receptors ERα and ERβ. ERRs have been identified in breast cancer tumors and ERRα has been associated with a negative response to treatment while ERRγ is considered an indicator of a positive response. We have recently demonstrated that ERRα and ERRγ are expressed in cell lines derived from astrocytoma and glioblastoma tumors. We have also demonstrated that there are different levels of expression of ERRα and ERRγ in the cell lines and this difference may be the source of the variation of these tumors to anticancer agents. MTT assays were performed to measure the metabolic activity of T98G cells treated with the ERRγ antagonists.
While ERRs share significant Ligand binding domain (LBD) homology with ERs, they have different responses to characterized ER agonists and antagonists. For example, the ER agonist estradiol is inactive in ERR expressing cell lines, the ER agonist diethyl stilbestrol is an antagonist of all three ERR subtypes, 4-hydroxytamoxifen does not affect ERRα and acts instead as an ERRγ antagonist. In addition, we have recently demonstrated that ERRα and ERRγ antagonists can effectively inhibit tumor growth. Based on this observation, we have created liquid chromatographic columns containing immobilized LBDs of the ERRα and ERRγ. In the initial study, the LBD of ERRγ was covalently immobilized onto the surface of an aminopropyl silica liquid chromatography stationary phase. The protein was immobilized via the N-terminus to create the ERRγ-column. The column was characterized using frontal chromatographic techniques and a series of tamoxifen derivatives were studied. Additionally, we have identified a second site to which diethylstilbestrol (DES) binds.
Acknowledgement
This research was supported entirely or in part by the Intramural Research Program of the NIH, National Institute on Aging.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5500.
Abstract 5503: Dual fluorescence high throughput screen detects MUC16 selective therapeutic candidates
Authors: Thapi Dharmarao , Normand Guillaume , Ivnitski-Steele Irena D. , Ostrovnaya Irina , Shum David , Spriggs David R. , Djaballah Hakim 
Published: February 2011

Abstract
BACKGROUND: The MUC16 gene encodes the CA125 ovarian cancer antigen. This glycoprotein is expressed in the majority of Epithelial Ovarian Cancers (EOC) but has limited expression elsewhere in the adult human. Expression of the carboxy-portion of MUC16 results in increased invasion, soft agar growth and in vivo growth.
OBJECTIVES: We sought to develop a suitable microscopy based assay system to screen for and identify small molecules which have a differential effects against MUC16+ cells compared to the isogenic MUC16 negative ovarian cancer cell lines. Using this assay we seek to identify MUC16 specific small molecules for development of potential chemotherapy drug candidates against ovarian cancer.
METHODS: Carboxy-terminus portions of the Human MUC16 gene were introduced into SK-Ov-3 and A2780 EOC cell lines and labeled with the red fluorescent protein (mCherry) vector. Control lines were labeled with green fluorescent protein (GFP) vector. Relative toxicity of tested compounds on MUC16+ (mCherry) and MUC16- (GFP) cell populations was assessed by cell viability using ratiometric fluorescence intensities of both co-cultured isogenic cells. Compounds with differential activity were selected, resupplied and tested in a 96 well plate viability assay for confirmation.
RESULTS: Both A2780 and SK-Ov-3 isogenic pairs had similar in vitro growth characteristics. A small scale pilot screen of 3,119 compounds was completed for each pair of isogenic cells. Six compounds, including Quinacrine, Ellipticine and Acriflavinium were relatively selective for MUC16+ A2780 cells and these three compounds were among the hits identified in the secondary SK-Ov-3 dual fluorescence screen. Caspase3 apoptosis assay was used to confirm Quinacrine as the most selective inhibitor in the MUC16+ cells.
CONCLUSIONS: MUC16 +/− dual fluorescence screen is an example of a high throughput screening strategy to identify potentially specific modulators of key oncogenic targets.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5503.
Abstract 5506: Novel toxin library for the discovery of oncology therapeutics
Authors: Zacny Valerie , Higley John S. , Cunningham Casey , Poma Eric E. 
Published: February 2011

Abstract
The A subunit of Shiga-like Toxin 1 (SLT1) is a ribosome-inactivating protein that works by catalyzing the depurination of the 28 S rRNA. The A subunit lacks the ability to bind and internalize into cells, and is normally dependent on the B subunit of SLT1 for this function. A toxin variant library has been created by inserting random amino acid stretches into an exposed loop structure of the A subunit. The inserts can confer binding and internalization properties to the SLT1 A variants but do not inhibit the A subunit's ability to intoxicate. This toxin library has been expressed in a phage display system that can be rapidly screened for variants with specific cell kill activity and predictable PK/PD properties. The library is being used to develop novel therapeutics for oncology.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5506.
Abstract 5512: Zirconium phosphate nanoparticle matrix as potential delivery system for cisplatin to cancer cells in vitro
Authors: González Millie , Díaz Agustín , Colón Jorge , Clearfield Abraham , Ortíz Mayra , Báez Adriana 
Published: February 2011

Abstract
Cisplatin (cis-diamminedichloroplatinum(II), have been shown to be active against ovarian and testicular cancer, Hodgkin's lymphoma, and certain other malignancies, however, its clinical effectiveness is limited by significant side effects and the emergence of drug resistance. Delivery of anticancer drugs such as cisplatin to the tumor cells without damaging healthy organs or tissues is highly difficult if not impossible. In the last several years nanoparticle-mediated drug delivery is being studied as a novel valuable mean for overcoming this problem. We developed a novel nanoparticle intercalated formulation of cisplatin on a layered matrix of zirconium phosphate (ZrP). It was expected that cisplatin, while being excluded from the external medium within ZrP layers will remain biologically inactive and that cisplatin will be released selectively in the tumor cell. The purpose of this study was to determine the cytotoxicity and effectiveness of ZrP nanoparticles in the delivery of cisplatin. Cell viability was measured by MTT assay. Flow cytometry analysis was used to determine cisplatin-induced apoptosis, and cell cycle distribution. ZrP alone did not affect the viability of breast cancer cells (MCF-7), human leukemic lymphoblast (CEM) and human peripheral blood lymphocytes (PBLs) after 24 hours in contact with different concentrations of ZrP. Cisplatin intercalated into ZrP at the highest concentration (30 μM cisplatin) reduced in 40% the cell viability of MCF-7 and CEM. The IC50 of cisplatin alone in MCF-7 was 10 μM. The side scatter plot showed that normal PBLs scatter plot is different from PBLs that have been in contact with cisplatin-intercalated ZrP for 18 hours. These PBLs carrying ZrP were displaced to the right quadrant suggesting an increased granularity that can be associated to the incorporation of ZrP nanoparticles. Moderate cisplatin-induced apoptosis was evident in cells treated with ZrP-intercalated cisplatin for 18 hours. The cell cycle analysis showed that ZrP alone does not cause cell cycle arrest. Our results suggest that application of ZrP nanoparticle-mediated drug transport in cancer chemotherapy is highly promising and more studies are warranted.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5512.
Abstract 5529: Silencing H-ferritin by siRNA delivered by cationic liposomes increases chemotherapeutic sensitivity for treating malignant tumor
Authors: Liu Xiaoli , Madhankumar A.b. , Slagle-Webb Becky , Sheehan Jonas M. , Surguladze Nodar , Connor James R. 
Published: February 2011

Abstract
Cancer cells have a robust appetite for iron to support their rapid growth and high metabolic rates. The high levels of iron uptake require a mechanism to limit iron induced oxidative damage. This function is the responsibility of ferritin; an intracellular iron storage protein. Ferritin has been found in the nucleus of tumor cells where it appears to protect DNA from iron induced oxidative damage and promote transcription. Therefore, we hypothesized that silencing H-ferritin gene expression could increase the sensitivity of malignant tumors to radiation and chemotoxins. Potentially targets of H-ferritin disrupted iron homeostasis in tumor cells result in proving effective at negatively affecting tumor growth. To test our hypothesis, H-ferritin siRNA was delivered to U251 human astrocytoma cells, malignant peripheral nerve sheath tumor (MPNST) cells and a breast cancer cell line (MCF-7) via our cationic liposomes. Western immunoblotting analysis demonstrated that the siRNA delivered in this manner decreased H-ferritin protein expression by 70 to 80% in all three cell lines within 48 hours. The results from SRB assay displayed decrease H-ferritin was associated with a decrease in the LD50 for carmustine (BCNU) from greater than 100 µM to 40 μM in the U251 cells, 30 µM in MCF7 cells and 18 µM in MPNST cells. In addition, the results of MTS assay revealed the presence of H-ferritin siRNA was associated with a 50% increase in cell death of U251 cells at 20 Gy of radiation. The in vivo efficacy of siRNA delivered by cationic-liposomes was tested in the Athymic nude mice subcutaneous glioma tumor model. Intratumoral injections of cationic liposomes containing H-ferritin siRNA reduced the required effective dose of BCNU for tumor suppression by more than 50%. The interaction of H-ferritin with DNA was investigated in vitro by supercoil relaxation assay, which demonstrated that H-ferritin maintained DNA in a relaxed form that would promote transcription of DNA even in the presence of radiation and chemotoxins. The outcome of study for Caspase-3 activity exposed that transfection with H-ferritin siRNA was associated with activation of apoptotic caspase pathway. Overall, our data demonstrated that silencing the H-ferritin gene appears to be an effective way to sensitize tumors to radiation and chemotherapy. As importantly, we have demonstrated that siRNA can be delivered effectively by cationic liposomes in an in vivo tumor model.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5529.
Abstract 553: Host cell OGR1 plays an important role in TRAMP cell tumorigenesis
Authors: Yan Libo , Wei Gang , Li Hui , Zhao Zhenwen , Singh Lisam Shanjukumar , Xu Yan 
Published: February 2011

Abstract
Ovarian cancer G protein coupled receptor 1 (OGR1) was previously cloned from an ovarian cancer cell line HEY. We have recently shown that OGR1 is a new tumor metastasis suppressor gene in prostate cancer. To further understand the physiological role of OGR1, we have generated OGR1 knockout (KO) mice. These mice are viable and grossly normal under regular physiological conditions. To exam the role of host cell OGR1 in prostate cancer development, TRAMP-C2 cells (a mouse prostate cancer cell line) were subcutaneously (s.c.) injected into OGR1 KO mice, significantly reduced tumorigenesis were observed when compared to that in wild type (WT) mice. 100% (20 of 20) of wild type mice developed tumors, while only 12.5% (4 of 32) of OGR1 deficient mice developed small tumors. To test whether OGR1 deficiency in host cells inhibits tumorigenesis of other mouse prostate tumor cells, RM-9 cells (an aggressive and poorly immunogenic mouse prostate tumor cells) were subcutaneously injected into OGR1 KO and WT mice (2 mice/group). All 4 mice developed big tumors at 10 days post tumor cell injection. No significant difference was observed between WT and OGR1 KO mice. Thus, host OGR1 deficiency differentially affected tumorigenesis of TRAMP and RM-9 cells, suggesting a tumor cell type specific effect. OGR1 are highly expressed in immune cells. We are currently testing whether these cells are functionally involved in host OGR1's effects. In addition, fibroblasts have a prominent role in the progression, growth and spread of cancers. The infiltration of fibroblasts in TRAMP-C2 tumors from WT and OGR1 KO mice were tested. Decreased fibroblast infiltration was observed in tumors from the KO vs. WT mice. The expression of OGR1 in mouse fibroblasts were confirmed by RT-PCR. Our results suggest that fibroblasts may be involved in OGR1's effect on tumorigenesis. Our new findings may lead to novel therapeutic strategies for prostate and/or other cancers.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 553.
Abstract 5531: Preparation and cellular delivery of doxorubicin nanoemulsion for cancer therapy
Authors: Pamujula Sarala , Chinta Dakshina Murthy Devanga , King Ashley , Kishore Vimal , Mandal Tarun K. 
Published: February 2011

Abstract
Purpose: The aim of this study is to formulate a novel nanoemulsion of doxorubicin to enhance the therapeutic efficacy of the drug.
Method: Four different multiple emulsions (W1/O/W2) were prepared (A, B, C, and D) by homogenization of a mixture of aqueous and oil phases. The resultant primary emulsions were later pass through a high pressure homogenization to further reduce the globules size to nanometer range. The formulation A is a control nanoemulsion, consisted of only deionized water as W1 phase. The W1 phase of the other three formulations (B, C, and D) consisted of aqueous solution of BSA (as an emulsion stabilizer), doxorubicin, and a mixture of BSA and doxorubicin, respectively. For all four formulations, the oil phase (O) consisted of a 2:1 mixture of soyabean oil and Span 80, and the outer aqueous phase (W2) consisted of prolaxamer P-85 and Vitamin E TPGS. The nanoemulsions were evaluated for stability, polydispersity index, size, and viscosity. Anticancer efficacy of these formulations was evaluated by Alamar Blue Assay and Colony Forming Assay using MCF-7 cells as a model malignant breast cancer cell line.
Results: The stability of nanoemulsions was found to be significantly better for the formulations containing BSA. Presence of BSA or doxorubicin resulted in a significant increase in viscosity from 2.09 to 10.6 cP. Cell culture studies showed that doxorubicin nanoemulsions had higher localization in cytoplasm as compared to the nucleus. In regard to cellular uptake (intracellular quantification of doxorubicin), the following results were obtained at 2h, 24h, 48h, and 120h post-incubation, respectively: 42%, 30%, 37%, and 22% with doxorubicin solution; 12%, 19%, 21%, 33% with formulation C; and 0.25%, 6%, 10%, 19% with formulation D. The results of Alamar Blue Assay showed IC50 values of 0.97μM (solution), 1.62μM (formulation C) and 1.91μM (formulation D). The results of Colony Forming Assay (12days-post-treatment with equivalent to 0.86 μM doxorubicin) showed 74% (solution), 86% (formulation C) and 92.6% (formulation D) growth inhibitions.
Conclusions: The presence of BSA within the aqueous phase enhances the stability of nanoemulsions. In comparison to doxorubicin solution, the nanoemulsions produce higher anticancer efficacy. This exploratory study will be followed by more detailed studies to confirm this preliminary finding.
Acknowledgements: This work was funded in part by the NASA grant # NCC3-946-S4, Louisiana Board of Regents RC/EEP (2007-10), NIH grant# 5P20CA118768-02, and Louisiana Cancer Research Consortium. We are thankful to Dr. Thomas Wiese of the College of Pharmacy, XULA, for providing the Cell culture lab facilities.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5531.