bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 272: Direct binding of neuregulin-1 to integrins is involved in neuregulin-1/ErbB signaling
Authors: Ieguchi Katsuaki , Ma Zi , Takada Yoko K. , Takada Yoshikazu 
Published: February 2011

Abstract
Crosstalk between growth factor receptors and integrins play a role in growth factor signaling. In current models of NRG/ErbB signaling, integrins and ErbB receptors transduce signals independently upon binding to their ligands (e.g., extracellular matrix and NRGs, respectively) and their signals are merged inside the cells. We discovered that the EGF-like domain of NRG1 binds directly to integrins αvβ3 and α6β4. Docking simulation predicted that three Lys residues at the amino terminus of NRG1 are involved in integrin binding. We mutated these residues to Glu (designated the 3KE mutant). Indeed the 3KE mutation markedly reduced the NRG1 binding to integrins, while the 3KE mutation did not affect the NRG1 binding to ErbB3. This is consistent with the prediction that the Lys residues critical for integrin binding are not involved in ErbB binding. We studied the effect of the 3KE mutation on NRG1/ErbB signaling in MCF-7 and T47D human breast cancer cells. The 3KE mutation markedly reduced the levels of ErbB3 phosphorylation, AKT and Erk1/2 activation, suggesting that direct integrin binding to NRG1 is critical for NRG1/ErbB signaling. Furthermore, stimulation of cells with WT NRG1 induced co-precipitation of ErbB3 with integrin α6β4. This suggests that WT NRG1 induces integrin-NRG1-ErbB3 ternary complex formation. WT NRG1 induced co-precipitation of ErbB3 with integrin αvβ3 to a less extent than with integrin α6β4. This probably reflects low levels of integrin αvβ3 expression compared to integrin α6β4 in these cells. In contrast, the 3KE mutation markedly reduced the levels of ternary complex formation, suggesting that this process depends on the ability of NRG1 to bind to integrins. Notably, the 3KE mutant acted as an antagonist to ErbB/NRG1 signaling in vitro and effectively suppressed proliferation of breast cancer cells in vivo. These results suggest that 1) NRG1-integrin interaction mediates crosstalk of integrins with ErbB receptors, 2) integrin α6β4 plays a major role in ErbB/NRG signaling in the cancer cells tested, and 3) the 3KE mutant has potential as a therapeutic in cancer.
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 272.
Abstract 2722: Development of automated methods of in-situ hybridization for the detection of ETS gene re-arrangements in FFPE biopsy and archived prostatectomy specimens
Authors: Nagy Dea , Santa- Cruz Ashley , Riley Janice P. , Sathyanarayana Ubaradka G. , Pommier C J. , Yun Steven , Ashworth-Sharpe Julia , Kosmeder Jerry , Nagle Raymond B. , Miller Phillip , Pestano Gary A. 
Published: February 2011

Abstract
Abstract
Prostate cancer (PCa) is the most common visceral neoplasm in men. In most cases, the disease is slowly progressive. Five year survival with organ-confined disease is 100%, while even distal spread is 32%. ETS gene re-arrangements in PCa have been shown to be associated with disease progression and have therapeutic and diagnostic implications.
The goal of this study was to develop methods of automated in-situ hybridization to detect the ETS gene re-arrangements in formalin fixed, paraffin-embedded (FFPE) prostate tissue. Methods were developed on the Ventana Benchmark®XT and Discovery®XT instruments for prostate cancer cell line xenografts, including VCap, LnCaP and H660, as well as for primary biopsy and archived prostatectomy specimens.
Key modules in the protocol have been evaluated and modified both in manual (offline) and automated (online) process of method development. Specific protocols were developed for deparaffinization, antigen retrieval, DNA denaturation, probe hybridization, sample washing, and detection with Quantum Dots (Qdots). Six different FISH probes specific for 3′ ERG, 5′ ERG, 5′ SLC45A3, and 3′ETV1 were used in assay development. Probe hybridization time was held to 8 hours, with overnight completion of the fully automated protocol.
Assay conditions on both instruments were similar for xenografts and in archived prostectomy specimens; however increased stringency of target retrieval was required for processing of the biopsy specimens. This adjustment was necessary to compensate for the tendency of biopsy specimens to become overly fixed during procurement and routine shipping and handling. In addition we observed that the presence of commonly used detergents in washing buffers post-hybridization may increase background staining with QDots.
In conclusion, we have developed fully automated protocols for the processing of clinically relevant tissue specimens. The xenograft cell line controls developed as a part of this study may also enable evaluation of assay performance in routine specimen processing for the detection of the ETS genes in biopsies and in prostatectomies.
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 2722.
Abstract 2729: Galectin-4 in colorectal cancer
Authors: Satelli Arun , Rao Prema S. , Rao U Subrahmanyeswara 
Published: February 2011

Abstract
Introduction: Colorectal cancer (CRC) is one of the leading diseases in the western world and is curable if diagnosed at an early stage. The normal-adenoma-carcinoma sequence concept suggests that the normal epithelia in the colorectal region transform into adenomas, which progress to invasive carcinomas. The role of galectin-4 (gal-4) a protein abundantly expressed in normal colonic epithelia, hereto considered a differentiation marker, on the onset or progression of colorectal cancer (CRC) is unknown. The main aim of this study is to evaluate gal-4 as a potential biomarker in detecting the onset of CRC and to determine the function of gal-4 in CRC development.
Methods: We analyzed the paraffin-embedded normal, adenomatous polyps and invasive carcinomas of the human colon biopsies and tissue microarrays for gal-4 expression by immunohistochemistry. Gal-4 expression in CRC cell lines was characterized by RT-PCR, western blotting, immunofluorescence microscopy and FACS analysis. The functional role of gal-4 was examined by cell proliferation, migration, motility, transfection and RNAi experiments.
Results: Biopsies immunostained for gal-4 were quantified using ImageJ software. The normal tissue as well as normal tissue adjacent to cancer contained highest amount of gal-4. Gal-4 intensity decreased in all the disease-states, which was found to be statistically significant (P &lt;0.05). The results suggested that as the normal colon develops adenomatous polyps, which transform subsequently into invasive carcinomas, the epithelial cells lining the crypts shut off gal-4 expression. Gal-4 negative cells showed increased cell proliferation, motility and migration, in comparison to gal-4 positive CRC cells. Transfection of the CRC cells (gal-4 -ve) with gal-4 led to decreased motility and cell cycle arrest. Externally added gal-4 on gal-4 -ve cells induced retarded cell migration. Gal-4 down-regulates cyclin D1 level and upregulates p15 and p21 levels.
Conclusions: We conclude that galectin-4 plays a key role the maintenance of epithelial cells in a differentiated state, and its loss allows the cells to de-differentiate, leading to increased cell proliferation, migration and motility, which are hallmark features of cancer. Galectin-4 brings about these changes through cell cycle regulation by modulating the cyclin D1, p15 and p21 levels. Finally, our findings that extracellular galectin-4 retards the migratory potential raise a possibility for it to serve as a therapeutic agent in controlling CRC progression. Thus beyond shedding light on the role of tumor-suppressor gal-4 in CRC, this study underscores the potential clinical value of gal-4 in prognosis and diagnosis of CRC.
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 2729.
Abstract 2733: Molecular markers for predicting progression of pure ductal carcinoma in situ of the breast
Authors: Abuázar Carolina Sens , Napolitano e Ferreira Elisa , de Toledo Cynthia Aparecida Bueno , Soares Fernando Augusto , Rocha Rafael Malagoli , Carraro Dirce Maria 
Published: February 2011

Abstract
INTRODUCTION: Ductal carcinoma in situ (DCIS) of the breast is a heterogeneous group of tumors which has an uncertain and poorly understood evolution. For this reason, the identification of molecular factors necessary for progression to invasive disease is one of the main concerns of the scientific community. We previously identified, by a microarray assay, a group of genes potentially involved in progression of DCIS, which are differentially expressed between pure DCIS and in situ component of lesions with co-existing invasive ductal carcinoma (DCIS-IDC). For this experiment, only the in situ components of pure DCIS and DCIS-IDC lesions were isolated using laser microdissection technique. AIM: Seek novel molecular markers to predict invasiveness potential in a DCIS group of breast carcinomas by qRT-PCR and Immunohistochemistry (IHC). METHODS: For qRT-PCR, 18 genes were selected, for which there were available antibodies. The qRT-PCR reactions were performed in ABIPrism 7700 Sequence Detection System (Applied Biosystems, Foster City, CA, USA) and the criterion of gene expression difference was equal or greater than 2-fold change. IHC was performed in a tissue microarray (TMA), using a biotin-free polymeric system for visualization (Flex+, DAKOTM). The distribution of IHC markers was analyzed using the Chi-square test for trend or Fisher's exact test. RESULTS: The expression of 4 genes (ADFP, ARHGAP19, ANAPC13 and CLTCL1) was confirmed by qRT-PCR, in the initial sample set used in the microarray experiment, presenting higher expression in the pure DCIS group. IHC was conducted to evaluate the corresponding proteins, using an independent group of 47 pure DCIS and 35 DCIS-IDC samples arrayed in TMA. Two genes, ANAPC13 (p=0.0379) and CLCTL1 (p=0.0471), confirmed the expression at protein level, showing significant higher cytoplasm staining in the pure DCIS group. CONCLUSION: ANAPC13 and CLTCL1 may be involved in progression of DCIS, being good candidates for predicting invasion of pure DCIS.
Financial support: CEPID and FAPESP.
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 2733.
Abstract 2738: Early detection of colorectal cancer applying a combination of serum markers
Authors: Wild Norbert , Andres Herbert , Rollinger Wolfgang , Krause Friedemann , Dilba Peter , Tacke Michael , Karl Johann 
Published: February 2011

Abstract
Background: Fecal occult blood testing (FOBT) is the recommended first line screening for the detection of colorectal cancer (CRC). To improve the detection of CRC we evaluated serum markers and combinations of serum markers as an alternative approach.
Methods: Applying Lasso Regression, a specialized form of penalized logistic regression, we selected six markers for an evaluation in a collective of 857 patients including 301 CRC patients, 143 patients with adenoma, 266 healthy controls and 147 disease controls. For each marker and marker combination the performance was assessed.
Results: We tested a total of 22 biomarkers for the detection of CRC from serum. Of these six markers were selected for a marker combination by Lasso Regression. Included were the well-known tumor markers CEA and CYFRA21-1 as well as novel markers or markers that are less routinely used for the detection of CRC: ferritin, osteopontin, anti-p53 and seprase. CEA showed the best sensitivity of all markers with 43.9 % at 95% specificity, followed by seprase (42.4%), CYFRA21-1 (35.5%), osteopontin (30.2%), ferritin (23.9%) and anti-p53 (20.0%). When these markers were combined a sensitivity of 72.3% was reached at a corresponding specificity of 95% and of 62.1% at 98% specificity. Focusing on more screening relevant stages, UICC stages 0-III, reduced the sensitivity slightly to 68.0% and 53.3%, respectively. In a sub-collective where matched stool samples were available (75 CRC cases and 234 controls) the sensitivity of the marker combination was comparable to fecal immunochemical testing (FIT) with 82.4% and 68.9% vs. 81.8% and 72.7% at 95% and 98% specificity, respectively.
Conclusion: When six markers were combined to detect CRC from serum, the combination reached a performance that was comparable to FIT. This provides a novel tool for CRC screening to trigger a follow-up colonoscopy for a final diagnosis.
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 2738.
Abstract 2740: Increased accuracy of TRAP-FISH combination in bladder cancer diagnosis in symptomatic patients
Authors: Bravaccini Sara , Casadio Valentina , Gunelli Roberta , Nanni Oriana , Arienti Chiara , Sacanna Emanuele , Zoli Wainer , Amadori Dino , Silvestrini Rosella , Calistri Daniele 
Published: February 2011

Abstract
Introduction: Previous case-control studies have shown the importance of urine telomerase activity determined by the telomerase repeat amplification protocol (TRAP) assay to detect early bladder cancer. However, its validation is needed in large and consecutive series of symptomatic patients before it can be considered for routine clinical use. The aim of the present study was to verify whether a multiple approach based on first-level TRAP assay analysis and successive evaluation with UroVysion test, limited to patients with TRAP-positive urine samples, could unmask false-positive results and improve diagnostic specificity.
Patients and methods: Three hundred individuals with urinary tract symptomatology (50 females, 250 males), but without prior history of bladder malignancy were consecutively enrolled between January 2007 and June 2008. All urine samples were processed for cytological examination and also for telomerase activity determination using a quantitative TRAP assay. FISH analysis was carried out only in TRAP-positive samples.
Results: At a cut-off of 50 arbitrary enzymatic units (AEUs), shown in previous case control-studies as the most accurate diagnostic discriminant, the positive predictive value (PPV) and negative predictive value (NPV) using TRAP assay were 48% and 86%, respectively, with an overall accuracy of 71%. With the two-step TRAP-FISH approach, PPV increased to 75% and overall accuracy to 83%.
Conclusion: Our results indicate that TRAP assay by itself could make a valid contribution to early bladder cancer diagnosis in high-risk patients with urinary tract symptomatology, and that its combination with FISH could improve specificity by unmasking false-positive TRAP results.
*Contributed equally to this work
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 2740.
Abstract 2742: Identification of tumor antigens in neu transgenic mice may provide biomarkers useful for the early detection and diagnosis of human breast cancer
Authors: Wu Mei , Marzbani Edmond , Mao Jianning , Shehata Mariam , Kim Rachel Y. J. , Lu Hailing , Disis Mary L. 
Published: February 2011

Abstract
Early detection of breast cancer is crucial for better prognosis and successful treatment of the patients. Development of novel methods for screening for breast cancer is needed for early detection and diagnosis. The human immune system responds to tumor-specific antigens in the pre-malignant stage of breast cancer and produces specific antibodies, which can be detected as potential breast cancer biomarkers. Due to the limited availability of well-annotated pre-cancer and post-cancer sera from human subjects, we performed serial serum collection in a neu transgenic mouse model from the age of 10 weeks to the terminal stage of disease. Using a technique called serological screening of cDNA expression library (SEREX), a high-throughput method to rapidly screen recombinant proteins expressed in a bacteriophage-based cDNA library, we have previously identified tumor antigens in the late stage of neu breast cancers. The mouse tumors have similar immunogenicity as their human counterparts due to the fact that some of these antigens are also immunogenic in human. Indeed, we have found that one mouse antigen called gelsolin, shares 93% homology with human gelsolin and has the highest response to human serum antibody among other antigens. In an ELISA analysis using the serum samples from 50 breast cancer cases and 50 normal donors, the area-under-curve (AUC) value of the receiver-operating-characteristic (ROC) curve of gelsolin ELISA detects more than 70% of cancer patients. Using the SEREX technique, we sought to further identify the early-stage serum antibody biomarkers by comparing the pre-cancer, early stage, and late stage sera from the same mouse. We have identified three early-stage biomarkers, A20/TNFaip3, Rpl5, and Pdhx, in neu mice. In summary, we conclude that identification of tumor antigens in the neu transgenic animal model is useful for the discovery of serum antibody biomarkers for early detection and diagnosis.
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 2742.
Abstract 2758: Population pharmacokinetics of cyclophosphamide in infants and young children
Authors: Tagen Michael , Panetta John C. , Gajjar Amar , Stewart Clinton F. 
Published: February 2011

Abstract
Cyclophosphamide (CTX) is an anticancer pro-drug used for induction therapy of brain tumors and other malignancies. An adequate pharmacokinetic study of CTX and its active metabolite, 4-hydroxy-cyclophosphamide (4OH-CTX), has never been performed in infants and young children. We are therefore examining the disposition of CTX, 4OH-CTX, and its inactivated metabolite, carboxyethylphosphoramide mustard (CEPM), in children &lt; 3 years of age with brain tumors. CTX was administered during the first course of treatment (1.5 g/m2 infused over 1 h), and serial samples were collected prior to infusion, at the end of infusion, and 3, 6, and 24 h after the end of infusion. CTX and CEPM were quantitated in plasma with a validated LC-MS/MS method and 4OH-CTX was stabilized by addition of phenylhydrazine to whole blood and quantitated with a validated LC-MS/MS method. A nonlinear mixed-effects model was fit to the data using NONMEM software and covariates were added to the model based on reduction in objective function value and improvement in diagnostic plots. Pharmacokinetic studies were performed with 28 patients. The median (range) age was 17.3 months (0.87 − 35.9 months) and median (range) BSA was 0.49 m2 (0.24 − 0.71 m2). A model with one compartment for CTX and each of its metabolites adequately described the data. BSA normalization of the dose reduced the inter-individual variability (IIV) in the volume of distribution of CTX (VCTX) from 32% to 13% and reduced the IIV in the elimination of 4OH-CTX through non-CEPM pathways (k4OH-CTX) from 60% to 38%. The covariate analysis showed that the BSA-normalized clearance of CTX to 4OH-CTX (CLCTX) increased slightly with age, whereas k4OH-CTX increased rapidly with age. The net effect was that exposure to 4OH-CTX decreased with age in a nonlinear fashion. These results demonstrate the importance of studying the pharmacokinetics of anticancer drugs in infants and young children, and will provide the basis for the rational design of CTX dosing strategies for infants and young children with brain tumors.
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 2758.
Abstract 276: Docosahexaenoic acid alters the spatio-temporal segregation and activation of EGF receptor
Authors: Turk Harmony F. , Lupton Joanne R. , Chapkin Robert S. 
Published: February 2011

Abstract
We have previously demonstrated that dietary docosahexaenoic acid (DHA, 22:6n-3), a component of fish oil, profoundly alters the biochemical make up of colonocyte lipid rafts/caveolae microdomains. Since epidermal growth factor receptor (EGFR, ErbB1) is considered a major target for treatment of colorectal cancer and its activation can be regulated by manipulating the lipid composition of the membrane, we further elucidated the effects of DHA with respect to the EGFR signaling axis. Immortalized mouse colonocytes (YAMC cells) were treated with fatty acids (50 μM) for 72 h and stimulated in the presence or absence of EGF (0-50 ng/mL) for 0-10 min and receptor stimulation was assessed by determining levels of phosphotyrosine-EGFR. Downstream signaling was evaluated by measuring Akt, ERK 1/2, and Stat3 phosphorylation. EGFR phosphorylation was increased &gt;2-fold (p&lt;0.05) in DHA treated cells compared to control. Erk 1/2 activation was significantly increased &gt;4-fold, while Stat 3 activation was decreased by 2-fold (p&lt;0.05). No change in activation of Akt was observed. In addition, chronically inflamed (DSS treated) and carcinogen (AOM) injected C57BL/6 mice were maintained on diets ± DHA (fish oil) for 18 wks. Protein from scraped colonic mucosa was analyzed for EGFR phosphorylation and downstream signaling. DHA-fed animals displayed &gt;2-fold (p&lt;0.05) increase in EGFR phosphorylation compared to corn oil fed animals. Total Erk 1/2 and activated Erk 1/2 were significantly increased in DHA-fed animals. Stat3 phosphorylation was decreased by ∼3-fold (p&lt;0.05). Akt activation showed no change. These data indicate that although DHA treatment results in an unexpected increase in EGFR phosphorylation, aspects of downstream signaling are suppressed. This could, in part, explain why fish oil feeding promotes colonocyte apoptosis and inhibits tumor growth. This work was supported in part by NIH grants CA59034, P30ES09106.
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 276.