bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1217: Detection of HER2 expression and phosphorylation in breast cancer CTC samples by CEER™
Authors: Hazra Saswati , Liu Limin , Webster Edvelyn , Fithian Andrew , Lee Tani , Magonova Katya , Barham Robert , Kim Phillip , Singh Sharat 
Published: August 2012

Abstract
Survival rates of metastatic breast cancers (mBCAs) are considerably low. Often, tumor cells at the primary site may not reflect the profile of the tumor cell population in recurrent disease. Circulating tumor cells (CTCs) isolated from peripheral blood offer a non-invasive disease monitoring modality. Identification of reliable molecular markers within (CTCs) from patients with recurrent disease may further improve breast cancer survival. Hence, we have developed a multiplexed immunoassay for monitoring activated HER2 protein and their expression levels using a multiplexed immuno-microarray platform. Collaborative Enzyme Enhanced Reactive-immunoassay (CEER™) was utilized to analyze HER2 profiles in CTCs isolated from 76 BCA patients (stages III to IV) with HER2 negative primary disease. The CEER technology utilizes the formation of a unique immuno-complex requiring co-localization of two detector antibodies around capture antibodies immobilized on immuno-array to profile HER2 protein expression and activation. The collaboration between two channeling-enzymes conjugated on two detection antibodies in proximity, enables the profiling of the target proteins with extreme sensitivity and specificity. Approximately 25% of HER2 negative BCA patients in this cohort showed varying levels of HER2 activation in CTCs isolated from the recurrent disease. About 8% of patients with HER2 activation in CTCs also showed significant HER2 over-expression. This suggests mechanisms of HER2 activation either through formation of receptor heterodimer (in samples without HER2 over-expression) or homodimer (in samples with HER2 overexpression) formation. The distinct HER2 discordance between primary tumor and recurrent disease demonstrates an urgent need for routine monitoring of HER2 status in CTCs found in mBCA patients. Incidence of HER2 alterations in CTCs should be considered in selecting effective treatment regimens for BCA patients with relapsed disease. Furthermore, CEER can be used for profiling other druggable targets for guiding effective clinical strategies including rational targeted agents.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1217. doi:1538-7445.AM2012-1217
Abstract 1223: Relief of feedback inhibition of RTK and RAS signaling in V600E BRAF melanomas exposed to RAF inhibitors buffers their effects on signaling
Authors: Pratilas Christine A. , Joseph Eric W. , Tadi Madhavi , Poulikakos Poulikos I. , Gabay Maytal , Lito Piro , Wong Wai , DeStanchina Elisa , Chandarlapaty Sarat , Rosen Neal 
Published: August 2012

Abstract
Activation of ERK signaling causes feedback inhibition of receptor tyrosine kinase and RAS signaling by multiple mechanisms and limits the steady-state output of the pathway. The V600E BRAF mutant does not depend on RAS-dependent dimerization and is not susceptible to feedback inhibition of RAS. In V600E BRAF melanomas, ERK pathway output and the expression of ERK genes such as DUSP6 and members of the SPROUTY family are markedly elevated. In these tumors, RTK signaling and RAS activation are actively suppressed. In the context of elevated DUSP6, RAF inhibitors rapidly inhibit ERK phosphorylation and output. This relieves the feedback inhibition of various RTKs and induces RAS activity and the formation of activated wild type RAF dimers that are insensitive to RAF-inhibitors. This results in a rebound in ERK phosphorylation and output that is sensitive to MEK, but not RAF inhibitors and suggests that the latter cannot maximally inhibit ERK signaling in these tumors. In support of this idea, combined treatment with MEK and RAF inhibitors reduces ERK output and the growth of V600E BRAF melanoma xenografts to a greater degree than either alone.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1223. doi:1538-7445.AM2012-1223
Abstract 1227: Modulation of EGF-receptors signaling by small peptidomimetic molecules in breast cancer cells
Authors: Banappagari Sashikanth , Satyanarayanajois Seetharama , Corti Miriam , Pincus Seth 
Published: August 2012

Abstract
Protein-protein interactions play a crucial role in regulating many biological processes as well as in disease pathology. Understanding the structural basis and modulating protein-protein interactions has a tremendous impact in developing new therapeutic strategies for many human diseases. Human epidermal growth factor receptor family of proteins (EGFR, HER2, 3 and 4) plays an important role in cell growth and proliferation. Aberrant interactions of EGFR family of proteins lead to tumor growth and metastasis. HER2, a member of EGFR family of proteins is overexpressed in approximately 25% of breast cancers. Deregulation of HER2 signaling pathways and overexpression of HER2 is known to occur in many cancers. HER2 is known to form heterodimers and is a major therapeutic target in breast cancer treatment. Formation of HER2 receptor heterodimerization with other EGFR family of proteins such as EGFR, HER3 is essential for intracellular kinase activation and is the basis for exploring direct inhibition of EGFR activation by blocking dimerization for therapeutic purposes. We have designed several small peptidomimetics to inhibit HER2-mediated signaling for cell growth. One of such peptidomimetics, Compound 5 (Arg-[3-amino-naphthyl-propionic acid]-Phe) designed, exhibited antiproliferative activity with IC50 values in the nM range against HER2 overexpressing breast cancer cell lines, SKBR-3 and BT-474. Binding studies using Florescence Microscopy, ELISA, Circular Dichroism (CD) and Surface Plasmon Resonance (SPR) confirmed that compound 5 binds specifically to HER2-extracellular domain. SPR studies revealed that compound 5 specifically binds to domain IV of HER2 extracellular region. Also, Compound 5 inhibited HER2-HER3 as well as HER2-EGFR heterodimerization evaluated by SPR. Pathhunter and proximity ligation assays confirmed the inhibition of HER2 heterodimerization with EGFR and HER3 by compound 5. Western blots revealed that compound 5 blocks the transphosphorylation of HER2 protein. Based on these observations small peptidomimetic inhibitor molecules can be potential therapeutic agents in disabling HER2 mediated heterodimerization.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1227. doi:1538-7445.AM2012-1227
Abstract 1235: p120ctn is a key effector of Ras-PKC∈-mediated oncogenic signaling
Authors: Dann Stephen G. , Golas Jonathon , Miranda Miriam , Shi Celine , Upeslacis Eric , Rosfjord Edward , Jin Guixian , Wu Jiang , Klippel Anke 
Published: August 2012

Abstract
Within the family of protein kinase C (PKC) molecules only the novel isoform member PRKCE (PKC∈) acts as a bona fide oncogene in in-vitro and in-vivo models of tumorigenesis. Previous studies have reported cancer-specific misexpression of PKC∈ at levels well above that of normal adjacent tissue in breast, prostate and lung tumors. We find that oncogenic Ras signaling promotes PKC∈ expression and results in hyperphosphorylation of CTNND1/p120-catenin (p120ctn). In this context, loss of PKC∈ by genetic or pharmacological means results in normalization of morphology and signaling responses. In a KRasD13-dependent breast cancer model loss of PKC∈ function results in growth inhibition in 2-dimensional (2D) and 3-dimensional (3D) culture systems as well as in orthotopic xenografts concomitant with the normalization of a subset of Ras-induced signaling responses. Using phospho-proteomic profiling analysis we observe that CTNND1 (p120ctn) phosphorylation at serine 268 (S268) occurs in a strictly PKC∈ dependent manner. Treatment with a specific PKC∈ inhibitor, PF-5263555, recapitulates the genetic loss of function phenotype and interferes with breast cancer cell growth in-vitro and in-vivo. We also show that PKC∈-mediated phosphorylation at S268 further stabilizes additional p120ctn phosphorylation sites and total protein levels of β-catenin. We demonstrate that p120ctn phosphorylation at S268 represents a specific readout for PKC∈ activity and as such can serve as a suitable biomarker for PKC∈ dysregulation in human cancer and for monitoring therapeutic response.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1235. doi:1538-7445.AM2012-1235
Abstract 1237: TGF-β signaling pathway analysis using MILLIPLEX MAP TGF-β multiplex panel
Authors: Johnston Randy , Warmke Tim , Wiese Rick , Hayes David , Hwang Joseph 
Published: August 2012

Abstract
Transforming growth factor beta (TGF-β) is a secreted protein that controls cellular proliferation, differentiation, apoptosis, migration, adhesion, immune response, and other functions in most cells. The binding of TGF-β to its receptor, TGFβRII, can lead to activation of both the SMAD-dependent canonical pathway as well as the SMAD-independent non-canonical pathways (such as PI3K/Akt, ERK, JNK, and p38). The TGF-β signaling pathway is coordinately regulated by phosphorylation and understanding the role of this pathway in normal physiological processes and in diseases such as cancer and diabetes requires the ability to simultaneously measure phosphorylation status of multiple protein targets. In recent years, bead-based assays, such as those using Luminex xMAP technology, have enabled the measurement of phosphorylation levels of up to 12 proteins simultaneously. Using the TGF-β Signaling Pathway multiplex panel, we observed increased levels of phosphorylated Smad2 and Smad3, but not phosphorylated ERK or Akt in TGF-β treated cells when compared to their unstimulated counterparts. Modifications in phosphorylation levels were confirmed by the treatment of cells with specific inhibitors against TGFβRI, which blocked phosphorylation of Smad2 and Smad3 in TGF-β treated HepG2 cells. Furthermore, significant increase in phosphorylated Smad3 was observed in human breast cancer versus matched breast normal tissues. The TGF-β Signaling Pathway multiplex panel allowed for simultaneous detection of multiple analytes in both the SMAD-dependent canonical pathway as well as the SMAD-independent non-canonical pathways in a specific, sensitive, and reproducible manner.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1237. doi:1538-7445.AM2012-1237
Abstract 124: Hypoxia-inducible miR-210 is involved in hypoxia-induced cell migration and invasion in human hepatocellular carcinoma
Authors: Kai Alan KL , Lo Regina CL , Lee Joyce MF , Wong Jack CM , Ng Irene OL 
Published: August 2012

Abstract
MicroRNA dysregulation is implicated in many human malignancies, yet little is known regarding the regulatory mechanisms of these small noncoding RNAs. Hypoxia is a prevalent tumor microenvironment in hepatocellular carcinoma (HCC) and plays a key role in modulating tumor aggressiveness. In the present study, we investigated the effects of hypoxia on microRNA expression in human HCC. First, we examined the miRNA expression profiles with TaqMan human microRNA Low-Density Array on HCC cell lines put under normoxic and hypoxic conditions to identify expression changes of microRNA. From the 664 microRNAs examined, miR-210 was overexpressed in SMMC-7721, Huh-7 and MHCC-97L HCC cell lines under 0.1% O2 hypoxic condition. The hypoxic induction of miR-210 was further validated in another two HCC cell lines (Hep3B and PLC/PRF/5) by individual real-time quantitative PCR (qPCR) and the same expression pattern was observed, suggesting that miR-210 is consistently induced in HCC cells under hypoxic conditions. The abundance of mature human miR-210 was also examined by qPCR in a human HCC cohort consisting of 48 pairs of primary HCC tissues and their corresponding non-tumorous livers. We found that miR-210 expression was significantly higher in the HCC tissues as compared with the corresponding non-tumorous livers (P=0.0003). Furthermore, we used the hypoxia marker carbonic anhydrase IX (CAIX) coupled with microdissection technique to isolate tissue samples in the peri-necrotic areas, which are considered pathologically hypoxic, in clinical formalin-fixed paraffin-embedded (FFPE) HCC samples. We extracted the microRNA from the tissues and compared the miR-210 expression with those from histologically preserved HCC areas away from necrotic areas. In line with the in vitro observations, with qPCR, miR-210 was remarkably overexpressed in hypoxic HCC regions. Furthermore, miR-210 overexpression significantly and inversely correlated with the overall survival and disease-free survival rates (P= 0.029 and 0.043, respectively) of HCC patients, indicating that overexpression of miR-210 in HCC was associated with a poorer prognosis. The biological function of the miR-210 induction in HCC was also investigated. Notably, with transwell and cell invasion assays, we demonstrated that the cell migratory and invasive abilities of SMMC-7721 were dramatically enhanced during hypoxia, and this enhancement was abolished upon miR-210 suppression using locked nucleic acid (LNA) inhibitor. Taken together, our present data indicate that miR-210 is a hypoxia-inducible microRNA in HCC, which may enhance HCC aggressiveness under hypoxic conditions.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 124. doi:1538-7445.AM2012-124
Abstract 1241: In vivo analysis of the role of Lyn tyrosine kinase in castrate-resistant LNCaP xenograft tumor growth
Authors: Zardan Anousheh , Cox Michael , Gleave Martin , Zoubeidi Amina 
Published: August 2012

Abstract
Introduction: It is proposed that advanced Prostate Cancer (PCa) tumors develop mechanisms, which re-activate the androgen receptor (AR) axis via oncogenic pathways, in which tyrosine kinases play a crucial role. Genetic manipulation of several tyrosine kinases in vivo revealed that only a knockout of Lyn tyrosine Kinase, compromised prostate gland development suggesting that Lyn plays a critical role in the regulation of AR in normal prostate development. Our results demonstrated that Lyn kinase enhances AR transcriptional activity and promotes prostate cancer cell survival. Thus, we hypothesized that Lyn kinase overexpression accelerates castrate-resistant LNCaP tumor growth and serum PSA relapse to pre-castration levels. Method: LNCaP cells stably overexpressing Lyn kinase (LNCaPLyn) or Empty vector (LNCapEmpty) were generated using clonal selection. 1 × 106 LNCaPLyn and LNCaPEmpty were injected into the male nude mice. Serum PSA level and tumor volumes were followed weekly and mice were castrated when serum PSA values reached 50ng/ml. Results: Interestingly, a higher rate of tumor growth and PSA relapse was observed after casteration in mice bearing the LNCaPLyn compare to the LNCapEmpty-bearing mice. Total protein was extracted from the xenograft tumors and levels of AR, PSA, FKBP52, AKT, pAKT were analyzed. Results of Protein analysis further confirmed the role of Lyn kinase in prostate cancer cell survival and progression to castrate resistant stage. Conclusion: Together, these results suggest that Lyn plays a key role in CRPC development and that inhibiting Lyn expression could be considered as a potential therapeutic target for CRPC.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1241. doi:1538-7445.AM2012-1241
Abstract 1262: Simple western and NanoPro analysis of key EGF signaling cascade proteins
Authors: Nguyen Uyen , Dermody Jessica , Ramirez Francisco , Kazakova Irina , Yang Tom , Gavin Robert , Boge Annegret 
Published: August 2012

Abstract
Aberrant expression and signaling in the EGF signaling pathway is a common occurrence in a variety of cancers including breast cancer. Understanding how EGF signaling impacts disease progression is key to the development of novel therapeutics. Detection of important protein targets in cancer samples frequently employs Western blot analysis. In-depth phosphorylation analysis often requires 2D gels which are extremely variable and labor intensive, followed by MS analysis. Each of these methods have their advantages and challenges. This study employs the use of two novel nano-scale capillary-based electrophoresis and immunodetection technologies to characterize and validate subtle changes in key protein targets involved in the regulation of disease progression. The Simon system performs size-based separation of proteins, known as Simple Western assays. Alternatively, NanoPro systems utilize charge-based separation via isoelectric focusing (IEF). Both methods utilize capillary electrophoresis (CE) to perform the separation events. Both Simon and NanoPro instruments provide automated workflow eliminating the need for manual processing of multiple steps traditionally needed to carry out either of these separation and detection processes. Proteins from a single sample preparation were analyzed on two platforms: either in a native conformation using charge-based separation (NanoPro assays) or in a denatured state followed by size-based separation (Simple Western assays). Both methods incorporate immunodetection to monitor both subtle and specific changes to key signaling protein targets. Utilizing both platforms allowed for detailed analysis of post-translational modifications (charge-based) and protein abundance (size-based) within a prepared sample. As a validation of synergy between the platforms, we investigated key members of the EGF signaling cascade in different phosphorylation states. We showed that the same cellular lysate preparation can be used for charge- or size-based separation. Advantages of Simple Western and NanoPro assays over gels (1D and 2D) and traditional Westerns include ease of use, minimal user intervention, automatic analysis and excellent reproducibility which ultimate results in better data quality to characterize key regulatory events involved with cancer progression.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1262. doi:1538-7445.AM2012-1262
Abstract 1266: Glioma biomarker discovery by mass spectrometry
Authors: Dekker Lennard , Kheirollahi Majid , Stockhammer Guenther , Luider Theo , Smitt Peter Sillevis 
Published: August 2012

Abstract
BACKGROUND Approximately 200.000 people are diagnosed with a primary brain tumor each year worldwide. The most common type of primary brain tumor (∼40%) are gliomas. Currently available prognostic markers require evaluation of tumor tissue, such as 1p and 19q choromosome co-deletion in oligodendroglial tumors, MGMT promoter methylation for glioblastomas (GBMs), and IDH mutations in several glioma subtypes. Serum biomarkers that correlate with tumor status have prognostic and predictive value and could significantly improve the care for glioma patients. In addition biomarkers could provide unique information related to longitudinal response to therapy. The only clinically validated serum marker of prognosis and disease status in high grade gliomas is serum YKL-40, a chitinase homolog also called human cartilage glycoprotein 39 or chitinase 3-like 1. DESIGN/METHODS For the discovery phase we used a set of 31 cerebrospinal fluid samples. This sample set included 5 CSF samples of patients with a GBM and 26 controls. These samples were digested by trypsin and subsequently measured by LCMS-Orbitrap. The acquired data were analyzed using a database search and a subsequent analyses of the spectral counts by Scaffold. The spectral counts for each individual identified protein are used as a semi quantitative measure for protein abundance. RESULTS / CONCLUSIONS From the total list of 771 identified proteins, candidate biomarker proteins were selected that showed a significant difference between the control and GBM (p&lt;0.01) and were at least 3 fold higher in spectral count than in the control group. Abundant serum proteins were removed to avoid inclusion of proteins that relate to the disruption of the blood-CSF barrier. Eventually, a list of eleven proteins was obtained that included 14-3-3 protein gamma, 72 kDa type IV collagenase, Alpha-enolase, Chitinase-3-like protein 1 (YKL-40), Fetuin-B, Glial fibrillary acidic protein, Insulin-like growth factor II, Metalloproteinase inhibitor 1, Serum paraoxonase/arylesterase 1, SPARC, IGF binding protein 2 and Metalloproteinase inhibitor 2. Many of the identified candidate markers have been previously reported in literature in relation to glioma including the clinically validated serum marker for glioma YKL-40. We are now planning a validation of these markers in a set of serum samples of glioma patients.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1266. doi:1538-7445.AM2012-1266