bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4161: Development of a real-time multiplex PCR assay for the detection of FGFR3 mutations in urine
Authors: Millholland John M. , Patel Sheryl G. , Fernandez Cecilia A. , Shuber Anthony P. 
Published: August 2014

Abstract
Background: We have recently reported the development of a Multi-Analyte Diagnostic Readout (MADR) non-invasive assay using urinary Matrix Metalloproteinases (MMPs) and FGFR3 as triage monitors in high-risk bladder cancer populations. This concept combines the marker performance characteristics of protein and DNA biomarkers into one assay for optimal performance. Eight common FGFR3 mutations in 3 exons have been associated with bladder cancer. Analysis of mutational status for each single mutation required 8 amplification steps, which were costly and time consuming. We have now developed a real-time multiplexed FGFR3 assay, generating a cost-effective, clinically applicable assay for the detection of FGFR3 mutations in urine.
Methods: Our approach involves a two-step PCR amplification process. The initial round generates exon specific PCR products, which are then used as template for real-time PCR mutation detection utilizing locked nucleic acid (LNA) oligonucleotides. These LNA suppress wild-type DNA amplification.
To convert our existing FGFR3 assay to a multiplex format, primary amplifications of exons 7, 10, and 15 were combined into a single real-time PCR assay for exon specific amplification and DNA quantitation. The LNA-mediated mutation detection was then converted from 4 reactions to 2 duplex amplifications. All multiplex assays were carried out on the Roche LC 480 real-time PCR platform.
Results: To validate the new the multiplex format, FGFR3 multiplex analysis was performed on DNA isolated from 50 Ta stage bladder tumors. FGFR3 mutations were detected in 90% (48/50) of the tumors. To directly compare performance with single mutation analysis, 40 urine samples previously analyzed using the singleplex format were again tested using the multiplex FGFR3 assay. 100% concordance was seen between the two assay formats.
Conclusions: By multiplexing the FGFR3 mutation analysis we reduced the number of amplification steps, improving assay turnaround time and throughput, without compromising assay performance. The FGFR3 multiplex analysis provides a robust, cost-effective DNA assay that in combination with MMP protein analysis delivers a clinically applicable assay with optimal performance.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4161. doi:10.1158/1538-7445.AM2011-4161
Abstract 417: α9β1 integrin, as a critical regulator of lymphatic metastasis and growth of human breast cancer cells
Authors: Ohta Daichi 
Published: August 2014

Abstract
Osteopontin (OPN) has been implicated in various stages of cancer progression such as malignant transformation, invasion and metastasis. Several groups independently demonstrated that plasma OPN level is a potential diagnostic and prognostic marker for several human malignancies (Bellahcene A et al, Nat Rev Cancer 8:212, 2008 and Anborgh PH, et al Clin Chem 55:895, 2009). Interestingly, a human breast cancer cell with an aggressive lymph node metastatic ability has been established, which demonstrated strong expression of α9β1 integrin and OPN (Vantyghem SA et al Clin & Exp Meta 22:351, 2005). Of note, α9β1 integrin is a receptor for not only OPN, but also VEGF-C and -D, known lymphatic vessel growth factors. To address whether and how α9β1 integrin is involved in tumorigenesis, growth, invasion and lymph node metastasis, we took advantage of using an inhibitory anti-human α9 integrin antibody (clone: K33N), which does not cross react with murine α9 integrin and an α9β1 integrin-positive human breast cancer cell line, MDA-MB-231 luc-D3H2LN (D3H2LN), in in vitro functional assays and an in vivo orthotopic xenotransplantation model.
We found that K33N inhibited in vitro migration and invasion of D3H2LN cells, indicating that the interaction of α9β1 integrin with its ligands was involved in the breast cancer cells motility. Next, we inoculated D3H2LN cells into the mammary fat pad of nude mice at day 0, and K33N or control IgG was given to them twice per week from day 15 to day 50 after tumor inoculation. K33N significantly reduced both primary tumor growth and draining lymph node metastasis. To gain an understanding of the mechanism by which K33N suppressed the primary tumor growth and lymph node metastasis, we examined the expression of various molecules which are associated with the cancer progression. We found significant matrix metalloproteinase (MMP) up-regulation in K33N-treated primary tumor tissues, consistent with a recent report demonstrating an indirect involvement of MMPs involved in tumor growth. Of note, upon orthotopic injection of human D3H2LN cells, plasma level of host-derived murine OPN, but not tumor-derived human OPN was significantly elevated. Surprisingly, host-derived murine OPN was significantly reduced in the mice treated with K33N. These results suggest that the interaction between α9β1 integrin on breast cancer cells and its ligands favors the generation of tissue microenvironments which contribute to primary tumor growth and lymph node metastasis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 417. doi:10.1158/1538-7445.AM2011-417
Abstract LB-200: Solution structure of c-Myc G-quadruplex complex with a quindoline compound
Authors: Dai Jixun , Carver Megan , Mathad Raveendra I. , Hurley Lawrence , Yang Danzhou 
Published: August 2014

Abstract
Overexpression of the c-Myc oncogene is associated with a broad spectrum of human cancers. Targeting transcriptional control of c-Myc represents an attractive means for cancer molecular therapeutics. A highly conserved nuclear hypersensitivity element III1 (NHE III1) of the c-Myc promoter controls 80–90% of the c-Myc transcription. It can form transcriptionally active and silenced forms, and the formation of DNA G-quadruplex structures has been shown to be critical for c-Myc transcriptional silencing. Small molecules that stabilize the c-Myc G-quadruplex have been shown to suppress c-Myc transcription and are anti-tumorigenic. We have determined the NMR structure of a 2:1 complex of the c-Myc quadruplex with a quindoline compound. This is the first molecular structure of a drug complex with a natural oncogene promoter quadruplex. The solution structure of this 2:1 quindoline-G-quadruplex complex shows unexpected features, including the drug-induced reorientation of the flanking sequences to form a new binding pocket. While both 3′ and 5′ complexes show overall similar features, there are identifiable differences that emphasize the importance of both stacking and electronic interactions. We identified the importance of the two flanking bases as well as the shape of the ligand in determining drug-binding specificity. The structural information will be crucial for structure-based rational drug design of molecules that specifically target the c-Myc silencer element. It also provides important implications for rational design of drugs that bind to unimolecular parallel G-quadruplexes commonly found in promoter elements.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-200. doi:10.1158/1538-7445.AM2011-LB-200
Abstract LB-201: New derivatives of etoposide show promise as more potent inhibitors of cancer cell growth
Authors: Gal Susannah , Young Rebecca , Slilaty Steve , Deliencourt-Godefroy Geraldine , Quirion Jean-Charles 
Published: August 2014

Abstract
The DNA repair enzyme Topoisomerase II (TopoII) has been recognized in the oncology field as a clinically important therapeutic target for a variety of cancers. The class of agents known as etoposides were specifically designed with this target in mind. Etoposides lacked the major problem of cardiac toxicity associated with the anthracyclines class of drugs (e.g. doxorubicin).
Sunshine Biopharma has recognized that the reduced efficacy and high toxicity of etoposide are due to instability of the molecule leading to a conversion of the drug which then functions as a tubulin inhibitor rather than a TopoII inhibitor (unpublished observations). Accordingly, the company set out to develop a true TopoII inhibitor and these efforts have yielded two compounds called Adva-27a and Adva-32a. These drugs and their derivatives have been described in US Patent Application Number: 20090318675. Using cell lines derived from a variety of human cancers, Adva-27a is almost always a more potent inhibitor of cell growth compared to etoposide, and cells expressing the MDR-drug resistance protein are very sensitive to this drug.
Furthermore, consistent with the literature, amplification of the TOP2α gene is associated with increased sensitivity to growth inhibition. The preliminary data show that in all but the brain tumor cell line, increased TOP2α copy number is associated with even greater percent inhibition by Adva-27a versus etoposide regardless of the tissue of origin of the tumor cell lines. This data and plans for further analysis of these drugs will be presented.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-201. doi:10.1158/1538-7445.AM2011-LB-201
Abstract LB-212: XGFR, an Fc-engineered dual signaling inhibitor targeting IGF-1R and EGFR
Authors: Wartha Katharina , Croasdale Rebecca , Schanzer Juergen , Brinkmann Ulrich , Hofmann Marco H. , Ries Carola , Rieder Natascha , Hoelzlwimmer Gabriele , Freytag Olivier , Herter Sylvia , Gerdes Christian , Kuenkele Klaus-Peter , Klein Christian 
Published: August 2014

Abstract
Background: Elevated signaling via the receptor tyrosine kinases IGF-1R and EGFR has been identified as common characteristic of multiple cancer type. IGF-1R and EGFR signal predominantly through the PI3K and MAPK signaling pathways and thereby mediate growth and survival signals crucial for the development and progression of cancer. There is strong cross talk on multiple levels between IGF-1R and EGFR dependent signaling pathways. Therefore, targeting IGF-1R and EGFR simultaneously is an attractive way to achieve maximal inhibition of signal transduction and to avoid resistance formation.
Methods: Bispecific IGF1R-EGFR antibodies were engineered by linking scFv domains of an EGFR Mab (GA201) via Serine-Glycine linkers to an IgG1 IGF-1R Mab (RG1507). The functional properties of the bispecific antibodies were evaluated in cellular in vitro assays (IGF-1R/EGFR phosphorylation, downregulation, 3D proliferation and ADCC assays) and in in vivo xenograft models for tumor growth inhibition and survival.
Results: Bispecific IGF-1R-EGFR antibodies (XGFR2, XGFR3, XGFR4) were successfully generated with yields and stability comparable to conventional IgG1 antibodies. XGFR antibodies effectively inhibited IGF-1R and EGFR phosphorylation and 3D proliferation in H322M tumor cells and induced strong downmodulation of IGF-1R and enhanced EGFR downmodulation compared to the parental EGFR antibody GA201. XGFR antibodies showed strong anti-tumor efficacy comparable to the combination of monospecific IGF-1R and EGFR Mabs in the BxPC3 and H322M xenograft models. To enhance the ADCC properties of XGFR, afucosylated, glycoengineered bispecific antibodies with enhanced affinity for FcγRIIIA were generated using the GlycoMab technology. Glycoengineered bispecific antibodies were shown to have superior ADCC properties in in vitro ADCC assays and XGFR4 significantly prolonged median and overall survival of mice in an ADCC competent in vivo model (A549 i.v.).
Conclusions: Bispecific IGF-1R-EGFR antibodies represent an attractive therapeutic strategy to simultaneously target two key components of multiple cancer types (IGF-1R and EGFR), resulting in effective inhibition of the PI3K and MAPK signaling pathway and to avoid the formation of resistance to therapy. Having overcome issues of stability and productivity, bispecific antibodies may become an advantageous way to reduce costs and infusion times in cancer therapy, while at the same time, achieving maximal anti-tumor effects through inhibition of multiple targets.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-212. doi:10.1158/1538-7445.AM2011-LB-212
Abstract LB-36: Circulating epithelial tumor cells are responsive to BMP-2
Authors: Bähring Franziska , Reber Elena , Müller Ute , Kilper Roland , Wagner Kathleen , Hochhaus Andreas , Clement Joachim H. 
Published: August 2014

Abstract
Aims: Bone morphogenetic proteins (BMP) play an important role in early embryonic development as well as in organ homeostasis and in tumorigenesis. We could show previously that BMP-2 enhance migratory and invasive properties of tumor cells. Furthermore, BMP-2 affects the expression of genes which are involved in the regulation of apoptosis. A crucial step in metastasis formation is the liberation of tumor cells from the primary site, the entrance into circulation and the long-lasting persistence in the peripheral blood. Therefore we asked whether BMPs might contribute to the vitality of circulating tumor cells.
Methods: Leukocytes and circulating epithelial cells from the peripheral blood of breast cancer patients were labelled with fluorescent antibodies and magnetically enriched with the MACS® system. After separation of single vital cells with the aureka® device, a single cell multiplex RT PCR was performed using the AmpliGrid system. The PCR products were analysed by urea-polyacrylamide gel electrophoresis. For real-time single cell PCR demonstrating the expression level of selected genes the Fastlane system (Qiagen, Hilden) was used. Localization of phosphorylated receptor Smads was analysed by laser scanning microscopy after one hour incubation with 100 ng/ml BMP-2.
Results: Circulating epithelial cells were identified by using the cell surface antigen EpCam. Vital EpCam-positive cells were picked and subjected to multiplex RT-PCR covering 5 components of the BMP signaling cascade as well as RPL13A for control. 121 of 265 selected cells expressed RPL13A. These cells were used for further analysis. Most of these cells expressed BMP-2 (54%), BMPR-IA (46%), BMPR-IB (82%). Transcripts of the type II receptor BMPR-II (35%) and the putative BMP antagonist BMP-3 (25%) were presen to a lower extend. The expression level of BMP-2 was comparable in individual EpCam-positive cells, thus at least an autocrine stimulation of the BMP signaling pathway can be assumed. For studying the responsiveness of circulating epithelial cells towards a BMP stimulus these cells were enriched from peripheral blood and incubated with BMP-2. Vital cells showed a predominant nuclear staining for pSmad1 demonstrating a functional BMP signalling network.
Conclusion: We show that key components of the BMP signaling pathway are expressed in leukocytes and epithelial cells in the peripheral blood of breast cancer patients. This indicates that the BMP signaling network potentially contributes to the activity and survival of circulating epithelial cells.
This work was supported by the Dr. Rainald-Stromeyer-Stiftung
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-36. doi:10.1158/1538-7445.AM2011-LB-36
Abstract LB-363: DNA-damage foci persistence and accelerated senescence as determinants of radiosensitization by PARP inhibitors
Authors: Efimova Elena V. , Mauceri Helena J. , Barreto Juan C. , Labay Edwardine , Sikora Jacek , Takahashi Satoe , Wolfgeher Don W. , Bindokas Vytautas P. , Darga Thomas E. , Beckett Michael A. , Sutton Harold G. , Vokes Everett E. , Posner Mitchell C. , Weichselbaum Ralph R. , Kron Stephen J. 
Published: August 2014

Abstract
Along with apoptosis and necrosis, accelerated senescence may be a desirable outcome when replicative cancer cells undergo unrepairable DNA double strand breaks (DSBs) after treatment with ionizing radiation (IR) or chemotherapy. In both normal and cancer cells, DSBs promote the assembly of Ionizing radiation induced foci (IRIF), domains of modified chromatin that serve a key role in DNA damage signaling and DNA repair. IRIF persistence is a critical determinant of accelerated senescence, making targeting IRIF resolution an attractive strategy to sensitize cancer cells to genotoxic therapy. As an IRIF reporter, we constructed an inducible GFP fusion to the IRIF-binding domain of 53BP1 (GFP-IBD) which we have expressed in breast, head and neck, prostate, glioma and pancreatic cancer cell lines. Typically, within minutes after IR exposure, GFP-IBD relocalizes to form fluorescent nuclear foci, which disperse over several hours. Anti-GFP immunoprecipitation and proteomic analysis of IRIF revealed chromatin modifiers and DNA damage response proteins at DSBs including the poly(ADP-ribose) polymerase PARP1. To block DSB repair in cancer cells and tumors, we treated cells with veliparib (ABT-888), one of several PARP inhibitors currently in clinical trials. Veliparib markedly enhanced IRIF persistence and increased cell senescence in the majority of cancer cell lines tested. Treatment of tumor xenografts in nude mice with veliparib and IR induced persistent foci, increased senescence in tumor and stroma, and delayed tumor regrowth. Our results suggest that the efficacy of PARP inhibitors may partially depend on a competent senescence response to accumulated DNA damage, validating this response as a desirable outcome of genotoxic therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-363. doi:10.1158/1538-7445.AM2011-LB-363
Abstract LB-365: Unfolded protein response regulator BIP/HSPA5 as a chemoresistance biomarker and molecular target in acute lymphoblastic leukemia
Authors: Uckun Fatih M. , Qazi Sanjive , Garner Amanda , Ma Hong , Ozer Zahide , Pitt Jason , Janda Kim 
Published: August 2014

Abstract
BIP/HSPA5, also known as glucose regulated protein (GRP) 78, is an endoplasmic reticulum (ER)-resident multifunctional molecular chaperone that represents a pivotal component of the pro-survival arm of the unfolded protein response (UPR) signaling pathway. We performed a comprehensive meta-analysis of 17 datasets and 668 leukemia/lymphoma cases using the Oncomine Clinical Research Database. 7 of 10 contrasts for high-grade B-lineage lymphoid malignancies exhibited signigficant up-regulation of BIP/HSPA5, including B-lineage acute lymphoblastic leukemia (ALL) (2-fold, P=5.9×10–10). Notably, the gene expression profiles of primary leukemia cells in matched-pair leukemic specimens obtained from 27 B-lineage ALL patients both at the time of initial diagnosis and at the time of relapse were extracted from the NCBI Gene Expression Omnibus (GSE18497) revealed a significant 1.4-fold up-regulation of the BIP8/HSPA5 gene in relapse clones (P=0.0016). Likewise, anti-BIP/HSPA5 Western blot analysis of whole cell lysates from primary leukemic cells from relapsed and newly diagnosed B-lineage ALL patients revealed higher levels of BIP/HSPA5 protein in relapse specimens. Confocal microscopy revealed abundant expression of BIP/HSPA5 protein in the plasma membrane of chemotherapy-resistant ALL cells. Pep42, a cyclic 13-mer cell-penetrating peptide that was identified by screening a pool of cyclic peptide phage display libraries using whole-cell panning against human melanoma cells, was found to not only specifically bind to surface expressed BIP/HSPA5 but also enter BIP/HSPA5+ cells by receptor-mediated endocytosis. We examined the ability of a doxorubicin conjugate of Pep42 (PEP-DOX) containing a cathepsin B-cleavable linker to target and induce apoptosis in surface BIP/HSPA5+ chemotherapy-resistant B-lineage ALL cells. By using a quantitative flow cytometric apoptosis assay, we documented that within 48 hours of treatment with 0.1–1.0 μM PEP-DOX (but not unconjugated drug-free Pep42, which was included as a control) apoptosis is induced in ≥99.9% of surface BIP/HSPA5+ ALL cells. As PEP-DOX specifically targets surface expressed BIP/HSPA5, 100-fold molar excess of drug-free Pep42 was able to compete with PEP-DOX for surface BIP/HSPA5 binding sites and thereby prevent PEP-DOX-induced apoptosis. Excess recombinant EGF, that was included as a negative control, did not affect PEP-DOX- induced apoptosis. The identification of the BIP/HSPA5 as a chemoresistance biomarker and molecular target for B-lineage ALL may lead to new anti-leukemic treatment strategies for relapsed disease that are much needed.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-365. doi:10.1158/1538-7445.AM2011-LB-365
Abstract LB-381: Inhibition of the hedgehog signaling pathway has significant preclinical activity and prevents development of androgen-independent prostate cancer
Authors: Mahalingam Devalingam , Liang Sitai , Buonamici Silvia , Sun LuZhe 
Published: August 2014

Abstract
Introduction: Prostate cancer (PCa) is the second leading cause of cancer death in American men. Patients with advance PCa are often treated first with anti-androgen therapy, but will invariably develop androgen-independent prostate cancer (AIPC). A recent study has shown that androgen deprivation can lead to increased expression of hedgehog signaling components. Hedgehog signaling is also known for normal and cancer stem cell self-renewal. Therefore, in this study, we tested the hypothesis that the inhibition of hedgehog pathway with a novel hedgehog inhibitor, LDE225 (Novartis®), will block the development and growth of AIPC.
Methods and Results: Two androgen-dependent PCa (ADPC) cell lines were adapted to grow in an androgen-free culture medium. The emergence of AIPC cells was found to be associated with increased expression of hedgehog ligands, and hedgehog signaling activity through the activation of its downstream Gli transcription factors as detected using a Gli-responsive promoter luciferase reporter. We also found that treatment with LDE225 was more effective in inhibiting the growth of AIPC cells than ADPC cells. Furthermore, the treatment with LDE225 also inhibited the emergence of AIPC cells when the ADPC cells were cultured in the androgen-free medium. Finally, treatment of orthotopic xenograft PCa model with LDE225 is underway to confirm our pre-clinical in-vitro activity in both ADPC and AIPC models.
Conclusion: These results support our hypothesis that the activation of hedgehog pathway contributes to the development of AIPC and provide the rationale and guidance for design of clinical trials utilizing LDE225 in PCa patients.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-381. doi:10.1158/1538-7445.AM2011-LB-381