bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4278: Spheroid growth in 3D alginate matrices
Authors: Dornish John Michael , Andersen Therese , Heier-Baardson Helene , Markussen Christine , Ward Carol , Mullen Peter , Langdon Simon , Kunkler Ian , Harrison David 
Published: July 2012

Abstract
The use of 3-dimensional (3D) matrices for cell culture is gaining popularity as a substitute for traditional 2D cell culture as it can approximate cell architecture and cell-cell contact found in tissues, organs and tumors. NovaMatrix-3D™ is an alginate-based cell culture system comprising an alginate foam matrix and an alginate immobilizing solution. In principle, cells are first suspended in a sodium alginate solution then the cell suspension is applied to the alginate foam. In situ gelation occurs when calcium ions are donated from the foam cross-linking the added alginate, effectively entrapping the cells within the pores throughout the foam. To show the utility of the NovaMatrix-3D™ culture system, various cell lines were first cultured in 2D then prepared as a suspension in 0.5 – 1% sodium alginate in cell culture medium. An aliquot of each cell suspension was added to γ-sterilized NovaMatrix-3DTM foams (12- or 24-well plate size). NHIK 3025 (cervix carcinoma), MCF7 (breast adenocarcinoma), ZR-75-1 (breast ductal carcinoma) and NIH:OVCAR-3 (ovarian adenocarcinoma) were used and single cells were immobilized in the alginate foams. Cell localization within the foam was visualized using confocal microscopy to identify fluorescently labeled (CellTrace™ CFSE) cells. Cell proliferation was measured by counting cells after de-gelling the foam using sodium citrate. In each case, cells were immobilized with or without RDG-coupled alginate to investigate the importance of the presence of cell attachment peptides within the alginate. For these cell lines, cell growth and multicellular spheroid formation was independent of the presence of RGD. Spheroids could also be removed intact by de-gelling the alginate matrix and further processed for histological staining. One example shows the selective staining of apoptotic cells within the spheroid. Use of alginate foams with concomitant in situ immobilization of cells results in a 3D cell culture model with the potential to approximate cell proliferation and architecture within tissues or tumors. The technology enables biomimetic approaches by varying e.g. matrix elasticity, gelling ions, attachment peptides and foam degradation making NovaMatrix-3D™ a versatile cell culture system. Portions of this work have been funded with the support from METOXIA project no.222741 under the 7th Research Framework Programme of the European Union.
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 4278. doi:10.1158/1538-7445.AM2011-4278
Abstract 4296: Activated ALK accelerates the onset of neuroblastoma in MYCN-transgenic zebrafish
Authors: Zhu Shizhen , Lee Jeong-Soo , Shin Jimann , Perez-Atayde Antonio R. , Kutok Jeffery L. , Rodig Scott J. , Neuberg Donna S. , Guo Feng , Helman Daniel , Feng Hui , Stewart Rodney A. , Wang Wenchao , George Rani E. , Kanki John P. , Look A. Thomas 
Published: July 2012

Abstract
Neuroblastoma is the most common extracranial solid tumor in children and originates during development of the peripheral sympathetic nervous system. Advanced-stage disease and a poor outcome are associated with amplification of the MYCN oncogene, often in combination with mutational activation or amplification of another oncogene ALK (anaplastic lymphoma kinase), suggesting that they cooperate in neuroblastoma pathogenesis. To investigate this possibility, we generated a transgenic zebrafish model of neuroblastoma in which human MYCN is expressed under the control of the dopamine-beta-hydroxylase promoter, and show that the resultant tumors recapitulate childhood neuroblastomas histologically, immunohistochemically, and ultrastructurally. Surprisingly, the tumors arise from a subpopulation of neuroblasts that migrate into the adrenal analogue (interrenal gland) in the zebrafish after organogenesis is complete. Coexpression of activated ALK with MYCN markedly increased the frequency of neuroblastoma and accelerated the time of onset, providing conclusive evidence for synergistic interplay between these two oncogenes in neuroblastoma pathogenesis.
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 4296. doi:10.1158/1538-7445.AM2011-4296
Abstract 4297: Morphology of MCF-7 colonies formed in agarose cell culture
Authors: Aulthouse Amy L. , Nino Elizabeth Di , Kinder David H. 
Published: July 2012

Abstract
Previously, we demonstrated that the MCF-7 cell line grows in a permissive 3-D agarose culture (Kinder and Aulthouse, Cancer Letters., 2004, 205(1), 49-53). Single cells form colonies over time; thus, mitotic activity can be monitored in response to various agents. We studied the morphology of the colonies that formed following growth from single cells at 3 time points: 1.5, 2, 2.5 weeks. The MCF-7 cells grew into clusters (solid) and hollow tube-like structures. These structures were confirmed by H&E stained serial sections of paraffin embedded cultures. H&E staining also revealed abundant mitotic figures. Because calcium foci are common in breast cancers, we also evaluated the cultures for calcium expression using the von Kossa reaction. Placenta was used as a control. At all time points, calcium was detected, but no particular pattern was noted in the calcium distribution between the clusters and the hollow tube-like structures. With morphology and calcium expression described, we further examined cell growth inhibition and cytotoxicity of estradiol and tamoxifen on the MCF-7 cells. Added estradiol enhanced the growth of the cells, while tamoxifen was cytostatic at concentrations below 1.3 × 10-4 nM. At high tamoxifen concentrations (&gt;2 × 10-4 nM) cytotoxicity was noted. This was likely due to sigma-2 receptor binding rather than estrogen receptor blocking. Combination of estradiol with tamoxifen had a more pronounced effect on the cytotoxicity caused by tamoxifen. These morphological and mechanistic findings support the use of this model for studying the MCF-7 cell line in agarose culture.
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 4297. doi:10.1158/1538-7445.AM2011-4297
Abstract 4310: Gene expression analysis of axial and appendicular osteosarcoma in the canine model
Authors: Dervisis Nikolaos , Klahn Shawna L. 
Published: July 2012

Abstract
Spontaneously occurring canine osteosarcoma (OSA) is considered an excellent model of the human disease. Canine axial and appendicular osteosarcoma appear histopathologically identical. The clinical course of the two forms, however, seems to diverge. Following surgery, the axial form may be associated with a lower metastatic rate and longer survival than the appendicular form. Identifying gene expression profiles associated with decreased metastatic potential may be crucial for the development of targeted therapies for the human disease. We hypothesized that there are distinct gene expression patterns for canine axial and appendicular OSA.
Six axial osteosarcoma tumor samples and six age, breed, and weight matched appendicular osteosarcoma clinical tumor samples were selected from the Center for Comparative Oncology, Michigan State University tissue bank. RNA extraction, amplification and labeling were performed using standard techniques. Raw gene expression values were obtained using the Affymetrix GeneChip Canine Genome 2.0 Array. Background correction, normalization, PM correction, summarization, and expression analysis were performed using the R software with the affy/limma/sva packages. Gene set enrichment analysis was performed using GSEA software.
All RNA samples passed quality control and were included in the data set. The expression analysis took into account RNA degradation, which was similar in all tumor samples. A total of 184 genes were differentially expressed by &gt;16-fold, between the two groups. Surrogate variable analysis did not result in any statistically significant differences in gene expression. Gene set enrichment analysis resulted in 9 gene sets enriched in axial, and 4 gene sets enriched in appendicular osteosarcoma samples at FDR &lt; 25%. Gene ranking was similar between the two methods of analysis, with MMP1, ATP6AP1, SLC6A8, and dysadherin expression increased in axial tumor samples, while ICAM2 expression increased in appendicular samples.
Expression analysis indicates no statistically significant differences in gene expression between the axial and appendicular OSA. Gene set enrichment analysis indicates potential differences in gene set enrichment between the two types of osteosarcoma. Further quantitative RT-PCR analysis is required to confirm our results. These data will guide hypothesis-driven translational studies, which may have direct clinical implications on the treatment of osteosarcoma.
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 4310. doi:10.1158/1538-7445.AM2011-4310
Abstract 4318: A constitutively active NF-κB2 mutant drives plasma cell tumor development in mice
Authors: Yang Liqun , McCarthy Brian , Ding Jane , Ding Han-Fei 
Published: July 2012

Abstract
NF-κB2 is a member of the NF-κB family of transcription factors. Genetic mutations that lead to the constitutive action of NF-κB2 signaling occur in approximately 20% human multiple myeloma, a common malignant tumor of plasma cells. Some of the mutations occur in the NF-κB2 gene, leading to the generation of C-terminally truncated NF-κB2 mutants that are constitutively active transcription factors. We investigated the pathogenic role of NF-κB2 mutation in the development of plasma cell tumors using a transgenic mouse line with targeted expression of a tumor-derived NF-κB2 mutant, p80HT, in lymphocytes. The mutant mice developed predominantly B cell-lineage lymphomas, with half of them being plasma cell tumors and producing monoclonal Ig. Some of the plasma cell tumors display key features of human multiple myeloma, including a marked increase in bone marrow plasma cell content, osteolytic bone lesions and/or diffuse osteoporosis. Gene expression profiling of splenic B cells from monoclonal Ig-producing p80HT transgenic mice and their wild-type littermates revealed 740 genes that were differentially expressed (575 upregulated and 165 downregulated) in p80HT B cells versus wild-type B cells. Among the genes selectively upregulated in p80HT B cells are those known to be relevant to human multiple myeloma pathogenesis, including cyclin D1, cyclin D2, CD27, CD80, IL10, and survivin. The upregulation of cyclin D1, IL10 and survivin was confirmed by immunoblotting and/or real-time PCR. Chromatin immunoprecipitation and quantitative PCR assays demonstrated direct interaction of p80HT with the surviving gene promoter in vivo. These findings provide direct evidence for a causal role of NF-κB2 mutation in the pathogenesis of plasma cell tumors in mice and suggest a molecular mechanism underlying the tumorigenic activity of NF-κB2 mutation.
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 4318. doi:10.1158/1538-7445.AM2011-4318
Abstract LB-163: Immunotherapy utilizing MUC-1-messangerRNA encoding DC and MUC-1 specific CTL in combination with Gemcitabine targeting unresectable or recurrent pancreatic cancer
Authors: Yoshimura Kiyoshi , Maeda Yoshinari , Shindo Yoshitaro , Watanabe Yusaku , Yoshino Shigefumi , Hazama Shoichi , Oka Masaaki 
Published: July 2012

Abstract
Background; Mucin1 (MUC1) is known as a mucin core protein expressing in inv asive ductal carcinomas of the pancreas. MUC1-specific cytotoxic T lymphocyt es (CTLs) recognize MUC1 moleculesin a HLA-unrestricted manner. We have prev iously reported that the adoptive immunotherapy (AIT) utilizing MUC1-CTLs stimulated with a MUC1-expressing human pancreatic cancer cell line, named Y PK-1, was useful and safe. We also reported the development of an AIT of MUC 1-CTLs in combination with dendritic cells (DCs) pulsed with MUC1-peptide as well.
Here we report that the therapy using DC encoding MUC1-mRNA and MUC1-CTLs targeting unresectable or recurrent pancreatic cancer is safe and effective.
Purpose; We assessed whether this immunotherapy is safe and effective.
Objective and Methods; Twenty patients with unresectable or recurrent pancreatic cancer had the therapy what we planned, one week later, peripheral blood mononuclear cells (PBMC) were harvested by leukapheresis. The PBMCs were separated and induced to be DCs and CTLs. Matured DCs received MUC-1-mRNA by electroporation, then administrated via subcutaneously. CTLs were co-cultured with YPK-1 pancreatic cancer cell line expressing MUC-1 under the stimulation with IL-2, then administrated via intravenously.
Results; Male/Female ratio was 1 to 1 (10/10), Mean age was 60.6 y.o. (37–78), Mean times of administration of cells was 3.35 (1.5–27.9), Mean number of c ells that were administrated was 46 million (DCs) and 1800 million (CTLs), Median observation period was 16.2 months (1.5–27.9). One-year survival rate was 39% (median survival time, 6.9 months). When we focus on the patients with unresectable cancer, their one-year survival rate was 54% (median surviva 1 time, 18.4 months). In addition, patients received DCs of more than 10 million had 58% of one-year survival rate, whereas no patient received less than 10 million's DCs did survive in one year (median survival time, 5.8 months) (p=0.048, Logrank). All of patients received the therapy as outpatients and showed no side effect.
Conclusion; First of all, this therapy is quit safe. Second, patients received DCs of more than 10 million had high one-year survival rate suggesting high volume DCs in this combination therapy had high potential.
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-163. doi:10.1158/1538-7445.AM2011-LB-163
Abstract LB-165: Low TCR diversity (divpenia) is a new prognosis factor of overall survival in metastatic breast cancer
Authors: Manuel Manuarii , Tredan Olivier , Bachelot Thomas , Rabeony Tioka , Chabaud Sylvie , Mouret Jean-François , Grives Audrey , Perez Solène , Puisieux Alain , Blay Jean-Yves , Caux Christophe , Pasqual Nicolas , Caux Christine Menetrier 
Published: July 2012

Abstract
Rationale
Lymphopenia (&lt;1000 Lymphocytes/μl) or CD4+ T cell lymphopenia (&lt;450/μl), detected before initiation of chemotherapy are predictive factors for toxicity and death in metastatic solid tumors (Borg et al 2004; Ray Coquard et al 2009).
The goal of the present study was to further identify the characteristics of the T cells in these lymphopenic patients. TCR diversity was investigated and tested as predictive factor for overall survival (OS).
Patients and methods
The ImmunTraCkeR® assay (ImmunID, Grenoble France), which analyzes, through semi quantitative multiplex PCR, the V-D-J combinatorial diversity of TCR-beta chain (TRB), was used to investigate diversity of T cell repertoire on cryopreserved blood samples from a prospective cohort of untreated metastatic breast cancer patients (SEMTOF study)(n=66). Univariate and multivariate analysis of prognostic factors for OS were investigated in this series as well as in a validation series.
Results
A severe T cell Divpenia (T cell diversity below 33%) (average diversity for healthy people is 70%) was associated with median survival of 9 months vs 24 months for the remaining patients (logrank p.value=0.0047). In a multivariate analysis, including haemoglobin level, polynuclear neutrophil count and liver metastasis, divpenia (defined at 30% threshold defined by ROC curve analysis) was identified as an independent prognostic factor. The NDL® score (Numeration Diversity Lymphocytes representation) that combines lymphocytes numeration with TRB diversity, demonstrated that lympho-divpenia (T cell diversity below 33% and lymphopenia at 1Giga/L threshold) was associated with a poor prognosis in term of patients survival compared to patients with either a good numeration or good diversity or both good numeration and diversity (p.value=0.0202). Prognostic value of NDL® score was then evaluated on a prospective validation cohort of 33 patients (logrank p.value=0.002).
Conclusion:
Divpenia, and a combined parameter NDL are predictive factors for survival in metastatic breast cancer patients. An open clinical study (LYMPHOS1) will investigate divpenia and NDL® score in a larger prospective cohort of metastatic breast patients.
Acknowledgments
M.MANUEL is a CIFRE recipient. This work (Proof of Concept “LYMPHOS1”) was financially supported by CLARA (LYON, France)
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-165. doi:10.1158/1538-7445.AM2011-LB-165
Abstract LB-169: Dissecting signaling pathways regulating translation in human glioblastoma
Authors: Grzmil Michal , Hemmings Brian 
Published: July 2012

Abstract
Glioblastoma multiform (GBM) is the most common and aggressive brain tumor entity, with a median survival of approximately one year. In the last decade, genetic profiling of brain tumors has improved our understanding of gliomagenesis and proposed many targeted therapies based on molecular interference with deregulated signaling networks. In a search for novel molecular targets, our kinome-focused microarray analysis identified overexpressed MAP kinase-interacting kinase 1 (MNK1) in primary GBM and its elevated protein level was confirmed in primary GBMs and in glioma cell lines. MNKs can bind to translation initiation factor eIF4G and phosphorylate the cap-binding protein eIF4E at Ser209 that is required for its oncogenic activity. Targeting MNK1 activity in GBM cells suppressed eIF4E phosphorylation, reduced proliferation and colony formation whereas, concomitant treatment with Mnk inhibitor CGP57380 and mTOR inhibitor rapamycin resulted in an additive effect on growth inhibition and cell cycle arrest. Analysis of polysomal profiles revealed inhibition of translation in CGP57380 and rapamycin treated cells. Microarray analysis of total and polysomal RNA from MNK1-depleted GBM cells identified mRNAs involved in regulation of TGF-β pathway. The translation of SMAD2 mRNA as well as TGF-β-induced cell motility was regulated by MNK1-signaling. TGF-β pathways regulate growth of glioma-initiating cells that are involved in tumorigenesis, resistance to cancer therapies and relapse. Our most recent analysis identified high activity of MNK1 pathway in human GBM-derived spheres. Furthermore, inhibition of MNK1 activity significantly reduced formation and growth of cancer spheres. Our model of gliomasphere growth regulation via MNK1-signaling pathways together with clinical implications will be presented and discussed.
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-169. doi:10.1158/1538-7445.AM2011-LB-169
Abstract LB-172: Role of post-translational modification of the RUNX2 transcription factor in osteosarcoma
Authors: Ito Tatsuo , Xinyang Zhao , Suehara Yoshiyuki , Kawai Akira , Nimer Stephen D. , Ladanyi Marc 
Published: July 2012

Abstract
Background-Several transcription factors or co-factors have recently been reported to be methylated at certain arginine residues, a post-translational modification mediated by two classes of protein arginine methyltransferases (PRMT). Moreover, some PRMTs have been shown to be key regulators of mesenchymal differentiation, and one of the PRMTs is involved in the epigenetic programming of early embryo development. RUNX2 is major transcription factor in osteoblast differentiation but little is known about its activity or function in osteosarcoma (OS). RUNX1, another member of RUNT family, is known to methylated at R206/R210, within the Runt domain, a region that has considerable homology with RUNX2. Here, we examined the possible role of PRMTs in OS in relation to the post-translational modification of RUNX2 and its possible impact on cell proliferation and differentiation.
Method-We generated arginine methylation-specific antibodies to RUNX2. The specificity of this antibody was confirmed by detecting immunoprecipitated RUNX2 from Saos2 cells by Western blot. We also confirmed the expression of RUNX2 and arginine methylated RUNX2 in Saos2 cell by immunofluorescence. We also examined the frequency of arginine methylation of RUNX2 in OS patient samples, compared to total RUNX2, by Western blot. Finally, we examined the influence of arginine methylation on the transcriptional activity of RUNX2 by luciferase assay using osteocalcin promoter plasmid.
Results-We found that PRMTs can methylate the arginine residue in the Runt domain of RUNX2. This arginine methylation of RUNX2 alters its transcriptional activity at the osteocalcin promoter.
Immunofluorescence with arginine methylation-specific antibody showed weak nuclear positivity in the SAOS-2 cell line. Patient tumor samples also show the expression of arginine methylated RUNX2, as well as total RUNX2.
Conclusion- Our results confirm that arginine methylation of RUNX2 affects its function as a transcription factor. Further study of the possible role of arginine methylation of RUNX2 in regulating its effect on osteosarcoma proliferation and differentiation could be of translational interest, given the therapeutic potential of PRMT inhibitors.
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-172. doi:10.1158/1538-7445.AM2011-LB-172