bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4793: MUC1 immunogenicity in a triple transgenic mouse model for epithelial ovarian cancer
Authors: Budiu Raluca A. , Mantia-Smaldone Gina M. , Brozick Joan , Edwards Robert P. , Vlad Anda M. 
Published: February 2011

Abstract
Background: Ovarian cancer continues to be associated with a very high mortality rate. Improvements on the development of new therapeutic approaches, including antigen-specific immune therapies, depend in part on the availability of adequate preclinical models for in vivo testing of treatment efficacy. MUC1 mucin is a tumor associated antigen and a potential target for cancer immunotherapy. Human and murine mucin 1 share little homology in the extracellular region and transgenic mouse models with de novo lesions expressing the human antigen are needed.
Objective: To identify MUC1 antigen-specific immune regulation in mice with de novo malignant ovarian lesions.
Methods: In order to generate a human MUC1 antigen expressing ovarian cancer mouse model, we crossed the MUC1 transgenic mice which express human MUC1 under the endogenous promoter with the previously described KrasG12D/+PtenloxP/loxP mice that develop orthotopic ovarian tumors.
Results: The newly generated MUC1+/−KrasG12D/+ PtenloxP/loxP triple transgenic mice constitute a valuable Cre-loxP conditional in vivo model for MUC1 expressing ovarian tumors. Upon injection of Cre-encoding adenovirus under the ovarian bursa, the triple transgenic mice develop aggressive ovarian tumors with clear cell histology. The lesions express high levels of MUC1 and are estrogen receptor and cytokeratin 7-positive, markers classically used to diagnose malignant epithelial proliferation. The tumors are accompanied by ascites and numerous peritoneal implants, mimicking late stage disease clinically seen in women. We identified increased accumulation of Treg cells in the para-aortic (draining) lymph nodes and presence of Th17 cells in ascites, mimicking closely the immune phenotype of the human disease. In addition, using quantitative PCR T cell gene profiler arrays, we identified 5 significantly up- and 16 down-regulated genes in the spleen T cells of tumor-bearing mice. One of the upregulated genes was osteopontin (Spp1), recently validated as a marker in detecting human recurrent ovarian cancer. Among the down-regulated genes we identified, Cd4, Cd40ligand (Cd40lg), Interferon-g (Ifnγ), IL-23 receptor (IL-23ra) all suggestive of a shift in immune effectors, some of which may promote the chronic inflammatory background.
Conclusions: Our results reveal important immune regulatory mechanisms in mice with de novo MUC1-expressing ovarian tumors and provide unique opportunities for future in vivo testing of MUC1 vaccines as ovarian cancer therapy and potentially immune prevention.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
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 4793.
Abstract 4795: Kupfer-type immunological synapse characteristics do not predict anti-brain tumor cytolytic T-cell function in vivo
Authors: Lowenstein Pedro R. , Yang Jieping , Sanderson Nicholas , Wawrowsky Kolja , Castro Maria G. 
Published: February 2011

Abstract
To analyze the in vivo structure of antigen-specific immunological synapses during an effective immune response, we established brain tumors expressing the surrogate tumor antigen ovalbumin, and labeled antigen-specific anti-glioma T cells using specific tetramers. Using these techniques we determined that a significant amount of antigen-specific T cells were localized to the brain tumor and surrounding brain tissue, and a large percentage could be induced to express IFNγ when exposed to the specific ovalbumin-derived peptide epitope SIINFEKL. Detailed morphological analysis of T cells immunoreactive for tetramers in direct physical contact with tumor cells expressing ovalbumin, indicated that the interface between T cells and target tumor cells displayed various morphologies, including Kupfer-type immunological synapses. Quantitative analysis of adjacent confocal optical sections was performed to determine if the higher frequency of antigen-specific anti-glioma T cells present in animals that developed an effective anti tumor immune response, could be correlated with a specific immunological synaptic morphology. Detailed in vivo quantitative analysis failed to detect an increased proportion of immunological synapses displaying the characteristic Kupfer-type morphology, in animals mounting a strong and effective anti-tumor immune response, compared to those experiencing a clinically ineffective one. We conclude that an effective cytolytic immune response is not dependent on an increased frequency of Kupfer-type immunological synapses
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 4795.
Abstract 4797: Dendritic cells loaded with autologous tumor lysate by electroporation effectively induce lysate specific IFN-γ producing T cells in glioblastoma patients
Authors: Takahara Masashi , Tomiyama Mai , Nieda Mie , Goto Shigenori , Peshwa Madhusudan V. , Kanemura Yonehiro , Maekawa Ryuji , Moriuchi Shusuke 
Published: February 2011

Abstract
Electroporation (EP) can facilitate antigen uptake by dendritic cells (DCs) with consequently enhanced antigen processing and presentation. Earlier we reported that DCs electroporated in the presence of tumor lysate are fivefold to 20-fold more effective in proliferation of tumor-lysate-specific T lymphocytes compared to controls, DCs incubated with the same lysate. In this study we examined the ability of EP-loaded DCs to induce tumor-lysate-specific IFN-γ producing T cells. DCs were prepared from peripheral CD14-positive monocytes by culturing and maturating in culture medium containing GM-CSF, IL-4, PGE2 and TNF-α. We electroloaded DCs from four glioblastoma patients who had been surgically treated. in the presence of autologous tumor lysates (final concentration of 2 mg/mL). In some experiments, we added zoledronate in the culture medium during DC maturating process in order to examine its enhanced effect on the induction of tumor-lysate-specific IFN-γ producing T cells. At 1.5 hrs after EP, we assessed such DCs’ ability to stimulate and proliferate T cells in comparison with DCs co-cultured with the same tumor lysate as control. We incubated autologous peripheral blood lymphocytes (1×106) with lysate-loaded DCs (1×105) for 14 days with IL-2 replenished every two to three days. Surface markers for T cell subsets were analyzed by flow cytometry on days 7 and 14 and IFN-γ secreting T cells by ELISPOT on day 14. EP-loaded DCs induced more IFN-γ producing T cells than control DCs (p&lt;0.001). In two patients (Patient #2 & #3) that expressed higher levels of HLA class I molecules in resected tumor tissue, especially, IFN-γ producing T cells were tumor specific because such T cells significantly higher induced compared with DCs without lysate (p&lt;0.001). Addition of zoledronate, further increased the induction of tumor lysate specific IFN-γ producing cells, concurrent to correlative expansion of the gamma-delta T cells in culture. We hypothesize that the gamma-delta T cells may play important roles in enhancement of tumor lysate specific IFN-γ producing cells. These results suggest that DCs electro-loaded with autologous tumor lysate in combination with zoledronate treatment induce more effective immune responses and may lead to development of more effective immunotherapies for treatment of glioblastoma patients.
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 4797.
Abstract 4798: Cytotoxic T cells kill tumor cells in a contact and antigen-dependent manner, but the interaction affects neighboring, non-target cells via interferon gamma secretion
Authors: Sanderson Nicholas SR , Puntel Mariana , Baker Greg , Castro Maria G. , Lowenstein Pedro R. 
Published: February 2011

Abstract
The interaction between cytolytic T cells and antigen presenting targets involves stable intercellular contacts known as immunological synapses. It has been postulated that one function of immunological synapses might be to restrict the effector functions of the T cell to the target cell, while protecting bystanders (non-target cells). We wished to test this hypothesis in the context of an anti-tumor immune response. To model the three-way interaction between T cells, target cells and bystanders, we used OT-I ovalbumin-specific T cells, GL26 murine glioma cells expressing ovalbumin, and wild type mouse astrocytes. We confirmed the formation of immunological synapses between T cells and GL26 cells, and assessed whether two effector functions of T cells, i.e., cytolysis and cytokine action, were limited to the post-synaptic target cells. We found that cytolysis was strictly restricted to the target tumor cells, but that the effect of interferon gamma, as measured by Stat1 phosphorylation, was more pronounced on the bystander astrocytes. We conclude that a likely model of cytotoxic T cell action in tumor immunity involves direct lysis of tumor cells, and interferon gamma action on the tumor stroma. Our data has implications for the development of improved anti-tumor imunotherapies which will target both the tumor cells and the tumor microenvironment.
This work is supported by National Institutes of Health/National Institute of Neurological Disorders & Stroke (NIH/NINDS) NIH/NINDS Grant 1R21-NS054143, 1UO1 NS052465 and 1RO1-NS057711 to M.G.C.; NIH/NINDS Grants 1 RO1 NS 054193, RO1 NS061107 and 1R21 NS047298 to P.R.L.
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 4798.
Abstract 480: Establishment of human metastatic hepatocellular carcinoma HCCLM3 nude rat model for drug evaluation
Authors: Zhang Jing , Wu Haiyan , Zhang Ruihong , Zhang Yanmei , Shi Mei-shu , Duan Jeff , Zhang Kevin 
Published: February 2011

Abstract
Currently most of antitumor pharmacology studies are performed in mice using either human xenograft or murine tumors. However, the utility of rat tumor models has not been fully explored. Some of the obvious advantages of using rats remain to be validated, for example, the easy combination of pharmacokinetic study with an efficacy study, the easy performance of animal procedures, the more options for local dosing or interventions, etc. HCCLM3 is one of the human hepatocellular carcinoma(HCC) lines that are originally derived from Chinese HCC primary tumor and well established in mouse model system over last 20 years. To realize the potential utilities of rats for tumor models, especially for the need of evaluating various local treatments for the HCC, we examined the growth and metastasis of HCCLM3 tumor following subcutaneous and orthotopic xenograft in immunodeficient rats (RNU nude rats, Charles River). Following either subcutaneous or orthotopic inoculation, HCCLM3 was able to successfully grow up with 100% take-up rate. The subcutaneous tumor grew more quickly in nude rats than in BALB/c nude mice and the tumor weight reached about 10% of body weight 28 days after inoculation. The treatment with Sutent (28mg/kg, p.o. 5 times per week) significantly inhibited the subcutaneous tumor growth by 93% (P&lt;0.05). In comparison, the orthotopic tumor grew rather slower but showed much stronger lung metastasis potential which was not observed in nude mice in 28 days. HCCLM3 in nude rats also demonstrated the growth pattern of the single nodule growth, different from the intrahepatic dissemination and multiple nodule development in the nude mice. Our established HCCLM3 subcutaneous and orthotopic xenograft rat models provided additional useful tools for evaluation of therapeutic profile of drug candidates and different interventional strategies for the treatment of human hepatocellular carcinoma.
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 480.
Abstract 481: METCAM promotes in vitro motility, invasiveness and colony formation, and in vivo tumorigenesis of human breast cancer cells
Authors: Wu Guang-Jer 
Published: February 2011

Abstract
METCAM (alternative names are MUC18, CD146, S-endo 1, MelCAM, and MCAM) is an integral membrane CAM in the Ig-like gene super-family because it has an immunoglobulin-like extra-cellular domain and a cytoplasmic domain, which contains five consensus sequences potentially to be phosphorylated by PKA, PKC, and CK2. METCAM, as predicted from the protein structure, is capable of performing typical functions of CAMs, such as cell-cell interaction, cell-extracellular matrix interaction, and cross-talk with many signaling pathways. Thus altered expression of METCAM should change cell motility and cellular invasiveness, alters angiogenesis, and affects cellular survival and growth. Consistent with this notion, aberrant expression of METCAM positively associates with the malignant progression of melanoma and prostate cancer. However, the role of METCAM in the progression of human breast cancer cells has been controversial. Two groups suggest that METCAM plays a tumor suppressor role in the development of breast cancer. Two other groups suggest the opposite role. To resolve this controversy, we reinvestigate the role of METCAM in the progression of breast cancer cells. We transfected the METCAM cDNA gene into two breast cancer cell lines, MCF-7 and SKBR3, and isolated G418-resistant clones, which expressed high levels of the protein, for testing the effect of enforced expression of the protein on their in vitro and in vivo cellular behaviors. We have found that METCAM is not expressed or very weakly expressed in five luminal breast cancer cell lines, MCF-7, T47D, BT-10, BT474, and SK-BR-3, but moderately expressed in two basal cell-like breast cancer cell lines, MDA-MB231 and 468. Enforced expression of METCAM in two breast cancer cell lines, MCF-7 and SKBR3, increased their in vitro motility and invasiveness and in vitro anchorage-independent colony growth, and promoted disorganized growth in 3D basement membrane culture. Enforce expression of METCAM also increased the in vitro motility and invasiveness of MDA-MB231 and MDA-MB-468 cells. Furthermore, the growth of MDA-MB231 and MDA-MB-468 in the 3D embedded basement membrane culture is inhibited by the anti-huMETCAM antibodies. Furthermore, enforced expression of METCAM dramatically promoted the tumor formation of SK-BR-3 cells in nude mice, but slightly promoted the tumor formation of MCF7 cells in SCID mice via orthotopic injection into mammary fat pads. Taken together, we concluded that over-expression of METCAM in breast cancer cell lines appeared to promote the tumorigenesis of breast cancer cells, suggesting its positive association with the progression of breast cancer cells, similar to melanoma and prostate 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 481.
Abstract 4810: Persistent migration of cancer cells in a 3-D mechanically confined microenvironment
Authors: Irimia Daniel 
Published: February 2011

Abstract
When cancer cells spread away from the primary tumour, they often follow the trajectories of lymphatic vessels, collagen fibres, white matter tracts, or other heterogeneous structures in tissues. To better understand this form of guided cell migration we designed a series of microfluidic devices that mechanically constrain migrating cancer cells inside microstructures with cross-section comparable to cell size and replicate critical 3D interactions that cancer cells would normally have inside tissues. We observed unexpectedly fast and persistent movement in one direction for several hours of cancer cells of different types. We observed the persistent motility in the absence of external gradients, suggesting the presence of intrinsic mechanisms driving cancer cell motility that are induced in conditions of mechanical confinement. To probe into the mechanisms responsible for this behaviour, we exposed cancer cells inside capillaries to drugs targeting the microtubules and measured significant reduction in the average migration speed in response. Surprisingly, a small numbers of cells appeared not to be affected by the treatment and displayed fast and persistent migration, comparable to untreated cells. The new matrix-free, 3D-confined motility assay is compatible with high-content, high-throughput analysis of cellular motility at single cell level and could provide useful insights into the biology of cancer cell migratory phenotype.
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 4810.
Abstract 4813: Modeling interactions between endothelial cells and metastatic breast cancer cells in a three-dimensional culture
Authors: Connor Yamicia , Harfouche Rania , Liu Cassie , Oh Michael , Sengupta Shiladitya 
Published: February 2011

Abstract
Standard two-dimensional model systems are limited in their ability to accurately capture the molecular mediators underlying breast cancer, and cannot be used to study interactions between multiple cell types. Previous studies have shown that three-dimensional systems provide a more physiologically relevant platform to study cancer progression in a structurally appropriate context 1. However, these systems do not capture the interactions between tumor cells and the angiogenic component, an essential step in tumor progression2. Here we describe, a novel three-dimensional co-culture system combining angiogenic endothelial cells and breast epithelial cells in a three-dimensional tumor matrix. Endothelial cells underwent angiogenic tubulogenesis on a matrigel matrix. Addition of metastatic green fluorescent protein+ve MDA-MB-231 resulted in a unique phenotype characterized by the alignment of the tumor cells on the endothelial tubes. We demonstrate FAK and B1 integrin signaling are important mediators of the observed phenotypes. Inhibiting FAK-B1 signaling disrupted the architecture, reducing the ability of the MDA-MB-231 cells to migrate and adhere to cultured vessels. Furthermore, treatment with small molecule inhibitors of phosphatidylinositol 3 kinase, LY294002 1.6uM, and mitogen activated protein kinase (MAPK), PD98059 20uM, also perturbed this architecture, indicating that these signaling pathways play a key role in controlling the observed phenotype. Furthermore, the inhibition of the PI3K and MAPK pathways, destroyed the tumor vasculature and inhibited progression and invasion of the MDA-MB-231 by 86% (p&lt;0.001) while enhancing the cells’ susceptibility to the cytotoxic agent doxorubicin. Our study shows that a three-dimensional co-culture system provides a powerful in vitro model system to study the mechanisms underlying tumor metastasis and is a powerful tool to screen combination therapies. Supported by DoD BCRP.
1 Lee GY, et al. Nat Methods. 2007 Apr;4(4):359-65.
2Sengupta S, et al. FASEB J. 2004 Oct;18(13):1565-7.
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 4813.
Abstract 4818: Validation and clinical utility of a human hepatocellular carcinoma metastasis signature as prognostic classifier
Authors: Roessler Stephanie K. , Jia Hu-Liang , Budhu Anuradha S. , Qin Lun-Xiu , Ye Qing-Hai , Thorgeirsson Snorri , Tang Zhao-You , Wang Xin W. 
Published: February 2011

Abstract
Hepatocellular carcinoma (HCC) is the most frequent malignant tumor in the liver and the third leading cause of cancer death worldwide. The high mortality of HCC is mainly caused by intra-hepatic metastasis or the high rate of recurrence after surgery. Despite potentially curative surgical resection, eligibility is scarce and patients suffer from a high recurrence rate. Recently, we developed a tumor-based 153-gene signature that is predictive of HCC portal venous metastasis and survival in 40 HCC patients (Ye et al, Nat Med, 9: 416-23, 2003). This signature was based on cDNA microarray profiling of 20 well defined HCC cases, in which ten developed tumor thrombi in the major branch of portal vein and ten had no metastasis at one year follow-up. In this study, we sought to validate this signature for its utility in predicting HCC survival using two independent cohorts. The first cohort consisted of 247 predominantly HBV+ HCC cases analyzed by the Affymetrix U133A 2.0 platform. The second cohort included 139 cases with mixed etiology analyzed by the Human Array-Ready Oligo Set (version 2) array platform. We employed a survival risk prediction algorithm with 10-fold cross validation and 1000 random permutation to test whether the 153-gene signature is associated with survival. We found that this gene signature was able to predict prognosis in both independent HCC cohorts. Importantly, the risk prediction was independent of the microarray platform and etiology. In addition, survival prediction was successful in patients with small (&lt;5 cm in diameter) and/or solitary tumor and the signature performed well on patients particularly with early recurrence (&lt;2 years). Thus, the 153-gene signature was validated in three independent cohorts to be related to HCC patient survival. Furthermore, the risk prediction performed independently of clinical characteristics and microarray platform. We recommend that this signture may serve as a molecular diagnostic test useful to assess the risk that an HCC patient will recur within 2 years after resection, particularly for those with tumors &lt;5cm and in a solitary presentation.
Supported by NIH intramural research funds
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 4818.