bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 510: Identification and characterization of immunogenic epitopes from CD133 and their potential for use to immunologically target cancer stem cells
Authors: Pinilla Clemencia , Judkowski Valeria , Bunying Alcinette , Ji Jianfei , Li Zhenhua , Bender James , Yu John S.
Published: August 2012

CD133 is a marker that identifies cancer stem cells (CSC) on many solid tumors and its expression has been correlated with shortened survival. Although expressed on many normal tissues and stem cells, CD133 is overexpressed on CSC and normal SC are negative or express low levels of MHC making them less sensitive to recognition by CD8+ cytotoxic T lymphocytes (CTL). Potential CTL epitopes were identified by algorithms to predict binding to HLA-A2. Binding studies showed high affinity binding and low off rates for four candidate epitopes. CTL induction studies using IL-12 secreting dendritic cells to stimulate CD8+ T cells evaluated the immunogenicity of the candidate peptides after three or four weekly in vitro stimulations. Multimer and IFNγ responses were observed with CD133-753, 405 and 708 epitopes. Subsequent studies demonstrated killing of T2 cells loaded with peptide. To evaluate the potential for autoimmunity, mouse homolog peptides of the 753 and 405 epitopes that were shown to have high affinity binding to human HLA-A2 were used to immunize HLA-A2 transgenic mice (9CB6F1-Tg (HLA-A*0201/H2-Kb) A*0201). Mice were immunized 3 times at 3 week intervals in IFA + HBV helper peptide and spleens were harvested 2 weeks after the last immunization and stimulated in vitro for one week with peptide pulsed LPS stimulated- APC. An additional group was sacrificed 6 weeks after the last immunization and studied. IFNγ ELISpot assays showed immune responses to the 753 and 405 peptides in 35% and 40% of mice. Organs, including heart, lung, liver, kidney, stomach, intestine, brain, bone marrow, gonads, and eyes from mice with immune responses were found to be negative for lymphocytic infiltrations supporting a lack of autoimmunity related to the immune response to these peptides. Together these studies support the safety and immunogenicity of these peptides as a potential vaccine to target CD133 cancer stem cells.

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 510. doi:1538-7445.AM2012-510
Abstract 5120: Functional genetic screens identify a rare isoform of RABL3 as a modulator of paclitaxel resistance in ovarian cancer
Authors: Zhang Qing , Ota Takayo , Ohta Tsuyoshi , Shridhar Viji , Chien Jeremy R.
Published: August 2012

Recent RNA-sequencing studies have identified variant transcripts, fusion transcripts and novel transcripts in cancer. However, little is known about the role of these variant transcripts in tumor development. To identify a specific role of these variant transcripts in tumor development toward chemotherapy resistance, we generated a custom retroviral cDNA expression library from 10 ovarian tumors with resistance to chemotherapy. Retrovirues were generated from the custom library, and they were used to transduced exogenous genes into ovarian cancer cell line A1847, a cell line chosen for its sensitivity to paclitaxel. A1847 cells transduced with retroviruses carrying alkaline phosphatase was used as a control. Transuced cells were selected with 25 nM paclitaxel. 87 clones survived the selection in library-tranduced cells whereas none survived the selection in alkaline phosphatase-tranduced control cells. Forty seven clones survive the secondary screens. Sequence analysis of transduced genes in these surviving clones indicate that 4 paclitaxel-resistant clones contain RABL3 splice variant. Although 195 accessions corresponding to wildtype transcripts are reported in AceView, there is a single accession corresponding to RABL splice variant that we identified in this screen. RNAi-mediated downregulation of RABL3 resulted in enhance sensitivity toward paclitaxel in these resistant clones. Ectopic expression of RABL3 splice variant resulted in increased resistant to paclitaxel. Collectively, these results demonstrate the feasibility of performing functional genetic screens to identify novel splice variants involved in tumor development.

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 5120. doi:1538-7445.AM2012-5120
Abstract 5125: Functional genomic strategies to identify oncogenes in breast cancer
Authors: Guest Stephen , Haddad Ramsi , Gray Joe , Ethier Stephen
Published: August 2012

During the process of tumorigenesis and tumor progression cancer cells acquire an array of genetic alterations. The theory of oncogene addiction proposes that a small portion of these alterations generate driver oncogenes that are critical for maintaining the transformed phenotype. Strong support for this theory comes from the identification and subsequent therapeutic targeting of individual driver oncogenes that has resulted in significant clinical responses. However, in solid tumors that contain multiple driver oncogenes, these responses are not durable and it is therefore likely that multiple oncogenes will need to be identified and targeted in these tumors in order to reverse the transformed phenotype and effect durable clinical responses. Identifying multiple driver oncogenes among the large number of mutated/amplified genes that are present in most solid tumors is a nontrivial task that will require novel approaches. In this study, we have developed an approach that combines exome sequencing, array comparative genomic hybridization, expression analysis and genome-wide, RNAi-based functional screening to identify driver oncogenes that are functioning in individual cell lines derived from human tumors. In a pilot study using this approach we have shown that of the 583 amplified genes in the SUM225 breast cancer cell line only 45 are hits in the RNAi-based functional screen. Similarly, of the 806 genes that are mutated in this cell line only 26 are hits in the RNAi-based functional screen. Interestingly, the SMPD3 gene that was both amplified and a hit in the functional screen also contained a point mutation and this point mutation was present in another HER2 positive breast cancer cell line, SUM190. The identification of SMPD3 as a likely driver oncogene in the SUM225 and SUM190 cell lines highlights the power of an approach that combines sequence and copy number data with functional screening to identify driver oncogenes. Future analysis will incorporate expression data to further refine this approach.

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 5125. doi:1538-7445.AM2012-5125
Abstract 5129: Identification of therapeutic targets for HPV-positive and HPV-negative head and neck cancers using high throughput RNAi screening
Authors: Azorsa David O. , Gonzales Irma M. , Henderson Meredith C. , Sima Chao , Arora Shilpi
Published: August 2012

Cancers of the head and neck account for three percent of all malignancies in the United States and is the sixth most common type of cancer worldwide. Historically, the two most common risk factors for head and neck squamous cell carcinomas (HNSCC) were tobacco use and/or heavy alcohol consumption. Recent growing evidence strongly supports a role for human papillomavirus (HPV) in a subset (15-30%) of HNSCC and there is currently a rise in HPV-related head and neck cancers. In order to improve treatment of head and neck cancers and identify etiology-specific therapeutic targets, we applied a functional genomics based approach utilizing high-throughput RNAi (HT-RNAi) to identify genes specifically involved in growth of either HPV-positive or HPV-negative HNSCC cells. A HT-RNAi assay was developed to quantify the effects of gene specific siRNA on growth inhibition of two HPV-positive (SCC-090 and UMSCC-047) and two HPV-negative (SCC-25 and CAL-27) HNSCC cell lines. For HT-RNAi screening, the four cell lines were reverse-transfected with siRNA from a library targeting 7000 druggable genes with four siRNA per gene and assessed for growth modulation at 96 hours post transfection. The data was normalized using Z-score analysis and statistical cut-offs were set. Following data analysis, we generated a hit list of gene targets that are involved in growth of HPV-positive or HPV-negative HNSCC cells. Validation of these genes will identify potential therapeutic targets for head and neck tumors depending on their HPV status. Furthermore, these findings demonstrate the effectiveness of using HT-RNAi as a tool for the identification of etiologic-specific targets.

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 5129. doi:1538-7445.AM2012-5129
Abstract 5160: Global metabolomic profiles distinguish malignant from non-malignant human mammary epithelial cell responses to endogenous p53 upregulation by MDM2 antagonism
Authors: Yau Christina , Gabriel Bianca , Scott Gary , Brand Martin , Benz Christopher C.
Published: August 2012

Wildtype (wt) p53 can alter metabolic pathways and mitochondrial function but it is unclear whether these responses are similar in normal and malignant cells, where it is also hypothesized that p53wt can modulate the Warburg effect. Therefore, we used a small molecule inhibitor of MDM2, MI-63 (Ding et al., J. Med. Chem. 2006; Ascenta/Sanofi), to induce ∼5-fold intracellular increases in p53wt protein levels within 24 h of treating malignant (MCF-7) or non-malignant (MCF10A) human mammary epithelial cells, whereupon global biochemical changes were assayed in replicate (x6) extracts using various mass spectrometry platforms (GC/MS, LC/MS/MS; Metabolon, Durham, NC). Extracts from similarly treated p53mut malignant SKBr3 cells were also assessed. Normalized (to total protein) and imputed data from 310 known and detected biochemicals were compared for treatment effects (Wilcoxon rank sum test, p<0.05). Treatment significantly altered levels of 93 biochemicals (58 up, 35 down) in the malignant MCF-7 cells, 62 (39 up, 23 down) in the non-malignant MCF10A cells, and 24 (20 up, 4 down) in the malignant p53mut SKBr3 cells. Common to both MCF-7 and MCF10A, p53wt upregulation increased NAD+ and NADP+ levels, responses not seen in SKBr3 cells. Upregulation of p53wt selectively suppressed polyamines (spermidine, putrescine) in MCF10A, whose response to MI-63 is cytostatic unlike the apoptotic response of MCF-7 or treatment unaffected growth of SKBr3 cells. Relative to MCF10A, MCF-7 cells showed opposing energy-related biochemical responses in glucose, glucose-6-phophosphate, 3-phosphoglycerate and pyruvate pools, along with increases in the glutaminolysis product succinate, suggesting that p53wt upregulation enhances glycolytic and mitochondrial (TCA) production of pyruvate in malignant MCF-7 cells yet decreases glycolysis and pyruvate production in non-malignant MCF10A cells. This conclusion that p53wt upregulation increases glycolysis, pyruvate production and mitochondrial ATP synthesis in MCF-7 was confirmed using live cell extracellular flux analysis of oxygen consumption and acid production (Seahorse XF), which showed opposing treatment responses by MCF10A and no treatment effects in SKBr3. Altogether, these findings point to opposing metabolic and bioenergetic responses induced by p53wt upregulation in malignant and non-malignant mammary epithelial cells, consistent with p53wt induced reversal of the Warburg effect, and offering new insights into pathways associated with different therapeutic and cell fate responses induced by novel small molecule MDM2 antagonists like MI-63.

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 5160. doi:1538-7445.AM2012-5160
Abstract 5163: Differential effects of IGF-2 on epithelial mesenchymal transition (EMT) in African- American and Caucasian breast cancer cell lines
Authors: Hamilton Nalo M. , Amis Louisa , Davis Willie L. , Saldivar April
Published: August 2012

Breast cancer is a multifactorial disease that disproportionately affects women. Breast cancer patients with the worst prognosis tend to be premenopausal, African-American (AA) and present with aggressive, estrogen-independent tumors unresponsive to standard endocrine therapy. Because of these factors, it is important to discover unique biomarkers for screening and treatment of breast cancer. One potential candidate is the insulin-like growth factor-II (IGF-2). Studies indicate that tumors that express high levels of IGF-2 are more aggressive, exhibit a basal-like phenotype and tend to metastasize. Suggesting that enhanced or altered IGF-2-mediated cell signaling pathways may be involved in the pathogenesis and disease progression of metastatic, ER-null breast cancer. We hypothesize that this may be due to the ability of IGF-2 to affect the mesenchymal epithelial transition (EMT) of these cells. To assess the potential effect(s) of proIGF-2 on EMT, we have over-expressed pro- and mature- forms of IGF-2 in human breast cancer cell lines: MCF-7, T47D, Hs578T, CRL-2329, CRL-2335 and Z75D-30. There are phenotypic changes that are characteristic of EMT, cellular morphology was evaluated by phase-contrast and confocal microscopy on days 5, 15, 20, and 30 post-transfection. In cells overexpressing proIGF-2 and mIGF-2, qRT-PCR and Western immunoblot analysis were used to assess expression and activation of E-cadherin, PI3K/AKT, SNAI1/2, RelB, Bcl-2, ERα and ERβ. This project will evaluate proIGF-2 as an initator of EMT and cancer metastisis. [Research support from the Robert Wood Johnson Foundation and Loma Linda University School of Pharmacy]

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 5163. doi:1538-7445.AM2012-5163
Abstract 5279: Over-expression of EGFRviii induces an angiogenic switch in infiltrative human glioblastomas
Authors: Røsland Gro V. , Talasila Krishna , Bjerkvig Rolf , Miletic Hrvoje
Published: August 2012

Glioblastoma multiforme (GBM) is the most common and most aggressive primary brain tumor where local cancer cell invasion into the normal brain represents a major obstacle preventing successful therapeutic interventions. A well known mediator of invasion, proliferation and survival, is the epidermal growth factor receptor (EGFR). Amplification and subsequent over-expression of EGFR protein is the most common genetic alteration in primary GBM, with a frequency of about 40%. Of the GBMs that have an amplification of EGFR, 63-75% also show genetic rearrangements of the gene, resulting in tumors that express both the wild type, as well as mutated forms of EGFR. The most common mutation in EGFR, is the EGFRviii, were the ligand binding part of the protein is missing (del exon 2-7). This mutation leads to a constitutive activation of the protein. In patients with amplified EGFR, 50-60% also express the EGFRviii. In this work we determined a functional role of EGFRviii within invasive human GBM cells in vivo. We cultured human GBM cells, lacking EGFR amplification and EGFRviii expression, in serum-free neuro-basal (NB) medium. By orthotopic transplantation into the brains of immunodeficient rats, tumors developed that showed a highly infiltrative non-angiogenic phenotype. By over-expressing EGFRviii in these tumors, we demonstrate a robust switch towards angiogenesis as shown by dynamic-contrast enhanced MRI and by histologic verification of angiogenic vessels. Molecular analysis confirmed an up-regulation of HIF1A as well as a panel of angiogenic factors. Our results demonstrate a distinct role of EGFRviii as a vital driver of angiogenic tumor growth within human GBMs in vivo. Thus, the presented observations should have important implications for therapeutic strategies targeting EGFRviii in 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 5279. doi:1538-7445.AM2012-5279
Abstract 5285: ADAM12: A novel mediator of tumor angiogenesis
Authors: Roy Roopali , Moses Marsha A.
Published: August 2012

ADAM12 (a disintegrin and metalloprotease) is a member of a family of integral membrane and secreted Zn-dependent metalloproteases that have diverse roles in normal and pathological states. Human ADAM12 is expressed as two alternatively spliced forms, a membrane-anchored long form (ADAM12-L), and a short secreted form (ADAM12-S). Increased levels of ADAM12 mRNA and protein have been reported in a variety of cancers including colon, hepatocellular, lung, gastric and glioblastoma. We have previously reported that urinary ADAM12 is predictive of disease status and stage in breast cancer. We have also demonstrated that expression of ADAM12 isoforms in breast tumor cells results in a higher rate of tumor take, increased tumor size and endocrine resistance. ADAM12-S expression stimulated migration and invasion of breast tumor cells in vitro and local and distant tumor invasion in vivo. In the course of these studies, we observed that ADAM12 is upregulated in the vessels of aggressive breast tumors as compared to normal adjacent breast tissue. ADAM12-overexpressing breast tumors also displayed a higher microvascular density. Furthermore, we found that angiogenic tumors express higher levels of ADAM12 compared to their preangiogenic counterparts. The goal of the present study was to determine whether ADAM12 regulates tumor angiogenesis. Basal levels of ADAM12-L expression in HUVEC and HMVEC are relatively low, however, both ADAM12 protein and mRNA expression is significantly increased when cells are stimulated with growth factors such as VEGF, bFGF or inflammatory cytokines such as TNFα, TGFβ or IL1α. Interestingly, VEGF stimulation of endothelial cells (EC) resulted in ADAM12 protein expression and/or localization specifically along the invadopodia of these cells. Consistent with these results, we found that ADAM12 overexpression in EC resulted in significantly increased rates of migration, capillary tube formation and increased adhesion to fibronectin and type IV collagen but not laminin. In vivo studies demonstrate that ADAM12 significantly increases bFGF-mediated angiogenesis in the mouse corneal pocket assay. Taken together, our data provide evidence supporting a novel role for ADAM12 in the regulation of endothelial cell function and tumor angiogenesis. Studies to investigate the mechanism/s by which ADAM12 may regulate angiogenesis are ongoing. [The authors acknowledge the support of NIH PO1 CA045548, The Breast Cancer Research Foundation and The Simeon J. Fortin Foundation]

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 5285. doi:1538-7445.AM2012-5285
Abstract 5305: Phosphorylations of Serine-10 and Tyrosine-198 residues in the cytoplasmic mislocalized p27 are critical to migration and invasion of osteosarcoma cells
Authors: Li Yiting , Flores Ricardo J. , Hicks John , Chintagumpala Murali , Lau Ching C. , Man Tsz Kwong
Published: August 2012

Osteosarcoma is the most common malignant bone tumor in adolescents and children. Metastasis is the major cause of death among osteosarcoma patients even with the advent of chemotherapy. Therefore, it is an urgent need to identify the key determinants of metastasis in osteosarcoma, so that new and effective therapeutic approaches can be developed to combat this deadly subtype of osteosarcoma. Recent studies have shown that the tumor suppressor protein p27 promotes metastasis when it is mislocalized from the nucleus to the cytoplasm in some solid tumors. However, the subcellular localization of the p27 protein in osteosarcoma and its role in metastasis are not known. In this study, we found that 82% human osteosarcoma cases harbored cytoplasmic staining of p27 using a tissue microarray. An immunohistochemistry analysis revealed that the p27 protein was localized in the nuclei of three pairs of non-metastatic osteosarcoma cell lines, but mislocalized in the cytoplasm of their metastatic sublines. Overexpression of a stable fusion construct of nuclear exporting sequence and p27 in a non-metastatic osteosarcoma cell line significantly increased tumor cell migration and invasion in vitro, suggesting that cytoplasmic p27 can promote metastatic potential in osteosarcoma cells. Site-directed mutagenesis on key phosphorylation sites of cytoplasmic p27 indicated that a mutation on Serine-10 or Tyrosine-198 residue abolished the promoting effects of p27 on cell motility and invasiveness, while a mutation on Tyrosine-157 had no effect. This result suggests that phosphorylations of these two residues are essential for tumor cell migration and invasion. Since mTOR signaling has been implicated in metastatic osteosarcoma, we tested if rapamycin, an mTOR inhibitor, can affect the subcellular localization of p27. We found that exogenous addition of the drug reversed the protein trafficking of p27 from the cytoplasm to the nuclei of metastatic osteosarcoma cells, suggesting that p27 may be a yet undiscovered player in the mTOR signaling of metastatic osteosarcoma. In summary, our results indicate that a novel relationship between p27 mislocalization and the metastatic potential of osteosarcoma cells. Therefore, further characterizations of the role of cytoplasmic p27 and its phosphorylations may lead to the development of a new therapeutic approach for metastatic osteosarcoma.

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 5305. doi:1538-7445.AM2012-5305
Abstract 5308: Id-1 gene and protein as novel therapeutic targets for metastatic cancer
Authors: Murase Ryuichi , Limbad Chandani , Murase Rumi , Soroceanu Liliana , McAllister Sean , Desprez Pierre Yves
Published: August 2012

During normal development and embryogenesis, helix-loop-helix Id proteins regulate the differentiation programs of many tissues. Moreover, recent studies have shown that dysregulated Id expression contributes to cancer progression and metastasis. Specifically, we found high levels of one of the Id family members, Id-1, in breast, prostate, brain and head and neck tumor cells which were highly aggressive, and low levels of Id-1 in non-invasive tumor cells. Ectopic expression of Id-1 in non-aggressive human cancer cells rendered them highly proliferative and invasive, and induced high levels of matrix metalloproteinase (MMP) expression. Conversely, human metastatic cancer cells became significantly less proliferative and invasive in culture when Id-1 protein expression was decreased by antisense RNA or shRNA directed against the Id-1 gene. Next, we asked whether Id-1 could be exploited as a therapeutic target to treat metastatic cancers. Using tumor-bearing animals and systemic gene targeting technique, we showed that reduction of Id-1 levels, and consequently MMP expression, could significantly decrease the metastatic spread of cancer cells. Further, several molecular pathways underlying Id-1 effects on epithelial to mesenchymal transition (EMT) and neoangiogenesis were also identified in certain types of cancer. These results suggested that Id-1 gene and/or protein could be a promising target for development of therapeutic strategies to reduce cancer metastasis. Since Id-1 expression is scarce in most mature adult tissues, a majority of normal cells would not be affected by systemic therapy targeting this gene. We determined that a cannabinoid compound, CBD, could specifically inhibit the expression of the Id-1 gene and, as a result, cancer cell proliferation and invasion in culture and tumor metastasis in vivo. In tumor-bearing mice, we observed significant anti-metastatic effects after daily treatment with CBD. Id-1 expression in tumor cells from lung foci was significantly down-regulated as well as Ki67, a marker of cell proliferation. Finally, CBD prolonged the survival of tumor-bearing mice even though the treatment was initiated after the establishment of metastases. Overall, our data suggest that Id proteins act as key regulators of tumor metastasis and suggest the use of Id genes as novel therapeutic targets.

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 5308. doi:1538-7445.AM2012-5308
Abstract 5374: Significance of ADCC for the action of trastuzumab quantified by novel procedures
Authors: Kute Timothy E. , Stehle John , Ornelles David , Walker Natalie , Vaughn Jim
Published: August 2012

Background: The killing of Her2 + breast cancer cells by the antibody trastuzumab is an attractive therapy because of its specificity and limited side effects. However, many Her2 + positive patients have innate resistants or will become resistant to this treatment. Although Her2 status can be determined by immunohisochemistry or FISH, still only 30% of patients positive for high Her2 status will respond to trastuzumab alone. Improved methods to predict sensitivity to trastuzumab are greatly needed. Since trastuzumab is believed to work by antibody-dependent cell-mediated cytotoxicity (ADCC), we have developed a novel and reproducible bioassay to measure trastuzumab-dependent ADCC which may be predictive of patient response. Methods: Using both an xCELLigence platform (Roche Applied Science) and a flow cytometry assay (PanToXiLux from OncoImmunin, Inc), we measured the ability of mononuclear cells (MNCs) to kill four different breast cancer cell lines in the presence or absence of trastuzumab. Two cell lines used have resistance to trastuzumab and two cell lines have lower Her-2 expression. The xCELLigence platform measures killing in real time while the flow cytometry assay measures the transfer of granzyme B and upstream caspase activity from mononuclear cells to the tumor cells and the activation of cell death processes. The ability of mononuclear cells to bind tumor cells in the presence and absence of trastuzumab was evaluated by microscopy. Using miltenyi beads and immunofluroescence, we have measured which MNC population is most effective in ADCC activity. MNCs from patients with breast cancer and from individuals without disease were analyzed. Results: These studies provide a simple and reproducible assay for ADCC activity. Effective ADCC killing is directly related to Her2 expression on the tumor target. The degree of killing is directly related to the number of mononuclear cells added in the study when trastuzumab is added. Addition of trastuzumab alone to these cell lines results in little killing. MNCs from non-diseased individuals as well as those obtained from breast cancer patients display a broad range of ADCC killing efficiency per cell. Finally, our results demonstrate that NK cells are the major MNC population of effect killing. Conclusion: The two assays presented here and other specific studies show that trastuzumab can work with the immune system to increase cell killing of Her2 overexpressing cells by ADCC. A correlation between the results of these assays and the clinical response to trastuzumab will be presented.

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 5374. doi:1538-7445.AM2012-5374
Abstract 5382: Vascular endotelial growth factor and transforming growth factor-beta are enhanced in plasma of breast cancer patients resistant to endocrine therapy
Authors: Zavadova Eva , Konopásek Bohuslav , Vocka Michal , Fucikova Terezie , Petruzelka Luboš
Published: August 2012

Although more than 2/3 of treated patients respond to the endocrine therapy, most patients with metastatic breast cancer will develop resistance. Estrogen modulate angiogenesis, in part, through the effects on VEGF. It appears that other factor contributing to the resistance may be a transforming factor-beta (TGF-beta). It is highly immunosuppressive factor that inhibits the natural and specific immunity against tumors and stimulates production of vascular endotelial growth factor /VEGF. Defects in the immune response correlated with poor prognosis and therefore in recent years, research focused on a precise description of the status and function of the immune system and its possible effect in cancer patients. The purpose of the study was to monitor immune responses in patients with hormone receptor positive breast cancer resistant to hormone therapy, particularly the examination of cellular (CD4, CD8, HLA-DR) as well as humoral immunity. TGF beta, and VEGF production was monitored and we analyzed the changes during hormonal treatment. Methods: 60 patients included in the research project were implemented routine cancer teratment with hormonal therapy. Basic parameters (histological type and grade, the degree of expression of ER and PR, HER2, and the proliferative marker) were established. Patients were evaluated by a cancer clinical immunologist to exclude immune disorders, allergic or autoimmune origin. TGF beta, VEGF were mesured by ELISA and anti-tumor cellular immunity (CD4, CD8, antigen presenting cells) will be measured by flow cytometry. Results In patients with resistance to hormone therapy mainly depression in cellular immunity was found.Immunglobulin plasma level was dicreased as well (mainly IgG4 subtype). Most patients have shown clinical symptoms of immunodeficiency (frequent infections of respiratory or urinary tract, herpetic infections) TGF beta as well as VEGF plasma level were increased. Significant increase of autoantibodies production was found, particulary in those, involved into blood coagulacion process/anti phospholipide and anti cardiolipin autoantibodies. Conclusion Correlation of these factors with resistance to hormonal therapy, and the state of anticancer immunity could help in the future with the the prediction of resistance to hotmonal therapy and contribute to the selection of targeted immune therapy in cancer patients. Dedication: This project was supported by grant IGA NT11168-3/2010

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 5382. doi:1538-7445.AM2012-5382
Abstract 5398: Direct interaction of inhibitory molecules LAG-3 and PD-1 in murine CD8+ T cell signaling
Authors: Huang Ruea-Yea , Eppolito Cheryl , Matsuzaki Junko , Odunsi Kunle
Published: August 2012

Chronic stimulation by tumor antigens results in exhaustion of antigen-specific T cells and loss of ability to produce key cytokines that are critical for the maintenance of T cell function. We have previously demonstrated that a subset of tumor antigen-specific CD8+ T cells coexpressing LAG-3 and PD-1 are impaired in IFN-γ and TNF-α production and that simultaneous blockade of LAG-3 and PD-1 restores effector function of ovarian tumor antigen-specific T cells in humans. Recently, others have reported that dual blocade of LAG-3 and PD-1 via either by antibodies or gene knockout leads to severe autoimmunity and enhanced immunity to tumors in mice. However, direct evidence of molecular interaction between LAG-3 and PD-1 in T cell signaling is lacking. The signaling molecules downstream of LAG3 are also unknown. Using a combination of ImageStream and confocal microscopy analyses, here we demonstrate that LAG-3 and PD-1 molecules co-localize in approximately 10-15% of activated murine antigen specific CD8+ T cells. Co-localization also occurred with trans-Golgi vesicles, microtubule organizing center, and early/recycling endosomes. Reciprocal immunoprecipitation with specific antibodies for LAG-3 and PD-1 revealed a physical association between LAG-3 and PD-1 and this interaction was abolished in T cells isolated from LAG3- and PD-1 knockout mice. The recruitment of Src family tyrosine kinases and phosphatases to the LAG-3 and PD-1 interaction complex are also implicated. Our data reveal a potential molecular mechanism by which LAG3 and PD-1 co-regulate T cell function.

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 5398. doi:1538-7445.AM2012-5398
Abstract 5472: Role of Harvey Ras (HRAS) mutations in head and neck squamous cell carcinoma (HNSCC)
Authors: Endhardt Katharina U. , Khattri Arun , Keck Michaela , Braegelmann Johannes , Mahmutoglu Derya , Leung Kelley , Dinali Mohamed El , Rieke Damian , Cohen Ezra , Seiwert Tanguy
Published: August 2012

Background: Harvey Ras (HRAS) was recently reported being mutated in head and neck squamous cell carcinomas (HNSCC) and likely plays an important role as an oncogene. The precise role of HRAS mutations for signaling and carcinogenesis of HNSCC remains to be determined. Methods: We completed mutational screening (Sanger Sequencing) for tissues and cell lines focused on the known hotspot mutations G12X and Q61X. Furthermore we performed viability testing for various cell lines and visualized the signaling-effects by itself, in presence of PI3K-, EGFR inhibitors and likewise in combination, by immunoblotting. After suppression of HRAS using siRNA, we determined the cell-viability. Results: In our study we sequenced 100 HNSCC tumor tissues and HNSCC cell lines and identified several canonical HRAS mutations. One cell line contained a G12D HRAS mutation and was further examined. Additional two cell lines with atypical HRAS variants were identified and compared to the classic hotspot mutated cell line. The viability for the mentioned cell lines were indicative of resistance to EGFR inhibition to different degrees. The protein activation levels in important signaling pathways (PI3K/MAPK) confirmed our viability data. HRAS signaling was primarily via PI3K/AKT. Silencing HRAS showed significantly decreased viability. Conclusions: Previous studies have shown that EGFR-targeting agents remain insufficient as single targeted therapy. HRAS appears to contribute to the EGFR-resistance of HNSCC. The canonical mutation G12D appears to signal primarily via PI3K and PI3K inhibitors may be effective. The G12D cell line model indicates a central role of mutated HRAS for signaling and viability consistent with role as a driver mutation.

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 5472. doi:1538-7445.AM2012-5472
Abstract 5482: Sun exposure and prostate cancer risk in New South Wales, Australia: A case control study
Authors: Nair-Shalliker Visalini , Smith David P. , Egger Sam , Hughes Ann Marie , Clements Mark , Kricker Anne , Armstrong Bruce K.
Published: August 2012

Background Ultraviolet-B radiation in sunlight may influence risk of developing prostate cancer. Method We examined the relationship between individual solar UV exposure and risk for prostate cancer in a case control study combining the New South Wales (Australia) Prostate Cancer Care and Outcome (cases) and Non-Hodgkin Lymphoma (controls) studies. Results There was evidence that increasing solar UV exposure increased prostate cancer risk (adjusted OR for highest vs. lowest quartiles of average weekly, warmer months sun exposure 2.07 95% CI: 1.36-3.16) and that this increased risk was most evident with weekend sun exposure (adjusted OR 5.54 95%CI: 2.94-10.43). Increased sun sensitivity was also associated with increased risk of prostate cancer (adjusted OR 1.63 95% CI: 1.09-2.44 for very high sun sensitivity). Conclusion Our results suggest that increasing solar UV exposure increases prostate cancer risk in a high ambient solar UV environment. Impact Given that previous studies conducted mainly in low solar UV environments found either no evidence of an association or evidence of a negative association; our findings suggest the possibility of a U-shaped dose effect relationship between exposure to solar UV and prostate cancer. Further studies are needed to test the hypothesis that high solar UV exposure is a risk factor for prostate cancer and to explore possible mechanisms for such an association.

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 5482. doi:1538-7445.AM2012-5482
Abstract 5499: A descriptive epidemiological study of lobular carcinoma of the breast and the effects of normal lobular regression at menopause
Authors: Yu Anna , Shah Amy , Henson Donald
Published: August 2012

Background: Lobular carcinoma (LC) is a distinctive neoplasm originating in the milk producing glands and constituting about 12% of all breast cancer cases. LC differs from the more common invasive ductal carcinoma (IDC) by morphology and biology. We report the results of a descriptive epidemiological study of LC and a comparison of the pathogenesis of lobular and ductal carcinoma of the breast. Methods: A total of 86,846 cases of invasive LC (ILC) from women, aged 20 to 85, were obtained from NCI's SEER Program for the years 1973-2008 in the United States. There were 20,512 cases of LC in-situ (LCIS) and 66,634 cases of ILC. For comparison, 692,201 cases of IDC and 61,569 cases of ductal carcinoma in situ (DCIS) were collected. The analysis compared trends, age specific rates, log-log plots, age frequencies and relative survival rates. Results: From 1973-2008, the age-adjusted rate of LC progressively increased from 5.8 to 16.4/ 100,000 women. In an age frequency analysis of age versus rate, LCIS increased to 0.22/ 100,000 women at age 50, and then declined to 0.02/100,000 women at age 80. ILC increased to 0.11/100,000 women at age 50, and then plateaued after menopause but did not decline. Log-log plots revealed parallel curves between ILC and IDC in both pre-menopausal and post-menopausal age groups. At age 50, the age specific rate of LCIS increased to 11.3/100,000 women then progressively decreased. In contrast, the age specific rate of DCIS was 22.1/100,000 women at age 50 and continued to increase after menopause. Conclusions: At menopause, the incidence of LCIS decreases as a result of lobular involution of the breast. In contrast, DCIS continues to increase. ILC does not decline but plateaus suggesting that the initiation of ILC occurs before menopause, but the tumor becomes clinically apparent only after menopause. Although morphologically different, parallel log-log plots of age specific rates and age frequency analysis suggest a similar pathogenesis for IDC and ILC even though epidemiological patterns are different.

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 5499. doi:1538-7445.AM2012-5499
Abstract 5519: Prevalence of three types of cancers in southern part of Nigeria
Authors: Nwosu Chinelo Jacinta , Anyangwe Njwen
Published: August 2012

There is an increase in the incidence of various cancers globally. It is estimated that ten (10) million new cases of cancers are diagnosed every year and fifty (50%) of this figure is from developing countries. As death toll from infectious diseases decline globally, cancer is becoming the most potent cause of death led by heart and cardiac diseases. This study was carried out to establish the prevalence of three common cancers that affect mainly women using the Southern parts of Nigeria as a case study. Three age groups of women were used for this study: those below forty (40) years, those between forty (40) and sixty (60) years and those from sixty one (61) and above. The review checked major government owned hospital records from 2006 to 2011 and the results are presented. It was observed that 15% of women from 61 years and above suffered from breast cancer within the period under review. About fifty five (55) % of breast cancer cases occurred in women of age forty (40) years and below while thirty (30) % of cases occurred in the age group forty (40) and sixty (60) years. About four (4) % of breast cancer cases occurred in men. Cervical cancer was more prevalent in women between ages 40-60 years followed by those in age group 61and above. The percentage of women who were diagnosed of cervical cancer in the age group 40-60 years was 37%, percentage of those in age bracket of forty (40) years and below was 16% and those in the age bracket of sixty one (61) years and above that suffered from cervical cancer were 22%. The prevalence of uterine cancer within the period under review was about nine (9) % and distributed in these order; age group forty (40) years and below was 2%, 40-60 years, 3% and 61 years and above was 4%. Thus the study shows that breast cancer was more prevalent in women of 40 years and below, cervical cancer was more prevalent in women of 40 years and above while uterine cancer was low in all age groups. Studies have shown that incidence of breast cancer is rising more in populations that hitherto enjoyed a low incidence rate like many parts of Africa. However there is significant under reporting of diagnosed cancer cases as well late reporting of cancer cases as a result of illiteracy. It was suggested that increase in cancer occurrences in the Southern part of Nigeria is largely due to lack of adequate education and awareness on early detection, and life style of patients. Early detection and treatment have been proven to be effective against cancer proliferation and helps in a better prognosis for 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 5519. doi:1538-7445.AM2012-5519
Abstract 5525: Influence of total dissected lymph node number on metastatic detection rate and survival in 10,382 esophageal cancer patients from the high incidence area for esophageal cancer in northern China
Authors: Wang Li Dong , Dong Jin Cheng , Zhang Peng , Zhou Fu You , Li Xue Min , Li Ai Li , Ji Ai Fang , Liu Zhi Cai , Song Xin , Zhao Xue Ke , Wang Xian Zeng
Published: August 2012

Lymph node (LN) metastasis has been recognized as one of the most important factors for poor prognosis of esophageal cancer (EC). However, the influence of dissected LN number on metastatic detection rate and survival in EC has not been well documented. The present study was undertaken to correlate the total number of dissected LN, metastatic detection rate and survival on 10,382 EC patients. All the patients had been performed surgical treatment and confirmed as esophageal squamous cell carcinoma (ESCC) by histopathological examination. The patients were from Henan, Hebei and Shanxi provinces, the high incidence areas for EC in northern China, including 6,375 males with a mean age of 58±9 and 4,007 females with a mean age of 59±9. Of the patients, 2,714 cases have been followed up by interview at home until July, 2010. Eight EC groups had been classified based on the total number of dissected LN (1-3, 4-5, 6-7, 8-9, 10-11, 12-13, 14-16 and ≥17). The results showed that the positive metastatic detection rate apparently increased with the total extended dissected LN number (P< 0.05) until reaching the level with a total of 17 or more dissected LN. 5-year survival analysis showed that the patients without LN metastasis had a higher survival rate than those with LN metastasis in all seven groups (P<0.05) except the group with ≥17. Interestingly, the survival rate was similar in the group of ≥17 between the patients with positive or negative LN (P>0.05). We concluded that the total dissected LN number could affect the survival rate for EC and that a total of 10 to 16 dissected LN should be recommended for EC surgical treatment. (supported by Innovation Scientists and Technicians Troop Construction Projects of Henan Province).

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 5525. doi:1538-7445.AM2012-5525
Abstract 5532: Anti-cancer activities of tea epigallocatechin-3-gallate in breast cancer patients under radiotherapy
Authors: Wu Erxi
Published: August 2012

Abstract: The purpose of this study was to test the hypothesis that administration of epigallocatechin-3-gallate (EGCG), a polyphenol present in abundance in widely consumed tea, inhibits cell proliferation, invasion, and angiogenesis in breast cancer patients. EGCG in 400 mg capsules was orally administered three times daily to breast cancer patients undergoing treatment by radiotherapy. Parameters related to cell proliferation, invasion, and angiogenesis were analyzed while blood samples were collected at different time points to determine efficacy of the EGCG treatment. Compared to patients who received radiotherapy alone, those given radiotherapy plus EGCG for an extended time period (two to eight weeks) showed significantly lower serum levels of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and reduced activation of metalloproteinase-9 and metalloproteinase-2 (MMP9/MMP2). Addition of sera obtained from patients treated by combination of radiotherapy and EGCG feeding for 2-8 weeks to in vitro cultures of highly-metastatic human MDA-MB-231 breast cancer cells resulted in the following significant changes: (1) suppression of cell proliferation and invasion; (2) arrest of cell cycles at the G0/G1 phase; (3) reduction of activation of MMP9/MMP2, expressions of Bcl-2/Bax, c-Met receptor, NF-κB, and the phosphorylation of Akt. MDA-MB-231 cells exposed to 5-10 µM EGCG also showed significant augmentation of the apoptosis inducing effects of γ-radiation, concomitant with reduced NF-κB protein level and AKT phosphorylation. These results provide hitherto unreported evidence that EGCG potentiated efficacy of radiotherapy in breast cancer patients, and raise the possibility that this tea polyphenol has potential to be a therapeutic adjuvant against human metastatic breast cancer.

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 5532. doi:1538-7445.AM2012-5532