bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1393: Chitinase-3-like-1 protein overexpression in lung epithelial cells enhances breast cancer metastasis to the lung
Authors: Libreros Stephania , Garcia-Areas Ramon , Onwuha-Ekpete Lilllian , Iragavarapu-Charyulu Vijaya
Published: August 2012

Breast cancer is the most common cancer in women in the US. Metastasis is responsible for a majority of these deaths in spite of significant improvements in diagnosis and treatments. Recently it was reported that under conditions of asthma, metastasis to the lung is increased. Patients with pulmonary inflammatory illnesses demonstrate higher levels of chitinase-3-like-1 protein (CHI3L1) in their lungs and in circulation. We have shown that there are elevated levels of CHI3L1 in circulation with increased tumor growth, higher levels of metastasis to the lung with shorter survival rates in mice with asthma. However, the direct role of CHI3L1 in promoting metastasis is not clearly delineated. It is also unknown if exposure to high levels of CHI3L1 previous to tumorigenesis directly predisposes to accelerated metastasis. We hypothesize that CHI3L1-induced pulmonary inflammation generates the proper environment for recruiting circulating breast cancer cells, thus increasing the rate of metastasis to the lung. To determine if inflammation associated with CHI3L1 in the lung alters the pulmonary environment to attract circulating breast tumor cells and accelerate metastatic growth, pulmonary inflammation is induced by either ragweed allergen sensitization or by locally expressing CHI3L1 in the lung using AAV-CC10-CHI3L1 vector. The role of CHI3L1 in pulmonary metastasis is delineated in CHI3L1 over-expressing lung epithelia by assessing the effects on the host immune system and the downstream effects. The studies proposed here will provide a greater understanding of the role of CHI3L1 in enhancing metastasis and the tissue-specific molecular signals that act to exacerbate metastasis under conditions of chronic inflammation

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 1393. doi:1538-7445.AM2012-1393
Abstract 1400: Mouse models for detection of circulating tumor cells from breast cancer
Authors: Navaraj Arunasalam , Dicker David T. , El-Deiry Wafik S.
Published: August 2012

Breast cancer is the most frequent malignancy among women, causing at least 373,000 deaths per year worldwide, and in United States alone an estimated 208,000 women were diagnosed with breast cancer in 2010. Recent studies indicate that breast cancer is initiated by breast cancer stem cells (BCSCs) that express CD44+/high CD24-/low surface markers. Mammospheres are non-adherent spherical cell clusters grown in selective culture conditions. Our experimental results using mammosphere cultures of different breast cancer cells such as MDA-MB-231 show drastic enrichment of breast cancer stem-like cells with the phenotype of CD44+/high CD24-/low by flow cytometry analysis. We have implanted mammosphere-derived cells into mammary fat pads of nude mice and sampled blood from such tumor-bearing animals to look for the presence of unique breast cancer cells with metastatic potential {circulating tumor cells (CTCs)}, by scoring for Epcam+/cytokeratin+/CD45- tumor cells using the FDA-approved Veridex CellSearch Sysytem. As these metastatic breast cancer cells stably express ZS-Green protein, isolated ZS-Green positive CTCs are being further subjected to breast cancer-specific multiplexed molecular marker analysis. When a mouse with the metastatic breast cancer cell line - 4T1, stained with the CellVue dye is implanted in mammary fat pads, high levels of metastasis are detected with the CellVue marker in different internal organs such as the lungs, liver, heart and lymph nodes. Fluorescence microscopy analysis of blood withdrawn from animals bearing orthotopic 4T1 breast xenograft tumors, could detect ZS-Green positive mouse breast cancer CTCs. These mouse animal models bearing xenograft tumors are facilitating our understanding of CTC detection as well as analysis methods and also in understanding of human breast cancer metastasis. Open questions we are addressing include heterogeneity of CTCs, relationships in stem cell markers and prognostic markers when CTCs are compared with the primary tumors, relationships between tumor burden and tumor aggressiveness with CTC numbers in this mouse model. In addition the model provides an opportunity for testing candidate therapeutic agents and drug combinations.

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 1400. doi:1538-7445.AM2012-1400
Abstract 1407: Sonic hedgehog signaling and Androgens are linked in tumor-stromal interaction through Epithelial-Mesenchymal transition (EMT) in prostate cancer progression
Authors: Shigemura Katsumi , Meligy Fatma , Behnsawy Hosny , Yamamichi Fukashi , Haung Wen-Chin , Li Xiangyan , Chung Leland , Yamanaka Kunito , Hanioka Keisuke , Yamashita Masuo , Shirakawa Toshiro , Tanaka Kazushi , Arakawa Soichi , Fujisawa Masato
Published: August 2012

Backgrounds: Sonic hedgehog (Shh) signaling is known to relate to cancer progression but the detail about how to correlate and link with another signaling is debating. The purpose of this study is to investigate the link between Shh signaling, androgens and epithelial-mesenchymal transition (EMT) in prostate tumor-stroma interaction during prostate cancer (PCa) progression. Materials and Methods: We investigated cell proliferation in LNCaP PCa cells, Normal Prostate Fibroblasts (NPF), and Cancer-associated Prostate Fibroblasts (CPF) in the presence of representative androgen, dihydrotestosterone (DHT) and/or Shh signaling inhibitor, cyclopamine (Cyc). With these stimulators or inhibitors, real time RT-PCR was performed to investigate downstream transcriptional factors of Shh signaling, Gli-1, EMT markers (E-cadherin, N-cadherin, and Vimentin), and Osteonectin (ON), a possible Shh target gene shown in our previous work. Expression levels of AR, Gli-1, Ki-67, Shh, and ON in specimens from PCa patients were measured by immunohistochemical (IHC) staining and correlated with clinical data. Results: While significant cell proliferation was induced by DHT in LNCaP, NPF, and CPF, Cyc inhibited these enhanced growths. CPF tended to react to DHT more but less to Cyc than LNCaP or NPF. RT-PCR results showed Shh induced EMT and DHT induced the expression of Gli-1, ON and EMT, and those inductions were blocked by Cyc. IHC staining showed that Stromal ON (ONstr) had significant correlation with serum PSA (p=0.031), high Gleason Scores had significant correlation with Ki-67, and PSA recurrence was related significantly to Gli-1 expression and lower expression of ONstr, suggesting PCa recurrence had a significant correlation with Shh-Gli-1 signaling. Conclusions: Shh signaling is involved in tumor-stromal interaction through a link with Androgens and EMT, and thus plays a pivotal role in PCa progression. Cyclopamine may therefore be a possible therapeutic agent for blocking PCa progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1407. doi:1538-7445.AM2012-1407
Abstract 1413: Rapamycin inhibits lymph node metastasis in an orthotopic model of head and neck squamous cell carcinoma: A possible role for a suppression of VEGFR-3 expression
Authors: Ekshyyan Oleksandr , Moore-Medlin Tara N. , Raley Matthew C. , Sonavane Kunal , Brodt Michael A. , Rong Xiaohua , Abreo Fleurette , Alexander Jonathan S. , Nathan Cherie-Ann O.
Published: August 2012

Background: Tumor cell dissemination to cervical lymph nodes via the lymphatic system represents the first step in the metastasis of head and neck squamous cell carcinoma (HNSCC) and is the most significant prognostic factor for tumor recurrence decreasing survival by 50%. The anti-lymphangiogenic properties of mTOR inhibitors are not yet well understood nor have the effects of rapalogues’ on tumor-associated lymphangiogenesis been investigated in HNSCC. Methods: The lymphatic suppressing effects of rapamycin were evaluated in vitro using lymphatic endothelial cell lines (SV-LEC; HMEC-1a). An orthotopic model of HNSCC was used to evaluate anti-lymphangiogenic effects of rapamycin in vivo. OSC-19 cells were injected into the basolateral tongue of SCID mice. After 5 days mice were given daily IP injections of vehicle (control group) or rapamycin at 5 mg/kg. On day 21 after OSC-19 administration mice were sacrificed and tongues, cervical lymph nodes and serum collected. The incidence of cervical lymph node metastasis, the number of tumor-free and tumor-associated lymphatic vessels in mouse lingual tissue, and expression of pro-lymphangiogenic markers were assessed. Results: We found that LEC proliferation was significantly reduced by rapamycin. LEC growth inhibition by rapamycin was linked with decreased expression of VEGFR-3 in both LECs, and in some HNSCC cell lines. Rapamycin significantly lowered the fraction of tumor-associated lymphatic vessels in mouse tongues (p=0.0167) and decreased the number of metastasis-positive lymph nodes (p=0.04). Rapamycin also significantly attenuated the extent of metastatic tumor cell spread within the lymph nodes (p<0.0001). Conclusion: The results of this study demonstrate anti-lymphangiogenic properties of mTOR inhibitors in HNSCC. These findings provide preclinical data showing that rapalogues exert anti-lymphatic effects in head and neck cancer via suppressing ofVEGFR-3 expression.

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 1413. doi:1538-7445.AM2012-1413
Abstract 1418: Dll4-Fc, an inhibitor of Dll4-notch signaling, suppresses liver metastasis of small cell lung cancer cells through down-regulation of NF-kappa-B activity
Authors: takuya kuramoto
Published: August 2012

Purpose: The Notch signaling regulates cell-fate decisions during development and postnatal life. Recent studies have demonstrated that the signaling through Notch and its ligand Dll4 in tumor endothelial cells plays an important role in cancer angiogenesis. Little is known, however, about the role of Dll4-Notch signaling between cancer cells, and how it affects on cancer metastasis. We therefore assessed the importance of cancer cell Dll4-Notch signaling in cancer metastasis produced by human small lung cancer cells. Experimental Design: On the basis of previous studies, we generated retroviral vectors encoding a soluble dimerized form of Dll4 fused to the human IgG1 Fc constant region (Dll4-Fc) that acts as a blocker of Dll4-Notch interactions. Dll4-Fc was introduced into human small cell lung cancer cell lines that express high level of Dll4 and Notch (SBC-3 and H1048 cells), and low level of Dll4 (SBC-5 cells). The cells were intravenously inoculated to natural killer cell-depleted SCID mice, and the multiple-organ metastases were evaluated. Result: The number of liver metastasis in the mice inoculated with SBC-3 and H1048 cells expressing Dll4-Fc were significantly lower than that of liver metastasis formed by control transfectant cells. However, the number of liver metastasis formed by SBC-5 cells was not significantly different between Dll4-Fc expressing and control cells, suggesting that Dll4-Notch signaling in cancer cells plays important role in liver metastasis of small cell lung cancer. Metastatic colonies in other organ such as lung, kidney and bone were not affected by Dll4-Fc overexpression in each cell lines. In order to study the molecular mechanism of the effect of Dll4-Fc on liver metastasis, genes associated with metastasis were screened by PCR array system. Seven genes (MMP7, IL1B, MMP1, S100A4, MMP10, CXCR4, SERPINE1) were down-regulated in Dll4-Fc-overexpressing SBC-3 and H1048, but not SBC-5 cells, compared with control transfectant cells. Since most of the selected genes were NF-kappa-B target genes, we explored whether NF-kappa-B is involved in cancer cell Dll4-Notch signaling. By using luciferase assay, we observed that NF-kappa-B activity was down-regulated in Dll4-Fc-overexpressing SBC-3 and H1048 cells, but not SBC-5 cells, compared with control transfectant cells, indicating that Dll4-Fc inhibits NF-kappa-B activity in small cell lung cancer cells by suppressing Dll4-Notch signaling. Conclusion: These results suggest that Dll4-Notch signaling in cancer cells plays a critical role in liver metastasis of small cell lung cancer, by regulating the NF-kappa-B signaling.

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 1418. doi:1538-7445.AM2012-1418
Abstract 157: Optimizing the flow cytometric analysis of cell signaling in heterogeneous samples
Authors: Gao Guojian , Coveney Christopher , Wang Xiao , Wu Xiaowei , Wei Ai-Li , Ji Xiang-dong , Anderson Lori , Kotecha Nikesh , Rohrer Jurg , O'Donnell Erika
Published: August 2012

Unlike conventional biochemical methods for cell signaling analysis, analyzing protein levels by flow cytometry allows measurements to be made at the single-cell level. Multiparameter flow cytometry provides a powerful tool for the analysis of protein expression and phosphorylation status within complex populations, such as those found in lymphoid tissues or blood. However, different intracellular and cell surface proteins often require different conditions for optimal detection, creating challenges for analyzing multiple protein types in a single sample. To facilitate the design of multiparameter staining panels for analysis of intracellular and cell surface proteins, we tested over 250 fluorescently conjugated antibodies in cells fixed and permeabilized under various conditions. Multiple fluorescent conjugates of each antibody were tested in human PBMCs, human whole blood, mouse spleen, and/or mouse bone marrow following fixation with BD Phosflow™ Lyse/Fix or BD Cytofix™ Fixation Buffer and permeabilization with one of the four BD Phosflow™ Perm buffers. These studies demonstrated that certain intracellular epitopes required harsh permeabilization conditions for optimal detection, whereas others were detectable under milder conditions. Surface marker stains often required optimization, particularly under harsh permeabilization conditions. Factors such as antibody clone, fluorophore, and concentration had a considerable impact on staining resolution, and alternative staining protocols were required for optimal staining of some surface markers. To facilitate the optimization of fixation, permeabilization, and staining conditions for multiparameter flow cytometry, experimental data and protocols from this study are available through an interactive database freely accessible at www.cytobank.org/facselect.

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 157. doi:1538-7445.AM2012-157
Abstract 1570: Frameshift peptides as prophylactic cancer vaccines antigens
Authors: Luhui Shen , Sykes Kathryn , Johnston Stephen Albert
Published: August 2012

Background: The goal of the project is to develop a universal, prophylactic cancer vaccine. We are integrating bioinformatics, genetics, proteomics and immunological methods to identify tumor DNA-encoded components for the development of an effective vaccine. In a previous study, we used bioinformatic and genetics analyses to identify a list of abberant transcripts associated with tumor cells that encode frameshift-derived (FS) neo-peptides. These FS peptides are a result of various gene mutations and abnormal splicings in tumor cells that would cause expression of gene sequences in alternate reading frames. The transcript encoding one of these vaccine component candidates, SMC1A (Structural Maintenance of Chromosomes 1A) FS was detected in numerous mouse and human tumor cell lines and primary human breast and pancreatic tumors. One dose genetic immunization with a plasmid expressing SMC1A FS significantly delayed tumor growth in C57BL/6 mice transplanted with the B16F10 melanoma cell line. To exam protection of SMC1A FS in a more realistic prophylactic model, we investigated the protection of SMC1A FS in two mouse spontaneous breast tumor models. Results: We similarly detected the transcript corresponding to the SMC1A FS peptide in tumors from spontaneous mouse breast tumor models: FVB/N-NeuT and BALB-NeuT. The SMC1A FS expression increases with the expression of Rat Her-2 in mouse mammary gland. Wild type and mutated Rat Her 2 are the oncogenes that induce spontaneously breast tumor development in FVB/N-NeuT and BALB-NeuT respectively. Both anti-SMC1A FS IgG and SMC1A FS peptide induced IFN-γ released splenocytes were detected in non-treated tumor bearing FVB/N-NeuT mice. One dose genetic immunization of mice with an SMC1A FS expression plasmid significantly reduced spontaneous breast tumor development in FVB/N-NeuT mouse as compared to that in non-treated mice. Genetic immunization followed by a protein boost (prime/boost regimen) in BALB-NeuT mice also significantly reduced tumor development and the reduction is associated with antibody reactivity to SMC1A FS. Discussion: The protection of SMC1A FS prophylactic immunization in spontaneous mouse tumor models indicates SMC1A S can be a potential prophylactic cancer vaccine candidate to prevent a variety of tumors types. We are applying the same basic approach to accumulate sufficient antigens to constitute a broadly protective, prophylactic cancer vaccine. (This research is supported by an Innovation Award from the DoD and The Keck Foundation to SAJ.)

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 1570. doi:1538-7445.AM2012-1570
Abstract 1571: Developing antigen-specific Th1 epitope based vaccines by excluding immunosuppressive peptide sequences
Authors: Cecil Denise L. , Holt Gregory E. , Herendeen Dan , Gad Ekram , Rastetter Lauren , Park Kyong Hwa , Disis Mary L.
Published: August 2012

We have determined that IGFBP-2 is immunogenic in ovarian cancer. Our aim is to develop a multi-epitope vaccine that will elicit Th1 immunity to IGFBP-2. Antigen specific Th1 can modulate the tumor microenvironment to enhance cross priming, supporting the proliferation of cytotoxic T cells which are capable of eradicating ovarian cancer cells. Although some clinical benefit has been demonstrated with this strategy, recent ex vivo analyses of human PBMC have described the presence of antigen-specific CD4+ T regulatory cell (Tregs) in cancer patients that were not detected in healthy individuals. Thus, we questioned if we could optimize our vaccine to include only Th1-stimulating epitopes. Using a combined scoring system from five algorithms for predicting class II binding to determine Th epitopes, we identified 14 IGFBP-2 peptides. Th1 immunogenicity (IFN-gamma) and potential immunosuppression (IL-10) was evaluated by ELISPOT for 40 different donors. Twenty-two percent of the donors only responded with IL-10 secretion to any peptide, 22% only responded with IFN-gamma and 53% of the patients had a mixed IFN-gamma and IL-10 response. To determine which peptides would induce a predominantly Th1 response in the greatest number of people, we used a ratio of IFN-gamma to IL-10 and analyzed both the magnitude and frequency of ELISPOT responses for each of peptides using the following algorithm: (corrected mean SPW) x (percent of responding donors). The peptides were then ranked from highest IL-10 response to highest IFN-gamma response. Interestingly, 6 of the 14 peptides demonstrated a preference to secrete IFN-gamma over IL-10 and are only located in the N-terminus (amino acids 1-163) of IGFBP-2; the remaining potentially immunosuppressive peptides are located in the C-terminus (amino acids 164-328). Vaccination with p1-163 in MMTV-neu mice demonstrated a robust Th1 response (p=0.03) and concomitant inhibition of tumor growth by 70% compared to adjuvant only control animals, p164-328-vaccinated or full length protein-vaccinated mice (p<0.001 for all). Vaccination with p164-328 or full length protein did not inhibit tumor growth. These data suggest that more effective vaccines can be designed when both Th1 epitopes and immunosuppressive epitopes are screened simultaneously and epitopes that are most likely to induce robust Th1 responses in the majority of individuals can be identified and included as vaccine components.

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 1571. doi:1538-7445.AM2012-1571
Abstract 1572: Development of novel and effective therapeutic peptide vaccines against HPV-induced tumors
Authors: Barrios Kelly , Celis Esteban
Published: August 2012

Our main goal is to generate effective therapeutic peptide-based vaccines against viral-induced cancers. We are first focusing on human papillomavirus (HPV) induced malignancies because of their high worldwide prevalence (e.g., cervical carcinoma and head & neck cancer). We have used an HPV-induced mouse cancer model to test vaccines composed of a CD8 T cell peptide epitope administered with potent adjuvants designed to generate vast numbers of high avidity cytotoxic T lymphocytes specific for the HPV16-E7 antigen. One vaccination strategy (TriVax) consists of intravenous administration of synthetic peptide HPV16-E749-57 administered together with a Poly-IC (a TLR3 agonist) and anti-CD40 monoclonal antibody (≥CD40 mAb) while the second more simple strategy (BiVax) comprises of peptide plus Poly-IC. We assessed both the immunogenicity and therapeutic anti-tumor effects of both vaccination strategies. While TriVax was clearly more immunogenic than BiVax, both vaccines showed remarkable anti tumor effects against the HPV16-E7 expressing tumors TC-1 and C3.43. In addition, we will present results suggesting that some peptides such as HPV16-E749-57 can be highly immunogenic in the absence of CD40 costimulation (i.e., in the BiVax format), due to their capacity to form complexes with Poly-IC. We hypothesize that peptide/Poly-IC complexes are highly immunogenic because they are targeted to professional antigen-presenting cells via some type of scavenger receptor specialized in capturing nucleic acids.

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 1572. doi:1538-7445.AM2012-1572
Abstract 1573: MAGED4B drives oral carcinogenesis and is a promising peptide vaccine target for the treatment of oral squamous cell carcinoma
Authors: Lim Kue Peng , Gan Chai Phei , Chong Chan Eng , Zain Rosnah Binti , Abraham Mannil Thomas , Rahman Zainal Ariff Abdul , Teo Soo-Hwang , Gutkind J Silvio , Patel Vyomesh , Ponniah Sathibalan , Cheong Sok Ching
Published: August 2012

MAGED4B is a member of the Melanoma Antigen Gene (MAGE) family. We demonstrated that MAGED4B is over-expressed in more than 50% of oral squamous cell carcinoma (OSCC) tissues and that the expression of MAGED4B is associated with lymph node metastasis and poor disease specific survival. OSCC cell lines over-expressing MAGED4B exhibited an increase in cell growth both in vitro and in vivo, and were significantly more resistant to UV-induced apoptosis compared to control cells, suggesting a role for MAGED4B in modulating cell death. Further, MAGED4B also promoted cell migration. MAGE proteins are good targets for cancer immunotherapy as they are highly immunogenic. We identified and synthesized MAGED4B peptides that were HLA-A2- specific and tested the ability of these peptides to generate an anti-tumour response using PBMCs from OSCC patients and healthy individuals. Firstly, the PBMC samples obtained from patients and healthy individuals were tested for HLA-A2 expression. Positive samples were analyzed for peptide-specific stimulation of IFN-γ and granzyme secretion ex-vivo via ELISPOT assay. Dendritic cells pulsed with these peptides were used to stimulate PBMC cultures, and were studied for IFN-γ and granzyme secretion. In parallel, the binding of the peptides to the MHC-Class I molecule was determined by the binding assay. Further, the ability of these bound peptides to interact with T cells was measured by the dimer assay. We demonstrated that all the peptides have good binding capacity with the MHC-Class I molecules. Moreover, the binding of these peptides on MHC-Class I molecules could attract and further interact with T cells. Notably, the PBMC stimulated once with peptide-pulsed dendritic cells or twice with peptide and peptide-pulsed dendritic cells showed peptide-specific cytotoxic activity against MAGED4B-expressing OSCC cell lines. In summary, we first demonstrated that MAGED4B drives oral carcinogenesis and second, showed that MAGED4B-specific peptides are apable of inducing anti-tumour specific immune responses. Taken together, this indicates that MAGED4B could be a promising peptide vaccine target for the treatment of OSCC.

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 1573. doi:1538-7445.AM2012-1573
Abstract 1575: A dual immunostimulatory strategy for pancreatic cancer treatment: MUC1 cytoplasmic tail peptide therapeutic vaccine with adjuvant TLR3 agonist poly(I:C12U)
Authors: Fink Darci M. , Tremayne Jarrod , Bunt Stephanie K. , Hollingsworth Michael A.
Published: August 2012

Immunotherapy directed against MUC1, a membrane-bound mucin, may be a treatment option for pancreatic cancer patients. MUC1 is overexpressed and differentially glycosylated and phosphorylated by pancreatic cancer cells and is thus a candidate antigenic target for a therapeutic vaccine. Vaccination studies using full length MUC1 cytoplasmic tail (MUC1-CT) peptide in vivo showed an increase in median survival of C57BL/6 mice transgenic for and immunologically tolerant to MUC1 (MUC1.Tg) challenged with a syngeneic murine pancreatic tumor cell line (Panc02.MUC1) as compared to nonvaccinated controls. In a recent study, we employed an immunostimulatory adjuvant in concert with MUC1-CT peptide to improve immune system mobilization in MUC1-tolerant animals thereby enhancing the therapeutic effect of vaccination. The chosen adjuvant, poly(I:C12U), is a clinically tested GMP-grade dsRNA molecule capable of activating Toll-like receptor 3 (TLR3) present on endocytic vesicles in antigen presenting cells (APC). Agonizing TLR3 induces dendritic cell maturation in vitro and in vivo, leading to increased antigen presentation and cytokine production that prompt an antigen-specific T cell response. We hypothesized that a dual immunostimulatory strategy pairing TLR3 agonist-mediated APC activation with tolerance-breaking MUC1-CT antigen-directed vaccination would lead to directed destruction of pancreatic cancer cells and improved overall survival. Treatment with poly(I:C12U) alone, peptide alone, or gemcitabine alone improved survival in this model system; however, co-administration of poly(I:C12U) and peptide did not provide additive or synergistic benefit in this protocol.

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 1575. doi:1538-7445.AM2012-1575
Abstract 1577: Lactobacilli secreting a tumor antigen and IL15 activates neutrophils and dendritic cells and generates cytotoxic T lymphocytes against cancer cells
Authors: Esuvaranathan Kesavan , Kandasamy Matheswaran , Bay Boon Huat , Lee Yuan Kun , Mahendran Ratha
Published: August 2012

Introduction and Objective Lactobacillus rhamnosus GG (LGG) are lactic acid bacteria, which were found to be better than BCG at inducing cell death in human bladder cancer cell lines. This study compares both LGG and BCG in vivo in an orthopic model and further evaluates the potential of recombinant LGG to induce a directed anti-tumor response. Methods MB49-PSA (murine bladder cancer cells secreting human PSA) cells were implanted orthotopically in C57BL/6 mice and either LGG or BCG was instilled weekly for 6 weeks. Tumor cure rates were noted and an immnohistochemical examination was performed on the tumors. The human prostate specific antigen (PSA) PSA was cloned into pLP500-slpAp vector under the control of S-layer promoter of L. acidophilus. The murine IL-15 cDNA minus its termination codon was cloned upstream of the PSA protein sequence. Lactobacilli were transformed with the plasmids to generate LGG-IL15-PSA, LGG-PSA and LGG-S (pLP500-slpAp) respectively. PSA and IL15 secretion was 16-19 ng/2x109 CFU/ml. Bone marrow cells were isolated from the tibia and femur of 8-10 weeks old female C57BL/6 mice. Neutrophils were isolated by positive selection with anti-Ly6G microbead kit (Miltenyi Biotech, Germany). To generate DCs, bone marrow derived cells were cultured for 9 days in media with 40ng/ml of GM-CSF (BD Bioscience) which was replaced every other day. Neutrophils and DC were exposed to the recombinant LGG for 2h and used to stimulate T cells to generate CTL against the murine bladder cancer cell lines expressing PSA (MB49-PSA). Results LGG was as effective as BCG in the mice. Analysis of tumors after repeated weekly LGG instillations showed increased infiltration of neutrophils and macrophages into the tumor mass after therapy. In vitro, recombinant LGG activated neutrophils (elevated MHC class I expression) induced DC maturation (increased expression of CD86, CD80, CD40, MHC II and CD83), T cell proliferation and PSA specific cytotoxic T lymphocytes (CTL) activity. Activated DC were more efficient than activated neutrophils in inducing T cell IL2 and IFNα production. DC activated with LGG for 2 h or with LGG treated neutrophils were re-isolated and plated with naïve T cells for 5 days to generate PSA specific T cells. Then PSA secreting or control MB49 cells were added to the wells. IL15 enhanced DC generation of CTL. Conclusions Intravesical instillation of LGG cured mice of tumors. LGG secreting tumor antigens and cytokines could be used to activate DC in vitro for DC therapy in patients or used as intravesical therapy.

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 1577. doi:1538-7445.AM2012-1577
Abstract 1580: LL-37-mediates tumor antigen-specific-T cell expansion and tumor regression
Authors: Singh Manisha , Liu Chengwen , Lou Yanyan , Gregorio Josh , Conrad Curdin , Hwu Patrick , Overwijk Willem W. , Gilliet Michel
Published: August 2012

We previously showed that the endogenous antimicrobial peptide LL-37, when coupled with self DNA, acts as a TLR-9 agonist and activates plasmacytoid dendritic cells (pDC) leading to type I IFN production. As tumor contains many dying cells releasing self DNA, we investigated whether intra-tumoral injection of LL-37 induces anti-tumor immunity through pDC activation. Mice bearing established subcutaneous B16-OVA melanoma were injected intratumorally with LL-37. LL-37 was found to induce IFN-γ expression in treated tumors. Furthermore, LL-37 induced an activation and expansion of OVA-antigen-specific CD8 T cells in draining lymph nodes as well as the tumor micro environment. This process was associated with delay in tumor growth. These findings suggest that LL-37 could induce anti-tumor immunity and provide a promising strategy for melanoma immunotherapy.

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 1580. doi:1538-7445.AM2012-1580
Abstract 1581: Identification of novel HLA-restricted T-cell epitopes specific for the breast cancer-associated tumor antigen NY-BR-1
Authors: Gardyan Adriane , Osen Wolfram , Jesiak Maria , Zörnig Inka , Jaeger Dirk , Eichmueller Stefan B.
Published: August 2012

Out of all malignancies, breast cancer resembles the most common malignant caner type (26%) of women in the western world and the second most frequent cancer related cause of death (15%) in women. Efficient treatment options are available for early stage breast cancers yielding five year survival rates of up to 85%, however for metastasized breast cancer types, five year survival rates are resided at much lower levels (20%) and therapy options are very limited. It is therefore of great importance to establish innovative therapy strategies which will affect the late stage breast cancer tumors. Immunotherapy approaches, particularly adoptive T cell transfer, constitute an attractive strategy to develop new therapy options for metastasized mammary carcinomas. The differentiation antigen NY-BR-1 was found to be expressed on mRNA level in breast tissue, testis, prostate, and most notably in 80% of tested breast cancers. Among normal tissues, expression of the NY-BR-1 protein was present solely in ductal epithelium of the breast. Compared to healthy breast tissue NY-BR-1 protein levels are highly elevated in invasive breast tumors and their corresponding metastasis. Thus, NY-BR-1 can be considered as a breast cancer associated tumor antigen that might represent a suitable target antigen for T cell based immunotherapy approaches. The main focus of this project is to identify novel HLA-restricted T cell epitopes that could be used in clinical trials. Therefore, various HLA-transgenic (tg) mouse strains (DR4tg, DR3tg and HHDtg mice), were immunized with the NY-BR-1- encoding expression plasmid pcDNA3.1-NY-BR-1 followed by ex vivo analyses of the NY-BR-1-specific T cell responses with a synthetic peptide library covering the entire NY-BR-1 protein. Applying this strategy, four potential HLA-DRB1*0301 restricted epitopes and three HLA-DRB*0401 restricted candidate epitopes could be determined. The relevance of these new epitopes for the human system will be analyzed by screening of PBMCs from tumor patients and healthy donors for the presence of cognate T cells. Such T cells will be expanded and used as donors for high affinity NY-BR-1-specific TCRs in order to establish autologous TCR-transduced T cells ready for adoptive transfer of breast cancer patients. Currently, blood-samples of NY-BR-1+ patients are being collected for subsequent in vitro analyses of HLA-restricted, NY-BR-1-specific T cell responses.

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 1581. doi:1538-7445.AM2012-1581
Abstract 1582: Therapeutic vaccine for pancreatic cancer
Authors: Khamar Bakulesh M. , Desai Nirav , Shukla Chandreswar
Published: August 2012

Cadi-05 (Heat killed mycobacterium w) is a potent TLR 2 agonist and induces pure potent Th1 response. In advanced Non small cell lung cancer, Cadi-05 is found to potentiate efficacy of chemotherapy (Paclitaxel + Cisplatin) with improvement in response rate and overall survival in a randomized controlled clinical trial. Cadi-05 is also a potent vaccine adjuvant for prophylactic vaccines providing faster, higher and sustained antibody titers in animals and humans. The therapeutic vaccine for pancreatic cancer, with killed pancreatic cancer cells as an antigen with Cadi-05 as an adjuvant was evaluated for its ability to induce cell mediated immune responses (by ELISPOT for IFN - α, IL-2 and Granzyme B) and effector function (By BrDu incorporation). The vaccine made using Mia PaCa-2 cell line induced cell mediated immune responses against the Mia PaCA-2 as well as against the other cell lines (Panc-1, ASPC-1, SW-1990). The vaccine also generated effector function of similar magnitude against all four cell lines. Effect of vaccine on tumor size and survival following intradermal immunization was also studied. For evaluating efficacy, C57BL/6 BYJ mice were injected Pan-02 cells (1x106) subcutaneously for developing pancreatic cancer model. At day 10 post tumor induction tumor size achieved was∼200-225 mm3. Mice were randomized to receive vaccine or placebo (18 mice each) on day 10. No other therapy was used in the experiment. In Vaccine arm tumor regression was seen in 61% of animals before progression. Tumor regression was not seen in placebo arm. Tumor progression was found to be slower in vaccinated animals with mean tumor size 35% and 59% of placebo arm on day7 (633 mm3 vs. 223 mm3) and day 12 (740 mm3 vs. 437 mm3) respectively resulting in survival benefit of 36% on day 40 following immunisation.

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 1582. doi:1538-7445.AM2012-1582
Abstract 1583: New strategy for the identification of breast cancer antigens that induce therapeutic immune responses in tumor-bearing mice
Authors: Cohen Edward P. , Kim Tae Sung , Chopra Amla , O-Sullivan InSug
Published: August 2012

This study describes a unique strategy designed to identify breast cancer antigens (TAA) that induce therapeutic immune responses in tumor-bearing mice. In a mouse model, the strategy led to the identification of growth factor receptor-bound protein 10 (Grb10) as a newly identified TAA. Grb10 is a signal transduction molecule associated with multiple transmembrane tyrosine kinase receptors. It was discovered by comparing microarrays of cellular breast cancer vaccines highly enriched for cells that induced breast cancer immunity with non-enriched vaccines. The vaccines were prepared by transferring a cDNA expression library derived from SB5b cells, a breast cancer cell line derived from a neoplasm that arose spontaneously in the mammary gland of a C3H/He mouse into LM cells, a mouse fibroblast cell line. As the transferred cDNA integrates spontaneously into the genome of the recipient cells, replicates as the cells divide, and is expressed, the vaccine could be prepared from a little as 10 micrograms of tumor tissue. Relatively few cells in the transfected cell population, however, incorporated cDNA fragments that included genes specifying TAA. (The vast majority specified normal cellular constituents.) A unique strategy was developed, therefore, to enrich the vaccine for immunotherapeutic cells. Comparative microarrays of enriched and non-enriched vaccines resulted in the identification of twenty overrepresented genes. One, the gene for Grb10, was approximately 100-fold overrepresented. To determine if Grb10 in the enriched vaccine was partly responsible for its therapeutic benefits, the gene was incorporated into an expression plasmid that was then transfected into the fibroblast cell line, which was then used as a vaccine. Mice with established breast cancer treated solely by immunization with the fibroblasts modified to express Grb10 developed robust immunity to the breast cancer cells, which, in some instances, was sufficient to result in tumor rejection.

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 1583. doi:1538-7445.AM2012-1583
Abstract 1592: Silibinin inhibits UVB-induced DNA-damage and apoptosis by enhancing interleukin-12 expression in JB6 cells and SKH-1 mouse skin
Authors: Narayanapillai Sreekanth C. , Gu Mallikarjuna , Agarwal Chapla , Agarwal Rajesh
Published: August 2012

Ultraviolet-B (UVB) radiation plays a pivotal role in non-melanoma human skin cancers (NMSCs) by inducing cellular DNA damage, mutations and immunosuppression. Decades of research has demonstrated silibinin efficacy against UVB-induced skin carcinogenesis by regulating different molecular pathways in various cell lines and animal models; however, the role of silibinin in regulating interleukin-12 (IL-12), an immunomodulatory cytokine with the capacity to reduce UVB-induced DNA damage and apoptosis, has not been studied. Here, we report that UVB irradiation caused a dose- and time-dependent caspase 3 and PARP cleavage as well as apoptosis in JB6 cells. Addition of recombinant IL-12 or silibinin immediately after UVB significantly protected UVB-induced apoptosis in JB6 cells, and antibody-mediated blocking of IL-12 activity compromised the protective effect of both IL-12 and silibinin. In addition, both silibinin and IL-12 accelerated the repair of UVB-caused cyclobutane-pyrimidine dimers (CPDs) in JB6 cells. Additional studies confirmed that indeed silibinin causes a significant increase in IL-12 levels in UVB-irradiated JB6 cells as well as in the SKH-1 hairless mouse epidermis, and that similar to in vitro findings, silibinin topical application immediately after UVB exposure causes a very strong protection against UVB-induced TUNEL positive cells in epidermis possibly through a significantly accelerated repair of UVB-caused CPDs in skin epidermis. Together, these findings for the first time identify an important insight regarding the pharmacological mechanism wherein silibinin induces endogenous IL-12 in its efficacy against UVB-caused skin damages. Considering the fact that millions of people get constantly exposed to solar UVB radiation and live with persistent UVB-induced DNA lesions, the post-damage use of silibinin as an inducer of endogenous IL-12 in UVB-damaged human skin for the rapid repair of DNA damage could be a more practical and translational approach in reducing sunlight-caused damages in human skin which eventually lead to NMSCs.

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 1592. doi:1538-7445.AM2012-1592
Abstract 16: Downregulation of CYLD leads to acquisition of mesenchymal state and increased migration via ligand-independent TGFβR1 activation in human oral squamous cell carcinoma cells
Authors: Shinriki Satoru , Nakamura Takuya , Hayashi Mitsuhiro , Guo Jianying , Atsuyama Eri , Nishimura Yasuharu , Yamamoto Yutaka , Sueyoshi Takanao , Ota Tomoko , Shinohara Masanori , Ando Yukio
Published: August 2012

The 5-year survival of patients with oral squamous cell carcinoma (OSCC) has not changed apparently for the past 30 years. Although cylindromatosis (CYLD) is thought as a potent tumor suppressor, its biological significances in malignancies are largely unknown. The aim of this study was to clarify the role of CYLD in OSCC progression. We investigated expression of CYLD in OSCC including intraepithelial neoplasia (IEN; n = 48) and invasive carcinomas (n = 133) tissues and normal oral mucosal tissues (n = 35) by immunohistochemistry. In addition, effects of CYLD knockdown by siRNA transfection on OSCC progression using 5 OSCC cell lines (SAS, Tu4, HSC3, Ca9-22, and SCC-NA) and HaCaT keratinocytes. Our immunohistochemical analyses revealed that CYLD expression was significantly reduced at invasive lesions in OSCC tissues whereas it was conserved in normal epithelium and IEN. In addition, lower CYLD expression was associated with the correlation with the increased tumor size, advanced clinical stage, and poor overall survival in invasive OSCC. Accordingly, CYLD knockdown led to acquisition of mesenchymal state and increased migratory activity in all the OSCC cell lines as well as HaCaT keratinocytes. Notably, such EMT-like changes were completely blocked by a TGFβR1 inhibitor in all the OSCC cell lines, but not in HaCaT keratinocytes. Furthermore, treatment with excess amount of anti-TGFβ antibody (1D11) did not inhibit the EMT-like changes induced by CYLD repression. These findings suggest that downregulation of CYLD promotes invasion through EMT-like changes via ligand-independent TGFβR1 activation in OSCC cells in mechanisms distinct from normal epithelium. Further studies for these mechanisms probably provide new insights into the biology including “TGFβ switch” and development of a novel therapy in OSCC.

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 16. doi:1538-7445.AM2012-16
Abstract 160: Targeting cysteine-mediated dimerization of the MUC1-C oncoprotein in human cancer cells
Authors: Raina Deepak , Ahmad Rehan , Rajabi Hasan , Panchamoorthy Govind , Kufe Donald W. , Kharbanda Surender
Published: August 2012

The MUC1 heterodimeric protein is aberrantly overexpressed in diverse human carcinomas and contributes to the malignant phenotype. The MUC1-C transmembrane subunit contains a CQC motif in the cytoplasmic domain that has been implicated in the formation of dimers and in its oncogenic function. The present studies demonstrate that MUC1-C forms dimers in human breast and lung cancer cells. MUC1-C dimerization was detectable in the cytoplasm and was independent of MUC1-N, the N-terminal mucin subunit that extends outside the cell. We show that the MUC1-C cytoplasmic domain forms dimers in vitro that are disrupted by reducing agents. Moreover, dimerization of the MUC1-C subunit in cancer cells was blocked by reducing agents and increased by oxidative stress, supporting involvement of the CQC motif in forming disulfide bonds. In support of these observations, mutation of the MUC1-C CQC motif to AQA completely blocked MUC1-C dimerization. Importantly, these studies were performed with MUC1-C devoid of fluorescent proteins, such as GFP, CFP and YFP. In this regard, we show that GFP, CFP and YFP themselves form dimers that are readily detectable with cross-linking agents. The present results further demonstrate that a cell-penetrating peptide that targets the MUC1-C CQC cysteines blocks MUC1-C dimerization in cancer cells. These findings provide definitive evidence that (i) the MUC1-C cytoplasmic domain cysteines are necessary and sufficient for MUC1-C dimerization and (ii) these CQC motif cysteine represent an Achilles’ heel for targeting MUC1-C 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 160. doi:1538-7445.AM2012-160