bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5179: Inhibition of U937-cell adhesion to human endothelial cells by glycosylated lysozyme mutants
Authors: Melcher Ralph , Kraus Ina , Demmig Stephanie , Moeller Dorothea , Kudlich Theodor , Rogoll Dorothee , Scheurlen Michael , Scheppach Wolfgang , Hasilik Andrej
Published: August 2012

The evascularization of cells is an important step in defense reactions and the spreading of metastases. During inflammation, the endothelium of the affected tissue becomes activated and interacts with cells bearing cognate structures such as sialyl Lewis X (sLex) substances. In this study, we present the sLex-antagonist hLysII/IV-FUCTVI by which a significant inhibition of adhesion is achieved at 1ng/ml (0.07 nM), making it the most potent known inhibitors of adhesion described to date. In our study we overexpressed the glycosylated human lysozyme mutant hLysII/IV in CHO cells stably expressing fucosyltransferases FUCIII, IV, V, VI, or VII. Digestional analysis with endo- and exoglycosidases showed that hLysII/IV-FUCTVI contains predominantly biantennary complex N-glycans with a variable number of lactosamine repeats; these were terminally sialylated and fucosylated, suggesting the presence of the sLex antigen. In contrast, hLysII/IV-FUCTIII and IV were not terminally sialylated and hLysII/IV-FUCV and VII were not terminally fucosylated. In a static adhesion assay, binding of U937 to activated HUVEC cells was exclusively blocked in the presence of medium supernatant containing hLysII/IV-FUCTVI and by affinity-purified hLysII/IV-FUCTVI.

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 5179. doi:1538-7445.AM2012-5179
Abstract 5260: Genomic gains in prostatic carcinoma and proliferative inflammatory atrophy in dogs
Authors: Alves Carlos E. Fonseca , Busso Ariane F. , Silveira Sara M. , Rogatto Silvia R. , Amorim Renee Laufer
Published: August 2012

The dog can spontaneously develop prostate cancer and consequently can be used as an experimental model for prostatic diseases associated with aging, including benign prostate hyperplasia (BPH) and prostate carcinoma (PCa). DNA copy number variations (CNVs) have been used to identify genes associated with cancer development and progression. DNA microarray based comparative genomic hybridization (aCGH) is a technique that allows to identify copy number of thousands of genes throughout the genome. aCGH was used to identify genomic regions with significantly different DNA copy number in three benign prostatic hyperplasia (BPH), four proliferative inflammatory atrophy (PIA), and 14 canine prostate carcinoma (PCa). Five histologically normal prostate tissue were used as reference. Genomic DNA was extracted from formalin fixed and paraffin embedded samples and CNVs data was evaluated in Canine Genome CGH Microarray 4x44K (G2519F, Design ID021193, Agilent). Data analysis was performed using Genomic Workbench Standard Edition 5.0.14 (Agilent). PCa showed higher number of altered genes related to canonical diseases process, cellular functions and molecular pathways as well as greater inter-relationship between genes, compared with PIA and BPH. In conclusion, PCa showed a more complex genotype, being losses the most frequent genomic changes. Some discrepancies between genomic alterations in human and canine carcinomas may indicate the different clinical behavior of these tumors in these two species. In addition, it was observed was an ascending pattern of genomic complexity in BPH, PIA and CA consistent with a model of multistep tumor 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 5260. doi:1538-7445.AM2012-5260
Abstract 5282: Leptin induces Notch in endothelial cells: Role of VEGFR-2 transactivation
Authors: Lanier Viola , Gillespie Corey , McGlothen Tanisha , Dickson Toi , Guo Shanchun , Gonzalez-Perez Ruben R.
Published: August 2012

Notch signaling is essential for breast cancer (BC) angiogenesis and its dysfunction is implicated in many cancers. Leptin (a pro-angiogenic, pro-inflammatory and mitogenic factor) induces the expression of VEGF/VEGFR-2 and Notch in BC cells. Moreover, the binding of leptin to its receptor (OB-Rb) in endothelial cells (EC) was shown to phosphorylate/transactivate VEGFR-2 inducing angiogenic features. We hypothesize that leptin is an inducer of Notch in EC and that this effect could occur upon leptin-mediated transactivation of VEGFR-2, thereby inducing angiogenic changes in EC. To this end, HUVEC cells were challenged with leptin, leptin inhibitor (LPrA2) VEGFR-2 kinase inhibitor (SU5416) and Notch signaling inhibitor (DAPT). Leptin-dose and time-course effects on expression of Notch (receptors: Notch1, Notch2, Notch3 and Notch4; ligands: JAG1, Dll-4 and targets: survivin and Hey2) were determined by WB and real time RT-PCR. The impact of LPrA2, SU5416 and DAPT on leptin-induced VEGFR-2 phosphorylation and Notch expression was also determined. The effects of leptin and inhibitors on EC proliferation, migration and tube formation were investigated. Results from our studies show for the first time that leptin is a potent inducer of Notch signaling in EC. Moreover, leptin-induced VEGFR-2 phosphorylation was closely related to leptin effects on Notch and the angiogenic features of EC. Therefore, leptin secreted either by adipose tissue or BC cells may contribute to tumor angiogenesis by acting directly on cancer cells inducing VEGF secretion or through EC inducing VEGFR-2/Notch crosstalk. Present results underline the notion that combinatory therapies targeting both Notch and leptin signaling could be a new strategy for BC treatment. [This work was supported in part by NIH/NCI1SC1CA138658-02; NIH/ARRA/3SC1CA138658-02S1 and the Georgia Cancer Coalition Distinguished Cancer Scholar Award (to RRGP)].

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 5282. doi:1538-7445.AM2012-5282
Abstract 5283: Autophagy regulation of angiogenesis in human neuroblastoma
Authors: Kim Kwang Woon , Chung Dai. H
Published: August 2012

Neuroblastoma is a progressive solid tumor in infants and children that originates from neural crest and it remains a clinical challenge to treat. It is a highly malignant tumor, in part, because angiogenesis is a key factor in tumor progression of neuroblastoma. We have previously identified that gastrin-releasing peptide (GRP)-receptor-mediated cell signaling is critical to its tumorigenicity. In this study, we hypothesized that increased autophagy via blockage of GRP receptor could potentially inhibit angiogenesis in human neuroblastomas. METHODS. Autophagic key molecular markers were detected with ATG5, Beclin-1 and LC3 antibodies. Phosphorylation of Akt, mTOR and S6R were detected by Western blotting. Angiogenesis were measured using tubule formation of human umbilical vein endothelial cells (HUVECs) grown on top of Matrigel Matrix Basement Membrane in vitro. They were also determined using a mouse model of the cornea pocket assay in vivo. Autophagy was determined by punctuate localization of GFP-LC3 fusion protein, and observed under a confocal fluorescence microscope in vitro. RESULTS. Levels of phospho-Akt, -mTOR and -S6R were increased in BE(2)-C cells treated with GRP in both time- as well as dose-dependent manner. However, the PI3K/Akt/mTOR pathway was inactivated in short-hairpin (sh) silenced targeted at GRP receptor in BE(2)-C cells when compared to control shLac cells, indicating that angiogenesis may be reduced. We also observed increased levels of ATG5, Beclin-1 and LC3 proteins, key autophagic mediators, in shGRP receptor-BE(2)-C cells when compared to controls (shLac cells). In particular, overexpression of ATG5 and Beclin-1 also significantly decreased tube formation on HUVECs when treated with exogenous GRP (100 nM). Consistent with the in vitro findings, treatment with GRP receptor antagonist (RC3095) showed a dramatically decrease in angiogenesis in a mouse model of the cornea pocket assay in vivo. CONCLUSIONS. Our results show that enhanced autophagy was associated with a downregulation of angiogenesis signaling pathways, and an increase in characteristic punctuate localization of GFP-LC3 as a marker for autophagy. A better understanding of how autophagic proteins can regulate the angiogenic process may lead to novel therapeutic strategies in the treatment of neuroblastomas or its vascular disorders.

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 5283. doi:1538-7445.AM2012-5283
Abstract 5304: RNA sequencing of circulating tumor cells implicates WNT signaling in pancreatic cancer metastasis
Authors: Yu Min , Ting David T. , Stott Shannon L. , Wittner Ben S. , Ozsolak Fatih , Paul Suchismita , Ciciliano Jordan C. , Smas Malgorzata E. , Gilman Anna J. , Ulman Matthew J. , Contino Gianmarco , Alagesan Brinda , Brannigan Brian W. , Milos Patrice M. , Ryan David P. , Sequist Lecia V. , Bardeesy Nabeel , Ramaswamy Sridhar , Toner Mehmet , Maheswaran Shyamala , Haber Daniel A.
Published: August 2012

Circulating tumor cells (CTCs) shed into blood from primary cancers include putative precursors that initiate distal metastases. While isolation and molecular analysis of these cells is technically challenging, they may identify cellular pathways that contribute to the blood-borne dissemination of cancer cells. Here, we adapted a microfluidic device for efficient capture of CTCs from an endogenous mouse pancreatic cancer model and subjected CTCs to digital gene expression (DGE) analysis using single molecule RNA sequencing. We identified Wnt2 as enriched in both CTCs and metastatic ascites cells, compared with primary tumors. Expression of Wnt2 in pancreatic cancer cells suppresses anoikis, enhances anchorage-independent sphere formation, and increases metastatic propensity in vivo. The effect of Wnt2 is correlated with fibronectin upregulation, and it is mediated in part through non-canonical Wnt signaling and suppressed by inhibition of the Map3k7 (Tak1) kinase, an integrator of Wnt, BMP and TGF-β signaling. DGE analysis of human pancreatic CTCs reveals significant enrichment for non-canonical Wnt signaling in 5 of 11 cases, with Wnt2 RNA detectable by in situ hybridization in a subset of CTCs. Identifying CTC-enriched transcripts such as Wnt2 may point to pathways that enhance haematogenous metastasis, thus providing novel therapeutic targets to prevent the distal spread of 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 5304. doi:1538-7445.AM2012-5304
Abstract 5309: Understanding the function of EZH2 as a determinant of breast cancer invasion and metastasis
Authors: Moore Heather M. , Gonzalez Maria E. , Daniels Christine N. , Li Xin , Toy Kathy , Kleer Celina G.
Published: August 2012

Breast cancer is the second most common cause of cancer-related deaths for women in the United States, and the mortality for those 20% of patients with recurrences and/or metastases is nearly 100%. EZH2 (Enhancer of Zeste Homolog 2), a Polycomb group protein that functions in transcriptional memory, is upregulated during progression from ductal carcinoma in situ to invasive carcinoma and distant metastasis. EZH2 protein is overexpressed in 55% of invasive breast carcinomas and women with tumors expressing high EZH2 have a worse disease free and overall survival than women with tumors expressing low EZH2. We hypothesize that EZH2 overexpression may give rise to breast cancers that are on a highly aggressive path from the outset and may enhance metastasis. To test our hypothesis, we have investigated the role of EZH2 in cell morphology, invasion and motility in vitro, and metastasis in vivo. Stable shRNA EZH2 knockdown was carried out in 3 breast cancer cell lines (MDA-MB-231, CAL51, SUM149) which normally express high levels of EZH2 and have motile and invasive abilities. EZH2 knockdown in all cell lines tested decreased their invasiveness using a modified Boyden chamber assay. Random motion motility assays using live cell imaging tracking on MDA-MB-231 vector and shEZH2 cells revealed that EZH2 knockdown significantly reduces the random motion average cell velocity of these cells. Concomitant with the effect of EZH2 knockdown in decreasing invasion and velocity of movement of breast cancer cells, we determined that EZH2 knockdown in MDA-MB-231 and SUM149 cells results in a morphological change and protein expression pattern consistent with a mesenchymal to epithelial transition (MET). Recent experiments have demonstrated that EZH2 knockdown in breast cancer cell lines decreases the levels of activated, phosphorylated p38 mitogen activated protein kinase (MAPK), a protein shown to regulate breast cancer cell motility and metastasis. The in vivo relevance is highlighted by our data showing that EZH2 knockdown in MDA-MB-231 cells significantly decreased the number of lung metastases per mouse, decreased the average size of lung metastases and decreased the proliferative index of the metastatic deposit as measured by decreased Ki67 immunostaining, when compared to controls. Decreased phosphorylated p38 staining has also been verified in the mouse xenograft lung metastases. Taken together, these data support a role for EZH2 in establishing rapid cell motion, invasion and metastasis in breast cancers. Experiments to further delineate the underlying mechanism and to determine the role of p38 in EZH2-mediated functions are underway.

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 5309. doi:1538-7445.AM2012-5309
Abstract 5310: Regulation of metastatic breast cancer by E2F transcription factors
Authors: Andrechek Eran R.
Published: August 2012

Metastasis is the leading cause of mortality in breast cancer and the purpose of this study was to examine the role of the E2F transcription factors in mediating the dissemination of the primary cancer. E2F transcription factors have previously been demonstrated to play a critical role in regulation of cell cycle progression. However, in prior work we have demonstrated that gene expression signatures for the E2Fs have variable levels of activation in a heterogeneous tumor model. Using predictive bioinformatic methods to assess the probability of gene pathway activation, we have predicted variable roles for the E2Fs in key mouse models of breast cancer, including Myc, PyMT and Neu. We then genetically tested these computational predictions by interbreeding E2F knockout mice to the various transgenic tumor models. These crosses have revealed that the E2Fs have unique roles in tumor initiation and progression depending upon the oncogene driving the initial tumor. There are differential effects on tumor latency with the loss of individual E2F alleles, including decreased latency with E2F2 loss in the PyMT background but increased latency and reduced incidence in the Myc background. Moreover, we have observed key differences in tumor progression, with loss of E2Fs affecting the heterogeneity of the resulting tumors. In addition to tumor initiation, we have observed striking differences in tumor progression with loss of E2Fs increasing metastasis in the Myc model but reducing metastasis in the PyMT model. In order to determine the mechanism of action for these effects, we have undertaken a gene expression analysis from both primary tumors and the metastases. Comparison of tumor models in the various backgrounds has allowed us to generate a list of E2F target genes that have effects on metastasis. Comparison of these genes to human breast cancer has revealed that the key genes also have critical roles in metastatic human breast cancer survival. These E2F targets include genes that have previously been identified as metastasis suppressors and are being validated in this process. Taken together, this work suggests a critical role for the E2F transcription factors, not only as mediators of cell cycle but in regulating the metastatic progression of metastatic breast cancer. These data also illustrate the power of combining various genomic and gene expression analysis techniques with both mouse models and clinical data associated with gene expression analysis in human breast cancer. Using this approach we are able to predict and test for the role of critical pathways, such as the E2Fs, in the progression of 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 5310. doi:1538-7445.AM2012-5310
Abstract 5375: Modulation of circulating regulatory T-cells and MDSCs in high risk melanoma patients treated with neoadjuvant ipilimumab
Authors: Tarhini Ahmad A. , Edington Howard , Butterfield Lisa H. , Moschos Stergios , Tawbi Hussein , Shuai Yongli , Shipe-Spotloe Janice , Simonetta Margaret , Johnson Jonas T. , Kirkwood John M.
Published: August 2012

Background: Melanoma regional lymph node and in-transit metastases (stage IIIB-C) carry a high relapse and mortality risk. Neoadjuvant ipilimumab may modulate the host suppressor immune response, enhance antitumor immunity and improve the clinical outcome. Methods: Patients with clinically palpable stage IIIB-C melanoma were treated with induction ipilimumab (10 mg/kg IV q3weeks x2doses) preoperatively and had definitive lymphadenectomy (week ≥ 6), followed by 2 maintenance doses of ipilimumab (q3 weeks). Tissue samples were obtained at baseline and at definitive surgery (after 2 doses of ipilimumab) and blood (serum/PBMC) collected at baseline, 6 weeks, then at 3, 6, 9, 12 months and/or progression. Circulating T-regulatory cells (T-regs) and myeloid derived suppressor cells (MDSC) were monitored utilizing multicolor flow cytometry comparing preoperative (6 weeks) and baseline samples. Results: Twenty nine patients (20 male, 9 female), age 40-87 (median 54) have been enrolled since February/2010 (24 cutaneous primary, 1 unknown and 4 mucosal). Five had AJCC stage IIIB (N2b, N2c) and 24 had IIIC (N3) melanoma. Ninety one cycles have been delivered (median 4). Grade 3 (worst) toxicities include diarrhea/colitis (5 patients; 17%), hepatic enzyme elevations (2; 7%), rash (2; 7%), lipase (1; 3%), all manageable. Median follow-up is 9.7 months and 18 patients (62%) continue disease free. Median PFS is 15.5 months, 95% CI = (8.1, -). The probability of 6 and 12 months PFS is 84.2% (95% CI=0.63, 0.94) and 54.1% (95% CI=0.31, 0.73) respectively. There is a significant increase in the frequency of circulating T-regs (CD4+CD25hi+ Foxp3+; p=0.023 CD4+CD25hi+CD39+; p=0.001) from baseline to 6 weeks. In parallel there is a significant decrease in circulating MDSCs: (1) monocytic: HLA-DR+/low/CD14+; p=<.0001 and less significantly for (2) other monocytic: lin1neg/HLA-DRneg/CD33+/CD11b+; p=0.198 and (3) lymphoid: Lin1neg/HLA-DR-/CD33+/CD11b+; p=0.338. Conclusions: Neoadjuvant ipilimumab is clinically promising and significantly downregulates MDSCs which appear to play a significant role in the clinical activity of ipilimumab. T-regs are upregulated as part of the overall CD4+ T-cell population, and functional studies of this population are ongoing.

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 5375. doi:1538-7445.AM2012-5375
Abstract 5378: Phase I trial of EUS-guided intratumoral vaccination with recombinant Panvac-F and systemic Panvac-V in patients with locally advanced pancreatic cancer
Authors: Poplin Elizabeth A. , August David A. , Ben-Menachem Tamir , Michael Hazar , Artymyshyn Renee , Gulley James L. , Schlom Jeffrey , DiPaola Robert S. , Lattime Edmund C.
Published: August 2012

Preclinical studies in our laboratory were the first to demonstrate the superiority of intratumoral poxvirus vaccines in overcoming immune escape and effectively treating murine solid tumor models. Murine models of bladder and orthotopic mammary tumor demonstrated anergy to systemic immunization using antigen encoding vaccinia recombinants while responding to immunization into the tumor microenvironment. We present here the translation of these findings to a Phase I trial in six patients with locally advanced inoperable adenocarcinoma of the pancreas (NCT00669734). The first dose cohort of patients were treated using a combination of EUS-guided intrapancreatic tumor injection of recombinant Panvac-F (Fowlpox encoding MUC-1, CEA, TRICOM, 108 PFU) in addition to systemic Panvac-V (vaccinia, 2 X 108 PFU) and Panvac-F (109 PFU) boosts (accompanied by subcutaneous rH-GM-CSF, 100 mcg X 4 days). Patients received a total of 2 intrapancreatic injections of Panvac-F (2 weeks apart) with systemic Panvac-V and Panvac-F boosts given with GMCSF extending to day 71 (total of 2 intrapancreatic Panvac-F, 1 sc Panvac-V, 4 sc Panvac-F). Patients were allowed to transition to standard care at day 35. Patients were evaluated for toxicity, tumor progression, and serum CA 19-9 and CEA levels. Two of six patients were removed from study after approximately two weeks due to rapid disease progression and died one and six months after trial initiation. Of the remaining four, one had received prior treatment with gemcitabine followed by capecitabine and radiation and received no further treatment after vaccination; three received post-vaccination standard treatment with gemcitabine. Of the 4 patients with followup of greater than 6 mos, all have stable disease with no progression to metastasis (19 mos, 17 mos, 16 mos, 13 mos). One pt had a DLT (pancreatitis) that resolved allowing completion of protocol therapy. Results demonstrate that the “first in human” intrapancreatic administration of recombinant poxvirus was well tolerated with the complete regimen suggesting an encouraging period of stable disease. The trial is currently accruing patients at intrapancreatic dose level 2 (109 PFU). This study is supported by the NCI Cancer Therapeutics Evaluation Program (CTEP) and by NCI U01-CA07031 and P30-CA72720.

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 5378. doi:1538-7445.AM2012-5378
Abstract 5379: Combination treatment with Bevacizumab, Lenalidomide, Docetaxel and Prednisone (ART-P) does not impact the immune response in patients with metastatic castration-resistant prostate cancer
Authors: Jochems Caroline , Tucker Jo A. , Poole Diane J. , Beatson Melony , Mulquin Marcia , Madan Ravi A. , Figg William D. , Dahut William L. , Gulley James L. , Schlom Jeffrey , Tsang Kwong-Yok
Published: August 2012

Purpose: Investigation of the effect of combination therapy for 5 cycles with bevacizumab, lenalidomide, docetaxel and prednisone (ART-P) on the immune response in patients with metastatic castration-resistant prostate cancer (mCRPC). The aim was to ascertain if treatment containing prednisone and docetaxel would impact immune responses, which could compromise the efficacy of subsequent immunotherapy. Experimental Design: We report here a study of immune responses in 12 patients enrolled in a phase II trial of ART-P at the National Cancer Institute (NCI). We compared peripheral blood mononuclear cells (PBMC) and serum samples collected at baseline and post 5 3-week cycles of ART-P treatment. PBMCs were analyzed using flow cytometry to characterize phenotypes of T-cells, regulatory T-cells, myeloid derived suppressor cells (MDSC) and natural killer (NK) cells, and T-cell proliferation as well as NK-cell functional activity were evaluated. Serum samples were analyzed for levels of cytokines, chemokines and vascular endothelial growth factor (VEGF) with ELISA assays. Results: The baseline characteristics for the 12 patients were: median age 65.7 years, Gleason score 8, and PSA 74 ng/ml. Greater than 50% decreases in PSA were seen in 9/12 patients, with a median decrease of 74%. Of the 12 patients 7 had a partial response, and 5 had stable disease by RECIST criteria. After 5 cycles of ART-P treatment we found no significant differences from baseline in T-cell proliferation and NK-cell function, or the frequencies of T-cells, regulatory T-cells, MDSC and NK-cells. There was also no change in the serum levels of cytokines and chemokines except for IL-6, which decreased. As expected, the serum levels of VEGF substantially decreased with therapy. Conclusions: Treatment of mCRPC patients with ART-P for 5 cycles did not alter the immune response in a way that would decrease the likelihood of successful immunotherapy, pre or post treatment with ART-P.

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 5379. doi:1538-7445.AM2012-5379
Abstract 5384: Frequency of gene allelic variants of the PAP prostate tumor antigen and association with immune response following antigen-specific genetic vaccination
Authors: Becker Jordan T. , McNeel Douglas G.
Published: August 2012

Prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) are target antigens in genetic vaccines (viral and DNA plasmid vaccines) being evaluated as treatments for prostate cancer. Amino acid allelic variants and different splicing variants of each protein have been identified as part of the human genome sequencing project. We hypothesized that allelic variants in the coding sequence of one of these target proteins in a vaccine recipient, different from the amino acid sequence of the gene encoded by the vaccine, could account for some immune responses observed following genetic vaccination. Such a finding would be essentially analogous to xenoantigen vaccination approaches using highly homologous antigens. In this report, we used direct Sanger sequencing to identify allelic variations in the genes encoding PAP and PSA from 33 patients with prostate cancer who had been previously immunized with a DNA vaccine encoding PAP. The goal of this study was to identify the frequency of variants encoding amino acid changes for two common tumor antigens, and to specifically determine if PAP-specific amino acid variants were more frequent in patients with evidence of immunological response following genetic vaccination and whether these responses localized to regions of allelic variance. Three synonymous coding mutations within the coding sequence of PAP were observed, one of which was observed in 10/33 patients (30.3%), another in 15/33 patients (45.5%), and another in 17/33 patients (51.5%). These variants appear to be linked, as 9 patients possessed all three. No non-synonymous coding mutations were identified. Thus, while a small and somewhat homogeneous patient population, these findings suggest that PAP amino acid variants may be uncommon and immune responses observed are indeed reactive to the host PAP amino acid sequence, not to antigen amino acid variants. Studies are currently underway to alternatively determine whether immune responses were elicited to peptides spanning four known splice variants of PAP. In contrast, we observed four synonymous coding mutations as well as three non-synonymous coding mutations among the five exons of the non-target antigen PSA. Synonymous or non-synonymous mutations were detected in 11/33 individuals (33.3 %). However, while more variations were observed, each occurred in a smaller frequency of patients and did not appear to be linked. The more heterogeneous sequence of PSA may warrant further investigation to determine whether immune responses elicited by genetic vaccines encoding PSA are reactive to host or vaccine-encoded epitopes.

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 5384. doi:1538-7445.AM2012-5384
Abstract 5385: Effect of lenalidomide on antileukemia activity in vitro against adult T-cell leukemia (ATL)
Authors: Kitagawa Yukiko , Matsuoka Masao , Mitani Shohei
Published: August 2012

Background: Lenalidomide is an FDA approved agent for del 5q myelodysplastic syndromes and in combination with dexamethasone for previously treated multiple myeloma. Recently, phase II clinical trial has shown its anti-tumor activity against recurrent and refractory T-cell lymphomas. However, the mechanism underlining its effect is poorly understood. Adult T-cell leukemia (ATL) is a mature T cell malignancy caused by Human T-cell leukemia virus type I (HTLV-I) that is endemic in southwestern Japan, the Caribbean islands and Central Africa, and it still remains as one of the most difficult malignancies to be cured. We have evaluated the anti-ATL activity of lenalidomide in vitro using HTLV-I positive cell lines. Methods: The effect of lenalidomide (10-3-102µM) and Fas antibody (25-400 ng/mL) was assessed on the proliferation of HTLV-I positive cell lines by CellTiter-Glo Luminescent Cell Viability Assay. The cytotoxic effect against HTLV-I positive cell lines of lenalidomide on IL-2 primed peripheral blood mononuclear cells (PBMNCs) as well as sorted CD56 positive and CD3 negative cells from IL-2 primed PBMNCs from healthy volunteers was studied by 51Cr releasing assay. Results: Lenalidomide alone did not inhibit cellular proliferation. Even 6 day incubation did not inhibit or stimulate proliferation of HTLV-1 positive cell lines. Cytotoxic activity of IL-2 primed PBMNCs from healthy volunteers against HTLV-I positive cell lines was enhanced by lenalidomide. Sorted NK cells have also shown enhanced cytotoxic activities on HTLV-1 positive cells. That effect was observed regardless of Tax or cellular FLICE-inhibitory protein (cFLIP) expression. Therefore, even Fas resistant HTLV-1 positive cell lines were succumbed to death by enhanced cytotoxicity by lenalidomide, whereas lenalidomide did not interact Fas inducing apoptotic pathways. Conclusions: These results have shown that lenalidomide enhances cytotoxic effects against HTLV-I positive cells, without direct anti-proliferative effect on those cell lines. That effect was mediated through CD56 positive and CD3 negative cells stimulated by lenalidomide. Further study is now on-going to elucidate the mechanism of those actions in an in vivo setting.

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 5385. doi:1538-7445.AM2012-5385
Abstract 5387: Exploring effects of nanosecond and microsecond pulsed electric field ablation in a rat orthotopic hepatocellular carcinoma model
Authors: Chen Ru , Sain Nova M. , Beebe Stephen J. , Long Gary , Shires Peter
Published: August 2012

Hepatocellular carcinoma (HCC) is the third most common cancer in the world. A diagnosis of HCC carries a guarded prognosis, especially when metastasis is documented. Reliable, curative treatment for very early, early, and intermediate stages is still an inconsistent ideal. Nanosecond (ns), or microsecond (μs) pulsed electric fields (PEFs) that produce irreversible electroporation (IRE) could be used to ablate local tumor masses through minimal invasive or open surgery delivery. Both nsPEFs and μsPEFs induce permanent permeabilization of cells resulting in cell death without any thermal effect. NsPEFs can also impact intracellular organelles and induce apoptosis. Previous studies indicated under certain condition, there was a protective tumor response following nsPEFs treatment in mouse subcutaneous HCC model. In these studies, we specifically test the efficacy of nsPEFs and μsPEFs in an orthotopic HCC rat tumor model. N1S1 cells stably transfected with luciferase were used to generate tumors. Orthotropic insertion of transfected N1S1 HCC cells in Sprague Dawley rat livers was used as the model. Different pulses numbers, rise times and pulse durations were applied. The effect on treated tumor tissue suggests that a single treatment of 500 or 1000 100ns pulses with either slow (50ns) or fast (10ns) rise times and 50 kV/cm can eliminate HCC tumors. Continuous delivery of eighty 100μs microsecond pulses at 2.5 kV/cm dramatically reverses lesion enlargement. 1000 pulses of 100 ns PEFs with fast rise times provoke an innate immune response. Immune responses in this orthotopic HCC model are still under investigation.

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 5387. doi:1538-7445.AM2012-5387
Abstract 5389: 3M-011, a toll-like-receptor-7/8 agonist, as immunological adjuvans in combined therapy of gastrointestinal tumors
Authors: Schoelch Sebastian , Rauber Conrad , Koch Moritz , Weitz Jürgen , Huber Peter E.
Published: August 2012

Background: Although there are reports about spontaneous regressions in advanced cancer stages, in most cases the immune response to solid tumors is unable to stop the progression. This immune response is mediated mainly by effectors of the cellular immune system, specifically NK cells and monocytes/macrophages, which are activated by the binding of antigens to toll-like-receptors. 3M-011, an agonist to TLR-7/8 of this sub-family of pattern recognition receptors (PRRs) may be able to boost the antitumoral immune response, especially in combination with other immunostimulatory treatments such as monoclonal antibodies or ionizing radiation. Materials/Methods: In vivo experiments comprised hetero- and orthotopic mouse models of colorectal cancer evaluating 3M-011, m225 (mouse IgG to EGFR) and photon irradiation alone or in combination. End-points were tumor growth, metastasis and tumor cell dissemination. Mice with complete regressions were re-challenged with tumor cells to evaluate the immunization effect of the therapeutic combination. The cytotoxic activity of various immune cell populations (NK cells, CD-8+ T-cells, macrophages) against gastrointestinal cancer cell lines was quantified in 51Cr release assays. Interactions between these subpopulations were investigated by co-culture experiments. NfkB-Activation was quantified by luciferase-based NfkB reporter assays. Results: A distinct growth retardation for subcutaneous tumors and reduction in metastastic activity of orthotopic tumors was demonstrated in vivo. The combination of the TLR-7/8 agonist and m225 or photon irradiation was shown to be over-additive. Mice with complete regressions after radio-immuntherapy showed distinct anti-tumor immunity when re-challenged with tumor cells as compared to control animals. In vitro assays revealed NK cells in combination with monocytes/macrophages as main effectors of the TLR-mediated early immune response followed by T-cell mediated adaptive immune response. Both effects are controlled by plasmacytoid dentritic cells. On a cellular level, NfkB was shown to be the pivotal pathway of the TLR-mediated immunostimulatory effects as well as the combined therapy effects. Conclusion: The addition of a TLR-7/8 agonist to radiotherapy and antibody therapy can achieve supra-additive therapeutic effects by boosting the immune response to the tumor. Therefore, the addition of a TLR-7/8 agonist to the treatment of advanced gastrointestinal cancers is a promising approach and merits further evaluation.

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 5389. doi:1538-7445.AM2012-5389
Abstract 5392: Angiocidin induces differentiation of acute myeloid leukemia (AML) cells
Authors: Tuszynski George P. , Rothman Vicki
Published: August 2012

Acute myeloid leukemia (AML) is a malignant proliferative disorder where blast stage cells fail to terminally differentiate and accumulate in the blood and bone marrow. AML represents a very significant health risk in the general population with rising incidence in aging adults. From 2004-2008 the median age at diagnosis was 67.8 years and it is estimated 12,950 adults will be diagnosed with 9,050 deaths from AML leukemia in 2011 (http://seer.cancer.gov/statfacts/html/amyl.html, accessed November 2011). Standard AML therapy requires intensive combination chemotherapy with a low rate of durable remissions often requiring stem cell transplantation and associated with significant treatment-related toxicity, especially in elderly patients. Due to this toxicity, the majority of elderly patients are not eligible for therapy. Therefore, new therapeutic options for the treatment of AML are urgently needed. In this study, we present evidence that the human protein angiocidin, discovered in our laboratory, has the potential to be used as a novel, non-toxic therapy for AML. We found that angiocidin can induce both AML cell lines and primary patient cells to increase expression of surface markers characteristic of a more normal differentiated phenotype. We evaluated expression of CD14, a major receptor for the innate immune system in macrophages, CD54 or ICAM-1, the major adhesion receptor used by macrophages for endothelial transmigration, and CD15, a neutrophil phagocytosis receptor. Expression of these receptors was measured by flow cytometry after cells were cultured for 24 hours in media alone or media containing 10 μg/ml angiocidin. Angiocidin up-regulated these receptors by more than 10 fold in 50% of the AML cells tested. The AML cells included the cell line THP-1 and 11 primary AML patient cells. Angiocidin up-regulated CD14 and CD54 in THP-1 cells and in 3 out of 5 M5 phenotype AML patient cells tested and 1 M4 AML phenotype out of 6 M4 samples tested. Angiocidin also up-regulated CD15 in 1 M4 AML patient phenotype out of 6 M4 patient cells tested. These results suggest that angiocidin can differentiate AML cells and therefore has the potential to be developed as differentiation therapy for AML.

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 5392. doi:1538-7445.AM2012-5392
Abstract 54: Cancer-associated fibroblasts induce epithelial-mesenchymal-transition and stemness in non-small-cell lung cancer
Authors: Shintani Yasushi , Abulaiti Abulimiti , Kimura Toru , Nakagiri Tomoyuki , Inoue Masayoshi , Sawabata Noriyoshi , Minami Masato , Okumura Menoshin
Published: August 2012

The tumor microenvironment is composed of different types of stromal cells and a key component of tumor progression. It has been hypothesized that these stromal components are functionally organized to promote the survival of cancer cells. A specific subset of stromal cells, termed cancer-associated fibroblasts (CAFs), secretes factors that promote tumorigensis. To investigate whether CAFs constitute a specific microenvironment conductive for lung cancer progression, we performed in vitro and in vivo experiments using CAFs derived from primary lung cancer in conjunction with the human lung adenocaricinoma cell lines. We first showed that conditioned medium (CM) from human CAFs was biologically active. When human lung adenocarcinoma cell lines (A549 and NCI-H358) were cultured with CM from CAFs, epithelial cells became spindle and scattered as well as decreased cell surface E-cadherin. Further CM increased N-cadherin and vimentin expression, which is a hallmark of an epithelial-mesenchymal-transition (EMT). Coculture experiments using traswell plates with inserts of pore size 3 μm also showed EMT change in cancer cells. CM led cancer cells to enhance expression of stem cell markers such as CD44 and CD133 mRNA levels by ten times. It also triggered the acquisitions of cancer stemness such the ability of resistance to cisplatin or gefitinib as well as sphere formation. These phenotype changes associated with EMT and stemness were inhibited by the addition of TGF-β inhibitor, SB431542 (10 μM). The s.c. coinjection of human lung adenocarcinoma cell lines and human CAFs to the mice result in a high rate of tumor formation compared with the injection of these cell lines without CAFs. A decreased expression of epithelial markers and upregulation of angiogenesis and proliferative index were detected in specimens from mice with coinjection of these cells. TGF β inhibitor attenuated the tumor formation enhanced by coinjection with CAFs. The compound was injected i.p. at 0.2 mg/kg (1 per day, 5 per week for 2 weeks) after coinjection. These results indicate that the paracrine interplay between CAFs and cancer cells mediated by TGF-β signaling leads to an EMT driven gain of cancer stem cell properties and suggest that the CAFs contribute a specific niche for tumor 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 54. doi:1538-7445.AM2012-54
Abstract 5477: Ambient exposure to air pollutants and risk of neuroblastoma
Authors: Heck Julia E. , Wilhelm Michelle , Lombardi Christina , Cockburn Myles , Ritz Beate
Published: August 2012

Background: Some reports have linked neuroblastoma to parental occupational exposure to hydrocarbons and other chemicals. Our aim was to assess the association between neuroblastoma and exposure in pregnancy and early life to criteria air pollutants (CO, O3, PM10, PM2.5, and NOx) and to certain air toxics which are established or suspected carcinogens (benzene, 1,3 butadiene, PAHs, xylenes, carbon tetrachloride, and others). Of the criteria air pollutants, CO and NOx arise primarily from motor vehicle exhaust and thus best represent traffic-related pollution, while O3 and PM arise from a variety of sources and should be considered as markers of regionally distributed background air pollution. Methods: The Air Pollution and Childhood Cancers study ascertained all cancers diagnosed 1988-2007 in the California Cancer Registry among children less than 6 years of age; we were able to match 89% to a California birth certificate. Controls were frequency-matched by year of birth and selected at random from California birthrolls. The final analysis included 1073 neuroblastoma cases and 209,700 controls. Home addresses, as listed on birth certificates, were geocoded and linked to the nearest California Air Resources Board air pollution monitor. For those living within 5 miles of an air pollution monitor, average measurements of criteria air pollutants and air toxics were determined for each trimester of pregnancy and for the first year of life. Multivariate regression models were used to assess the relation between air pollutants and neuroblastoma, with adjustment for maternal race, socioeconomic status, and birth year. Odds ratios compare children in the highest quartile of exposure to those in the lowest exposure group. Results: Exposure to ozone in the 1st (OR =1.27, 95% CI 1.01, 1.60) and 2nd (OR=1.30, 95% CI 1.02, 1.64) trimesters was associated with an increased risk of neuroblastoma (n cases=565 within 5 miles of a monitor). Neuroblastoma was also associated with exposure to carbon tetrachloride in the first trimester (OR=2.62, 95% CI 1.06, 6.46), although this result was based upon 60 cases living within 5 miles of an air toxics monitor. We found no evidence of associations between neuroblastoma and other criteria air pollutants nor other air toxics, including no evidence of associations with traffic-related pollution. Conclusions: This is the first study to suggest a relation between neuroblastoma and ambient levels of air pollution.

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 5477. doi:1538-7445.AM2012-5477
Abstract 5480: Consumption of Lake Ontario sport fish and the incidence of melanoma in the New York State Angler Cohort Study (NYSACS)
Authors: Green Joseph W. , Swanson Mya K. , Olson James R. , Bonner Matthew R.
Published: August 2012

Exposure to polychlorinated biphenyls (PCBs) and other persistent organic pollutants (e.g., toxaphene) has been linked to an increased risk for melanoma in occupational settings. Great Lakes fish bioaccumulate PCBs and organochlorine pesticides, thus consumption is thought to be an important non-occupational exposure route. We investigated the consumption of sport-caught fish from Lake Ontario and the incidence of melanoma in the New York State Angler Cohort Study (NYSACS), a prospective cohort of 17,097 anglers and their spouses aged 18 to 40 years old at enrollment. Participants completed a mailed self-administered questionnaire upon enrollment in 1991. The questionnaire queried for the number of years that fish from L. Ontario were consumed, frequency of species specific fish consumption as well as demographic factors, smoking history and other selected potential confounders. As of December 31, 2008, fifty-one histologically confirmed, first primary, incident melanoma cases were identified via the New York State Cancer Registry. Vital status was determined by linkage with the Social Security Administration Death file. Prevalent cancer cases (n = 196) were excluded. Of the remaining 16,901cohort members, 10% (n=1,672) were lost to follow-up and censored in the year they left New York State. Analyses were restricted to Caucasians (n= 16,326). Fish consumption was dichotomized into never vs. ever. Duration of consumption was categorized into tertiles based on the distribution among the cases reporting eating Lake Ontario sport-caught fish. Poisson regression was used to calculate rate ratios (RR) and 95% confidence intervals, adjusting for age, income, years of education and number of years fishing in New York State. Lake Ontario sport-caught fish consumption was weakly associated with melanoma risk RR=1.33(0.6-2.99) as compared with non-consumers. Duration of Lake Ontario fish consumption was not convincingly associated with melanoma incidence (non-eaters RR = 1.0 (ref); 1st tertile RR = 1.07 (95% CI = 0.38-3.06); 2nd tertile RR = 1.41 (95% CI = 0.49-4.03); 3rd tertile RR = 2.16 (95% CI = 0.79-5.89); p-for trend =0.10. Although interpretation of our preliminary results is complicated by a small number of cases (n=52) and inability to control for potential confounding from sun exposure, they suggest that consumption of Great Lakes fish contaminated with PCBs and other organochlorine compounds was not associated with incidence of melanoma.

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 5480. doi:1538-7445.AM2012-5480