bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5328: Interleukin-4 receptor regulates metastasis of mammary epithelial cancer cells
Authors: Venmar Katherine T. , Hwang Daniel G. , Dozier Ashley , Carter Kathy J. , Gillani Sareena , Fingleton Barbara
Published: August 2012

Interleukin-4 (IL-4), a cytokine produced mainly by immune cells, may promote cancer cell survival by mediating apoptosis resistance in epithelial tumors. The IL-4 Receptor (IL-4R) is overexpressed in a variety of human solid cancer cell lines and primary cell lines, and may be a promising target for anti-tumor therapy. Here, we show that loss of IL-4Rα, a component of the IL-4 receptor, in two murine mammary tumor cell lines (R221A and 4T1), reduces tumor cell survival and results in attenuated metastatic growth in secondary sites. Proliferation and direct cell counting analyses, following knockdown of IL-4Rα with RNA interference, show that R221A clones with reduced IL-4Rα expression proliferate at a comparable rate to control clones, but the number of cells is significantly reduced. This data correlates with the finding that reduced IL-4Rα expression in R221A cells almost completely abolishes tumor take after orthotopic mammary injection in vivo. Furthermore, reduced IL-4Rα expression in R221A and 4T1 cell lines results in a decrease [2.4 fold] in lung metastatic growth in vivo. Reduced expression of IL-4Rα in 4T1 cells results in a similar decrease of liver metastasis. In support of this data, proliferation assays following serum starvation and a modified clonogenic survival assay show that both 4T1 and R221A knockdown clones have a decrease [2 fold] in survival ability in comparison to control clones. Taken together, our results indicate that the IL-4 receptor promotes survival of breast cancer cells at metastatic sites, and suggests that drugs targeting this pathway, currently in clinical trials for respiratory ailments, may have potential for limiting metastatic disease.

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 5328. doi:1538-7445.AM2012-5328
Abstract 533: Dendritic cells are professional educators of Natural Killer cells
Authors: Wiernik Andres , Miller Jeffrey S.
Published: August 2012

Dendritic cells (DCs) and Natural Killer (NK) cells are important components of innate immunity. NK cells are the principal effectors of the innate immune system and play an important role in tumor surveillance and anti-viral immunity. It has been commonly accepted that CD56bright NK cells are the main source of cytokine production while CD56dim are mostly responsible for cytolytic activity and target cell killing. CD56dim NK cells acquire KIR (Killer cell Immunoglobulin-like Receptors or KIR) receptors by a process called NK cell education or licensing. NK cell education has been defined as the mechanism by which NK cell acquire inhibitory KIR and how signaling by specific cognate ligands leads to gain of NK cell function. DCs are antigen presenting cells which act as sentinels for the immune system by engulfing pathogens, processing and presenting them to CD4 and CD8 T cells. Interactions between dendritic cells and natural killer cells have been extensively studied but the role that DCs might play in NK cell education remains elusive and poorly understood. We evaluated the in-vitro interaction between adult monocyte-derived dendritic cells and natural killers cells from healthy donors. Mature monocyte-derived dendritic cells (mdDCs) were co-cultured with KIR negative and KIR positive NK cells in addition to different concentrations of IL-15. After 5 days of co-culture; the KIR negative population (0.8% KIR positive after 5 days of co-cultured) acquired up to 3.8% KIR expression when co-cultured with mdDCs (1:1 ratio) and 0.5 ng/mL of IL-15. In addition, a population of NK cells which expressed 28% KIR after 5 days of culture expressed up to 38% KIR when mdDCs and 0.5 ng/mL of IL-15 were added to the culture. Of note, enhanced KIR expression (as a marker of maturation) required both IL-15 and mdDcs as no effect was seen when mdDcs or IL-15 were added alone. Aside from the differences noted in KIR expression, a population of CD56dim NK cells became CD56bright when both KIR negative (8.44% to 23.6%) and KIR positive (0.8% to 24.9%) NK cells were co-cultured with mdDCs and IL-15. In conclusion, our studies suggest that dendritic cells have the ability to educate NK cells and that this process is dependent on IL-15, which can be trans-presented through IL-15Rα on DCs. Dendritic cells in addition to IL-15 may also revert a population of mature CD56dim NK cells to a CD56bright NK cells, which are considered less differentiated, suggesting that NK maturation can be bidirectional and may exhibit more plasticity than previously recognized.

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 533. doi:1538-7445.AM2012-533
Abstract 5344: Subtyping of glioblastoma stem-like cancer cells with neural lineage markers predicts invasiveness and correlates with survival
Authors: Zorniak Michael , Clark Paul A. , Leeper Heather E. , Tipping Matthew D. , Francis David M. , Kozak Kevin R. , Salamat M. Shahriar , Kuo John S.
Published: August 2012

Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinically useful molecular prognostic markers. GBM gene expression and microRNA profiling recently revealed developmental subtypes that potentially correlates with prognosis and treatment response. However, the clinically significant subset of glioblastoma stem-like cells (GSC) still lacks detailed molecular characterization and correlation to patient survival. We report that molecular classification of patient-derived GSCs using neural lineage markers show association with differential xenograft invasiveness, and also demonstrate significant correlation to survival in both the mouse model and human patients. Immunoblot array of neural lineage markers classified five independently isolated human GSC lines into three classes exhibiting differential expression of oligodendrocyte progenitor cells (OPC), astrocyte progenitor cells (APC), and neural progenitor cells (NPC) markers. After orthotopic injection into immunodeficient mice, Ki-67 proliferative index independent xenograft infiltration was observed: class I GSCs (OPC and NPC positive) created focal lesions, class II GSCs (NPC positive) formed minimally invasive lesions, and class III GSCs (APC positive) established highly infiltrative lesions. The OPC marker, cyclic nucleotide phosphodiesterase (CNP), exhibited high expression in focal xenografts versus low expression in invasive xenografts. Differential CNP expression significantly correlated with mouse model survival, and CNP assay of a large clinically annotated human GBM tissue microarray also showed differential patient survival. Therefore, GSC molecular characterization with neural lineage markers revealed a novel and potentially useful clinical prognosis marker, and suggests clinical importance for patient-specific GSC subtyping.

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 5344. doi:1538-7445.AM2012-5344
Abstract 5353: Characterization of breast cancer stem cells (BCSCs) in Egyptian breast cancer patients through genetic profiling using the RT-PCR and quantitative real time microarray
Authors: Zekri Abdel-Rahman N. , Bahnassy Abeer A. , Ahmed Ola S. , Mourad Magda
Published: August 2012

Background: Stem cells (SC) are cells that can differentiate along distinct lineages through systemic differentiation steps. The concept of self renewal is crucial to understand cancer stem cells (CSC) and the mechanisms by which current therapies might evade the available treatment modalities. The aim of the present study was to characterize breast cancer stem cells (BCSC) by genetic profiling of properly isolated BCSCs at different stages using the recently developed quantitative real time microarray technology to understand the origin and contribution BCSCs to breast cancer development and progression. Material and Methods: 60 cases of BC were subjected to enzymatic digestion to obtain single cell suspension for: 1) flowcytometric characterization of cells, 2) mammosphere culture 3) magnetic separation of different subsets of BCSCs (CD44 and CD24), and 4) RNA extraction for RT-PCR and microarray using the SABiosciences's QC PCR profiler Arrays with SC panel of 84 genes (SC specific markers, SC differentiation markers, and signaling pathway markers for SC). Samples for the QC PCR profiler Arrays were categorized into groups: 1) normal breast tissue, 2) BC cells obtained from tumor samples, 3) cells from mammosphere cultures, 4)CD44+/CD24low/EpCAM- cells, 5) CD44+/ CD24low cells. ER, PR and Her-2 assessment was also performed. Results: Discrete mammospheres of different sizes were obtained from 10 cases and IHC of these samples confirmed the SC lineage. By FCM, cells expressing CD44+/CD24low ranged from 0.1% - 79%. MYC, MYST2, DHH, BTRC AND CXCL12 genes showed over-expression in the studied groups in comparison to normal breast tissue. Comparing all studied groups, we identified 5 candidate genes DVL1 and EP300 (Notch pathway), BTRC (Wnt Pathway), DHH (Chro-Chro Modulation), MYST2 (self Renewal), NCAM1 (Cell Adhesion Molecules) which could be used for characterization and molecular target therapy for breast cancerl. Conclusions: Our QC PCR profiler Arrays showed significant differences between the studied groups shedding light on the nature and helping in proper characterization of BCSCs. Correlation between prognostic factors of BC and the number of BCSCs in tumor biopsies together with the probability of mammoshpere formation in culture denotes the contribution of CSCS to the progression of BC.

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 5353. doi:1538-7445.AM2012-5353
Abstract 5368: A phase II study of ipilumimab plus androgen deprivation therapy in castration-sensitive prostate carcinoma
Authors: Aparicio Ana , Sun Jingjing , Tang Derek Ng , Arap Wadih , Araujo John , Corn Paul , Tannir Nizar , Wang Xuemei , Siefker-Radtke Arlene , Sharma Padmanee
Published: August 2012

Anti-CTLA-4 therapy (Ipilimumab, Yervoy™) was recently FDA-approved for the treatment of metastatic melanoma based on improved median overall survival observed in Phase III clinical trials. Since anti-CTLA-4 targets a T cell specific molecule, as opposed to a tumor specific molecule, it is being investigated as treatment for multiple malignancies, including prostate cancer. Given that anti-CTLA-4 mediates anti-tumor responses by enhancing T cell responses, it has been postulated that standard agents that enable tumor cell death may allow for priming of a T cell immune response that can be augmented by combination with CTLA-4 blockade. Therefore, we hypothesized that adding ipilimumab to standard androgen ablation (AA) therapy in men with metastatic prostate cancer (mPC) would improve immune and clinical responses. We designed a clinical trial whereby men with castrate-sensitive mPC, ECOG performance status ≤ 2 and normal organ and marrow function are registered ≤1 month of initiation of an LHRH agonist or antagonist (LHRHa). Men with autoimmune diseases, known brain metastases, small cell carcinoma morphology, known HIV, Hepatitis B or C infection, untreated symptomatic spinal cord compressions or chronic immunosuppressive therapies are excluded. Starting on day 29, patients receive 4 monthly doses of 10mg/kg ipilimumab. The primary endpoint of the trial is to estimate the rate of PSA ≤ 0.2ng/mL at 7 months, shown to be a powerful predictor of survival (Hussain et al. J Clin Oncol. 2006; 24:3984). Secondary endpoints include: immune responses, time to testosterone recovery (>50ng/mL), time to disease progression off AA, overall survival and safety of the combination.Accrual is ongoing. Data from 8 patients accrued to date show a significant increase in the frequency of T cells expressing the inducible costimulator (ICOS), as we have previously shown with ipilimumab monotherapy (Liakou et al. Proc Natl Acad Sci U S A. 2008;105:14987; Carthon et al. Clin Cancer Res. 2010;16:2861), and significant increases in memory T cell markers. ICOS+ T cells play an important role in anti-tumor responses mediated by anti-CTLA-4 therapy (Fu et al. Cancer Res. 2011;71:5445). Significant changes in dendritic, natural killer and macrophage cell populations have not been observed. To date, one patient developed an asymptomatic grade 3 transaminitis after 2 doses of ipilimumab that resolved with high dose steroids. Assessment of immune responses induced by the combination of ipilimumab and AA, and evaluation of clinical endpoints are ongoing. Updated data will be presented.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5368. doi:1538-7445.AM2012-5368
Abstract 5370: Adjuvant breast cancer vaccine improves overall survival of patients with depressed immunity
Authors: Head Jonathan F. , Elliott Robert L.
Published: August 2012

Breast cancer patients were vaccinated in the adjuvant setting with a whole cell vaccine, containing autologous and allogeneic breast cancer cells, to evaluate the effect on host lymphocyte immunity and overall survival. We began preparing whole cell preparations for a breast cancer vaccine clinical trial in 1995. Stage I and II breast cancer patients had host lymphocyte immunity against tumors associated antigens evaluated before standard therapy. Those patients with depressed immunity determined by a lymphocyte blastogenesis assay (LBA) were offered the whole cell vaccine. Patients were given six intradermal injections (three weekly followed by three monthly). Ten weeks after the last injection the LBA was repeated. Thirty-seven patients were vaccinated in the adjuvant setting with the whole cell vaccine, containing autologous and allogeneic breast cancer cells. Some patients experienced slight pain and swelling at the injection site. There were no severe toxicities and the vaccine was well tolerated. There has been a ten year follow-up for all vaccinated patients. The survival data of the vaccinated patients with depressed immunity (79% survival at 10 years) compared to an historic control of unvaccinated patients with depressed immunity to their tumor associated antigens (59% survival at 10 years), demonstrates an improvement in overall survival of the vaccinated patients. This study confirms the importance of maintaining good host lymphocyte immunity after completion of standard therapy, and validates cancer immunotherapy in the adjuvant 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 5370. doi:1538-7445.AM2012-5370
Abstract 5404: Up-regulation of proliferative and migratory genes in regulatory T cells from patients with metastatic castration-resistant prostate cancer
Authors: Tucker Jo A. , Huen Ngar-Yee , Pang Alan LY , lee Tin-Lap , Vergati Matteo , Intrivici Chiara , Cereda Vittore , Jochems Caroline , Chan Wai-Yee , Rennert Owen , Gulley James L. , Schlom Jeffrey , Tsang Kwong-Yok
Published: August 2012

Immune suppression by regulatory T cells (Tregs) is associated with tumor evasion. An increase in the frequency and suppressive function of Tregs has been shown in a variety of solid tumors. The purpose of this study was to determine if the enhanced number and activity of these cells in prostate cancer patients could be due to tumor-induced changes in gene expression. CD4+CD25hiCD127lo Tregs were isolated from the peripheral blood of healthy donors and metastatic castration-resistant prostate cancer (mCRPC) patients. These samples were selected based on similar suppressive activity in a functional assay. A genome-wide expression array of 38, 500 genes was then performed to detect any effects in expression potentially induced by tumor. We found 384 genes had a three-fold or greater difference in expression between the groups. Differentially expressed genes were involved in cell cycle processes, cellular growth and proliferation, immune response, hematological system development and function, as well as the IL-2 and TGF-β pathways. Genes most up-regulated in the Tregs of mCRPC patients included C-FOS, C-JUN, DUSP1 and RGS1, which are crucial for regulation of T cell proliferation, activation and migration. We also observed increased expression of c-Jun protein in patient Tregs by flow cytometry. Patients had a significantly higher percentage of Tregs in their CD4+ population than the healthy donors, but interestingly, there was no difference in FoxP3 expression. These results indicate that tumor-derived factors may contribute to Treg mediated immune suppression by up-regulating genes that increase their proliferative and migratory capacities, allowing them to better home to the tumor and inhibit the host response.

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 5404. doi:1538-7445.AM2012-5404
Abstract 5411: Rb1 controls the differentiation of myeloid-derived suppressor cells subsets
Authors: Youn Je-In , Collazo Michelle , Kumar Vinit , Gabrilovich Dmitry I.
Published: August 2012

Myeloid derived suppressor cells (MDSC) consist of two distinct subsets, polymorphonuclear/granulocytic-MDSC (PMN-MDSC), and monocytic-MDSC (M-MDSC) that differ in morphology, phenotype and preferred suppression mechanisms. PMN-MDSC is the predominant subset of MDSC that accumulates in spleen, bone marrow, blood, and tumor tissues in various mouse tumor models. M-MDSC produced higher level of cytokines and chemokines including G-CSF, IL-1b, IL-6, TNF-a, MCP-1 and RANTES compared to PMN-MDSC. The current paradigm considers PMN-MDSC and M-MDSC to differentiate from different progenitors alongside different lineages (granulocytic and monocytic, respectively). M-MDSC differentiate into dendritic cells and macrophages, whereas PMN-MDSC are terminally differentiated cells. Our study demonstrated that in cancer M-MDSC represent highly proliferative population of cells that differentiate not only to DC and macrophages but also to PMN-MDSC. The retinoblastoma gene (Rb1), a central regulator of the cell cycle and cell differentiation, is known to be involved in monocytic and granulocytic lineage commitment. When examining Rb1 levels in freshly isolated cells, M-MDSC expressed high levels of Rb1 similar to those in dendritic cells, macrophages, and neutrophils isolated from naïve mice. In sharp contrast, PMN-MDSC had undetectable Rb1. Lack of Rb1 in mice with conditional deletion of the gene promoted an expansion of splenic CD11b+Ly6G+Ly6Clo cells, similar to the phenotype of PMN-MDSC in tumor-bearing mice. Lack of Rb1 in PMN-MDSC was caused by transcriptional repression of the gene. In culture with GM-CSF, Rb1 expression in M-MDSC remained the same while in PMN-MDSC it was increased over time to the level seen in control neutrophils. Inhibition of rb1 expression in PMN-MDSC correlated with the level of histone acetylation of rb1 promoter. Treatment of PMN-MDSC with inhibitors of histone deacetilases (HDAC) resulted in the increase in rb1 expression suggesting that it may be controlled by histone modification. HDAC inhibitors abrogated differentiation of M-MDSC to PMN-MDSC in the presence of tumor explants conditioned medium. These results suggest that down-regulation of Rb1 plays a major role in accumulation of MDSC in cancer by regulating PMN-MDSC differentiation from M-MDSC. HDAC may be considered as potential targets for therapeutic regulation of MDSC.

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 5411. doi:1538-7445.AM2012-5411
Abstract 5419: Targeting colon cancer stem cells using sulforaphane
Authors: Rangarajan Parthasarathy , Subramaniam Dharmaligam , Anant Shrikant , Dhar Animesh
Published: August 2012

Colon cancer is a leading cause of cancer death in the United States, accounting for approximately 51,000 deaths each year. For patients with advanced colon cancer, the 5-year survival rate is less than 10%. Recent evidence suggests the existence of a small population of tumorigenic stem cells responsible for tumor initiation, metastasis and resistance to chemotherapy and radiation. Identification of the regulatory mechanisms and signalling pathways involved in cancer stem cells will help in designing novel agents to target this refractory cell population in colon cancers. We are currently investigating the mechanism by which sulforaphane, an isothiocyanate present in cruciferous vegetables such as broccoli, Brussels sprouts and cabbage inhibits growth of colon cancer cells. Our studies demonstrate that the compound had significant cytotoxicity against HCT116 colon cancer cells in both a dose- and time-dependent manner. Colon CSCs can be allowed to divide and grow in ultra-low binding tissue culture dishes to form multicellular spheroids called colospheres. Treatment with sulforaphane decreased the number and size of the primary and secondary colospheres in a dose dependent manner, suggesting that sulforaphane targets CSCs. To understand the mechanism of inhibition of colospheres, the signaling pathways affected by sulforaphane were dissected. Aberrant activation of Sonic Hedgehog signaling pathway has been associated with renewal of cancer stem cells, and in the development of several solid cancers. Sonic hedgehog (Shh) upon binding to its cognate receptor patched, allows smoothened to accumulate and activate Gli transcription factor. Treatment with sulforaphane inhibited the expression of both Shh and smoothened in the colon CSCs. Furthermore, it inhibited the expression of patched-1 and Gli-1, downstream targets of the hedgehog signalling pathway. Taken together, these data suggest that sulforaphane effectively inhibits colon cancer stem cells by down regulating the sonic hedgehog pathway, thereby inhibiting tumorigenesis (supported in-part by the US PHS grants CA 109269 and CA 135569 to SA and CA 151727 to AD).

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 5419. doi:1538-7445.AM2012-5419
Abstract 5433: Dietary factors and the risk of esophageal cancer: A population-based cohort study in Korea
Authors: Nam Byung-Ho , Bak Jeongin , Kim Jeongseon
Published: August 2012

Objective: To date, there have been few prospective cohort studies that have investigated the association between dietary factors and the risk of esophageal cancer (EC). A large population-based cohort study was conducted to assess the effect of dietary factors on the risk of EC in Korean adults. Methods: The data for this study came from the National Health Insurance Corporation Study (NHICS), Korean Central Cancer Registry. The participants aged 30-80 years were 822,745 Korean men and 368,918 women, cancer-free at baseline in 1996 and 1997, and were followed up to December 31, 2007. The hazard ratio (HR) and 95% confidence interval (CI) of the HR were estimated using the Cox proportional hazards regression model. Results: During the follow-up period, crude incidence rates per 100,000 person-years for EC were 8.1 in overall total population, 11.1 in men, and 1.2 in women. In the total population, the estimated HRs (95% CI) for meat consumption of more than 4 times per week compared with consumption of less than once per week were 1.267 (1.018-1.577) with a p value of 0.0337. In men only groups, the HRs (95% CI) for those who had irregular meal patterns compared with those with regular meal patterns were 1.359 (1.087-1.699) with a p value of 0.0071. Conclusion: The present findings suggest that dietary factors may be associated with the risk of EC.

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 5433. doi:1538-7445.AM2012-5433
Abstract 5450: H-RASG12V transformation modulates antioxidant activities and enhances genotoxicity to arsenite in human urothelial cells
Authors: Liao Yi-Chun , Lai Kuo-chu , Lee Te-Chang
Published: August 2012

Inorganic arsenite (iAs) is a human carcinogen. Epidemiological evidence has revealed the strong association of chronic exposure to arsenic with increased incidences of cancers, including skin, lung, liver, bladder, etc. However, the exact carcinogenic mechanisms of iAs are not completely understood. Numerous studies have shown that mutation-activated H-Ras is frequently observed (about 30 ∼ 50%) in human urothelial carcinoma. In this study, we explored the gentoxic effects of iAs in human urothelial cells ectopically expressing H-RasG12V, an activated H-Ras oncogene. Our results showed that H-RasG12V-transformed human uroepithelial cells (HUC-RAS) were more susceptible to arsenite-induced cell death, DNA damage, micronuclei formation, and anchorage independent growth than control cells (HUC-neo). Furthermore, we found that iAs treatment induced higher intracellular levels of reactive oxygen species (ROS) in HUC-RAS cells than HUC-neo. In addition, iAs-induced genetic damages could be effectively suppressed by N-acetyl-L-cysteine (NAC). We therefore further determined the effects of iAs on antioxidant activities in these 2 cell lines. Our results showed that glutathione (GSH) and superoxide dismutase (SOD) activity were significantly elevated by treatment of HUC-neo cells with iAs, but not in HUC-RAS cells.Alternatively, catalase activity, 2-fold increase in HUC-RAS cells as compared to HUC-neo cells, was significantly suppressed by iAs. Taken together, we may infer that H-RasG12V expression disturbs redox homeostasis by increasing catalase activity. Since catalase activity is sensitive to iAs, it may account for the susceptibility of H-RAS transformed cells to iAs.

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 5450. doi:1538-7445.AM2012-5450
Abstract 5462: Single dose dermal irritation study of apaziquone in rabbits
Authors: Reddy Guru , Chawla Shanta , Fruchtman Steven
Published: August 2012

Introduction and Objectives: Apaziquone is a novel bioreductive alkylating agent in development for intravesical therapy of nonmuscle invasive bladder cancer (NMIBC). The standard dose of apaziquone in all ongoing clinical studies is 4 mg/ 40 mL (0.1mg/mL). For clinical use, lyophilized apaziquone is reconstituted with diluent and instilled in the bladder. During the preparation and handling of the product, there is a possibility of accidental dermal contact for the healthcare professionals. To date, dermal irritation potential of apaziquone has not been investigated. The purpose of this study was to evaluate the dermal irritation potential of apaziquone in rabbits. Materials & Methods: Apaziquone was administered topically on the skin (non-occlusive) to New Zealand White (NZW) rabbits as a single dose for 4 hours followed by removal of excess material, and skin examination for up to 72 hours after removal of excess material. Each male NZW rabbits (3/group; one group) had three identified treatment sites on the back: (1) untreated (2) vehicle (diluent for apaziquone) and (3) apaziquone (0.1 mg/mL). Animals were treated with a single non-occlusive dermal dose of apaziquone (0.5 mL of 0.1 mg/mL) and vehicle for 4 hours followed by removal of excess material. No test material or vehicle was applied to the site designated as untreated. The treatment sites were observed and scored at baseline and at various times (4, 24, 48, and 72 hours) after removal of excess test article and vehicle. The following parameters were evaluated: viability, clinical observations, and dermal observations/scoring by modified Draize method. Results: No clinical signs of toxicity were observed during the study and no signs of dermal irritation related to test article or vehicle/diluent were observed at any of the dose sites in any of animals. The Primary Dermal Irritation Index for apaziquone, administered at clinically relevant exposures, had a value of zero (0) indicating no irritation at the concentration and for the duration of exposure. Conclusion: Apaziquone did not cause any dermal irritation in rabbits at a clinically relevant concentration.

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 5462. doi:1538-7445.AM2012-5462
Abstract 5546: UVC irradiation inhibits superficial tumor growth in the brain of nude mice
Authors: Momiyama Masashi , Hiroshima Yukihiko , Suetsugu Atsushi , Tome Yasunori , Mii Sumiyuki , Yano Shuya , Chishima Takashi , Bouvet Michael , Endo Itaru , Hoffman Robert M.
Published: August 2012

Temozolomide and UVC light were used for induction of cancer cell apoptosis of cancer cells which was imaged in real time at the subcellular level in the brain. Temozolomide inhibited brain tumor growth (p < 0.001). UVC irradiation inhibited superficial brain tumor growth (4.7-fold) (p < 0.0001) and improved survival (39.0%) (p = 0.002). This model can be used to evaluate new treatment modalities of brain metastasis or brain cancer at the subcellular level. In order to visualize single cancer cell nuclear-cytoplasmic dynamics of cancer cells in the brain in real time, including mitosis and apoptosis, Lewis lung cancer cells or human U87 glioma cells were injected into the brain either stereotactically or via the carotid artery of nude mice. The cancer cells were labeled with GFP in the nucleus and RFP in the cytoplasm, such that mitosis and apoptosis could be clearly imaged. A craniotomy open window was made for real-time fluorescence imaging of the cancer cells growing in the brain. Mitosis as well as treatment-induced apoptosis of the individual cancer cells were imaged in real time. UVC irradiation was initiated 1 day after the cancer cell inoculation and tumors were irradiated for 120 seconds on days 1, 4, 7, and 10. The tumor volume and survival were compared with untreated mice. UVC irradiation inhibited 4.7-fold changes in the brain tumor growth (p < 0.0001) and improved 39% higher survival (p = 0.002).

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 5546. doi:1538-7445.AM2012-5546
Abstract 5547: Oral delivery of tumor-targeting Salmonella typhimurium A1-R is effective therapeutically
Authors: Tome Yasunori , Zhang Yong , Momiyama Masashi , Maehara Hiroki , Kanaya Fuminori , Tomita Katsuro , Bouvet Michael , Hoffman Robert M. , Zhao Ming
Published: August 2012

The S. typhimurium A1-R mutant, which is auxotrophic for leu-arg to attenuate growth in normal tissue, and selected for high antitumor virulence, has been shown to be effective as monotherapy against human prostate, breast, pancreatic, and fibrosarcoma tumors. In the present study, we first administered A1-R to nude with subcutaneous murine colon cancer Colon-26 which expressed red fluorescent protein (RFP). S. typhimurium A1-R (1×107 cfu) was administered intravenously (i.v.) or intratumorally (i.t.) to the subcutaneous tumor as a positive control. The size of tumor was measured by weekly fluorescence imaging. Both i.v.- and i.t.-treated mice had smaller tumors and survived longer (p<0.05) compared to the control group treated with PBS. When S. typhimurium A1-R was administered orally to nude mice with orthotopically-transplanted Colon-26 RFP, tumor growth was inhibited (p<0.05) and animals survived longer than untreated controls (p<0.05). Furthermore, orally-treated mice had no observable side effects suggesting oral delivery of tumor-targeting S. typhimurium A1-R is a promising route for bacterial therapy 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 5547. doi:1538-7445.AM2012-5547
Abstract 5553: Real-time imaging of αv integrin-GFP expression in osteosarcoma cells trafficking in blood vessels of nude mice
Authors: Tome Yasunori , Sugimoto Naotoshi , Yano Shuya , Momiyama Masashi , Maehara Hiroki , Yamamoto Norio , Tsuchiya Hiroyuki , Tomita Katsuro , Kanaya Fuminori , Bouvet Michael , Hoffman Robert M.
Published: August 2012

We report here real-time imaging of αv integrin-GFP expression during osteosarcoma cell trafficking in blood vessels in nude mice. 143B osteosarcoma cells were transfected with an αv integrin-GFP vector. Confocal microscopy demonstrated that αv integrin immunofluorescent staining colocalized with αv integrin-GFP. Osteosarcoma 143B cells expressing αv integrin-GFP (1×106) were injected in the epigastric cranialis vein. Subsequently, round osteosarcoma cells diffusely expressing αv integrin-GFP were observed in larger veins. In contrast, punctuated αv integrin-GFP was imaged in stretched cells attached to narrow vessel walls. Osteosarcoma cells, arrested in venous valves, were observed to strongly express αv integrin-GFP. αv integrin-GFP allows the imaging of αv integrin expression during cancer cell trafficking and arrest in blood vessels which will enable the further understanding of its role in this process.

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 5553. doi:1538-7445.AM2012-5553
Abstract 5565: Circulating tumor cells in the peripheral blood and cerebrospinal fluid of patients with central nervous system metastases
Authors: Allen Joshua E. , Patel Akshal S. , Dicker David T. , Sheehan Jonas M. , Glantz Michael J. , El-Deiry Wafik S.
Published: August 2012

The enumeration of circulating tumor cells in peripheral blood is a validated method of monitoring disease status in patients with epithelial malignancies involving the breast, colon, prostate, or lung. As cancer therapies improve and aim to decrease metastatic burden, the central nervous system (CNS) is emerging as an important sanctuary site for metastasis. Current surveillance and clinical markers of metastatic disease progression involving the CNS are lacking adequate sensitivity, reproducibility, and specificity. One of these clinical markers is cerebrospinal fluid (CSF) cytology, which lacks clarity, is insensitive, requires high sample volumes and is subject to interpretation bias by the pathologist. To overcome these limitations, we previously adapted the Cellsearch system (Veridex) to isolate and enumerate tumor cells in the CSF of breast or non-small cell lung cancer patients with CNS metastases. We recently reported that the number of cells in the CSF directly correlated with the subject's clinical condition and Karnofsky performance status (KPS) in metastatic breast cancer. Furthermore, imaging of the neuroaxis in these patients revealed disease presence, however, was not indicative of disease burden. A sharp decline in the number of CSF tumor cells (CSFTCs) was universally noted after the onset and maintenance of such ventricular intrathecal chemotherapy. We have continued to follow these patients and have expanded our patient population to include non-small cell lung cancer. This includes a patient that we have followed from a baseline CSFTC count of >10,000 to zero over the course of intrathecal treatment, which as been accompanied by drastically reduced disease by and complete resolution of clinical symptoms. We have also begun to perform concomitant CTC enumeration from peripheral blood in conjunction with CSFTC analysis. Interestingly, while there appears to be no strong correlation between the two subpopulations, tumor cells seemed to be compartmentalized to either the blood or the CSF. Together, the current data indicates that CSFTCs may serve as viable markers for diagnosis and prognostication in the setting of CNS metastases. This novel method of isolating CSFTCs provides a semi-automated platform that greatly increases sensitivity, accuracy and reliability over the standard CSF cytology. Furthermore, this platform enables studies to elucidate the significance and biology of this unique subpopulation of tumor cells, including its relationship with CTCs.

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 5565. doi:1538-7445.AM2012-5565
Abstract 5577: HER2 cytoplasmic staining and penile cancer
Authors: Silva Alice M. , Cunha Isabela W. , Guimarães Gustavo C. , Cubilla Antonio , Lopes Ademar , Soarea Fernando A.
Published: August 2012

BACKGROUND: Although rare in developed regions, penile carcinoma is a serious concern for undeveloped countries such as Brazil, where it can represent up to 10% of all malignancies. HER2 is a well-known growth factor membrane receptor that has its expression level usually associated with aggressive disease. Recently, some studies have shown that soluble fractions of HER2 intracellular domain are found in tumors cytoplasm and they still activate intracellular pathways related to cell proliferation and survival. MATERIAL AND METHODS: Immunohistochemistry against HER2 was carried out in 195 penile carcinoma samples selected from the files of AC Camargo Hospital, Brazil, using the monoclonal CB11 antibody (Novocastra™), which recognizes specifically HER2 intracellular domain. Cases were classified as positive or negative according to membrane and to cytoplasmic staining. All statistical analyses were performed by the SPSS for Windows 18.0, SPSS Inc. Comparison between category variables was performed by the Pearson chi-square test or the Fisher's exact test. Survival rates were calculated using the Kaplan-Meier method and the curves were compared by means of the log-rank test. In all statistical tests, the alpha error was set at 5%. RESULTS: None of the cases showed membrane staining for HER2. However, a clear and strong cytoplasm staining was observed in 28 cases (14,9%), as shown in figure 1. The staining was limited to the tumor cells and does not represent background or artifact. Univariate analyses showed that overexpression is associated with higher histologic grade (p <0,001) and lower overall survival (p=0,007). Multivariate analyses showed that HER2 in the cytoplasm is an independent risk factor for death (RR=2,966; IC 95% [1,6-5,2]; p <0,001). CONCLUSIONS: Corroborating with recent findings that cytoplasmic fragments of HER2 can be active kinases, the present study suggests that the presence of HER2 in tumor cytoplasm is a prognostic marker in penile carcinoma. We have also demonstrated that, although no membrane staining was seen, presence of HER2 in cytoplasm is an independent risk factor for death in this neoplasm.

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 5577. doi:1538-7445.AM2012-5577
Abstract 5591: A phase I study of bavituximab and sorafenib in patients with advanced hepatocellular carcinoma (HCC)
Authors: Yopp Adam , Arriega Yull , Singal Amit , Mansour John , Balch Glen , Thorpe Phillip
Published: August 2012

Background: HCC is the fastest growing cause of cancer-related deaths in the US, with the majority of patients presenting with advanced stage disease. Despite sorafenib being the only systemic therapy with proven efficacy in advanced HCC, the median survival for these patients is still less than one year. Sorafenib, in pre-clinical HCC models, upregulates phosphatidylserine (PS) exposure on tumor vascular endothelium. The phosphatidylserine (PS)-targeting antibody, bavituximab, given in combination with sorafenib has greater efficacy in pre-clinical HCC models than either therapy alone. Bavituximab acts by directing immune cells to destroy the PS-expressing tumor vasculature, and by reactivating tumor immunity.The aim of our study was to evaluate the maximum tolerated dose (MTD) of bavituximab in combination with 400 mg bid of sorafenib in patients with advanced HCC. Methods: In this single institution, phase I study (NCT01264705), sorafenib-naive adult patients with measurable, advanced HCC, ECOG PS ≤ 2, and Child-Pugh (CP) class A cirrhosis received escalating doses of bavituximab weekly (0.3, 1.0, and 3.0 mg/kg) and sorafenib 400 mg bid for 28 days. Results: Median age of patients was 61(range 43-76) years and seven patients were male. Five patients had CP A5 and 4 patients had CP A6 cirrhosis. The majority of patients had underlying HCV (7/9), one patient had HBV and one patient had NASH. Five patients (4 TACE, 1 resection) received prior treatment. Four patients had metastatic disease at initiation of treatment. No dose limiting toxicities were seen at any dose level. No grade 3/4 treatment-related events in the first 28 days were noted. Common toxicities at all grades were related to sorafenib and included hand-foot syndrome (33% grade I, 22% grade II), fatigue (33% grade I) and anorexia (22% grade I). Conclusions: Bavituximab and sorafenib can be given safely in patients with advanced HCC at 3.0 mg/kg weekly and 400 mg bid, respectively. Hand-foot syndrome, fatigue and anorexia were the most common adverse events and are consistent with known adverse events with sorafenib therapy. These doses are currently being evaluated in an ongoing phase II study in this population.

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 5591. doi:1538-7445.AM2012-5591
Abstract 5618: Tumor suppressor genes? The status of genes PAX 5&6 in primary and secondary gliomas and glioblastoma multiforme tumors
Authors: Newman Mary B. , Wallace David , Mitchell Brian , Byrne Richard W.
Published: August 2012

Background: Gliomas are the most common brain tumors, and glioblastoma multiforme (GBM) tumors have the poorest prognosis, and due to their molecular heterogeneity, these tumors remain incurable and highly resistant to treatments, in most cases. The PAX (paired box) family of genes is comprised of are nine members that are subdivided based on the products of their protein domain structures. PAX5 and PAX6 are developmental genes that are vital during embryonic stages of development, and their transcription factors have supportive, if not, key functions in tumorigenesis. Interestingly, gene PAX6 has been reported to be over and under expressed in high grade gliomas, GBMs, and glioma cell lines. However, these results could be an effect of upstream or downstream manipulation of other transcription factor(s), the origin of the tissue (PAX6 expression is restricted to the forebrain, optic cup, hindbrain, and spinal cord, lens placode, and nasal epithelium), and the epigenetic regulation of some other gene. In addition, there are no reported mutations in either gene PAX5 or PAX6 in the cells from GBMs. Therefore, confusion exists as to role these genes have in tumorigenesis, and whether either gene has the properties or characteristics of a tumor suppressor gene. Study Objective: The overall study objective was to determine if either PAX5 or PAX6 are tumor suppressor genes, and can be silenced or over-expressed to inhibit the progression of cultured human adult primary glioma tissue samples, by employing small interfering RNA (siRNA) methodology. Methods: Primary cell cultures were incubated in reduced serum conditions for at least 1 month prior to transfections and cells were passed every 7 to 10 days or when flask was 90% confluent. To investigate and measure true inhibition of cell proliferation or progression of the cells, several techniques were used, such as, real-time quantitative reverse transcription PCR, enzyme-linked immunosorbent assays, and immunohistochemistry. Results: The high grade (IV) primary glioma tissue samples and their respective cell cultures showed relatively high expression levels for the gene PAX6 and lower levels of expression for gene PAX5 when compared to GAPDH gene and standard controls. Our initial transfections for knocking down of PAX6 using siRNA (TriFECTA, IDT) showed 90% of control samples were transfected, and our primary cells were knockdown by 82%, 70%, and 65% for each of PAX6 isoforms (N000280.21.1-S, 2-S, and 3-S) respectively. We are currently analyzing the rest of the PAX6 data and all of the PAX5 data.

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 5618. doi:1538-7445.AM2012-5618