bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2607: Genetic variants in the vitamin D pathway and breast cancer disease free survival
Authors: Pande Mala , Thompson Patricia A. , Yoo Suk-Young , Do Kim-Anh , Sahin Aysegul A. , Amos Christopher I. , Bondy Melissa , Brewster Abenaa
Published: August 2012

Objective. Vitamin D deficiency has been associated with poor outcomes in patients with breast cancer. The objective of this study was to investigate the association between single nucleotide polymorphisms (SNPs) in 7 Vitamin D-related genes and disease free survival (DFS). We hypothesized that vitamin D-related genes may modify breast cancer DFS. Methods. A total of 102 SNPs in Vitamin D pathway genes were explored as predictors of DFS among 1030 stage I/II breast cancer patients treated at MD Anderson Cancer Center between 1985 and 2000. Genotyping was performed using the Illumina GoldenGate array. We applied recursive partitioning tree analysis (RPART) to explore interactions of variables predictive of DFS including vitamin D-related SNPs, age at diagnosis, race, stage, nuclear grade, estrogen (ER) and progesterone (PR) receptor status, chemotherapy, hormone therapy and BMI. Results. Results from partitioning identified chemotherapy as the optimum first split for DFS where patients receiving chemotherapy experienced longer DFS. In patients receiving chemotherapy (n=540), SNP rs1118569 (RXRA) followed by rs10881583 (RXRA) were associated with greater DFS. Patients with two major alleles of rs1118569 had the best outcome, whereas those with two minor alleles of rs10881583 and one or two minor rs1118569 alleles had poorer DFS. In patients who did not receive chemotherapy (n=482), stage I patients had longer DFS than stage II. Further, stage I patients who were homozygous for any allele of rs2248098 (VDR) had longer DFS. Among the stage II patients, those with ER/PR negative tumors had poorer DFS with heterozygous carriers of rs344781 in PLAUR having the worst DFS. Conclusion. While exploratory, we show that chemotherapy, stage, ER and PR status and vitamin D-related pathway polymorphisms but not race/ethnicity modified disease free survival in early stage breast cancer patients. Validation of vitamin D-related pathway genes as genetic determinants of patient outcomes would offer insights on alternative approaches for the secondary prevention of breast cancer recurrence considering clinical pathological and treatment characteristics of the patient 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 2607. doi:1538-7445.AM2012-2607
Abstract 2613: Racial/Ethnic specific polygenic models of breast cancer risk
Authors: Thomas Venetta , Liu Zhi , Srinivasan Sathish K. , Chung Ren-Hua , Martin Eden , Levine Edward A. , Freimanis Rita I. , Hu Jennifer J.
Published: August 2012

Breast cancer is the most frequently diagnosed cancer and the second leading cause of cancer death in American women. Earlier family linkage studies have identified high-penetrance breast cancer genes, such as BRCA1, BRCA2, and TP53. Moreover, genes involved in DNA repair, such as CHEK2, ATM, BRIP and PALB2, have been associated with moderate breast cancer risk. Recent Genome-wide Association Studies (GWAS) have identified multiple single nucleotide polymorphisms (SNPs) associated with breast cancer risk. We evaluated whether breast cancer risk is associated with multiple SNPs in DNA damage signaling and repair and immune and inflammatory responses in addition to SNPs identified from GWAS. We hypothesize that targeting critical candidate pathways may yield important information regarding SNP-SNP interactions in breast cancer risk. We selected SNPs from DNA damage signaling and cell cycle control, base excision repair, nucleotide excision repair, double-strand break repair, mismatch repair, and immune and inflammatory responses. We have genotype results of 3,754 SNPs in 910 controls (763 Whites and 147 African Americans) and 793 breast cancer cases (667 Whites and 116 African Americans). 16.2% and 20.6% of controls and cases (respectively) had a first degree relative with breast cancer. 48% and 55.7% of the controls and cases (respectively) had their first child before or at the age of 24. 40.5% and 42.2% of the controls and cases (respectively) had a history of smoking. Our study results suggest that 6 SNPs were associated with breast cancer risk in Whites: (1) 2 SNPs from GWAS - rs3817198 in LSP1 (p<0.001) and rs3803662 in TNRC9 (p<0.001); (2) 2 SNPs from inflammation - rs1801157 in SDF-1 (p<0.001) and rs12720356 in Tyk2 (p<0.001) and (3) 2 SNPs from DNA repair and apoptosis - rs2992 in CHAF1A (p<0.0001) and rs2231301 in BCL2l2 (p<0.001). Only 1 SNP, rs1990760 in IFIH1 (p<0.01) was associated with breast cancer risk in African Americans. The IFIH1 gene was previously linked to Type 1 diabetes but not in cancer risk. There was significant trend in breast cancer risk with increasing numbers of risk genotypes for rs3817198, rs3803662, rs1801157, rs12720356, rs2992, and rs2231301 (p test for trend <0.0001) in Whites after adjustment for potential confounders. Our results suggest that multiple SNPs contribute to breast cancer risk in a racial/ethnic specific manner and larger studies are warranted to further evaluate our findings. More importantly, functional analysis is under way to corroborate the association of multiple SNPs with breast cancer risk.

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 2613. doi:1538-7445.AM2012-2613
Abstract 2630: DNA repair single nucleotide polymorphisms interact with indoor tanning and melanoma
Authors: Luo Li , Lilyquist Jenna M. , Stidley Chris , Flores Kristina , White Kirsten , Paine Susan , Gonazales Melissa , Erdei Esther , Vogel Rachel , Lazovich DeAnn , Berwick Marianne
Published: August 2012

Development of melanoma has previously been associated with compromised DNA repair which can be analyzed by genotyping SNPs in DNA repair genes. The Minnesota Skin Health Study collected DNA samples to investigate the hypothesis that DNA repair SNPs are associated with increased risk for melanoma among tanning bed users. 92 SNPs, with minor allele frequencies greater than 0.05, were analyzed in 20 genes for 1659 samples_893 melanoma cases and 766 controls. The SNPs were genotyped on the Illumina BeadExpress GoldenGate platform. The SNPs were then analyzed for individual SNP associations with melanoma, haplotype associations with melanoma, individual SNP interactions with ever/never indoor tanning, and haplotype interactions with ever/never indoor tanning after adjustments for age, gender, and phenotypic index. There were several SNPs that had an association with melanoma including: six SNPs in three nucleotide excision repair genes ERCC4, ERCC6 (rs4253190, adjusted OR=1.19, 95%CI: 1.04-1.37; rs4838518, adjusted OR=0.86, 95%CI: 0.75-0.99; rs4253121, adjusted OR=0.84, 95%CI:0.71-0.99), and XPC, one SNP in base excision repair gene RFC1, one SNP in direct reversal DNA repair gene MGMT, and one SNP in a novel gene FBRSL1(rs4883571, adjusted OR=0.87, 95%CI: 0.75-1.00). There were four haplotype associations with melanoma including one ERCC6 block (rs10745261, rs6537537, rs4253121: GGA, adjusted OR: 0.84, 95%CI: 0.71-0.99), two MGMT blocks, and one XPC block. Additionally, there was one SNP in FBRSL1 (rs4883557, p=0.0056) and one SNP in ERCC6 (rs10745261, p=0.025) that had an interaction with indoor tanning along with two haplotypes including one block in ERCC6 (rs10745261, rs6537537, rs4253121: AAG, p=0.0355) and one block in MGMT. While the interaction analyses were suggestive, the study is limited because none of the SNPs or haplotypes were only significant (p<0.05) before Bonferroni correction. Further analyses of the function of these SNPs will be necessary, particularly for ERCC6, which had SNPs that were significant in every analysis, and FBRSL1, which is a novel gene whose function is unknown.

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 2630. doi:1538-7445.AM2012-2630
Abstract 264: ENC1, a BTB/Kelch protein, links DNA-damage signals to apoptosis by ubiquitination of retinoblastoma protein
Authors: Yu Rong , Ma Quan
Published: August 2012

ENC1 (ectodermal neuronal cortex 1) is a BTB/Kelch domain-containing protein, whose gene was originally identified in a p53-overexpressing cell line, and was later found to show abnormal expression in various tumor tissues. Here, we report that ENC1 is a new regulator that links DNA-damage signals to apoptosis. ENC1 is induced in different cancer cells by DNA-damage signals. The induced ENC1 protein then interacted with cullin-3 and retinoblastoma protein (Rb), forming an E3 ubiquitin ligase complex and ubiquitinating Rb protein. Upon ubiquitination, Rb protein failed to interact with E2F, allowing for transcriptional activation of E2F-dependent genes. Inhibition of ENC1 by RNA interference or by a dominant-negative mutant attenuated Rb ubiquitination, E2F-depedent gene activation, as well as apoptosis induced by DNA-damage signals. However, epitopic overexpression of ENC1 gene activated E2F-dependent genes, but failed to trigger apoptosis. Deletion and mutation analysis shows that a conserved motif located within Kelch domain at the C-terminus is essential for regulation of Rb function and apoptosis in response to DNA-damage signals.

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 264. doi:1538-7445.AM2012-264
Abstract 2644: No evidence for increased risk of cancers other than testicular cancer among first-degree relatives of testicular germ cell tumor (TGCT) patients from multiple-case TGCT families
Authors: McMaster Mary L. , Heimdal Ketil R. , Greene Mark H.
Published: August 2012

Familial aggregation of testicular germ cell tumors (TGCT) has been reported, but it is unclear whether other cancers co-aggregate in multiple-case TGCT families. METHODS: We performed an observed-to-expected (O/E) analysis in a cohort of TGCT families with documented family cancer history assembled in Norway and the U.S. All bloodline first-degree relatives of TGCT cases were eligible. Relatives with missing vital status, gender, or dates of birth and/or death were excluded. We used population-based, age-adjusted cancer incidence rates from Norway and the Surveillance, Epidemiology and End Results (SEER) Program of the National Cancer Institute, respectively, to calculate expected numbers of cancer cases. Only non-TGCT cancers were analyzed. RESULTS: A total of 130 TGCT families with 84 reported cancers and 31,556 person-years at risk were included in the analysis. Overall, there was no excess cancer risk, all non-TGCT sites combined, observed among first-degree relatives of TGCT cases (O/E = 0.89; 95%CI 0.71 - 1.10), either collectively or stratified by center. Although numbers were small, site-specific increased risks were observed for soft tissue cancers (n=4; O/E=7.14; 95%CI 1.95 - 18.29) and leukemia (n=9; O/E=6.48; 95%CI 2.96 - 12.29), and site-specific decreased risk was observed for breast cancer (n=6; O/E=0.44; 95%CI 0.16 - 0.95). These risks were statistically significant overall and in Norway, but not in the U.S. CONCLUSION: In this, the largest study of site-specific cancer risk within multiple-case TGCT families yet performed, we found no excess risk of non-TGCT cancers overall among first-degree relatives of TGCT cases. There was limited evidence supporting altered site-specific risks for soft tissue cancer, leukemia and breast cancer; however, cancer numbers were small, and the results were not consistent between centers, suggesting that differences in cancer reporting among families, case validation methods, or other factors, rather than an etiologic association, explain these findings. Familial testicular cancer appears to be a site-specific syndrome.

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 2644. doi:1538-7445.AM2012-2644
Abstract 2651: Association of p53 codon72 polymorphism, BCCIP & CDKN2A with melanoma risk. A p53-dependent mechanism
Authors: Andavolu Radhika G. , Rubakovic Svetlana , Rubin Jacob , Bir Jaski , Shu Shinger , Andavolu Murthy , Murray James Mac
Published: August 2012

Cancers arise due to accumulation of mutations in genes critical for cell proliferation, differentiation, and cell death. Direct interaction with cell cycle regulators characterizes growth suppression by several tumor suppressor genes. Mutation or polymorphisms that occur in the promoter or coding regions may affect gene expression and may thus have the potential to be of phenotypic or even of pathological significance. In our previous studies, we showed the differential gene expression patterns and gene interactions/ association of of BCCIP and/or CDKN2A gene1, TNF and/or CCR52 with respect to genotypes, melanoma types and family history. We reported a significant interaction between BCCIP and CDKN2A genotypes among melanoma cases and a significant difference between cases and controls for the BCCIP/CDKN2A haplotype, when adjusted with age and sex, possibly confering different levels of susceptibility to melanoma. In the same report we also had a finding of CDKN2A mutant genotype showing significant association with Melanoma type 1 followed by type 2 suggesting this gene to be critical only in cases with family history and /or with other malignancy. Previous reports have shown that BCCIP as an important component in maintaining the tumor suppression function of wild type p53.Without BCCIP, p53 fails to form tetramers, cannot bind with its target promoter sequences, and is defective in transactivation activity. The p53 pathway responds to cellular stresses that threaten the fidelity of DNA replication, genome stability, chromosome separation and cell division. Many of the cellular stresses can be oncogenic in nature and in skin, the most damaging stresses in carcinogenesis is UV radiation. In this follow up study we show a strong association of p53 codon72 polymorphism (p<0.003) with melanoma (N=126) as compared to controls (N=172). Positive correlations of p53 gene with BCCIP polymorphism (p<.013), BCCIP(p<0.010) & CDKN2A(p<0.032) gene expressions from tumor tissues from melanoma patients suggest the possible functional role of these reported gene interactions in causation or in disease progression. A 12 gene panel (TNF, CCR5 and other CDK pathway genes) and their possible interactions showing clinical relevance of these markers in melanoma development and/or progression through p53-dependent mechanism are discussed.

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 2651. doi:1538-7445.AM2012-2651
Abstract 27: Chemotherapy-induced Akt survival signaling is regulated by CD44- Ezrin/Radaxin Moesin (ERM) scaffolding, dependent on EGFR activity
Authors: Goldman Aaron J. , Shunmugam Ishwarya , Kulkarni Ashish , Rivera Fernando , Sengupta Shiladitya
Published: August 2012

Akt survival signaling has emerged as a predictor of poor prognosis in tumors resistant to chemotherapy. Despite the firm correlation between enhanced Akt survival signaling in resistant tumor tissue, there is a poor understanding of the exact mechanisms which elicit this robust pathway activation. In the present study we sought to uncover the full pathway of Akt activation in response to chemotherapy in order to fully elucidate mechanisms and proteins involved in chemotherapy resistance. Using breast and ovarian cancer cells, immunoprecipitations revealed that the CD44 glycoprotein initiates a unique scaffolding effect with the Ezrin/Radixin/Moesin (ERM) proteins and Akt in response to the anti-tubulin chemotherapies, docetaxel and vincrisine. Knockdown of CD44 using siRNA revealed that the activation of Akt(ser473) and ERM(tyr353) by chemotherapy are abolished in a time and dose dependent manner. Flow cytometry, caspase activation and cell viability assays confirmed that loss of Akt signaling sensitizes cancer cells to chemotherapy-induced apoptosis and a similar sensitivity was observed in the absence of CD44 expression. Furthermore, immunoprecipitations revealed that inhibition of EGFR using the small molecule inhibitor, Erlotinib, prevented CD44/ERM/Akt scaffolding in response to chemotherapy. In summary, CD44 and ERM scaffolding is responsible for the activation of Akt survival signaling induced by chemotherapy, in-vitro. Loss of CD44 sensitizes cancer cells to apoptosis. This scaffolding effect is dependent on the activity of EGFR, suggesting that a unique interaction exists between this ErbB1 receptor tyrosine kinase and the CD44 glyco-protein. This is the first evidence for cell surface events which precipitate survival signaling in cancer cells as a consequence of chemotherapy. These findings elucidate novel biomarker interactions for translation in the clinical 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 27. doi:1538-7445.AM2012-27
Abstract 2833: The protein ps20 increases tumourigenic potential in prostate cancer epithelial cells
Authors: Hickman Oliver , Vyakarnam Annapurna , Smith Richard A.G. , Dalgleish Angus G. , Dasgupta Prokar , Galustian Christine
Published: August 2012

Introduction The protein ps20 encoded by the WFDC1 gene contains a highly conserved whey 4-disulphide core (WFDC) domain and is a member of a protein family that regulates inflammation. Ps20/WFDC1 is expressed by prostate cancer stromal cells but its expression by cancerous prostate epithelium is associated with tumour progression in patients. The aim of this study was to determine how WFDC1expression by prostate cancer cells alters their tumourigenic properties. Methods The cell-lines LNCAP and PC3 were transfected with WFDC1 vector or empty vector (EV) and their growth, adhesive and migratory characteristics were investigated by MTT proliferation, transwell migration and ICAM-1 adhesion assays. Results PC3 and LNCAP cells transfected with WFDC1 increased in numbers by 3148 +/−151% and 523+/−311% after 48 hours respectively compared to EV transfected cell lines (p <0.01 by unpaired T test). WFDC1-transfected cells became smaller and tightly clustered with greater cell-cell contact (See Figure 1). WFDC1 transfected cell migration through 8µM transwells was increased by 73% in PC3s and 57% in LNCaP compared to EV cells (p<0.05 by paired t-test). ICAM-1 expression increased by 67% in WFDC1-LNCAP and by 50% in WFDC1-PC3s compared to EV-cells (p<0.05 by paired t-test). VEGF (vascular endothelial growth factor) was also strongly detectable in the WFDC1-PC3 and LNCaP cells by western-blot whereas none was detectable in EV cells. Conclusions WFDC1 transfection of prostate cancer epithelial cells increases proliferation, migration and ICAM-1 expression, and also induces VEGF, which is a potent angiogenic growth factor. WFDC-1/ps20 may therefore be a critical target for prostate cancer therapeutics.

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 2833. doi:1538-7445.AM2012-2833
Abstract 2844: Inhibiting androgen receptor and JunD protein interaction to prevent prostate cancer progression
Authors: Schmidt Joseph , Kegel Stacy , Mehraein-Ghomi Farideh , Saphner Elizabeth , Reuter Quentin , Church Dawn , Hoffmann F Michael , Basu Hirak S. , Wilding George
Published: August 2012

Development of an effective therapy to prevent prostate cancer (PCa) progression to castrate-resistant PCa (CRPCa) remains an unmet medical need, mainly due to a poor understanding of the mechanism of PCa progression. Reactive oxygen species (ROS) are produced in high amounts in PCa cells. ROS play a major role in the development and progression of PCa. We have published that the activated androgen receptor (AR)-JunD complex induces spermidine/spermine N1-acetyl transferase (SSAT). SSAT is the first, and a regulatory, enzyme in a major polyamine catabolism pathway that leads to an over-production of ROS, specifically in the polyamine-rich PCa cells. A focus of our current research is to identify compounds that specifically target and block steps in this newly discovered ROS production pathway downstream to AR activation. Such compounds have the potential to become new targeted therapeutic agents offering minimal side effects for preventing progression to CRPCa in patients with early stage progressing PCa. Here we present data on drug-like small molecule inhibitors of the AR-JunD interaction that initiates this ROS-generating pathway in PCa. A novel high throughput assay based on Gaussia Luciferase enzyme reconstitution via protein-protein interaction was used to screen NCI Diversity Set and Life Chemicals libraries of drug-like small molecules to identify inhibitors of the AR-JunD interaction. The 27 selected hits were categorized as antiandrogens or non-antiandrogens through the use of a published fluorescence polarization assay. Sixteen non-antiandrogens, acting downstream of AR activation, were chosen for further study. In vitro studies entailed the measurement of the ability of the compounds to inhibit androgen-independent and dependent growth as well as androgen-induced ROS in human PCa cells. The top four compounds exhibited significant inhibition of growth and androgen-induced ROS at concentrations of 5 uM or less. Immunocytochemistry (ICC) studies were performed to determine the ability of the selected compounds to specifically inhibit the AR-JunD co-translocation to the nuclei. Based on the cell culture and ICC data, lead compounds were selected for pharmacokinetics studies to determine oral bioavailability. Detailed pharmacokinetic data of selected top compounds showing their bioavailability after intra venous and oral administration 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 2844. doi:1538-7445.AM2012-2844
Abstract 2847: PECAM-1 targeting extends overall survival in various advanced metastatic mouse tumor models
Authors: Handumrongkul Chakkrapong , Collins Chris , DeLisser Horace , Kashani-Sabet Mohammed , Nadel Cory , Nosrati Mehdi , Keyt Bruce , Haqq Christopher , Fong Sylvia , Debs Robert J.
Published: August 2012

Objectives: Patients who die from cancer succumb to advanced metastatic progression (AMP). While targeted therapies are available that address tumor cell derived regulators of metastasis, few therapeutics are being developed to address the molecular mechanisms of AMP. Our group has previously shown that vascular endothelial cell (VEC) PECAM-1 is a critical molecular master switch which regulates AMP by controlling the release of VEC-derived, paracrine growth factors that drive its progression. Anti-PECAM-1 monoclonal antibody (mAb) effectively and safely treats even pre-terminal cancers in AMP-bearing mice. The current experiments were performed to assess whether, to what extent and how widely targeting PECAM-1 can improve overall survival in mice bearing advanced metastases. Methods: Groups of wildtype (WT) C57Bl/6 mice, together with age and gender matched PECAM-1 knockout (KO) mice were intravenously injected with 25,000 murine B16-F10 melanoma cells, 30,000 murine EL4 lymphoma cells or 1,000,000 murine MC38 colon cancer cells. We then injected matched groups of PECAM-1 KO vs WT mice with 1,000,000 B16-F10 cells, to model maximally aggressive AMP. All groups were assessed for overall survival, using IACUC approved guidelines. Results: In all cases, overall survival (OS) was significantly prolonged in PECAM-1 KO vs WT controls (Kaplan-Meier ranged from p = 0.018 to p = 0.0002). Targeting microenvironmental PECAM-1 was as effective in increasing OS in mice receiving the very high tumor burden of 1,000,000 B16-F10 cells as in mice receiving 25,000 cells, further confirming the efficacy of targeting PECAM-1 against the most advanced models of metastasis that can be achieved preclinically. Furthermore, anti-PECAM-1 mAb-based therapy of B16-F10 metastase was as effective as the PECAM-1 KO genotype in increasing OS. Conclusion: To date, many approved mAb- and small molecule-inhibitor based targeted-therapies have shown limited abilities to either prolong OS as single agents, or to effectively treat multiple different solid tumor types. These results suggest that anti-PECAM-1 mAb therapy, which specifically targets VEC and paracrine factors they produce, has the potential to significantly increase OS in the setting of lethal advanced metastatic progression, independent of tumor type. Anti-PECAM-1 mAb therapy is as effective as a PECAM-1 knockout in improving OS. Phase 1 clinical trials are planned within 2 years.

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 2847. doi:1538-7445.AM2012-2847
Abstract 2867: A novel WT1-derived peptide induces cellular senescence and inhibits tumor growth in a human melanoma cell line and xenograft model
Authors: Massaoka Mariana Hiromi , Matsuo Alisson Leonardo , Pereira Felipe Valença , Figueiredo Carlos Rogerio , Scutti Jorge Augusto , Girola Natália , Travassos Luiz Rodolpho
Published: August 2012

Malignant melanoma is the most aggressive and deadly type of skin cancer. The Wilms tumor protein 1 (WT1) is a transcription factor that has been associated with melanoma oncogenesis and proliferation, hence could be a target of tumor control procedures. A lysine-arginine rich peptide (WT1 pTj) based on WT1 C-terminal zinc finger domain was synthesized and tested for anti-proliferative effects on human and murine melanoma cell lines (A2058 and B16F10-Nex2, respectively), including inhibition of cell growth and colony formation, cell cycle arrest and induction of cell senescence. Internalization of WT1 pTj into A2058 cells was established by confocal microscopy, using biotinylated peptide and staining by streptoavidin-FITC. An effect on p53 activity was measured by a luciferase reporter assay. Competitive ELISA assay was performed to verify whether the peptide interferes with p53-WT1 interaction. In vivo, athymic nude mice and C57Bl/6 were inoculated with A2058 and B16F10-Nex2 cells, respectively, and treated for 5 consecutive days with C-amidated WT1 pTj. Here, we show that WT1 pTj decreased, in a dose and time dependent manner, the number of melanoma viable cells. The anti-proliferative activity of WT1 pTj was linked to an irreversible G1 cell cycle arrest and induction of senescence as suggested by positive staining for beta-galactosidase. Treatment with WT1 pTj reduced the total number of soft-agar colonies by A2058 melanoma cells. The peptide entered the cell within 15 minutes and was diffusely distributed throughout the cell, including the nucleus. Moreover, WT1 pTj enhanced p53 activity and competed with WT1 protein for binding to p53. In vivo, WT1 pTj reduced the number of lung nodules in the B16F10-Nex2 syngeneic model of metastatic melanoma and prolonged survival of mice in the A2058 subcutaneous xenograft model. In conclusion, WT1 pTj peptide entered cancer cells and promoted antitumor activity in vitro and in vivo. The data support the hypothesis that WT1 pTj may prevent important interactions between WT1 protein and its partners via a zinc finger motif, inhibiting melanoma development. Therefore, WT1 pTj is a promising agent against WT1-expressing malignancies (e.g. MCF-7 but not SK-BR-3 breast cancer or normal human fibroblasts; and also HL60 acute myeloid leukemia). WT1 pTj may act also as a trojan carrier peptide for potential drug delivery. Supported by FAPESP grant 2010/51423-0 and the Brazilian National Research Council (CNPq)

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 2867. doi:1538-7445.AM2012-2867
Abstract 2877: Perifosine alone and in combination with antimetabolites interferes with NF-kB pathway activation in colon cancer cell lines
Authors: Aicher Babette , Schmidt Peter , Engel Jürgen , Guenther Eckhard G. , Teifel Michael
Published: August 2012

Background: Perifosine, a novel, first-in-class, oral Akt inhibitor, in combination with the antimetabolite Capecitabine is currently in Phase III clinical trials for the treatment of refractory advanced colorectal cancer. We have shown previously that Perifosine in combination with antimetabolites displayed synergistic cytotoxicity and apoptosis in colon cancer and other cell lines [1]. Constitutive NF-kB activation is frequently observed in colorectal cancer and, moreover, is associated with resistance to chemotherapy [2]. Hence we analysed, wether downregulation of the NF-kB pathway contributes to Perifosine mechanism of action and to the observed synergism with antimetabolites in colon cancer cell lines. Methods: Cellular NF-kB activation in nuclear extracts was measured by the TransAm NF-kB p65 ELISA (Active Motif, USA) monitoring binding of NF-kB to its response element. To allow for higher throughput of NF-kB pathway analysis a monoclonal SW620 colon cancer cell line ectopically expressing a NF-kB response element driven reporter gene (Promega, USA) was established. Cellular cytotoxicity and apoptosis assays as well as western blotting were performed according to standard procedures. The analysis of drug combinations was conducted by CalcuSyn software (Biosoft, UK) [3]. Results: Perifosine pretreatment decreased TNFα-induced NF-kB activation in SW620 cells in both independent assay sytems with IC50 values of about 7-10 µM. This effect seems to be specific, since the results were not mimicked by cytotoxic effects of Perifosine. Some of the antimetabolites alone upregulated NF-kB activity in the reporter gene assay. In the presence of these antimetabolites Perifosine was still capable to downregulate NF-kB activity. Conclusions: In summary, these results demonstrate that downregulation of the NF-kB pathway may contribute to the mechanism of action of Perifosine. Moreover, cotreatment with Perifosine has the potential to interfere with the development of antimetabolite resistance due to NF-kB activation in colon cancer. Further experiments addressing Perifosine's mechanism of action in combination with antimetabolites in gastric cancer cell lines are ongoing. References [1] Aicher et al. (2010), Perifosine in combination with antimetabolites induces synergistic effects on cytotoxicity and apoptosis in human colon, multiple myeloma, breast, renal, and liver tumor cell lines. AACR-NCI-EORTC International Conference, Berlin, Germany. [2] Sakamoto and Maeda (2010), Expert. Opin. Ther. Targets 14, 593-601 [3] Chou (2006), Pharmacological Reviews 58, 621-681

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 2877. doi:1538-7445.AM2012-2877
Abstract 289: Hydrogen bond surrogate (HBS) helices as orthosteric regulator of hypoxia inducible transcription
Authors: Kushal Swati , Bullock Brooke , Henchey Laura , Arora Paramjit , Olenyuk Bogdan
Published: August 2012

Hypoxia-Inducible Factor (HIF-1) is a heterodimeric transcriptional activator which plays a critical role in tumorigenesis and therefore is an important therapeutic target. Transcriptional activation of HIF-1 is known to involve the interaction between cysteine-histidine rich region1 (CH1) of a coactivator protein p300 or Creb Binding Protein (CBP) and C-terminal activation domain (C-TAD) of its α-subunit (HIF-1α C-TAD). Based on the hydrogen bond surrogate (HBS) approach, we have rationally designed stabilized alpha helix that can disrupt the binding interface between C-TAD domain of HIF-1α and CH1 domain of p300/CBP. We have shown by fluorescence polarization that such stabilized alpha helix mimetics can directly bind to the CH1 domain of p300 and also can disrupt the HIF-1α/p300 complex. HBS helix mimetics are also shown to selectively downregulate the hypoxia inducible genes in cell culture. Thus, orthosteric inhibition of the HIF-1 transcriptional complex by rationally designed helix mimetics offers a novel approach to downregulate the expression of key hypoxia inducible genes responsible 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 289. doi:1538-7445.AM2012-289
Abstract 3125: Relative distribution of folate species is associated with global DNA methylation in human colorectal mucosa
Authors: Liu Jia , Hesson Luke B. , Meagher Alan P. , Bourke Michael J. , Hawkins Nicholas J. , Rand Keith N. , Molloy Peter L. , Pimanda John E. , Ward Robyn L.
Published: August 2012

Folate, an important cellular methyl donor, exists as functionally diverse species within cells. Folate deficiency is thought to contribute to DNA hypomethylation in colorectal cancer (CRC) however findings on the relationship between total folate concentration and global DNA methylation have been inconsistent. The aim of this study was to determine if global DNA methylation in blood and colorectal mucosa from healthy and CRC patients is related to folate species distribution. Blood and colorectal mucosa was collected from 112 CRC patients, 114 healthy individuals and 82 low folate individuals. Global methylation and folate was determined using liquid chromatography tandem mass spectrometry and repeat element methylation was determined using end-specific polymerase chain reaction. In colorectal mucosa mean global methylation was 8% lower compared to blood while LINE-1 and Alu elements were 3.3-fold and 1.9-fold hypomethylated respectively (P <0.0001). After adjusting for age and smoking, statistically significant but small (2.2%) differences in global but not repeat methylation were found in normal colorectal mucosa from CRC patients compared to healthy individuals. Low folate patients had 18% lower blood 5-methyltetrahydrofolate distribution (P <0.0001) and global (P =0.001) and LINE-1 demethylation (P <0.0001) compared to healthy individuals. The colorectal mucosa from cancer and healthy patients had an altered distribution of folate species (lower 5-methyltetrahydrofolate) similar to the folate distribution found in the blood of low folate individuals. On a background of tissue specific hypomethylation, global methylation of the normal mucosa of CRC patients is similar to healthy individuals. The level of global and repeat element hypomethylation may reflect the underlying distribution of folate species rather than total folate concentration. Future studies on the relationship between methylation and folate should consider folate species distribution, and how genetic or other factors may alter this balance in different tissue types and in neoplasia.

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 3125. doi:1538-7445.AM2012-3125
Abstract 314: The interaction between chronic alcohol consumption and melanoma skews the iNKT cell cytokine profile towards IL-4 dominance
Authors: Zhang Hui , Zhu Zhaohui , Meadows Gary G.
Published: August 2012

Despite the growing evidence that alcohol consumption is associated with the incidence of many different types of cancer, there is little research regarding the effect of continued alcohol consumption on growth, progression and subsequent survival of cancer patients. Using a well-established mouse model we previously found that chronic alcohol consumption impairs the anti-melanoma functions of CD8+T cells and NK cells, and this leads to decreased survival of melanoma-bearing mice. The underlying mechanism for these effects is not known. Herein we examined the relationship of NKT cells to regulation of anti-tumor responses in alcohol consuming mice since NKT cells play important roles in the regulation of the anti-tumor immune response. The phenotype and function of invariant (i)NKT cells were examined in female B16BL6-bearing C57BL6 mice given 20% alcohol for three months. We found that alcohol consumption increased iNKT cells in the liver of mice not inoculated with melanoma, but did not alter iNKT cells in other organs. iNKT cells were dramatically increased in the blood and liver of chronic alcohol consuming, melanoma-bearing mice; however, these cells were not increased in the spleen or draining lymph nodes. Immature NK1.1- iNKT cells decreased 2-4 fold in the blood, spleen and liver of the alcohol-consuming, melanoma-bearing mice, suggesting that alcohol interacting with the melanoma tumors stimulates iNKT cells maturation. Upon activation, fewer iNKT cells produced IFN-gamma in these mice compared to water-drinking, melanoma-bearing mice. The ratio of IL-4:IFN-gamma-producing iNKT cells increased 2-fold as a result of the alcohol/tumor interaction, thus skewing the cytokine profile toward Th2 dominance. The Th2 dominant cytokine profile could negatively regulate the anti-tumor activities of CD8+ T cell and NK cells as well as increase myeloid derived suppressor cells, which we previously reported. Collectively, these data support an important role for iNKT cells in regulation of melanoma progression. (Supported by NIH grants R01AA07293 and K05AA017149, and WSU Alcohol and Drug Abuse Research Program grant 13B-2957-1246).

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 314. doi:1538-7445.AM2012-314
Abstract 3243: Dysfunctional of TGF-β signaling via loss of β2SP activates SOX9 and C-MYC via Notch signaling in Barrett's esophageal adenocarcinoma
Authors: Song Shumei , Maru Dipen M. , Piera-Velazquez Sonsoles , Stroehlein John R. , Davila Marta , Mishra Lopa
Published: August 2012

Background: Esophageal adenocarcinoma (EA) is often considered to arise from a clonal stem like population of cells, potentially responsible for its poor prognosis. TGF-β and Notch signaling pathways play important roles in regulating self-renewal of stem cells and gastrointestinal carcinogenesis. We have demonstrated loss of TGF-β signaling components and activation of Notch signaling in Barrett's esophageal adenocarcinoma; however, their contributions to EA and the mechanisms of their action remain unclear. Methods: Immunoblotting and immunofluorescence were used to evaluate protein expression and localization. Notch targets Hes-1, SOX9 and C-MYC transcription were assayed using their luciferase reporters and Q-PCR. Functional studies were performed in β2SP wild type and mutant MEFs cells and in EA cell lines genetically engineered to express high or low levels of β2SP. Results: Increased levels of Notch signaling Hes1 and Jagged1 occurred in EA tissues and cell lines, compared to normal tissues. Loss of β2SP in MEFs cells increased Hes1 expression by 40 fold. In addition, SOX9, a target gene of Notch signaling and a documented stem cell marker was highly up-regulated in EA tumor cells and tissues. Expression of SOX9 was increased in the absence of β2SP in MEFs cells by 9 fold. Down-regulation of β2SP in SKTG-4, FLO-1 and BE3 EA cells by lentivirus shRNA led to increased SOX9 expression and enhanced nuclear localization of both active intracellular Notch1 domain (ICN1) and SOX9. Reintroducing β2SP into EA cells with knock down β2SP decreased SOX9 promoter activity by 10 fold. Concomitantly, an increased proportion of stem cells in β2SP knock down cells were identified using stem cell marker OCT3/4, indicating expansion of putative cancer stem cells in the absence of β2SP. Most interestingly, we observed a direct interaction between Smad3 and ICN1 via Smad3 MH1 domain by GST-pull down assays and that loss of β2SP increases the binding of Smad3 with ICN1 and induces Notch targets SOX9 and C-MYC transcription. Conclusions: Our findings suggests that loss of TGF-β signaling adaptor β2SP may switch Smad3 function from tumor suppression to tumor promotion by binding intracellular Notch1 domain (ICN1) and activating Notch signaling. Thereby, a potential therapeutic value for targeted therapy in EA is in the setting of loss of β2SP/TGF-β with active Notch signaling.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3243. doi:1538-7445.AM2012-3243
Abstract 3245: The Golgi protein GLG1 participates in tumor progression
Authors: Planche Anne , Bacac Marina , Stamenkovic Ivan
Published: August 2012

Increasing evidence suggests that endocytosis and membrane trafficking are deregulated in cancer cells. Because of their dependence on selected signaling pathways, tumor cells may need to rely on accelerated receptor recycling and increased secretion of a variety of molecules such as matrix components, adhesion molecules and growth factors. Mass spectrometry analysis of proteins associated with separase, recently shown to be an important regulator of membrane trafficking, identified the Golgi apparatus protein 1 (GLG1). GLG1 is primarily known as an E-selectin ligand that mediates the initial step of tethering and rolling of leukocytes on vascular endothelium. We therefore addressed the possibility that GLG1 could play a similar role in circulating tumor cells, facilitating arrest and extravasation into adjacent tissues. Consistent with this possibility, tail vein injection of stably GLG1-depleted tumor cells into NOD/SCID mice resulted in reduced numbers of metastatic nodules. Moreover, metastatic tumor growth was almost exclusively intravascular, indicating impaired extravasation of tumor cells. Further investigation, however, showed that, in tumor cells, GLG1 is almost exclusively localized to the Golgi apparatus, challenging the notion that it plays a direct role in mediating tumor cell adhesion to endothelium as in leukocytes. To elucidate the molecular mechanisms by which GLG1 could affect tumor cell migration through interactions at the Golgi level, we conducted experiments using GLG1-depleted human tumor cell lines. Transient silencing of GLG1 by siRNA was observed to modify cell shape (elongated morphology), to markedly alter Golgi architecture, to inhibit migration of a range of tumor cell lines, and to decrease β1 integrin protein level and impair its internalization/recycling. In addition, subsequent experiments led to the identification of BIG1, an Arf-GEF required to initiate membrane vesicle formation by promoting guanine-nucleotide exchange on Arf1 and Arf3, as a new interactor of GLG1. Taken together, our observations provide insight into a novel and specific role of GLG1 in tumor progression by affecting, thanks to its privileged position in the secretory pathway, trafficking of key molecules implicated in cell migration.

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 3245. doi:1538-7445.AM2012-3245
Abstract 3436: Suppression of the epithelial-mesenchymal transition (EMT) by a wound-healing gene, Grainyhead-like-2
Authors: Cieply Benjamin , Riley Philip A. , Ivanov Alexey , Pifer Phillip M. , Addison Joseph B. , Widmeyer Joseph , Denvir James , Frisch Steven M.
Published: August 2012

Grainyhead genes are involved in wound healing and developmental neural tube closure. In light of the high degree of similarity between the epithelial-mesenchymal transitions (EMT) occurring in wound healing processes and the cancer stem cell-like compartment of tumors, including TGF-β-dependence, we investigated the role of a Grainyhead gene (GRHL2) in oncogenic EMT. Grainyhead was specifically down-regulated in the claudin-low subclass of mammary tumors and in the basal-B subclass of breast cancer cell lines. Functionally, GRHL2 suppressed TGF-β-induced, Twist-induced or spontaneous EMT, enhanced anoikis-sensitivity, and suppressed mammosphere generation in mammary epithelial cells. These effects were mediated in part by its suppression of ZEB1 expression, by direct repression of the ZEB1 promoter. GRHL2 also inhibited Smad-mediated transcription, and up-regulated mir200b/c as well as the TGF-β receptor antagonist, BMP2. The expression of GRHL2 in the breast cancer cell line MDA-MB-231 triggered a mesenchymal-to-epithelial transition and sensitized the cells to anoikis. These results indicate that GRHL2 is a suppressor of the oncogenic EMT, and provide a novel molecular focus for the wound-healing/oncogenic EMT relationship.

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 3436. doi:1538-7445.AM2012-3436