bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3449: Development of small molecule Bcl-XL inhibitors for treatment of lung cancer.
Authors: Park Dongkyoo , Magis Andrew T. , Li Rui , Owonikoko Taofeek K. K. , Sica Gabriel L. , Sun Shi-Yong , Ramalingam Suresh S. , Khuri Fadlo R. , Curran Walter J. , Deng Xingming
Published: August 2014

Bcl-XL is a major anti-apoptotic protein in the Bcl-2 family whose overexpression is more widely observed in human lung cancer cells than that of Bcl-2, suggesting that Bcl-XL is more biologically relevant and therefore a better therapeutic target for lung cancer. Here, we screened small molecules that selectively target the BH3 domain (aa 90-98) binding pocket of

Bcl-XL using the UCSF DOCK 6.1 program suite and the NCI chemical library database.

We identified two new Bcl-XL inhibitors (BXI-61 and BXI-72) that exhibit selective toxicity against lung cancer cells compared with normal human bronchial epithelial cells. Fluorescence polarization assay reveals that BXI-61 and BXI-72 preferentially bind to Bcl-XL protein but not

Bcl2 in vitro with high binding affinities. Treatment of cells with BXI-72 results in disruption of Bcl-XL/Bak interaction, oligomerization of Bak and cytochrome c release from mitochondria.

These two BXI compounds potently repress lung cancer xenografts in vivo without significant normal tissue toxicity within effective doses. Importantly, BXI-72 can also overcome acquired radioresistance of lung cancer. Based on our findings, the development of BXI(s) as a new class of anticancer agents is warranted and represents a novel strategy for improving lung cancer outcome.

Citation Format: Dongkyoo Park, Andrew T. Magis, Rui Li, Taofeek K. K. Owonikoko, Gabriel L. Sica, Shi-Yong Sun, Suresh S. Ramalingam, Fadlo R. Khuri, Walter J. Curran, Xingming Deng. Development of small molecule Bcl-XL inhibitors for treatment of lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3449. doi:10.1158/1538-7445.AM2013-3449
Abstract 3472: Excision repair cross complementing group 1 (ERCC1) and regulatory subunit of Ribonucleotide Reductase (RRM1) as prognostic and predictive markers for ovarian cancer.
Authors: Zhou Jun , Ahmed Quratulain , Chen Wei , Li Xin , Morris Robert , Ali-Fehmi Rouba , Bepler Gerold
Published: August 2014

Background: Ninety percent of ovarian cancers are epithelial in nature (EOC); the standard approach for treatment is surgery followed by platinum and taxane based chemotherapy. Gemcitabine has been evaluated recently in combination with platinum and taxane for platinum sensitive recurrence and as a single agent for platinum refractory or resistant recurrent EOC. ERCC1 and RRM1 are involved in DNA synthesis and repair derived from genotoxic stress, and they have also been reported as predictive markers for therapeutic efficacy in patients with lung and pancreatic cancers. Their expression, association and clinical significance in ovarian carcinoma remain uncertain.

Design: 198 archived cases of epithelial cell ovarian cancer were reviewed and divided into three subtypes: borderline tumor (BLT), Type I and Type II. 2mm TMAs were developed with duplicate cores per case. Immunofluorescence combined with Automated Quantitative Analysis (AQUA) was used to assess the expression of ERCC1 and RRM1. The average of duplicate cores was used for analysis and Greedy search was used for establishing the high and low score cutoff points.

Results: 44 cases (22%) were BLT, 29 (15%) were Type I tumors, 121 (61%) were

Type II tumors, and 4 cases (4%) were unclassified. Type II tumors were significantly higher among elderly Caucasian women (p<0.001) as compared to African American women. Kaplan Meier curves showed no statistically significant difference in overall survival (OS) between Type I and Type II cancers. Low ERCC1 scores were significantly associated with lower OS rates (hazard ratio (HR) = 1.46), when compared with high ERCC1 scores. In contrast, low RRM1 scores were associated with better overall survival (HR = 0.36). Scores for both markers were not significantly associated with tumor type, histology grade or stage. On multivariate analysis including ERCC1, RRM1, tumor stage, tumor type, age and race; only age and tumor type (BLT I&II) persisted as independent prognostic factors for poor overall survival.

Conclusion: Our study suggests that low expression of ERCC1 and high expression of RRM1 are associated with poor survival in ovarian cancer patients. Large studies are warranted to elucidate whether or not the expression of ERCC1 and RRM1 can be used as prognostic or predictive markers.

Citation Format: Jun Zhou, Quratulain Ahmed, Wei Chen, Xin Li, Robert Morris, Rouba Ali-Fehmi, Gerold Bepler. Excision repair cross complementing group 1 (ERCC1) and regulatory subunit of Ribonucleotide Reductase (RRM1) as prognostic and predictive markers for ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3472. doi:10.1158/1538-7445.AM2013-3472
Abstract 3499: Anti-adenoma Adnab-9 antibody recognizes a serum GI cancer candidate biomarker.
Authors: Tobi Martin , Weinstein Douglas , Vizgoft Emma , Pedersen Susanne
Published: August 2014

Population-based serum banks may allow for the early detection of shed markers of common gastrointestinal cancers. Few GI pan-biomarkers are available for serum testing and those that are lack specificity and universal sensitivity for colorectal, gastric and pancreatic cancers. The p87 antigen recognized by the Adnab-9 antibody may be an ideal candidate for such a biomarker. It has both diagnostic and prognostic biomarker ability for these GI cancers but its ability to detect the antigen in serum is unknown. Methods: In order to gauge feasibility of serum testing we conducted a literature search to determine the existence of model established serum banks internationally. An indirect Adnab-9 ELISA was performed on a validation set of sera from 20 CRC patients and 10 controls. A known positive stool extract was used as the positive control. Results: Six countries were found to possess serum banks with the numbers of specimens ranging from 1,982 to 2,132,215. Of these 6 countries, age-adjusted mortality from CRC was highest in the Netherlands at 21/100,000, from gastric cancer highest in Japan 35/100,000 and from pancreatic cancer highest in the USA 11/100,000. The validation sera set Australian patient demographics were similar for gender and ethnicity. The controls were significantly younger than the cancer patients (64±10.3 versus 52.2±3.4 years [p<0.002]) but there was no direct linear correlation between OD450 and age (r2=0.104;p=0.102). Dilution of serum at both 1:1 and 1:3 ratios showed significantly different mean OD450 values between cancer and control patients of 0.076±0.007 (x±STDEV) versus 0.069±0.006 (p<0.025) and 0.083±0.008 versus 0.074±0.004 (p<0.02) respectively. Eighty percent of cancer patients’ serum were positive versus 30% of controls (OR 9.3; CI 1.6-53.2;p<0.015). Signal to noise ratios of the positive control was satisfactory with the lowest OD450 reading at 0.085 obtained at a 1:6,400 dilution with a linear dilution-reaction curve. Conclusions: National serum banks already exist that could allow for graduated targeted screening of a specific disease based on relative mortality rates. The Adnab-9 antibody is a promising candidate for a pan-biomarker based on the outcome of the indirect ELISA. Since sensitivity of Adnab-9 stool binding is lowest for CRC compared to gastric and pancreatic, it is likely that serum testing for the other 2 cancers would yield comparable results. In order to elucidate this further, testing in prospective studies should be undertaken.

Citation Format: Martin Tobi, Douglas Weinstein, Emma Vizgoft, Susanne Pedersen. Anti-adenoma Adnab-9 antibody recognizes a serum GI cancer candidate biomarker. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3499. doi:10.1158/1538-7445.AM2013-3499
Abstract 350: Inhibition of SHP2 impairs oncogenic activity of EGFR mutants in non-small cell lung cancer.
Authors: Schneeberger Valentina E. , Ren Yuan , Luetteke Noreen , Berns Hartmut , Chen Liwei , Lawrence Harshani R. , Lawrence Nicholas J. , Lopez Alex S. , Haura Eric B. , Coppola Domenico , Wu Jie
Published: August 2014

Activating mutations of epidermal growth factor receptor (EGFR) are found in 5-15% of non small cell lung carcinomas (NSCLC) and are targets of NSCLC treatment. However, most patients treated with EGFR tyrosine kinase inhibitors eventually acquire drug resistance, pointing to the need of evaluating other molecules as potential targets for developing new therapy for NSCLC. The protein tyrosine phosphatase (PTP) SHP2 mediates signaling of growth factor receptors including that of EGFR. SHP2 is activated by EGFR mutants in lung adenocarcinoma. Inhibition of SHP2 with the SHP2 PTP inhibitor SPI-112Me or SHP2 knockdown suppressed proliferation of EGFR-dependent HCC827 and H1975 cells. To assess the role of Shp2 in the transgenic mouse model of EGFR mutant-driven lung cancer, we generated transgenic mice carrying a doxycycline (dox)-inducible, PTP-defective SHP2 (tetO-SHP2CSDA). Bitransgenic CCSP-rtTA/tetO-SHP2CSDA mice were crossed with tetO-EGFRL858R mice. F1 offspring of dox-induced transgenic mice were analyzed. Erk1/2 and Src were activated by EGFRL858R in bitransgenic CCSP-rtTA/tetO-EGFRL858R mice. Both of them were suppressed by SHP2CSDA in CCSP-rtTA/tetO-EGFRL858R/tetO-SHP2CSDA tritransgenic mice. Importantly, expression of SHP2CSDA significantly inhibited EGFRL858R-induced lung tumor in the tritransgenic mice. These results suggest that SHP2 is critical for lung tumorigenesis mediated by EGFR mutations.

Citation Format: Valentina E. Schneeberger, Yuan Ren, Noreen Luetteke, Hartmut Berns, Liwei Chen, Harshani R. Lawrence, Nicholas J. Lawrence, Alex S. Lopez, Eric B. Haura, Domenico Coppola, Jie Wu. Inhibition of SHP2 impairs oncogenic activity of EGFR mutants in non-small cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 350. doi:10.1158/1538-7445.AM2013-350
Abstract 352: Murine model of chordoma: Sonic Hedgehog promoter-driven Cre activation of Brachyury (T) expression induces spinal disk abnormalities and perinatal lethal developmental defects.
Authors: Alcorta David A. , Maier Jennifer A. , Harfe Brian D. , Kelley Michael J.
Published: August 2014

Introduction: Chordoma is a rare malignancy arising in the axial skeleton from notochord remnants. The T-box transcription factor, Brachyury (T), is associated with familial chordoma, is expressed in over 90% of chordomas, and is a driver of cell growth in cultured chordoma cells. Suppression of T expression or function in vitro results in down-regulation of cell growth pathways and reduction in matrix formation. To further understand the mechanisms of T's oncogenic properties, we sought to generate a murine model of constitutive, notochord-directed T expression.

Methods: To produce elevated expression of T in the notochord-derived (ND) cells, we generated a murine transgenic animal in which a cassette (T/GFP) containing the β-actin promoter, a transcriptional stop sequence flanked by LoxP sites (FS), the murine T cDNA, an intragenic ribosomal entry sequence and a GFP cDNA was homologously recombined into the ROSA26 locus. We termed these animals FS-T/GFP mice. FS-T/GFP mice were mated to mice with Cre expression regulated by the developmentally-timed and tissue-specific control of the Sonic Hedgehog promoter (Shh-Cre mice).

Results: Excision of the FS sequence by Cre was confirmed to result in constitutive expression of T and GFP in 293 cells prior to transgenic creation, and subsequently in cultured fibroblasts from ear punches of the transgenic animals. Mice containing homozygous FS-T/GFP at ROSA26 were healthy with no apparent physiologic or developmental defects and with the expected Mendelian ratios of sex and genotype. A cross of FS-T/GFP with Shh-Cre mice resulted in T/Cre pups that died perinatally with grossly evident heart, lung and kidney defects. In addition, these animals showed incomplete fusion of the vertebral bodies, and dysmorphic spinal disks. Elevated T protein levels and GFP co-expression in disk cells was demonstrated by immunofluorescent detection in tissue sections.

Summary: Constitutive expression of T in ND lineage cells results in alteration of the morphology of intervertebral disks possibly indicative of a proliferative effect of Brachyury on ND cells. Expression of T in “off-target” tissues that express Sonic Hedgehog likely results in perinatal death.

Citation Format: David A. Alcorta, Jennifer A. Maier, Brian D. Harfe, Michael J. Kelley. Murine model of chordoma: Sonic Hedgehog promoter-driven Cre activation of Brachyury (T) expression induces spinal disk abnormalities and perinatal lethal developmental defects. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 352. doi:10.1158/1538-7445.AM2013-352

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 3532A: Development of gene therapy for pancreatic carcinoma: from experimental models to phase I clinical trial.
Authors: Buscail Louis , Bournet Barbara , Vernejoul Fabienne , Lulka Hubert , Cambois Gilles , Hanoun Naima , Gross Fabian , Tiraby Jean , Cordelier Pierre
Published: August 2014

Background: the median survival of patients diagnosed with locally advanced or metastatic pancreatic adenocarcinoma (PDA) is 4 to 6 months. Chemotherapies (gemcitabine, FOLFIRINOX) and molecular targeted therapies only offer a marginal survival benefit. Consequently, we developed new strategies based on therapeutic gene transfer to help alleviate the dismal prognosis of PDA. We demonstrated that transferring SSTR2 gene in PDA experimental models strongly inhibited tumor progression. We generated DCK::UMK fusion gene to chemosensitize tumor cells to gemcitabine to provoke tumor shrinkage in immune competent animals. Based on these promising results, we asked whether combining SSTR2 and DCK::UMK genes delivered by a non-viral vector may impede PDA growth in patients. Methods: in this first-in-man clinical trial, 22 patients (12 first line, 10 second line) were included in the CHU of Toulouse from 12/2010 to 9/2012. Four different doses (125, 250, 500 and 1000μg) of the gene therapy product (GTP) were injected twice using endoscopic ultrasound (EUS) (day 1 and day 28). Patients concomitantly received gemcitabine for 2 months. Results: the feasibility of this approach is excellent, as all the patients were successfully injected despite differences in the site and the size of the primary tumor, and pre-existing treatments (biliary stent, surgical bypass). No serious adverse events related to the GTP were recorded. We found that less than 0.001% of GTP was detected in blood, while urine was negative in all cases. We found that CA 19.19 levels decreased in 68% (15/22) of patients following gene therapy. In first line patients, we demonstrated partial tumor shrinkage in 2 cases (16%), stable disease in 9 cases (75%), and tumor progression in 1 case (8%). In these latter patients, progression-free survival and median survival following gene therapy reached 7 and 11.3 months, respectively. Conclusions: all together, we demonstrated for the first time that delivering antitumoral genes using non-viral vectors by EUS is feasible and safe in patients with advanced PDA. As our preliminary results tend to demonstrate therapeutic efficacy, we are currently designing a phase II trial based on this approach.

Citation Format: Louis Buscail, Barbara Bournet, Fabienne Vernejoul, Hubert Lulka, Gilles Cambois, Naima Hanoun, Fabian Gross, Jean Tiraby, Pierre Cordelier. Development of gene therapy for pancreatic carcinoma: from experimental models to phase I clinical trial. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3532A. doi:10.1158/1538-7445.AM2013-3532A
Abstract 3542: Effects of single nucleotide polymorphisms on treatment outcomes and toxicity in patients treated with sunitinib for biliary tract and gastric cancers.
Authors: MAENG Chi Hoon , Lee Jun Ho , Lee Jeeyun , Hong Jung Yong , Choi Moon Ki , Kim Young Saing , Jung Hyun Ae , Park Joon Oh , Park Se Hoon , Park Young Suk , Kang Won Ki , Lim Ho Yeong
Published: August 2014

BACKGROUND: We analyzed the efficacy and toxicity profile of sunitinib according to single nucleotide polymorphisms (SNPs) of VEGFA and KDR among the patients with gastric or biliary tract cancer.

METHODS: We examined 8 known SNPs of VEGFA and 5 SNPs of KDR among patients with gastric or biliary tract cancer who were treated with sunitinib in our two previous phase II studies. We assessed progression free survival (PFS), overall survival (OS), and toxicity and their relationships to these SNPs.

RESULTS: A total of 63 patients were evaluable. Among candidate SNPs, rs2010963, rs833068, and rs1870377 were associated with poor PFS (P = .009, .002, and .029, respectively), while rs1870377 and rs7692791 were associated with poor OS (P = .001 and 0.03, respectively). Multivariate analysis showed that only rs1870377 had significant effects on both PFS and OS. Toxicity evaluation indicated that rs1531289 was associated with grade 3-4 anemia. (P = .03).

CONCLUSIONS: Certain SNPs of KDR may affect treatment outcome and toxicity of patients treated with sunitinib.

Citation Format: Chi Hoon MAENG, Jun Ho Lee, Jeeyun Lee, Jung Yong Hong, Moon Ki Choi, Young Saing Kim, Hyun Ae Jung, Joon Oh Park, Se Hoon Park, Young Suk Park, Won Ki Kang, Ho Yeong Lim. Effects of single nucleotide polymorphisms on treatment outcomes and toxicity in patients treated with sunitinib for biliary tract and gastric cancers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3542. doi:10.1158/1538-7445.AM2013-3542
Abstract 3551: CD44 expression in Ewing sarcoma is associated with reduced chemosensitivity in vitro.
Authors: Dioufa Nikolina , Li Guang , Tsokos Maria
Published: August 2014

CD44 is the primary hyaluronate (HA) receptor in a variety of cell types and has been implicated in several processes, such as cell-to-cell and cell-to-extracellular matrix interactions, cell migration and invasion, as well as metastasis and drug resistance. Even though CD44 expression has been studied in various types of cancer, such as prostate and lung cancer, little is known about its role in Ewing sarcoma, the second most common malignancy of bone and soft tissue in children and adolescents. We assessed the expression of CD44 in 25 cases of Ewing sarcoma by immunohistochemistry and found that CD44 is expressed in 40% of them. Furthermore, we noticed that in 4 out of 10 patients CD44 expression increased in one or more subsequent recurrent or metastatic biopsy samples. Therefore, we hypothesized that CD44+ cells in Ewing sarcoma represent cell clones, which are resistant to treatment and lead to tumor relapses. In order to study the role of CD44 in Ewing sarcoma, we cultured tumor cells for 24 to 72 hours in anchorage-independent sphere forming culture conditions, (low attachment plates), which are known to induce CD44 expression and compared them to adherent cells. CD44 mRNA and protein expression was induced in the anchorage-independent group as early as 24h in all 7 Ewing sarcoma cell lines. We then measured mRNA levels of ABCB5 and ABCC1, members of the superfamily of ATP-binding cassette (ABC) transporters related to drug resistance by real time PCR and found them increased. These data were corroborated by our finding of significantly increased efflux of the fluorescent DNA binding dye, Hoechst 33342, in the anchorage-independent group, indicating that the cells that overexpress CD44 are more effective in effluxing the chemotherapeutic drugs, thus becoming more resistant to chemotherapy. Finally, changes in the expression of efflux proteins were associated with increased resistance to doxorubicin-induced apoptotic cell death documented by increased expression of annexin V. In summary, our data indicate that CD44 is involved in drug resistance in Ewing sarcoma and may be clinically useful in making therapeutic decisions.

Citation Format: Nikolina Dioufa, Guang Li, Maria Tsokos. CD44 expression in Ewing sarcoma is associated with reduced chemosensitivity in vitro. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3551. doi:10.1158/1538-7445.AM2013-3551
Abstract 3555: Identification of candidate epigenetic markers for early detection of nasopharyngeal carcinoma.
Authors: Yang Xuesong , Szeto Ying Ying , Cheng Yue , Kwong Dora Lai-wan , Lung Maria Li
Published: August 2014

Nasopharyngeal carcinoma (NPC) is a human malignancy that is particularly prevalent in Southern China and Southeast Asia, and is strongly related to Epstein-Barr virus (EBV) infection. Since early detection of NPC will greatly improve the overall survival, screening becomes important for early NPC diagnosis. In this study, the methylation sensitive-high resolution melting (MS-HRM) method was established for detection of promoter DNA methylation using a panel of five tumor suppressor genes (RASSF1A, WIF1, DAPK1, RARβ2, p16), which were top candidates selected from 13 genes that had been tested in seven NPC cell lines and one immortalized NP cell line and which, showed highly different methylation status between cancer and immortalized cells. Biopsy samples from 52 NPC patients showed methylation for individual genes ranging from 30% to 96%. The combination of WIF1-RASSF1A, WIF1-DAPK1, and WIF1-RARβ2 all reached detection rates of 98% or above in biopsy samples, validating the frequent methylation status of these genes in NPC patients. WIF1, an important secreted Wnt inhibitor, is the most frequently methylated gene in our panel, as reported in many other cancers. In addition, the WIF1-RASSF1A combination showed the highest detection rate, reflecting that both early genetic events in chromosome 3p and the Wnt pathway play important roles in NPC development. To develop non-invasive methods for NPC high-risk screening, early diagnosis and prognosis, more NP brushings and cell-free plasma samples from NPC patients and healthy individuals have been tested for their methylation status of these candidate markers. Preliminary data indicate that the sensitivity and specificity reached more than 90% in certain gene combinations with NP brushings from patients of early NPC stage. The detection rate of methylation in cell-free plasma from NPC patients is even higher than the traditional EBV DNA test, which together show great potential for non-invasive high-risk screening for early NPC and early detection of local recurrence after therapy. In conclusion, a MS-HRM panel of multiple methylation markers is proposed as a complementary test for NPC early detection in combination with more established EBV-based markers.

Citation Format: Xuesong Yang, Ying Ying Szeto, Yue Cheng, Dora Lai-wan Kwong, Maria Li Lung. Identification of candidate epigenetic markers for early detection of nasopharyngeal carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3555. doi:10.1158/1538-7445.AM2013-3555
Abstract 3560: The chromatin architecture at gucocorticoid receptor binding sites.
Authors: Burd Craig J. , Ward James M. , Burkholder Adam B. , Fargo David C. , Archer Trevor K.
Published: August 2014

Steroid hormone receptors are transcription factors that are implicated in the development and progression of several cancer types. It is clear that receptor activity is deregulated in these disease states. Using the glucocorticoid receptor (GR) as a model factor for transcriptional initiation, we aimed to understand the role chromatin structure plays in defining the specific genetic programs initiated by receptor activation. Using Formaldehyde Assisted Isolation of Regulatory Elements (FAIRE), we examined the chromatin state at GR binding locations. FAIRE-seq data indicated an enrichment of signal, suggesting a more open chromatin state, surrounding the GR binding sites even in the absence of hormone. This enrichment is only seen at binding sites and not at unutilized response elements. This data is in line with recent developments indicating accessibility as a determinant of receptor binding. However, further characterization of binding regions suggests consistent nucleosome occupancy at these binding locations. Taken together, these data suggest that receptor binding may be determined by a very specific chromatin architecture that is defined by a transitional accessibility state.

Citation Format: Craig J. Burd, James M. Ward, Adam B. Burkholder, David C. Fargo, Trevor K. Archer. The chromatin architecture at gucocorticoid receptor binding sites. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3560. doi:10.1158/1538-7445.AM2013-3560
Abstract 3573: 17β-estradiol enhances hPygopus2 expression via ERα-SP1 promoter complexes.
Authors: Tzenov Youlian R. , Andrews Phillip G. , Kao Kenneth R.
Published: August 2014

Background: The chromatin remodeling protein Human Pygopus2 (hPYGO2) is widely overexpressed in, and required for the proliferation of several cancer cell lines derived from a variety of tumors. Because of the requirement of hPYGO2 for the G1/S cell cycle transition, its cell cycle dependent expression and its regulation by the Retinoblastoma protein in breast cancer (BC) cells, we hypothesized a fundamental link to the cell cycle. Thus, due to their predominant role in BC, we tested if 17β-estradiol (E2) would cause upregulation of hPYGO2 through Estrogen receptor alpha-SP1 transcription factor (ERα-SP1) promoter complexes.Methods: Eight ERα positive (ER+) and negative (ER-) BC cell lines were grown in low serum conditions and treated with ERα agonists and/or antagonists. mRNA levels were analyzed by Q-PCR and protein by immunoblot. ERα and SP1 promoter binding sites and DNA binding domains were functionally inactivated by site-directed mutagenesis. ERα and SP1 occupancy at the hPYGO2 promoter after treatment with various activators/inhibitors was analyzed by chromatin immunoprecipitation assays. Cell cycle progression was assessed by flow cytometry.Results: hPygopus2 mRNA and protein levels were higher in ER+ BC cells relative to ER- cells. ERα agonists induced, while antagonists inhibited, hPygopus2 mRNA and protein expression. ERα and SP1 occupancy at the hPYGO2 promoter required intact promoter binding sites and functional DNA binding domains. E2 increased, while Fulvestrant decreased, ERα binding to the hPYGO2 promoter while having no effect on SP1 binding. SP1 directed siRNA reduced SP1 occupancy at the hPYGO2 promoter and decreased hPygopus2 mRNA and protein expression in both ER+ and ER- cells. Treatment with betulinic acid, an SP1 inhibitor, resulted in cell cycle arrest in ER- BC cell lines.Conclusion: Our findings indicate that ERα and SP1 regulate hPygopus2 expression via promoter binding and that SP1 continues to play a role in hPYGO2 regulation in ER- cells, thereby revealing the potential efficacy of betulinic acid as a treatment for endocrine-disruptor resistant BC.

Citation Format: Youlian R. Tzenov, Phillip G. Andrews, Kenneth R. Kao. 17β-estradiol enhances hPygopus2 expression via ERα-SP1 promoter complexes. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3573. doi:10.1158/1538-7445.AM2013-3573
Abstract 3578: Androgen receptor transcriptionally targets the ErbB3 regulator Nrdp1 in the presence of nuclear Filamin A.
Authors: Savoy Rosalinda M. , Chen Liqun , Siddiqui Salma , Melgoza Frank U. , Durbin-Johnson Blythe , Roy Mohana , Jathal Maitreyee K. , Bose Swagata , Wang Yu , Mooso Benjamin , D'Abronzo Leandro , Fry William H. , Carraway Kermit L. , Ghosh Paramita M.
Published: August 2014

Despite availability of improved androgen receptor (AR) inhibitors for the treatment of castration-resistant prostate cancer (CRPC), resistance to treatment develops, and has been traced to activation of multiple signaling pathways suppressed by the AR. We previously showed that the AR maintained castration sensitivity in prostate cancer (PCa) by transcriptional regulation of the E3 ubiquitin ligase Nrdp1, which degrades the receptor tyrosine kinase (RTK) ErbB3; whereas AR inhibition suppressed Nrdp1 levels, thereby activating ErbB3. This resulted in CRPC growth and AR stimulation, but in CRPC, the AR was unable to regulate Nrdp1 or suppress ErbB3, causing uncontrolled progression. Here, we investigate the mechanism by which the AR regulates Nrdp1 transcription and why this regulation is lost in CRPC. Immunohistochemical studies in human PCa tissue and in PCa mouse models demonstrated Nrdp1 localization in both nucleus and cytoplasm. In vitro studies determined cytoplasmic Nrdp1 as 36kDa while nuclear Nrdp1 is 28kDa, the 36kDa form, but not the 28kDa form, negatively correlated with ErbB3 levels, but both forms positively correlated with AR. We demonstrate that Nrdp1 is a direct transcriptional target of the AR in androgen-dependent cells expressing a truncated form of the structural protein Filamin A (FlnA) in the nucleus, but in CRPC cells which have lost nuclear FlnA expression, the AR is no longer able to bind to the Nrdp1 promoter. Restoration of nuclear FlnA restored the ability of AR to regulate Nrdp1 transcription. Thus dual targeting of ErbB3 and AR will be effective in patients whose tumors express nuclear FlnA.

Citation Format: Rosalinda M. Savoy, Liqun Chen, Salma Siddiqui, Frank U. Melgoza, Blythe Durbin-Johnson, Mohana Roy, Maitreyee K. Jathal, Swagata Bose, Yu Wang, Benjamin Mooso, Leandro D'Abronzo, William H. Fry, Kermit L. Carraway, Paramita M. Ghosh. Androgen receptor transcriptionally targets the ErbB3 regulator Nrdp1 in the presence of nuclear Filamin A. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3578. doi:10.1158/1538-7445.AM2013-3578
Abstract 3580: Acetaldehyde and drug hypersensitivities of Fanconi anemia defects: Implications for cancer initiation, prevention, and therapy.
Authors: Ghosh Soma , Sur Surojit , Yerram Sashidhar R. , Rago Carlo , Bhunia Anil K. , Hossain M. Zulfiquer , Paun Bogdan C. , Ren Yunzhao R. , Iacobuzio-Donahue Christine A. , Azad Nilofer A. , Kern Scott E.
Published: August 2014

Studies of cells harboring Fanconi anemia (FA) pathway defects have aided clinical understanding of inherited cancer risks and therapeutic strategies. Here, we observed a novel and large (27X) hypersensitivity of BRCA2- and PALB2-null genotypes to the epidemiologically important ethanol metabolite, acetaldehyde. This prominent acetaldehyde sensitivity may hold evolutionary and clinical significance. Interrogation of a novel panel of cells engineered to be null for various FA genes also revealed two classes of chemical hypersensitivities: the shared and divergent phenotypes. Prominent chemical hypersensitivities to various interstrand crosslinking (ICL) agents in vitro (melphalan, mitomycin C, and cisplatin), to γ-radiation in vitro, and to mitomycin C in vivo were essentially similar among the tested genotypes. A large divergence of responsiveness existed, however, between the cell lines when using the PARP inhibitor KU0058948, the topoisomerase II inhibitor etoposide, and acetaldehyde. These results indicate that, toward some agents, not all FA defects are necessarily equivalent; this divergence among phenotypes may dissect functions differing among FA genes and may presage differing clinical and epidemiological implications. We additionally present the first engineered PALB2-null human cancer cells. The results suggest new applications in cancer epidemiology, prevention, and targeted therapy.

Citation Format: Soma Ghosh, Surojit Sur, Sashidhar R. Yerram, Carlo Rago, Anil K. Bhunia, M. Zulfiquer Hossain, Bogdan C. Paun, Yunzhao R. Ren, Christine A. Iacobuzio-Donahue, Nilofer A. Azad, Scott E. Kern. Acetaldehyde and drug hypersensitivities of Fanconi anemia defects: Implications for cancer initiation, prevention, and therapy. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3580. doi:10.1158/1538-7445.AM2013-3580
Abstract 3585: Nicotine contributes to oral cancer by promotion of cell invasion through ASAP1 signaling pathway.
Authors: Chiang Chang Hsu
Published: August 2014

Oral cancer is characterized as prolonged exposure to potential risk factors, including tobacco use or cigarette smoking. However, the underlined mechanism of tobacco leads to cancer is still largely unknown. To shed light on the molecular basis of tobacco induced oral carcinogenesis, we established three sublines of oral cancer cells chronically treated with nicotine (IC30 dose for 3 months). Phenotypic characterization showed that the sublines were tolerant to nicotine, increased cell invasion and migration abilities, along with the altered expressions of epithelial-mesenchymal transition markers. The transcriptomic profiling analysis was performed between parental and the subline cells to global survey and identify nicotine induced genes. Heretical clustering analysis reveals that 1004 genes were differentially expressed, in which 28% of the genes associated with cell mobility. Total of 13 genes were confirmed by RT-PCR assay that over-expressed in the nicotine sublines. ASAP1 was significantly increased in all 3 cell lines, and it was selected for further studied. Knockdown ASAP1 expressions by shRNA reduced nicotine induced cell migration (to 60% ∼ 80%) and invasion (to 55% ∼ 60%) abilities. This ASAP1 silencing further inhibited nicotine- induced MMP2 enzyme activity. Clinical study also supported this finding that ASAP1 was significantly elevated in the oral cancer tissues from patients with smoking habits (P=0.042) but not in the cancer patients without smoking (P=0.714). We therefore concluded that tobacco contributes to oral cancer by the promotion of cell migration and invasion, which was at least in part, through ASAP1 associated signaling pathway.

Citation Format: Chang Hsu Chiang. Nicotine contributes to oral cancer by promotion of cell invasion through ASAP1 signaling pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3585. doi:10.1158/1538-7445.AM2013-3585
Abstract 3596: Short-term effect of nose-only cigarette smoke exposure on redox homeostasis and mitochondrial metabolism in mice.
Authors: Raza Haider , John Annie , Nemmar Abderrahim
Published: August 2014

The components of cigarette smoke have been implicated in the development of cancer as well as in cardiovascular diseases. We have previously reported oxidative stress in rat tissues induced by tobacco-specific toxins nicotine and 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK). Recently, we have also shown increased oxidative stress and associated inflammatory responses in various tissues after exposure to cigarette smoke. In this study, we have further investigated the effects of nose-only cigarette smoke exposure on mitochondrial functions and glutathione-dependent redox metabolism in tissues of BALB/C mice. Liver, kidney, heart and lung tissues were analyzed for oxidative stress, glutathione (GSH) and cytochrome P450 dependent enzyme activities and mitochondrial functions after exposure to smoke generated by 9 cigarettes/day for 4 days. Control mice were exposed to air only. An increase in oxidative stress as observed by an increased production of reactive oxygen species (ROS) and altered GSH metabolism was apparent in all the tissues, but lung and heart appeared to be the main targets affected. Increased expression and activity of CYP450 1A1 and 1A2 were also observed in the tissues after exposure to cigarette smoke. Mitochondrial respiratory functions in the tissues, as observed by alterations in the activities of Complex I and IV enzymes, was also observed after exposure to cigarette smoke when compared with air only exposed mice. SDS-PAGE and Western blot results also indicate that alterations in the expression of enzyme proteins were in accordance with the changes in their catalytic functions. These results suggest that even short term exposure of cigarette smoke have adverse effects on mitochondrial functions and redox homeostasis in tissues which may progress to further complications associated with chronic smoking. (The study is supported by Terry Fox Cancer Research Funds and by the Research Committee Grants from College of Medicine and Health Sciences)

Citation Format: Haider Raza, Annie John, Abderrahim Nemmar. Short-term effect of nose-only cigarette smoke exposure on redox homeostasis and mitochondrial metabolism in mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3596. doi:10.1158/1538-7445.AM2013-3596
Abstract 3621: Dietary zinc and prostate cancer: a case-control study in African Americans men.
Authors: Dabbous Firas , Al-alem Umaima , Springfield Sparkle , Mahmoud Abeer M. , Batai Ken , Shah Ebony , Ahaghotu Chiledum , Kittles Rick A.
Published: August 2014

Introduction: Prostate cancer is a solid malignancy that disproportionately affects men of African descent. Zinc, an essential dietary trace mineral, has been implicated in the pathogenesis of prostate cancer. Total zinc intake is lower in older African American men compared to European Americans (NHANES III). Despite the high prevalence of prostate cancer and decreased levels of zinc intake among African Americans, very little is known about the relationship between zinc intake and prostate cancer in this group. This is significant because dietary factors such as zinc intake are potentially modifiable risk factors. Here, we examine the association between self-reported zinc intake and prostate cancer in a hospital-based case-control study.

Methods: Newly diagnosed African American men with histologically confirmed prostate cancer (n=141) were recruited from an urban academic urology clinic in Washington, DC. Controls (n=109) were unaffected volunteers. Usual dietary intake was assessed at the time of recruitment (2000 to 2004) with the use of a food-frequency questionnaire. Two tailed T-Test was used to compare characteristic means between cases and controls. Unconditional logistic regression was used to calculate the unadjusted and adjusted odds ratio (OR) and 95% confidence intervals (95%CI).

Results: In this population, cases were older and had higher levels of PSA. Controls had higher zinc intake, with a mean of 13.23 mg/day (SD±13.9) versus 10.67 mg/day (SD±7.3) for cases. An increase of 1 mg/day of zinc decreased the odds of prostate cancer by 3% (OR=0.97 and 95% CI: 0.95-1.002). Controlling for age, body mass index (BMI), PSA and total food energy intake did not affect the observed association.

Conclusions: Overall, these results suggest that zinc intake may have slight protective effect on prostate cancer. These analyses should be repeated in large studies to confirm these findings.

Citation Format: Firas Dabbous, Umaima Al-alem, Sparkle Springfield, Abeer M. Mahmoud, Ken Batai, Ebony Shah, Chiledum Ahaghotu, Rick A. Kittles. Dietary zinc and prostate cancer: a case-control study in African Americans men. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3621. doi:10.1158/1538-7445.AM2013-3621
Abstract 3640: Age-related DNA methylation in normal breast tissues.
Authors: Johnson Kevin C. , Jensen Amanda K. , Murphy Megan A. , Koestler Devin C. , Christensen Brock C.
Published: August 2014

Age-related DNA methylation in normal breast tissues

It is well established that age is a key risk factor for breast cancer, with increasing incidence associated with older age. Epigenetic mechanisms, such as DNA methylation alterations, may contribute to age-related increases in breast cancer risk. Altered patterns of DNA methylation can modify the regulation of gene expression or expression potential, and DNA methylation alterations are frequently observed in the early stages of breast tumorigenesis. However, genome-wide interindividual variation in normal breast DNA methylation has not been well characterized for its relation with age and other key breast cancer risk factors. To investigate the relationship between age and DNA methylation in normal breast tissues, we first performed a meta-analysis using two independent data sets GSE32393 (n=23) and GSE31979 (n=15), from the Gene Expression Omnibus (GEO) database. For each of the considered datasets, genome-wide DNA methylation was assessed using the Illumina Infinium HumanMethylation27 Beadchip, which profiles the methylation status of ∼27,000 CpG loci, associated with >14,000 genes. Associations between DNA methylation and age were examined by fitting a series of linear regression models to arcsine square root transformed average beta values for all CpG loci in both data sets. Between these data sets, 204 CpG loci were significantly and consistently associated (P < 0.05) with subject age. Relative to the distribution of CpG loci on the array, we observed a significant enrichment of age-related DNA methylation patterns at CpG loci residing in CpG islands (P = 8.7e-6). Further, compared to non-island CpGs, loci in CpG islands were significantly more likely to exhibit increased age-related methylation (P = 0.008). To validate observed age-related DNA methylation, we performed bisulfite pyrosequencing of selected CpGs in an independent set of normal breast tissues (n=20) obtained through the National Disease Research Interchange. Our findings demonstrate consistent age-related methylation changes in normal breast tissue that may represent early events contributing to breast carcinogenesis and additional investigation for enrichment of local sequence and bioinformatic features near CpGs with age-related methylation is ongoing. This work contributes to the understanding of epigenetic mechanisms by which age serves as a major risk factor in the development of breast cancer.

Citation Format: Kevin C. Johnson, Amanda K. Jensen, Megan A. Murphy, Devin C. Koestler, Brock C. Christensen. Age-related DNA methylation in normal breast tissues. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3640. doi:10.1158/1538-7445.AM2013-3640
Abstract 3676: Selective modulation of CHOP/GADD153 in prostate cancer cells by alpha-mangostin promotes cell death.
Authors: Li Gongbo , Petiwala Sakina , Pierce Dana , Johnson Jeremy
Published: August 2014

Background: It is well-established that during endoplasmic reticulum (ER) stress cells utilize several cell signal pathways collectively named “unfolded protein reaction (UPR)” to re-establish homeostasis and promote cell survival. The upregulation of UPR proteins to an extent is thought to be a growth advantage for cancer cells. However, chronic ER stress or prolonged UPR will lead to apoptosis. Alpha-mangostin, a xanthone from mangosteen fruit, has been reported to effectively induce prostate cancer cell apoptosis in vitro and suppress xenograft tumor growth. Thus, we investigated the alteration of ER stress-related proteins in mangosteen fruit extract (MFE)- or α-Mangostin-treated prostate cancer cells as well as its correlation with apoptosis.

Methods: Two human prostate cancer cell lines, 22Rv1 and LNCaP, and a prostate epithelial cell line were treated with MFE or α-mangostin. Flow cytometry, MTT and BrdU were used to evaluate cell apoptosis, viability and proliferation. Western blot and fluorescent microscopy were used to detect proteins involved in apoptosis or ER stress. We also evaluated the anti-cancer activity of MFE and α-mangostin in nude mice inoculated with 22Rv1 cells. In addition, pharmacokinetics data of α-mangostin was collected on wild type C57BL6 mice for toxicity concern.

Results: MFE increased apoptosis, decreased viability and inhibited proliferation in prostate cancer cell lines. Both MFE and α-mangostin significantly upregulated the expression of ER stress proteins, such as PERK, CHOP and IRE1α. Both MFE and α-mangostin also effectively suppressed tumor growth in a xenograft tumor model without affecting body weight. Interestingly, CHOP knockdown enhanced apoptosis in α-mangostin-treated prostate cancer cells; both MFE and α-mangostin decreased PERK expression and did not induce ER stress in prostate epithelial cells (PrECs) from prostate cancer patients.

Conclusions: Both MFE and α-mangostin selectively promote ER stress in prostate cancer cells and inhibit prostate cancer cell growth in a xenograft mouse model without observable toxicity, suggesting further research is needed to evaluate its potential as an anti-cancer agent.

Citation Format: Gongbo Li, Sakina Petiwala, Dana Pierce, Jeremy Johnson. Selective modulation of CHOP/GADD153 in prostate cancer cells by alpha-mangostin promotes cell death. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3676. doi:10.1158/1538-7445.AM2013-3676