bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2704: A role of the ADCC in the antibody therapy for gastrointestinal carcinoma
Authors: Matsuyama Tatsuzo , Kokura Satoshi , Matsumoto Tsuguhiro , Okajima Manabu , Okajima Manabu , Ishikawa Takeshi , Funaki Jun , Adachi Satoko , Matsuura Yuko , Sakamoto Naoyuki , Naito Yuji , Yosikawa Toshikazu
Published: June 2012

(Background and Aim) Both cetuximab and trastuzumab inhibit growth signal by binding to growth factor receptors, and also have an alternative function regarded as ADCC. In this study, we examined cetuximab-mediated ADCC against EGFR-positive colon cancer cells and trastuzumab-mediated ADCC against HER2-positive gastric cancer cells. Moreover, we investigated the new methods to enhance those ADCC activities.

(Materials and Methods) We used human gastric cancer cell lines (MKN1, MKN45) and colon cancer cells (HT-29), cetuximab and trastuzumab as an antibody which potentially had ADCC, and non-specific IgG1 antibody as a control. ADCC was measured by Europium release assay: First, we used peripheral blood mononuclear cells (PBMCs) from healthy volunteers as effector, and we prepared those cancer cell lines combined with those kinds of antibodies as target. Next, we used PBMCs activated by IL-2 instead of normal PBMCs. At same time, CD16, perforin, granzyneB on PBMCs were evaluated by Flow Cytometer and PCR. NK activity of PBMN and activated PBMC against for K562 cell and ADCC activity of PBMN and activated PBMC were evaluated.

(Results) Cetuximab and trastuzumab didn't kill cancer cells by themselves, but the cytotoxicity of PBMCs was increased when the target cells were treated with these antibodies in advance. This increase was further strengthened by activated PBMCs with IL-2. But IL-2 didn't enhance CD16 expression on PBMCs. Finally PBMC, which have high potential of NK activity shows high ADCC activity,

(Conclusion) These two antibody therapy shows ADCC in vitro, As a next step, we have to start in vivo study

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2704. doi:10.1158/1538-7445.AM2011-2704
Abstract 2733: KITENIN and ErbB4 contribute to further malignant changes in colorectal cancer through c-Jun/AP-1 activation
Authors: Bae Jeong A , Sun Eun Gene , Kim Hangun , Park So Yeon , Lee Hyun Jong , Ryu Hee Won , Kim Kyung Keun
Published: June 2012

KITENIN (KAI1 C-terminal interacting tetraspanin, Vangl1) is not only a major component of PCP pathway but also a functional protein promoting colorectal cancer (CRC) cell motility and invasiveness. Here, we report the mechanism of which KITENIN leads further malignant changes in CRC progression after loss of tumor suppressor Adenomatous polyposis coli (APC). KITENIN increases the level of AP-1 activity through interaction with Dvl2, which is subsequently being degraded. Decreasing Dvl2 via interaction with KITENIN or via treatment of si-RNA to Dvl2 significantly increases the AP-1 activity through elevating the level of c-jun and c-jun phosphorylation. EGF acts upstream of KITENIN/AP-1 signaling and induces synergic AP-1 activation in the presence of elevated KITENIN, which requires the interaction of KITENIN with ErbB4. The morphological characteristics of adenocarcinoma were observed in intestinal tumor tissues from compound villin-KITENIN transgenic/APCmin/+ mice but not from APCmin/+ mice, both of which express the elevated level of ErbB4. This result demonstrates that further malignant phenotype of intestinal tumor can be induced by elevated KITENIN and ErbB4 via promoting c-Jun/AP-1 activation within an APC loss-associated tumor microenvironment. Our present results suggest the possibility of the addition of anti-KITENIN therapy to the use of EGFR inhibitors in the treatment of CRC patients with APC mutation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2733. doi:10.1158/1538-7445.AM2011-2733
Abstract 2734: Genome-wide analysis of direct Nkx3.1 target genes
Authors: Logan Monica , Anderson Philip , Abdulkadir Sarki
Published: June 2012

NKX3.1 is an androgen-regulated, prostate-specific tumor suppressor gene that is essential for development and maintenance of the prostate gland. Loss of Nkx3.1 expression results in the formation of premalignant lesions called prostatic intraepithelial neoplasia lesions (PIN). With additional mutations, these PIN lesions will progress to invasive carcinoma and eventually to metastatic disease in which there is currently no effective treatment. The specific mechanism(s) by which Nkx3.1 suppresses prostate tumorigenesis remains unknown. Previous gene expression profiling studies in Nkx3.1-deficient mouse prostates have shown aberrant expression of many genes that may be involved in prostate tumorigenesis. However, it is unknown whether these genes are direct targets of Nkx3.1 in vivo. To identify direct Nkx3.1 target gens, we used chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-Seq) to identify Nkx3.1 binding sites in the genomes of a human prostate cancer cell line (LNCaP) as well as the normal mouse prostate. We obtained 7 to 15 million uniquely mapped tags each from Nkx3.1 ChIP-seq using LNCaP cells or Nkx3.1+/+, Nkx3.1+/−, and Nkx3.1−/− mouse prostates. This led to the identification of about 8000 Nkx3.1 binding sites in the genome (the Nkx3.1 cistrome). The binding site data were integrated with gene expression profiling results from Nkx3.1 mutant mice to yield a core set of “direct” Nkx3.1 target genes. One gene in particular that is upregulated in Nkx3.1 mutant mice and in human prostate tumors and bound by Nkx3.1 in our ChIP-Seq studies is the receptor activity modifying protein 1 (RAMP1) gene. Ramp1 is a single transmembrane protein that is required for trafficking of the calcitonin gene-related peptide (CGRP) receptor to the plasma membrane. The CGRP receptor binds CGRP, a neuropeptide that has been shown to promote cancer progression. Current studies are focused on whether RAMP1 expression in human prostate cancer cells function to promote tumorigenesis. Given that Ramp1 is localized to the plasma membrane, data from my project may provide support for investigating Ramp1 as a therapeutic target for prostate cancer or for imaging prostate lesions.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2734. doi:10.1158/1538-7445.AM2011-2734
Abstract 2740: Decision tree-based modeling of androgen pathway genes and prostate cancer risk
Authors: Barnholtz-Sloan Jill S. , Guan Xiaowei , Zeigler-Johnson Charnita , Meropol Neal J. , Rebbeck Timothy
Published: June 2012

Background: Inherited variability in genes that influence androgen metabolism has been implicated in the etiology of prostate cancer. However, joint effects of these genes have not been fully defined in terms of risk.

Objective: The objective of this analysis was to evaluate gene-gene and gene-environment interactions for prostate cancer risk using classification and regression tree (CART) models (i.e. decision trees), and to evaluate whether these interactive effects add information about prostate cancer risk prediction beyond that of “traditional” risk factors.

Methods: We compared CART models to traditional logistic regression models for associations of factors with prostate cancer risk using 1084 prostate cancer cases and 941 controls. All analyses were stratified by race. We used unconditional logistic regression (LR) to complement and compare to the race-stratified CART results using the area under curve (AUC) for the receiver operating characteristic (ROC) curves.

Results: In European Americans, single SNP or haplotype associations were found with CYP3A5 (OR=2.11 (1.13, 3.97)); increasing number of CAG repeats in the AR gene (OR=0.90 (0.83, 0.98)); and the CYP3A4/CYP3A5 AG haplotype (OR=2.86 (1.15, 7.12)). No significant single SNP or haplotype associations were found in African Americans. The CART modeling of prostate cancer risk showed different interaction profiles by race. For European Americans, interactions among CYP3A43 genotype, history of BPH, family history of prostate cancer and age at consent revealed a distinct hierarchy of gene-environment and gene-gene interactions. While for African Americans, interactions among family history of prostate cancer, individual proportion of European ancestry, number of GGC AR repeats and CYP3A4/CYP3A5 haplotype revealed distinct interaction effects from those found in European Americans. For European Americans the CART model had the highest AUC while for African Americans, the LR model with the CART discovered factors had the largest AUC.

Conclusion: These results show novel gene-gene and gene-environment interactions specific for European Americans and African Americans discovered using CART as compared to standard LR techniques.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2740. doi:10.1158/1538-7445.AM2011-2740
Abstract 2746: Genomic data mining for selecting informative markers for ancestry
Authors: Baye Tesfaye Mersha
Published: June 2012

Given the huge amount of single nucleotide polymorphism (SNP) data available from high-throughput sources such as HapMap, data mining using various methods is a reasonable approach to identify SNPs that are informative for genetic ancestry. We extensively investigated the distribution and density of the SNPs across the genome of African and European populations available within the framework HapMap database to prioritize potential candidate SNPs useful for ancestry mapping in an admixed population. About 4 million SNPs were compared between Africans and Europeans using various measures of ancestry informativeness in use today viz. absolute allele frequency differences (Δ), Fisher Information Content (FIC), Shannon Information Content (SIC), informativeness for assignment (I), and Fixation Index (FST). Each method provides different sets of candidate ancestry informative markers (AIMs). The selected SNPs represent valuable resources for both controlling population structure and gene mapping studies. The overlap and non-overlap between selected AIMs by different measures of informativeness, data filtering strategies and the accuracy of each measure in classifying ancestral populations and applications in cancer genomics are discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2746. doi:10.1158/1538-7445.AM2011-2746
Abstract 2758: Genetic variability in tobacco-specific nitrosamine NNK to NNAL metabolism
Authors: Ter-Minassian Monica , Asomaning Kofi , Zhao Yang , Chen Feng , Su Li , Carmella Steven G. , Lin Xihong , Hecht Stephen S. , Christiani David C.
Published: June 2012

Introduction: Tobacco-specific nitrosamine (TSNA) NNK metabolites NNAL and NNAL-gluc) together, (total NNAL), have recently been shown to be good predictors of lung cancer risk, years prior to diagnosis. However, the inter-individual variability in the metabolism of NNK evaluated by the total NNAL output could be explained in part by genetic variability in the genes of NNK metabolism. We sought to determine the contribution of several genetic polymorphisms to total NNAL output by a regression analysis of NNAL levels on genotyping results, controlling for measures of smoking, age and sex.

Methods: The study subjects were derived from the Harvard/ Massachusetts General Hospital Lung cancer case-control study. We selected 92 subjects (39 lung cancer cases and 53 controls) smokers at time of diagnosis and urine collection (1992-1996). We tested 77 tagging SNPs in 16 genes related to NNK to NNAL metabolism. Using case status as a mediator, we tested for the association of each SNP with square-root (sqrt) transformed NNAL (pmol per mg creatinine), controlling for age, sex, sqrt packyears and sqrt nicotine (ng per mg creatinine).

Results: After a sqrt transformation, nicotine significantly predicted a 0.02 (0.015, 0.023) pmol/mg creatinine unit increase in NNAL for every ng/mg creatinine increase in nicotine at p = 2.85E-12. Three HSD11B1 SNPs, located in a 14kb region flanked by rs2235543 and rs3753519 in intron 1, were significantly associated with decreasing NNAL levels. Two of these SNPs, rs2235543 (p = 4.84E-08) and rs3753519 (p = 0.0017), passed multiple testing correction at FDR q value=1.07E-05 and 0.06 respectively. AKR1C4 rs7083869 G>A was also to be significantly associated with decreasing NNAL levels, p=0.019, FDR q=0.48.

Conclusions: HSD11B1 and AKR1C4 enzymes are carbonyl reductases directly involved in the single step reduction of NNK to NNAL. The HSD11B1 SNPs may be correlated with the functional variant rs13306401 and the AKR1C4 SNP correlated with the enzyme activity reducing variant rs17134592, L311V.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2758. doi:10.1158/1538-7445.AM2011-2758
Abstract 2765: Genetic variants in immunoregulaion and DNA repair related genes and risk of lymphoma among children in Korea
Authors: Song Nan , Kang Daehee
Published: June 2012

Although lymphoma is recognized as a cancer of the immune system, little is known regarding a role for genes involved immune system. Genetic susceptibility studies of lymphoma may serve to identify at risk populations and to elucidate important disease mechanisms. In order to estimate the genetic susceptibility for childhood lymphoma, we investigated whether genetic variation 1536 tag single nucleotide polymorphisms (SNPs) selected in 138 candidate gene regions related to immune response, apoptosis, cell cycle, and DNA repair. We used the GoldenGate (Illumina) oligonucleotide pool assay (OPA) in 40 cases patients and 254 controls among Korean children. The association study was estimated as odds ratios and 95% confidence intervals adjusted for age. The minP test was used to identify statistically significant association at gene level. Five genes (IL1RA, IL2, IL12RB1, JAK3, and TNFRSF13B) in immune response and one gene (XRCC3) in DNA repair had statistical significance (minP < 0.05). In these regions, total twelve SNPs (rs11677140, rs10857092, rs2069762, rs4833248, rs365179, rs3212701, rs3212711, rs7250423, rs3751987, rs3751991, rs4343329, rs861547) were significantly associated the risk of lymphoma (Ptrend < 0.05). The most significant association with the risk of lymphoma was seen for IL2 rs2069762 (ORCC/AA = 10.1, 95% CI = 2.4-43.1, Ptrend = 0.002, minP = 0.005). These findings suggest that common genetic variants in immune response and DNA repair related genes might play a role in the genesis of childhood lymphoma with limited biological evidence. Additional larger studies are needed to identify these findings and understand the mechanisms of these genes in childhood lymphoma patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2765. doi:10.1158/1538-7445.AM2011-2765
Abstract 277: Genetic re-programming of the acute myeloid leukemia cell line HL-60
Authors: Roberts Michael , Bittner David , Brnich Sarah , Conner Bryan , Cox Carla , Filiberti Julia , Grant Michael , Mansuy Michael , Forrester Jeffrey
Published: June 2012

HL-60 cells represent an established line of human acute myeloid leukemia cells that can be induced to differentiate along several distinct myeloid lineages. Phorbol esters activate a signaling network, which includes the protein kinase C (PKC) signaling pathway, causing the leukemia cells to differentiate into macrophage-like cells. As the leukemia genetic program is converted to the macrophage program, the cells exit the cell cycle, undergo morphological changes, become adherent, display other macrophage properties, and ultimately initiate apoptosis. This study provides a detailed DNA microarray analysis of the genetic network of the leukemia cell during first 24 hours post-induction of differentiation. Approximately 1,000 genes were identified that exhibit significant changes in mRNA levels from the leukemia expression profile. A cluster analysis was performed to group these genes by their pattern of expression resulting in a collection of distinct expression profiles. Expression levels of selected genes within each cluster were validated by real time PCR. Among the most significant changes observed were dramatic increases in the expression of CCL chemokine family genes and a variety of transcription factor genes including members of the AP-1 and NF-κB families. Expression level changes in genes involved in cell cycle progression (ex. MYC and p21) were also observed along with genes involved in the regulation of apoptosis (ex. BCL2A1, TNF, IER2 and OLR1). Functional analysis at the protein level using the electrophoretic mobility shift assay (EMSA), western blotting and immunofluorescence imaging will also be presented. This study provides a comprehensive view of the re-programming events that occur in phorbol ester induced macrophage differentiation of HL-60 cells with regard to the inflammatory response, cell cycle exit, differentiation and initiation of apoptosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 277. doi:10.1158/1538-7445.AM2011-277
Abstract 2772: 2’-5’oligoadenylate synthetase polymorphism is associated with prostate cancer: Effect modified by age and race
Authors: Mandal Sanjay K. , Abebe Fisseha , Chaudhary Jaideep
Published: June 2012

Introduction: 2’ -5’ -oligoadenylate synthetase (2-5 OAS1) is an antiviral gene that converts ATP to a series of (2-5A). 2-5A promotes dimerization of latent ribonuclease (RNASE L) to form catalytically active RNASEL, a candidate hereditary prostate cancer gene. Thus a defect in OAS1 can not only attenuate RNASEL function, but can also alter cell growth and apoptosis independent of RNASEL. OAS1 polymorphism (rs2660) has been associated with increased disease susceptibility and viral infection like, HCV, West Nile virus infection, SARS, diabetes and MS. The AA genotype, known to have lower enzymatic activity promotes disease susceptibility whereas GG genotype with higher enzyme activity confers protection. In the present study we investigated this functional polymorphism (rs2660) in OAS1 in a large prostate cohort.

Methods: Genomic DNA from Buffy coat samples was used from age matched controls (n=140) and prostate cancer patients (n=164) who had undergone prostatectomy for genotyping. Information regarding age, race, and cancer stage were available for all samples. PCR-RFLP, followed by direct sequencing was used for genotyping. SNPs rs2660, rs1131454 and rs34137742 were genotyped on all samples. Statistical analysis was performed using chi2 test and logistic regression model with NCSS software.

Results: The rs2660 genotype distribution is summarized in table 1. Genotype AA increases the risk of prostate cancer by 61%. Prostate cancer incidence increases with age, however when combined with rs2660 genotype AA the odds of prostate cancer incidence are higher.SNPs rs1131454 and rs34137742 were not associated with prostate cancer.

Conclusion: OAS1 SNP rs2660 AA genotype is significantly associated with prostate cancer whereas GG genotype protects against prostate cancer. The effect of this genotype distribution was more profound in African American population as compared to Caucasians.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2772. doi:10.1158/1538-7445.AM2011-2772
Abstract 2779: Plasma carotenoids and DNA methylation status modulate prostate cancer aggressiveness
Authors: Su L Joseph , Zabaleta Jovanny , Chiang Tung-Chin
Published: June 2012

Introduction: Prostate cancer is the most frequently diagnosed malignancy in men. Many low grade prostate cancer would never progress further through the life of the patients. Identifying nutritional factors that may be associated with epigenetic factors associated with high aggressive prostate cancer could potentially prevent the progression of prostate cancer.

Methods: A pilot study was conducted to examine the association between plasma carotenoids concentrations and DNA methylation status previously found to be associated with prostate cancer aggressiveness, defined using clinical criteria abstracted from medical records at time of prostate cancer diagnosis. Among 88 newly diagnosed prostate cancer patients (47 African Americans and 41 Caucasians), cancer on 18 subjects were classified as high aggressive and 70 subjects as low or intermediate aggressive. The GoldenGate Methylation Cancer Panel I was used to obtain DNA Methylation data. Plasma carotenoid concentrations were measured using High-performance liquid chromatography. Analysis of variance adjusting for age, race, and body mass index was used to evaluate the relationship between plasma carotenoids concentration and DNA methylation status among the low and the intermediate aggressive prostate cancer cases.

Results and Conclusions: This study found that EYA4, IGF1, PALM2AKAP2, and SNRPN were less likely to be hypomethylated (p < 0.05) among subjects with higher plasma lutein-zeaxanthin level. Similarly, Elevated plasma lycopene is inversely associated with hypomethylation at EYA4, FYN, and SNRPN. Both EYA4 and IGF1 genes have previously demonstrated with anti-apoptotic function; while SNRPN have tumor suppressor function. The result of this pilot study suggests that carotenoids may modulate DNA methylation, possibility through their antioxidant property, that affects the aggressiveness of prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2779. doi:10.1158/1538-7445.AM2011-2779
Abstract 2783: A novel functional variant is associated with regulation of the prostate stem cell antigen (PSCA) gene and bladder cancer risk in males
Authors: Fu Yi-Ping , Kohaar Indu , Muchmore Brian , Mumy Adam , Tang Wei , Porter-Gill Patricia , Garcia-Closas Montserrat , Chanock Stephen , Figueroa Jonine , Baris Dalsu , Purdue Mark , Thun Michael , Albanes Demetrius , Malats Nuria , Real Francisco X. , Kogevinas Manolis , Johnson Alison , Schwenn Molly , Rothman Nathaniel , Silverman Debra , Prokunina-Olsson Ludmila
Published: June 2012

Genome-wide association studies (GWAS) identified allele T of SNP rs2294008 within the prostate stem cell antigen (PSCA) gene as a risk factor for diffuse gastric cancer (OR=1.67, p=2.2×10-15) (The Study Group of Millennium Genome Project for Cancer, Nat Genet, 2008) and bladder cancer (OR=1.13, p=4.4×10-11) (Rothman et al, Nat Genet, 2010). In the present study we used data from the Stage 1 bladder cancer GWAS, which was performed on 8,801 individuals of European ancestry (3,529 cases and 5,115 controls), and information from the 1000 Genomes and HapMap 3 projects to impute all other SNPs within +/- 50kb region surrounding PSCA. Association analysis on the combined set of all genotyped and imputed SNPs (n=375) revealed a novel bladder cancer association signal for a variant genotyped in all samples. The association was found only in males (n=7,359, crude OR =1.14, p=1.15×10-4; adjusted for all covariates and rs2294008, OR=1.13, p=1.32×10-3), while no association was observed in females (n=1,442, crude OR=0.98, p=0.81; adjusted for all covariates and rs2294008, OR=0.98, p=0.82). The low linkage disequilibrium between these two SNPs (D’=0.202 and r2=0.019 in 8801 GWAS samples) suggests these variants are independent. A joint analysis showed that these two SNPs have a compound association with bladder cancer dependent on the number of risk genotypes (0, 1 or 2) in males (adjOR=1.18, p=3.93×10-6) but not in females (adjOR=1.01, p=0.95).

The risk allele T of rs2294008 is functional as it introduces a novel translation start site that creates a PSCA protein with a leader peptide extended by 11 amino acids. The novel SNP that was found 10 Kb upstream of rs2294008, lies within an alternative untranslated first exon of PSCA that is marked by a H3K4me3 signal and is in a vicinity of an androgen receptor binding site, suggesting a possible role for this variant in regulation of PSCA promoter activity. In conclusion, a novel independent functional variant associated with bladder cancer risk was found within the PSCA gene, which might be important for male-specific regulation of PSCA expression and carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2783. doi:10.1158/1538-7445.AM2011-2783
Abstract 2786: Identification and preclinical characterization of EMD 1204831 – A selective c-Met kinase inhibitor in clinical phase 1
Authors: Bladt Friedhelm , Blaukat Andree , Dorsch Dieter , Fittschen Claus , Friese-Hamim Manja , Graedler Ulrich , Knuehl Christine , Meyring Michael , Schadt Oliver , Stieber Frank
Published: June 2012

The involvement of the mesenchymal endothelial transition factor (c-Met) in the primary event of oncogenic transformation and the secondary ability to mediate metastatic spread has been convincingly demonstrated in preclinical and early clinical settings. The clinical benefits of c-Met kinase inhibitors with various modes of actions and selectivity profiles are currently under investigation hoping that inhibitors of c-Met might emerge as valuable cancer therapeutics in the future. During an HTS run 3-(diethylamino)propyl N-[3-[[5-(3,4-dimethoxyphenyl)-2-oxo-6H-1,3,4-thiadiazin-3-yl]methyl]phenyl]carbamate was identified as an attractive lead structure with an interesting overall profile (clogD (7.4): 2.5, S (pH 7.4): >100 µg/ml, IC50 (cMet enzyme): 30 nM, IC50 (cMet A549): 800 nM) providing a valid starting point. The co-crystal structure of 1 revealed the binding mode and the essential structural features. The initial HTS hit was bound in a DFG-in conformation interacting with the main chain nitrogen atom of Met1160 within the hinge region and with the main chain nitrogen of Asp1222. After subsequent optimization of potency, efficacy, PK properties and the safety profile of thiadiazinone 1, EMD1204831 was identified as development compound. EMD1204831 is currently being investigated in a phase 1 clinical trial. The pyridazinone EMD1204831 inhibits enzymatic and cellular c-Met kinase activity with IC50 values of 12 nM and 15 nM, respectively. EMD1204831 displayed an exquisite selectivity when tested in vitro against a panel of more than 400 potential off-targets, including kinases, GPCRs, ion channels, transporters and various enzymes. EMD 1204831 demonstrated excellent anti-tumor activity in vivo in a variety of xenograft models, including U87-MG glioblastoma cells (autocrine HGF expression), TPR-Met-transformed mouse fibroblasts (oncogenic Met fusion protein) or Hs746T gastric cancer cells (c-Met gene amplification and HGF-independent activation). Depending on the particular model, complete regressions were observed with doses as low as 6 mg/kg/d administered per os. PK/PD analysis revealed efficient, dose- and time-dependent inhibition of c-Met phosphorylation, reduction of IL-8 and cyclin D1 expression as well as an induction of the cell cycle inhibitor p27. The overall profile of EMD1204831 including first time structural disclosure, some structure activity relationships, in vitro potency, selectivity profile and in-vivo data will be presented.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2786. doi:10.1158/1538-7445.AM2011-2786
Abstract 2787: Discovery of GDC-0980, a selective PI3K/mTOR inhibitor in clinical trials
Authors: Sutherlin Daniel P. , Belvin Marcia , Bao Linda , Berry Leanne , Berry Megan , Castanedo Georgette , Edgar Kyle , Folkes Adrian , Friedman Lori , Heffron Tim , Patel Sonal , Olivero Alan , Lesnick John , Lewis Cristina , Marsters James , Nonomiya Jim , Pang Jodie , Prior Wei Wei , Salphati Laurent , Sampath Deepak , Tsui Vickie , Wallin Jeff , Wei BinQing , Weismann Christian , Zhu Bing-Yan
Published: June 2012

PI3 Kinase and mTOR have been identified as promising targets for the treatment of cancer. These enzymes participate in related, but not redundant, signaling networks to transmit cellular growth and survival signals, which are hallmarks of tumor growth. An interest in targeting both of these two important points along this critical signaling pathway, and the ability to leverage the high degree of structural similarity in the active sites of PI3K and mTOR kinase, has resulted in the discovery of GDC-0980 as a Class I PI3K and mTOR kinase inhibitor for oncology indications. The structure, efficacy, and medicinal chemistry behind the discovery of this compound is described.

Beginning with the morpholin-4-yl-thieno[3,2-d]pyrimidine core of the Class I PI3K inhibitor GDC-0941, structural substitutions were made external to the core that added mTOR potency, improved the metabolic stability in vitro and in vivo, and lowered the plasma protein binding of the scaffold. Homology models of mTOR using PI3Kγ structures with bound inhibitors provided hypotheses for increasing mTOR potency relative to previous compounds. The solubility of the modified compounds was improved through the addition of polar functionality in the solvent exposed region of the scaffold, resulting in GDC-0980.

GDC-0980 is potent across Class I isoforms with IC50's of 5, 27, 7, and 14 nM for PI3Kα, β, Δ, and γ, and inhibits mTOR with a Ki of 17 nM. The compound is highly selective versus a large panel of kinases including others in the PIK family. Based on the excellent PK profile, linear increase in exposure, strong potency in a broad range of cancer cells, and high free fraction, GDC-0980 is efficacious in animal models of cancer when dosed orally at low doses. Furthermore, this compound is efficacious when dosed intermittently as well as on a daily schedule. These preclinical data provide compelling support for GDC-0980 as a clinical candidate, and early stage clinical trials are underway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2787. doi:10.1158/1538-7445.AM2011-2787
Abstract 279: Use of GFP-STAT3βtc for EMSA analysis of protein-protein and protein-DNA interactions in tumorigenic signalling pathways
Authors: Wilderspin Andrew F. , Nkansah Edwin , Zinzalla Giovanna , Stephenson F. Anne , Thurston David E.
Published: June 2012

STAT3 (Signal transducer and activator of transcription 3) is a homodimeric transcription factor that binds to the response element of a number of genes that promote tumorigenesis when over-expressed. Tumorigenesis, in many cases, correlates with an aberrant signalling cascade that leads to uncontrolled activation of STAT3. Activation is widely understood to be a result of phosphorylation at tyrosine 705 which leads to dimerisation and DNA binding.

We have investigated the dimerisation and DNA binding of STAT3βtc in an isolated cell-free system in order to understand the conditions which modulate these interactions, and to facilitate the discovery of small molecules capable of interfering with these interactions. We designed a GFP-STAT3βtc construct in which the N-terminal 126 amino acids of STAT3 were replaced with spectral variants of the GFP (green fluorescent protein) to act as a reporter. The constructs also lacked the unstructured C-terminal activation domain of STAT3 to avoid aggregation of the purified protein. Phosphorylated and non-phosphorylated GFP-STAT3βtc proteins were purified from BL21(DE3)TKB1 and Rosetta strains of E. Coli, respectively. Soluble expression was observed when tagged with GFP at the N-terminus but not the C-terminus.

Using both conventional ELISA and fluorescence-based microtiter plate assays, we demonstrated that both active (phosphorylated) and inactive (non-phosphorylated) fluorescent STAT3βtc can bind to immobilised dsDNA (M67; a modified c-fos sis inducible enhancer) or an immobilised surrogate receptor (a phosphotyrosyl peptide derived from the interleukin-6 receptor subunit gp130), respectively. Furthermore, we were able to demonstrate disruption of a pre-formed non-phosphorylated GFP-STAT3βtc/DNA complex with a phosphotyrosyl peptide at an IC50 of 30 µM. In contrast, 500 µM peptide appeared to have no effect on phosphorylated GFP-STAT3βtc bound to M67.

In summary, we have developed an EMSA assay for the STAT3/STAT3 transcription factor based on a protein mobility shift rather than a DNA mobility shift, and which utilizes fluorescent tagged STATs. Both non-phosphorylated and phosphorylated fluorescent STAT3 proteins form a stable complex with M67 DNA, but only the non-phosphorylated STAT3/DNA complex can be disrupted with a phosphotyrosyl peptide in the EMSA assay. This assay is now being utilized for the discovery of novel small molecule inhibitors of the STAT3/STAT3 interaction.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 279. doi:10.1158/1538-7445.AM2011-279
Abstract 2804: Increased metastasis linked to defective mitochondria and elevated ROS in BNip3 deficient mammary tumorigenesis
Authors: Tracy Kristin , Chen Hong , Barth Sandra , Macleod Kay F.
Published: June 2012

BNIP3 is a hypoxia-inducible mitochondrial protein that has been implicated in mitochondrial fragmentation and subsequent mitophagy. BNIP3 protein has also been shown to be up-regulated at pre-malignant stages of various different human cancers, including breast cancer and down-regulated later through a variety of mechanisms, including promoter methylation and altered sub-cellular localization. With an established role for hypoxia in promoting invasiveness and progression to metastasis in breast cancer, we were interested to determine how BNIP3 (as a HIF target gene) acted in tumor progression. To do this, we crossed mice carrying a targeted deletion in the mouse BNip3 gene to the MMTV-PyMT mouse model of breast cancer. All analyses were performed on mice backcrossed onto a pure FVB/N genetic background. We show that while loss of BNip3 has only a modest effect on primary tumor growth, there is a highly significant increase in the numbers of lung metastases and a reduced latency to metastasis in the MMTV-PyMTl;BNip3-/- mice compared to control MMTV-PyMT;BNip3+/+ mice. BNip3 null tumors showed increased invasive properties and nuclear grade at earlier stages that were transplantable to wild-type host mice arguing that the phenotype was cell-intrinsic to BNip3 null tumor cells. This invasive pathology was also associated with increased tumor cell proliferation, increased HIF-1α levels, increased CD31-positive blood vessel formation and elevated MMPase activity. BNip3 null tumor cells exhibited abnormal mitochondria, reduced mitochondrial membrane potential and increased ROS levels. In vitro, quenching ROS limited the increased invasion/migration properties of BNip3 null tumor cells under hypoxia. These results suggest that BNip3 is a metastasis suppressor by virtue of its role in maintaining mitochondrial integrity that mitigates against the metastasis promoting activity of hypoxia.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2804. doi:10.1158/1538-7445.AM2011-2804
Abstract 2824: PENAO: A small molecule tumor metabolism inhibitor
Authors: Dilda Pierre J. , Decollogne Stephanie , Weerakoon Lakmini , Hogg Philip J.
Published: June 2012

Tumors reorganize the metabolic steps used by normal tissues for glucose utilization and ATP production. Normal tissues rely heavily on oxidative phosphorylation for ATP, while tumor cells employ a route that includes a much greater dependency on glycolysis. This aberrant metabolism allows for tumor survival when oxygen is limiting while providing a mechanism to poison the extracellular environment with acid, which facilitates invasion and metastasis.

In particular, tumors harness a crucial enzyme, hexokinase II (HKII), to entrap and channel glucose toward glycolysis. HKII is bound to mitochondria via interaction with the voltage dependent anion channel (VDAC). This affords HKII preferential access to the mitochondrial ATP via inner-membrane adenine nucleotide translocase (ANT) to rapidly phosphorylate and trap the incoming glucose, effectively initiating and regulating the glycolytic flux across the tumor cytosol.

We have developed a novel covalent inhibitor of ANT called PENAO, (4-(N-(S-penicillaminylacetyl)amino)phenylarsonous acid) (1). This molecule blocks ANT delivery of ATP to VDAC-bound HKII, thus inhibiting tumor cell metabolism, and triggers the mitochondrial permeability transition pore, resulting in proliferation arrest and apoptotic cell death. PENAO rapidly enters the cell via an organic ion transporter and reacts with Cys160 and Cys257 on the matrix face of ANT, which leads to arrest of proliferation of the cell and then apoptosis. PENAO targets both proliferating endothelial and tumor cells. It is very well tolerated in mice and has potent anti-tumor activity in murine tumor models when administered either subcutaneously or intravenously.

PENAO is the most potent anti-tumor compound of its type so far reported and will enter a Phase I/IIa clinical trial in adults with solid tumors refractory to standard therapy in 2011.

1. Dilda PJ, Decollogne S, Weerakoon L, et al. Optimization of the antitumor efficacy of a synthetic mitochondrial toxin by increasing the residence time in the cytosol. J Med Chem 2009;52(20):6209-16.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2824. doi:10.1158/1538-7445.AM2011-2824
Abstract 283: Vascular ageing as a modulator of tumor angiogenesis, antiangiogenesis and metastasis
Authors: Meehan Brian , Kadlubowska Dorota , Dombrovsky Alexander , Rak Janusz W.
Published: June 2012

Ageing profoundly influences vascular homeostasis and impacts angiogenesis, including in cancer. This is rarely considered in preclinical studies, as mouse tumor models usually rely on animals between 1-6 months of age (5-25% of life expectancy), while the majority of adult cancer patients are diagnosed in later decades of life. Thus a better characterization of a pediatric, adult and geriatric state of the vascular system is badly needed to properly design age-appropriate antiangiogenic anticancer therapies.

Using aged (over 12 months old), aged and atherosclerotic (ApoE-/-) and aged and diet induced obese (DIO) C57BL/6 mice we have demonstrated that model tumors (Lewis Lung Carcinoma) exhibit profoundly different angiogenic and metastatic patterns in those settings then in conventional syngeneic recipients. We documented age-related molecular differences in tumor blood vessels and have shown that the antitumor effects of several therapeutic agents (L-NAME, Sutent, AG28262, metronomic cyclophosphamide) are affected by vascular age and comorbidities. Age also changes the pattern of metastasis as aged mice tend to exhibit less aggressive metastatic growth. On the other hand metastasis following treatment with antiangiogenic agents (Sutent without chemotherapy) is more accentuated in young mice then in their aged counterparts. Finally, we report age-related changes in vascular characteristics of human kidney tumours, including a significantly higher expression of eNOS in tumours of younger patients.

Overall, these observations suggest that age may play a significant role in modulation of angiogenesis, antiangiogenesis and metastasis in various tumor settings and agents more effective in adult patients may not have the same impact in the pediatric population.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 283. doi:10.1158/1538-7445.AM2011-283
Abstract 2831: Novel antibody-drug conjugates for selective tumor activation
Authors: Setter Jocelyn R. , Doronina Svetlana O. , Bovee Tim D. , Anderson Marti E. , Senter Peter D. , Benjamin Dennis R. , Lyon Robert P.
Published: June 2012

Auristatin based antibody-drug conjugates (ADCs) have demonstrated profound activites, both preclinically and clinically. Upon internalization, the active auristatin is released through proteolytic degradation of the linker, where it induces cell kill through the inhibition of tubulin polymerization. Once released from the antibody, clearance of the auristatin proceeds via the hepatobiliary route. This finding prompted us to explore new auristatin-peptide drug linkers that had the capacity to release the active form of the drug in tumors, but an inactive form in the liver. To this end, we designed a novel in vivo screen in which multiple ADCs were cassette dosed in mice. LC-MS/MS was used to determine which sequences amongst approximately 800 new auristatin-peptide derivatives allowed for differential drug activation. Sequences were identified that allowed for more selective intratumoral release of the active drug.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2831. doi:10.1158/1538-7445.AM2011-2831
Abstract 2847: Host angiopoietin-2 inhibits tumor growth and angiogenesis in the liver
Authors: Im Jae Hong , Balathasan Lukxmi , Yameen Sabira , Tapmeier Thomas , Hill Sally , Bernhard Eric , Gorden D.Lee , Kruse Karoline , Augustin Hellmut , Muschel Ruth J.
Published: June 2012

For growth and survival in metastasis, tumor colonies must develop new intratumoral vasculature for supply of nutrients and oxygen. Since angiopoietin-2 (Ang2) resulted in a burst of angiogenesis in the glioma model, we examined the effects of host Ang2 on colony growth and neo-angiogenesis during the liver colonization.

Liver colonization was induced through the intrasplenic injection of MC38-GFP (murin colorectal carcinoma cells) into two genotype mice of the ang2−/− mice and the wild type mice. When colonies were established, the livers were isolated and analyzed using flow cytometry, immunoflorescent histochemistry and ELISA. In addition, the blood was collected from vena cava for measurement of MCP-1 at 14 days.

Flow cytometry analyses of whole livers showed that the tumor cell population in the ang2−/− mice was approximately 2 times larger than in the wild type mice. Immunoflorescent histochemistry of the liver tissue using antibodies for CD31, CD34, CD146, VE-cadherin and NG2 showed that intratumoral vascular density and vascular maturity in the ang2−/− mice were approximately 2 times higher than the wild type mice. Intratumoral vessels were significantly positive for CD146 and CD34, markers more frequently associated with neo-angiogenesis, but much less for CD31. There was no significant difference in VEGF level which is known to mediate vascular response in combination with Ang2. CD11b positive cells and MCP-1 concentration in the livers were equivalent in two genotypes. Notably, there were approximately 10 times more Tie2+/CD11b+ cells in the ang2−/− mice than the wild type mice. CD11c+/CD45+ cell populations were slightly different in two genotypes. These results suggested that host Ang2 would inhibit tumor growth and neo-angiogenesis in the liver, and that the inhibition is correlated with the decreased recruitment of TEMs, one of myeloid cells, into the liver.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2847. doi:10.1158/1538-7445.AM2011-2847