bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3747: Polymorphisms of ERCC1 C118T and ERCC2 Lys751Gln predict response and prognosis of platinum-treated gastroenterology cancer patients: A finding of pooled meta-analysis
Authors: Yin Ming , Yan Jingrong , Abad Albert , Graziano Francesco , Lenz Heinz-Josef , Kim Hyo-Jin , Robert Jacques , Im Seock-Ah , Wang Wei-Shu , Wei Qingyi
Published: February 2011

The nucleotide excision repair (NER) pathway modulates platinum-based chemotherapeutic efficacy by removal of drug-produced damage to DNA. To determine if NER genes (ERCC1 and ERCC2) influence responses to platinum-based chemotherapies, we performed a meta-analysis in gastric and colorectal cancer for polymorphisms of ERCC1 C118T and ERCC2 Lys751Gln. The analysis included 1413 (451 Asians and 962 Caucasians) cancer patients, who were treated with the platinum-based regimen. The primary outcomes included therapeutic response (TR) as evaluated by complete response + partial response vs. stable disease + progressive disease, progression-free survival (PFS) and overall survival (OS). We calculated odds ratio [OR] or hazard ratio [HR] with 95% confidence intervals [CI] to estimate the risk or hazard in the dominant model. We found consistent and clinically substantial risk or hazard estimates for TR, PFS and OS in the platinum-treated gastroenterology cancer patients. For ERCC1 C118T, we found an ethnical discrepancy in the prognostic values, favoring a pronounced effect in Asians (TR: OR, 0.53 and 95% CI, 0.35-0.81 for four studies of 378 patients; PFS: HR, 1.69 and 95% CI, 1.05-2.70 for five studies of 504 patients; and OS: HR, 1.85 and 95% CI, 1.37-2.48 for four studies of 405 patients, compared with Caucasians: TR: OR, 0.98 and 95% CI, 0.44-2.19 for five studies of 638 patients; PFS: HR, 1.15 and 95% CI, 0.72-1.86 for five studies of 609 patients; and OS: HR, 1.22 and 95% CI, 0.49-3.05 for three studies of 347 patients). For ERCC2 Lys751Gln, the prognostic effect was observed in colorectal cancer for both ethnicities (overall patients: TR: OR, 0.41 and 95% CI, 0.26-0.66 for four studies of 437 patients; PFS: HR, 1.63 and 95% CI, 1.19-2.22 for six studies of 741 patients; and OS: HR, 2.08 and 95% CI, 1.29-3.34 for four studies of 479 patients). This significance remained in Caucasians (TR: OR, 0.41 and 95% CI, 0.23-0.73 for three studies of 249 patients; PFS: HR, 1.55 and 95% CI, 1.08-2.21 for five studies of 553 patients; and OS: HR, 1.64 and 95% CI, 1.23-2.18 for three studies of 291 patients). Based on these results, we concluded that ERCC1 C118T and ERCC2 Lys751Gln polymorphisms might be useful prognostic factors in platinum-treated gastroenterology cancer patients. Further clinical trials are warranted to confirm these findings. (Supported by NIH grants R01CA131274 and R01ES011470)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3747.
Abstract 3750: Functional analysis of the Fanconi Anemia pathway in primary lung tumors
Authors: Duan Wenrui , Gao Li , Wu Xin , Zhao Weiqiang , Leon Marino , Otterson Gregory A. , Villalona-Calero Miguel A.
Published: February 2011

The Fanconi Anemia (FA) pathway is a major mechanism of homologous recombination DNA repair in response to genotoxic insults. Lack of formation of FANCD2 foci has been reported as a predictor of cisplatin and mitomycin C (MMC) sensitivity in cancer cells. Given that this pathway plays essential roles in response to the DNA damage, cancers with defective FA pathway are expected to be more sensitive to DNA cross-link based therapy, or to treatments in which additional repair mechanisms are targeted.

We have developed a test to identify tumors with defective FA pathway using paraffin embedded tissues. Paraffin embedded lung primary cancer specimens were subjected to immunofluorescence microscopy (IF) analysis to evaluate the status of FancD2 subnuclear foci formation, Ki67 protein expression and DAPI staining of nuclei simultaneously in order to eliminate false negative tumor specimens due to absence of tumor cells or low proliferation. Additional validation using FA defective PD220 and PD20 cell-lines as well as fresh tumor tissues was done; IF analysis and Western blotting analysis of FDACD2 protein mono-ubiqutination were performed.

The ratio of FANCD2 foci negative tumor was 20% among 20 paraffin embedded non-small cell lung cancer samples analyzed. Histological characteristics of the defective tumors include large cell carcinoma and adenocarcinoma. The proposed immunofluorecence staining method evaluates functional status of FA pathway using paraffin embedded tissues even after long term storage. Given the clinical availability of agents targeting alternative mechanisms of repair, the status of FA pathway may be an important biomarker for personalized therapeutic treatment in patients with lung cancers. A clinical trial to validate this hypothesis is under way.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3750.
Abstract 3756: Prediction of xenograft response to MM-121, an anti-ErbB3 inhibitor, using computational modeling and measurements of five biomarkers
Authors: Onsum Matthew , Burenkova Olga , Fulgham Aaron , Nie Lin , Kalra Ashish , Xiao Dongmei , Grantcharova Viara , Adams Sharlene , Luus Lia , Paragas Violette , Bukhalid Raghida , Moulis Sharon , Wille Lucia , Garcia Gabriela , Moyo Victor , Schoeberl Birgit , Kubasek Bill , Nielsen Ulrik
Published: February 2011

One of the challenges faced by targeted therapeutics currently in the clinic is the relatively small population of patients who derive significant benefit from their use. We report the development of a preclinical classifier which can correctly predict xenograft response to MM-121, an anti-ErbB3 antibody, based on the measurement of a few key biomarkers in cell lysates.

Deregulation of the ErbB family receptors is common in many cancers. Using a combination of computational modeling and quantitative experiments we identified ErbB3 as a key mediator of mitogenic signaling downstream of the ErbB receptors. Based on these results, we developed MM-121, a first in class anti-ErbB3 monoclonal antibody that blocks heregulin-induced signaling and inhibits tumor growth in multiple xenograft models of human cancer.

Here we present our efforts to derive a predictive biomarker signature that identifies tumors that are responsive to MM-121. Using our computational model of the ErbB signaling pathway we identified the five most critical proteins for predicting activation of phospho-AKT - a key mediator of cell survival and apoptosis. These proteins include MM-121's target, ErbB3, and its ligand, heregulin. We profiled these biomarkers in a large panel of cancer cell lines, and using the measured effect of MM-121 on inhibiting tumor growth in eight xenograft tumor models, we determined a classification rule for predicting xenograft response. We subsequently used this classification rule to correctly predict a priori MM-121 response in 11 xenograft models.

These results suggest that our computationally-derived biomarker signature is sufficient for predicting response to MM-121 in xenografts, and could offer significant clinical benefit by helping select patients for MM-121 treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3756.
Abstract 3762: Automated analysis of breast cancer tissue microarrays to investigate etiologic heterogeneity in the Breast Cancer Association Consortium
Authors: Sherman Mark E. , Howatt William , Bolton Kelly , Pharoah Paul , Garcia-Closas Montserrat
Published: February 2011

Background: Data suggest that breast cancer risk factor associations vary by tumor marker expression. However, individual studies often lack the power required to assess these relationships and pooled analyses are limited by non-standardized scoring of tumor markers. To address these limitations, the Breast Cancer Association Consortium is assessing whether automated image analysis of immunohistochemically stained tissue microarrays (TMAs) can permit standardized scoring of tumor markers.

Methods: TMA sections prepared in 10 case-control studies containing up to 9,298 breast cancers stained for estrogen receptor (ER); progesterone receptor (PR); human epidermal growth factor receptor 2 (HER 2); epidermal growth factor receptor (EGFR) and cytokeratin 5/6 (CK 5/6) were scanned as digital images. Automated algorithms were used to score markers in tumor cells using the Ariol system. We are analyzing consistency of automated scores within individual studies; between studies; and against visual reads.

Results: Preliminarily, distributions of automated scores were generally similar across different TMA blocks within a study; however isolated blocks yielded disparate results related to low or high intensity staining for a marker or the counterstain across the entire slide. Distributions of scores varied between studies, suggesting the need for study-specific cutpoints for negative vs. positive. Analysis of receiver operator characteristic curves comparing automated analysis to independently recorded visual reads in two studies demonstrated areas under the curves of 0.90 and 0.93 for tumors classified visually as ER+/PR+ and 0.91 and 0.88 for tumors scored visually as ER-/PR-. Cores scored as negative for EGFR or CK 5/6 by automated methods agreed well with visual scores, but many tumors scored as positive by automated analysis were negative by visual review, irrespective of ER / PR status. Thus, restricting analyses to ER-/PR- tumors (a subset of interest) reduced the number of discrepancies. Ongoing analyses are comparing automated and visual scores for all markers / studies and are assessing risk factor associations for breast cancer by tumor marker expression (negative vs. positive and strength of expression) based on automated scores.

Conclusions: Automated analysis of immunohistochemically stained TMAs of breast cancers may be useful in standardizing high throughout characterization of cancer markers in multicenter studies and in consortia, although additional methodologic work is needed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3762.
Abstract 3766: Development of a model using genetics and PSA to reduce unnecessary prostate biopsies
Authors: Cross Deanna S. , McCarty Catherine , Reding Douglas J.
Published: February 2011

Background: Prostate cancer screening with PSA has decreased the average age of prostate cancer detection and increased the number of men who undergo prostate biopsies. Currently two thirds of the prostate biopsies performed due to a high PSA test are negative for prostate cancer leading to unnecessary expense and distress for the individual. One way to decrease the unnecessary procedures associated with PSA screening would be to identify individuals who are at higher or lower risk of prostate cancer using newly discovered genetic and environmental risk factors.

Aims: Using the males in the Personalized Medicine Research Project (PMRP) population that have had a PSA test, determine if genetic and environmental factors can be incorporated into the PSA screening model to better discriminate individuals who will have a diagnosis of prostate cancer from those with negative biopsies.

Methods: All men in PMRP who have had a PSA test (N=4659) were genotyped for 12 polymorphisms that have been associated with prostate cancer in GWAS studies and an additional 52 polymorphisms that were associated with a number of diseases. We also included dietary history, particularly dairy intake if available for the individual.

Results: A total of 935 of the 4659 men in the study have had a prostate biopsy with a positive biopsy rate of 34%. Using a panel of 12 previously identified polymorphisms; seven were significantly associated with prostate cancer in our population. Another three polymorphisms were found to be associated in our population (LPL rs328, NOS3 rs1799983, and PON2 rs7493). None of the tested polymorphisms were associated with the biopsy procedure. Individuals with 4 or fewer of the 10 associated risk alleles (17.33% of the population) were more likely to have a negative biopsy (OR 1.87 p=0.0017) while those individuals who have 8 or more associated risk alleles (11% of the population) were more likely to have a positive biopsy (OR 2.68).

Conclusions: The PMRP population that has experienced at least one PSA screen is a viable population for testing new screening algorithms that incorporate genetic and environmental factors for better identification of prostate cancer risk.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3766.
Abstract 3772: Lifestyle factors and global methylation in peripheral leukocyte DNA
Authors: Zhang Fang Fang , Morabia Alfredo , Carroll Joan , Gonzalez Karina , Fulda Kimberly , Kaur Manleen , Santella Regina M. , Cardarelli Roberto
Published: February 2011

Global hypomethylation is associated with genomic instability and may mediate the association between environmental and lifestyle factors and cancer risk. We examined the association between lifestyle factors and levels of genomic DNA methylation in the peripheral blood leukocytes of 160 participants in the North Texas Healthy Heart Study aged 45-75. We used in-person interviews for demographics and lifestyle factors, a self-administrated Block food frequency questionnaire for dietary data, bioelectrical impedance analysis (BIA) and CT-scan for body composition including fat mass % and areas of visceral and truncal subcutaneous adipose tissues at the L4L5 level. We measured genomic DNA methylation using bisulfite conversion of DNA and real time PCR (MethyLight) for SAT2-M1. The median level of global methylation was 61.3% in this cancer-free population. Male gender was positively associated with global hypomethylation (i.e. < median) (OR=3.7, 95%CI: 1.4-9.7). Compared to Hispanics (n=58), non-Hispanic whites (n=33) and non-Hispanics blacks (n=68) had a significantly higher level of global hypomethylation (OR=8.4, 95%CI: 2.4-29.6 and OR=4.1, 95%CI: 1.5-11.2 respectively). Although the area of L4L5 subcutaneous adipose tissue was positively associated with DNA methylation in the univariate model (342 cm2 among subjects with methylation < median vs. 395 cm2 among subjects with methylation ≥ median, p=0.04), none of the body composition measures were associated with DNA methylation in the multivariate model after adjusting for age, gender and ethnicity, nor were cigarette smoking, alcohol drinking, dietary folate intake and perceived stress. Although there was no association of lifestyle factors with genomic methylation in the peripheral leukocyte DNA, the strong association with gender and race/ethnicity indicates that interactions between these variables and lifestyle factors need to be explored in a larger sample before firm conclusions can be reached.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3772.
Abstract 3773: Ability to predict breast cancer in asian women using a polygenic susceptibility model
Authors: Hartman Mikael , Chen Suo , Miao Hui , Lim Wei-Yen , Koh Woon Puay , Teo Yik-Ying , Chia Kee Seng
Published: February 2011

Ability to predict breast cancer in Asian women using a polygenic susceptibility model

Background Recently identified susceptibility alleles from Genome wide Association Studies (GWAS) for breast cancer suggest a new venue to both prevention and treatment of the disease. Given variations in allele frequency between populations this ability may vary.

Methods We examined the possibility for individualized disease prevention and for population screening in Asian women based on common breast cancer risk alleles. We tested using simulations and available allele frequencies from the Singapore Genome Variation Project how these markers perform together with known environmental risk factors for the disease.

Results Our analysis suggests that the risk profile generated by 13 single nucleotide polymorphisms (SNP) as identified thru GWAS does not provide sufficient discrimination to warrant individualized prevention. They may however be useful in risk stratification for screening programs especially together with a few reproductive risk factors for disease in breast cancer prevention in the general population (Table 1).

Conclusions The clinical use of single, common, low-penetrance genes is limited, but 13 validated SNPs in combination with a few readily available reproductive factors may distinguish women who are at high risk for breast cancer from those who are at low risk, particularly in the context of population screening.

Table 1: Proportion of cases of breast cancer explained by the proportion of the population at highest risk for the disease. Estimates based on 13 currently known susceptibility alleles by ethnic group and number of SNPS.Proportion of general population at risk above a given level  Proportion of Cases at risk above a given level Chinese 13 SNPsMalay 13 SNPsIndian 13 SNPsChinese 13 SNPs & Environment 20%30.6%30.9%32.2%33.5% 40%53.1%53.5%54.9%56.0% 60%71.9%72.4%73.5%74.2% 80%87.8%88.1%88.6%89.0% 

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3773.
Abstract 3777: Diesel motor exhaust and lung cancer risk in a pooled analysis from case-control studies in Europe and Canada
Authors: Olsson Ann C. , Gustavsson Per , Kromhout Hans , Peters Susan , Vermeulen Roel C. H. , Brüske Irene , Pesch Beate , Brüning Tomas , Siemiatycki Jack , Pintos Javier , Wichmann Heinz-Erich , Consonni Dario , Plato Nils , Merletti Franco , Mirabelli Dario , Richiardi Lorenzo , Jöckel Karl-Heinz , Ahrens Wolfgang , Pohlabeln Hermann , Jolanta Lissowska , Szeszenia-Dabrowska Neonila , Adrian Cassidy , Zaridze David , Stücker Isabelle , Benhamou Simone , Bencko Vladimir , Foretova Lenka , Janout Vladimir , Rudnai Peter , Fabianova Eleonora , Mates Dana , Bueno-de-Mesquita Bas , Gross Isabelle , Benhaim-Luzon Veronique , Boffetta Paolo , Straif Kurt
Published: February 2011

Introduction: Diesel-motor exhaust (DME) is classified by IARC as probably carcinogenic to humans. The epidemiological evidence is evaluated as limited since many studies lack adequate information on tobacco smoking and only few studies reported on exposure-response relationships. Our objective is to investigate the risk of lung cancer following occupational exposure to DME, while controlling for smoking and potential occupational confounders.

Methods: The SYNERGY project pooled information on lifetime work histories and tobacco smoking from more than 13300 cases and 16300 controls from case-control studies conducted in 12 European countries and Canada. A job exposure matrix based on ISCO codes, assigning no (0), low (1) or high (2) exposure to DME was applied to determine level of exposure to DME. Cumulative exposure was defined as: ∑(level2 * duration). Odds ratios (OR) of lung cancer and 95% confidence intervals (CI) were estimated by unconditional logistic regression, adjusted for age, sex, study, pack-years and time since quitting smoking, and ever employment in a “Group A” job, i.e. occupations with established lung cancer risk.

Results: Workers exposed to low levels of occupational DME exposure had an increased risk of lung cancer after 40 years of exposure OR 1.31 (95% CI 1.14-1.51), while workers exposed to high DME levels experienced a similar risk already after short duration (<10 years) OR 1.29, 95% CI 1.15-1.46. Cumulative DME exposure showed a significant exposure-response trend (p-value <0.000) with an OR of 1.31 (95% CI 1.20-1.44) in the highest quartile. These results were similar in workers never employed in “Group A“ jobs, lending support to the assumption that confounding due to other occupational exposures was not responsible for the observed risk. Analyses in sub-populations of women and never-smokers also indicated an increased risk of lung cancer following occupational DME exposure.

Conclusion: Our results indicate that occupational exposure to DME is associated with an increased risk of lung cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3777.
Abstract 3781: 3,3′-diindolylmethane(DIM)-induced apoptosis in ovarian cancer cells is mediated by inhibition of EGFR-ERK pathway
Authors: Kandala Prabodh K. , Pramanik Kartick C. , Boreddy Srinivas Reddy , Srivastava Sanjay K.
Published: February 2011

Ovarian cancer is one of the leading causes of gynaecological deaths in the United States and several other countries. Genetic alterations including overexpression of EGFR (about 70% of ovarian tumors) play a crucial role in the signal transduction pathways that regulate key cellular functions such as cell survival and proliferation and are responsible for comprising traditional chemotherapy. Several epidemiological studies suggested a decreased risk of ovarian cancer in the population consuming Brassica vegetables. 3,3′-diindolylmethane (DIM) is an indole compound present in Brassica vegetables. In our previous studies, we have demonstrated that BR-DIM, a formulated version of DIM suppress the growth of ovarian cancer cells by causing cell cycle arrest. However, the exact mechanism by which DIM suppress the growth of ovarian cancer cells was not clear. In the present study, we delineated the mechanism of DIM in SKOV3, OVCAR-3 and TOV-21G human ovarian cancer cells. Our results show that DIM treatment induces apoptosis in all the cell lines in a dose-dependent manner, as analyzed by flow cytometry. Our western blot analysis reveals that DIM treatment causes significant down regulation of constitutive EGFR and HER2 protein level as well as phosphorylation of EGFR at Tyr992, Tyr845 and Tyr1173 in SKOV-3 and OVCAR-3 cells. To determine whether DIM can suppress the activation of EGFR by activating phosphorylation, cells were treated with EGF after DIM treatment. Our results reveal that DIM treatment block the activation of EGFR induced by EGF in OVCAR-3 and TOV-21G cells. In addition, DIM treatment drastically reduces the phosphorylation of ERK without affecting the protein level. DIM treatment also inhibits the kinase activity of ERK as observed by the down regulation of p-ELK in all the three ovarian cancer cell lines. ERK is known to be regulated by EGFR. Taken together our results indicate that DIM induces apoptosis in ovarian cancer cells by inhibiting EGFR-ERK pathway. Further mechanistic studies are in progress. [Supported in part by RO1 grants CA 106953 and CA129038 (to S.K.S) awarded by the National Cancer Institute].

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3781.
Abstract 3787: D, L-Sulforaphane inhibits constitutive and interleukin 6-induced activation of signal transducer and activator of transcription 3 in human prostate cancer cells
Authors: Hahm Eun-Ryeong , Singh Shivendra V.
Published: February 2011

D, L-Sulforaphane (SFN), a synthetic analogue of broccoli-derived L-isomer, inhibits viability of human prostate cancer cells in culture and in vivo. We also showed recently that oral administration of SFN prevents development of prostate cancer and pulmonary metastasis in a transgenic mouse model (TRAMP mice) by reducing cell proliferation and augmenting activity of natural killer cells. However, the mechanism underlying anticancer effect of SFN is not fully understood. We now demonstrate that SFN inhibits constitutive and interleukin 6 (IL6)-induced activation of signal transducer and activator of transcription 3 (STAT3), which is implicated in development and progression of prostate cancer. Growth suppressive concentrations of SFN (20 and 40 μmol/L) inhibited constitutive (DU145 cells) and IL6-induced (LNCaP cells) phosphorylation of STAT3 (Tyr705) as well as its upstream regulator Janus-activated kinase 2 (JAK2; Tyr1007/1008). Treatment of DU145 and LNCaP cells with SFN resulted in suppression of IL6-induced transcriptional activity of STAT3 as revealed by luciferase reporter assay and nuclear translocation of STAT3 as judged by immunofluorescence microscopy. The SFN-mediated inhibition of STAT3 activation was due to down-regulation of JAK2 mRNA expression in both cell lines. Levels of many STAT3-regulated gene products including Bcl-2, cyclin D1, and survivin were also down-regulated in SFN-treated cells. The IL6-mediated activation of STAT3 conferred partial but marked protection against SFN-induced apoptosis in LNCaP cells as evidenced by cleavage of poly-(ADP-ribose)-polymerase and caspase-3 and cytoplasmic histone-associated DNA fragmentation. In addition, siRNA knockdown of STAT3 in DU145 cells resulted in a modest yet statistically significant increase in SFN-induced apoptotic DNA fragmentation. The results of the present study demonstrate SFN-mediated inhibition of STAT3 activation in human prostate cancer cells. This investigation was supported by the National Cancer Institute grant CA115498-05.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3787.
Abstract 3791: Sulphoraphane inhibits STAT3 and its related genes in Du 145 cancer cells
Authors: Koh Wonil , Ahn Kwangseok , Jeong Soojin , Kwon Taerin , kim Jihyun , Shin Jimin , Yun Sunmi , Jung Deokbeom , Jung Jihoon , Lee Eunok , Lee Hyojeong , Lee Hyojung , Lu Junxuan , Kim Sung Hoon
Published: February 2011

Sulforaphane is a naturally occurring isothiocyanate compound mainly found in cruciferous vegetables. Sulforaphane has demonstrated cancer chemopreventive activities in a number of organ sites, including prostate. Mechanistic investigations have shown that SFN induces caspase-mediated apoptosis that is mediated through a rapid intracellular ROS generation. Since STAT3 signaling is crucial for cancer cell survival and ROS has been linked to inactivation of this transcriptional factor, we hypothesized that STAT3 may be an important target for SFN. Here, we investigated the effect of sulforaphane on the regulation of STAT3 in association of ROS in Du145 prostate cancer cells. We first confirmed that sulforaphane induced apoptosis by activation of caspase 8, 9, and 3, PARP cleavage and increase of TUNEL positive cells in Du145 cells, which were preceded for many hours by the increase of intracellular ROS, peaking at 30 minutes of exposure. We demonstrate for the first time that sulforaphane suppressed constitutive STAT3 activation and DNA-binding of STAT3 in a time- and dose-dependent manner in Du145 cells. These actions were also found in U266 multiple myeloma cells and U937 lymphoma cells. In addition, sulforaphane down-regulated the expression of STAT3-related genes such as bcl-2, bcl-xL, survivin, cyclin D1, and VEGF in Du145 cells. Furthermore, sulforaphane cross-linked JAK2, the upstream kinase of STAT3, in a time- and dose-dependent manner. Thiol-reducing agent dithiothreitol (DTT) reversed the inactivation of STAT3 and JAK2 induced by sulforaphane. Interestingly, N-acetyl cysteine (NAC), which leads to increased intracellular GSH synthesis, reversed the molecular patterns of PARP cleavage and inactivation of JAK2 and STAT3 induced by sulphoraphane. Taken together, these findings suggest that sulforaphane exposure leads to a rapid generation of ROS in Du145 cells and inhibition of JAK2/STAT3 pathway, followed by caspase-mediated apoptosis. Our data suggest that the JAK2/STAT3 pathway may be a novel target of SFN contributing to apoptosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3791.
Abstract 3795: STAT-3: A Novel Target of Diindolylmethane in Ovarian Cancer
Authors: Kandala Prabodh K. , Srivastava Sanjay K.
Published: February 2011

Signal transducers and activation of transcription (STATs) are the important for growth and development. However defects in STAT's may lead to malignant transformation. STAT-3 is an oncogene which is over expressed in several cancers including ovarian cancer. We have previously demonstrated that Diindolylmethane (DIM), an active metabolite of Indole-3-Carbinol present in cruciferous vegetables, is effective in suppressing the proliferation of human ovarian cancer cells, however, the mechanism through which DIM reduces the growth of cancer cells was not clear. We hypothesized that DIM inhibits the growth of ovarian cancer cells by inhibiting STAT-3 activation. Our present studies demonstrate that DIM treatment inhibit the phosphorylation of STAT-3 at Tyr-705 and Ser-727 in a concentration-dependent manner in SKOV-3 and OVCAR-3 cells. Interestingly, DIM treatment also reduced the constitutive protein expression of STAT-3 in both the cell lines. In a time dependent study, we observed that STAT-3 inhibition was as early as 1h after DIM treatment. Next, we wanted to see whether DIM can block IL-6 mediated activation of STAT-3. Treatment of SKOV-3 or OVCAR-3 cells with 5ng/ml IL-6 for 15min increase the activation of STAT-3 by phosphorylation at Tyr705 by almost twenty fold. Our results reveal that DIM treatment almost completely blocked IL-6 induced phosphorylation of STAT-3 at Tyr705 in both SKOV-3 and OVCAR-3 cells and completely in TOV-21G cells. STAT-3 after activation translocate to nucleus, bind to DNA and is involved in the transcription of responsive genes such as Mcl-1, survivin and Bcl-2. Our results demonstrate that DIM inhibits the DNA binding activity of STAT-3 resulting in the reduced expression of Mcl-1, survivin and Bcl-2. Overexpression of STAT-3 by gene transfection abrogates the cell growth inhibitory effects of DIM. Taken together, our cellular results establish STAT-3 as a therapeutic target of DIM in ovarian cancer. Furthermore, our just completed in vivo studies indicate that oral administration of 2mg DIM per day for 6 weeks significantly suppressed the growth of SKOV-3 tumor xenografts in athymic nude mice. Whether SKOV-3 ovarian tumor growth suppression is through inhibition of STAT-3 in vivo is currently under investigation. [Supported in part by RO1 CA 106953 and CA 129038 awarded to S.K.S by NIH]

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3795.
Abstract 3807: Gugulipid-induced apoptosis in human prostate cancer cells is regulated by Bax/Bak and Bcl-xL
Authors: Zeng Yan , Badmaev Vladmir , Singh Shivendra V. , Xiao Dong
Published: February 2011

Indian Ayurvedic medicinal plant Commiphora mukul has been safely used for treatment of different disorders for thousands of years. Our previous studies have shown that the extract of Commiphora mukul, gugulipid (GL), inhibits growth of human prostate cancer (Pca) cells, but not a normal prostate epithelial cell line (PrEC), by causing apoptosis. We also showed that GL-mediated inhibition of Pca cell proliferation correlated with reactive oxygen species (ROS) production. The present study was designed to gain insight into the mechanism of GL-induced apoptosis downstream of ROS production. We found that the GL-induced apoptosis correlated with induction of proapoptotic proteins Bax and Bak and down-regulation of antiapoptotic proteins Bcl-2 and Bcl-xL in LNCaP and PC-3 cells. The SV40-immortalized mouse embryonic fibroblasts derived from Bax and Bak double knockout mice were significantly more resistant to GL-induced apoptotic cell death (DNA fragmentation and cleavage of PARP) compared with wild-type mouse embryonic fibroblasts. Bcl-xL overexpression in PC-3 cells conferred protection against GL-induced apoptotic cell death. The GL treatment caused activation of c-Jun-N-terminal kinase (JNK), which functions upstream of Bax activation in apoptotic response to various stimuli. Pharmacological inhibition of JNK conferred partial yet significant protection against GL-induced apoptosis. The GL-mediated apoptosis and conformational change of Bax was markedly suppressed by ectopic expression of catalytically inactive mutant of JNK kinase JNKK2 (AA) in LNCaP cells. In conclusion, the present study indicates that Bax/Bak and Bcl-xL regulate GL-induced apoptosis in human prostate cancer cells. This investigation was supported by USPHS grant R21 CA143104-01.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3807.
Abstract 3809: Tumor-released GPI-anchored proteins bind to mannose receptor and polarize tumor-infiltrating macrophages
Authors: Dangaj Denarda , Zhao Aizhi , Nunez-Cruz Selene , Sandaltzopoulos Raphael , Scholler Nathalie
Published: February 2011

Tumors polarize their microenvironment to escape immune-mediated destruction and to promote their growth. Macrophages respond to microenvironmental signals and can be activated toward a classical (M1) or an alternative (M2) phenotype. Tumor-associated macrophages (TAM) resemble to M2 macrophages with a phenotype characterized by the overexpression of IL10, scavenger receptor SR-A, CD68 and MR phenotype but hampered levels of IL-12. M2, in contrast to M1 macrophages, promote TH2 immune responses as well as tissue remodelling and M2 infiltration in solid tumors has been shown to significantly correlate with poor prognosis. Recent evidences suggest that macrophage differentiation towards TAM involves a “chemical conversation” via exchange of unknown soluble extracellular mediators between tumor cells and macrophages. In addition, previous work showed that the cross-linking mannose receptor (MR) with an anti-MR mAb could cause dendritic cell differentiation into APCs promoting T-cell anergy.

We hypothesized that MR expressed by macrophages could be bound by tumor-released GPI-anchored glycoproteins through the GPI-anchor mannose residues freed by cleavage. We also hypothesized that MR binding by GPI-anchor could trigger macrophage polarization toward TAMs. To test our hypotheses, we established an in vitro model system of cell co-culture in transwell allowing chemical exchanges between human monocyte-derived macrophages and human ovarian cancer cell line Ovcar3 that sheds mesothelin, a cancer biomarker and a GPI-anchored glycoprotein. Macrophage phenotype changes were monitored by flow cytometry and qRT-PCR. We also isolated three novel recombinant antibodies (scFv) against MR by magnetic and flow sortings from a novel yeast-display human scFv library. We describe here for the first time that tumor-released mesothelin bound to macrophage cell surface in presence of calcium, and that mesothelin binding to macrophages could be blocked by the anti-MR scFv. Furthermore, macrophage phenotype switch toward M2 during coculture with tumor cells could be prevented by one of the anti-MR scFv. Finally, we demonstrated that M1 phenotype could be rescued during coculture with tumor cells with one of the anti-MR scFv. To further characterize the role of MR in vivo, we have implanted mannose receptor knock-out mice intraperitoneally (IP) with luminescent-labeled ovarian cancer cells that overexpress mesothelin. We are now monitoring the tumor growth by in vivo imaging.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3809.
Abstract 3813: Recruitment of macrophages, NK cells, and dendritic cells with metronomic cyclophosphamide-induced regression of gliosarcoma xenografts
Authors: Doloff Joshua C. , Waxman David J.
Published: February 2011

Metronomic chemotherapy displays improved therapeutic activity over conventional schedules for several malignancies in both preclinical and clinical studies. In particular, metronomic cyclophosphamide treatment (140 mg/kg BW every 6 days) induces dramatic, near-complete regression of established xenografts of human U251 and rat 9L gliosarcomas in s.c. scid mouse models, whereas maximum tolerated dose cyclophosphamide effects only moderate tumor growth delays. Furthermore, metronomic cyclophosphamide-induced regression of 9L tumors is accompanied by reduced microvessel density, induction of thrombospondin-1 (TSP1) and impaired drug uptake, all of which are consistent with the prevailing view that anti-angiogenesis is a major factor in the therapeutic effectiveness of metronomic chemotherapy. However, VEGF receptor inhibition induces substantially greater anti-angiogenesis in these tumor models but does not induce tumor regression. Therefore, we sought to investigate other non-anti-angiogenic mechanisms to explain the substantial regression and have found that it is strongly associated with recruitment of the host innate immune system. In regressing 9L and U251 tumors, host (mouse) PEDF (pigment epithelium-derived factor; Serpinf1) and PAI-1 (plasminogen activation inhibitor-1; Serpine1) expression is increased, with the levels of PEDF RNA being strongly correlated (r=0.89) with TSP1 RNA in large sets of 9L and U251 tumors isolated various times after initiation of metronomic cyclophosphamide therapy. Notably, PAI-1 and TSP1 are both expressed by tumor-associated macrophages. Metronomic cyclophosphamide also increased Fas, which stimulates macrophage activation, and the macrophage marker CD68, supporting the involvement of tumor-associated macrophages. Fas is also associated with cytotoxic lymphocytes, such as NK cells, and NK cell markers and cytotoxic effectors were dramatically increased in metronomic cyclophosphamide-treated tumors. Furthermore, dendritic cell markers important for cell-target interactions and antigen presentation were increased by metronomic cyclophosphamide treatment. Cytokines and chemokines were also up regulated in the metronomic cyclophosphamide-treated tumors, suggesting that chemokine and cytokine gradients help mobilize the host innate immune system in regressing 9L and U251 tumors. These findings highlight the ability of metronomic chemotherapy to induce macrophage, natural killer cell, and dendritic cell recruitment, thereby activating innate immunity leading to regression of established gliosarcomas and potentially other tumor types. Supported by NIH grant CA49248 (to DJW).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3813.
Abstract 3818: CD11c+CD123+ human myeloid dendritic cells generated using IL4, IL13 and IL3 show characteristics of myeloid derived suppressor cells
Authors: Edwards Jeremy V. , Zhang Hua , Mackall Crystal L.
Published: February 2011

Dendritic cells (DCs) are critical for initiation of adaptive immune responses, but they also play important roles in self-tolerance, and in tumor bearing hosts, some DC subsets may contribute to immune evasion. Myeloid derived suppressor cells (MDSC) are also important mediators of tumor immune evasion. The relationship between myeloid DC and MDSCs in humans remains unclear. Human myeloid DCs are defined as CD11c+CD123-, although some have reported dim CD123 expression on human myeloid DCs. We describe a CD11c+CD123+ myeloid DC subset that shows characteristics of a MDSC.

Monocytes elutriated from normal donors were incubated for 48hr with IL4 ± IL13 ± IL3. Monocytes cultured with all three cytokines acquired strong expression of both CD123 and CD11c, with 79.5% of cells CD123+CD11c+. CD11c+CD123+ cells demonstrated over-expression of both arginase and iNOS by flow cytometry and/or real-time PCR in comparison to myeloid derived DCs generated with IL4, GM-CSF and LPS (mean fluorescence intensity of arginase in the IL4, IL13 and IL3 generated cells of 5973 vs. 1961 in IL4, GM-CSF and LPS generated cells). CD11c+CD123+ DCs also coexpress IL-4Rα and IL7Rα, characteristics of MDSC and regulatory DC subsets respectively. IL3 appears to be a potent inducer of CD123 expression on CD11c+ DCs, since myeloid DCs generated with only IL4 and IL13, in combination or separately, showed a less pronounced phenotype (38-52.7% CD11c+CD123+). Moreover, CD123 expression on myeloid DCs appears to be a marker for arginase production since CD123+ cells within the IL4, IL13 and IL3 cultures show higher arginase expression than CD123- cells within the same cultures. Functional studies are ongoing.

In conclusion, human myeloid DCs generated by incubating monocytes with IL4, IL13 and IL3 have a novel CD11c+CD123+ phenotype and show characteristics of myeloid derived suppressor cells, raising the prospect that IL3 plays an important role in generating myeloid derived suppressor cells in humans.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3818.
Abstract 382: Platelets are implicated in the rapid host responses induced by cytotoxic chemotherapy which limit its anti-tumor efficacy
Authors: Milsom Chlöe C. , Shaked Yuval , Lee Christina R. , Man Shan , Kerbel Robert S.
Published: February 2011

Previous studies from our lab have shown that the overall anti-tumor impact of certain cytotoxic drugs such as paclitaxel (PTX) given at maximum tolerated doses (MTDs) or OXi-4503, a microtubule inhibiting vascular disrupting agent (VDA), can be reduced by a rapid systemic counter-acting host response (Y Shaked et al., Science 2006 and Cancer Cell 2008). The aforementioned host response includes rapid induction of elevated plasma levels of multiple cytokines or chemokines such as G-CSF and SDF-1, which can promote mobilization and tumor-homing/colonization of proangiogenic and tumor-growth promoting bone marrow-derived cells (BMDCs), including circulating endothelial progenitor cells. This reactive host response promotes recovery/repopulation of the drug treated tumor. Here we asked what might be the source(s) of the elevated levels of plasma chemokines and cytokines induced by MTD PTX. We found platelets - known major sources of SDF-1 and VEGF - become activated and release the contents of their alpha granules into the circulation (plasma), such that decreases in platelet associated SDF-1 (and VEGF) are observed accompanied by increases in their plasma levels. SDF-1 is known to mobilize a variety of BMDCs including CEPs. Because platelets are not known to be a source of G-CSF - which is also elevated in the plasma by MTD PTX - other cellular and/or non-cellular sources are likely affected as well by the drug treatment. We also found that pharmacologic-mediated reductions in platelet numbers or activation results in reduced expression of the counter-acting host cytokine/chemokine response induced by MTD PTX treatment, thus providing further support for a contribution of platelets. Finally we have found low-dose metronomic chemotherapy does not induce the counter acting host response which is likely due in part to a lack of effect on platelets. In summary, our results implicate a new way in which platelets can impact tumor biology and response to therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 382.
Abstract 3827: Interferons induce CXCR3-cognate chemokine production by melanoma
Authors: Lucht Amanda T. , Clancy-Thompson Eleanor , Dengel Lynn T. , Strieter Robert M. , Slingluff Craig L. , Mullins David W.
Published: February 2011

Immune-mediated cancer regression requires tumor infiltration by Ag-specific effector T cells, but lymphocytes are commonly sparse in melanoma metastases. Activated T cells express CXCR3, whose cognate chemokines are CXCL9/MIG, CXCL10/IP-10 and CXCL11/I-TAC. Little is known about expression of these chemokines in lymph node (LN) metastases of melanoma. We evaluated whether metastatic melanoma induces these CXCR3-cognate chemokines in human LN-derived tissues. Also, because these chemokines can be induced by interferon (IFN), we evaluated whether type I or II IFNs (IFN-α or IFN-γ, respectively) can modulate chemokine expression in an in vitro model of the human tumor microenvironment. Production of CXCL9-11 by melanoma-infiltrated nodes (MIN) was no different than tumor-free nodes (TFN); both produced less than activated LN (sentinel immunized nodes, SIN). These data suggest melanoma infiltration into LN neither induces nor reduces CXCL9-11. Stimulation with IFN-α or IFN-γ increased production of CXCL10-11 from MIN, but not TFN or SIN. IFN-γ also increased production of CXCL9 in MIN. In IFN-treated MIN, the primary source of CXCL9-11 was melanoma cells themselves, whereas CD14+ cells were the source of these molecules in SIN. Melanoma cell lines expressed IFN receptors and produced CXCL9-11 in response to IFN treatment. Treatment of melanoma cell lines or primary melanomas in vitro with IFN-γ induced the migration of CXCR3+ T cells in transwell assays, whereas T cell did not significantly migrate in response to resting melanoma cells. Thus, melanoma infiltration of LN is insufficient to induce the production of CXCL9-11, but melanoma may be a significant source of IFN-induced chemokines. Collectively, these data suggest that IFN-α or IFN-γ may act in the tumor microenvironment to increase the chemotactic gradient for CXCR3+ T cells, thus enhancing the capacity of therapeutic vaccines to mediate clinical efficacy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3827.