bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5556: Evaluation of maspin expression in primary tumors and metastasis of melanoma patients: Evidences for prognostic significance
Authors: Martinoli Chiara , Gandini Sara , Luise Chiara , Mazzarol Giovanni , Confalonieri Stefano , Testori Alessandro , Ferrucci Pier Francesco
Published: August 2012

Background: Maspin is a member of the serpin family of protease inhibitors that is thought to act as a tumor suppressor in several cancer types, including melanoma. Recent studies have shown that maspin sub-cellular distribution may impact its effect on disease outcome. Similar studies are missing in melanoma. Methods: Using 3 different tissue microarrays (TMA), Maspin expression was evaluated by two pathologists in a blinded manner on 150 primary lesions and 106 metastasis obtained by melanoma patients. Maspin expression in nuclear and cytoplasmic compartments were evaluated separately and correlated with clinicopathological characteristics (logistic model) and disease-free survival (Cox model) of patients included in the study. Results: Maspin was expressed in nuclei of 52% and in cytoplasms of 17% samples of primary melanomas. Nuclear maspin expression was significantly associated with melanoma thickness (p<0.0001), mitotic rate (p<0.0001), ulceration (p=0.003), LN involvement (p=0.0005) and worse prognosis (p=0.02). On the contrary, higher cytoplasmic expression was observed in patients with thin lesions (p=0.007), with mitotic rate <1/mm2 (p<0.005), no LN involvement (p=0.012), and better prognosis (p=0.05). Metastatic samples showed Maspin expression in the nuclei of 33% cases and in the cytoplasms of 10% cases, showing a progressive lost of expression with disease spreading. Conclusion: Our study shows a compartment-dependent effect of maspin expression in primary and metastatic melanomas. Further studies will be aimed to dissect the biological significance of maspin expression and localization during disease progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5556. doi:1538-7445.AM2012-5556
Abstract 5557: Mutation analysis of melanoma tumor samples from ECOG 2603 clinical trial
Authors: Wilson Melissa Ann , Zhao Fengmin , Letrero Richard , D'Andrea Kurt , Rimm David , Kirkwood John , Kluger Harriet , Lee Sandra , Flaherty Keith , Nathanson Katherine
Published: August 2012

The BRAF kinase, a member of the MAP kinase pathway, has emerged as an attractive molecule to target in melanoma. In an initial attempt to target BRAF in melanoma, a randomized, phase III trial (ECOG 2603) of carboplatin, paclitaxel, and sorafenib versus carboplatin, paclitaxel, and placebo was performed. Patients enrolled on the trial had either unresectable, locally advanced Stage III or Stage IV melanoma. We analyzed patients’ tumor samples for 74 mutations in 13 genes using a custom iPlex (Sequenom). We have preliminarily analyzed 157 tumor samples from patients enrolled in this clinical trial. Analysis of the initial data set demonstrates that 62 samples (42%, 62/148, 95% CI 34%,50%) carried BRAF mutations - 60 samples with V600 and two samples having a BRAF V601 mutation. Thirty eight samples (26%, 38/148, 95% CI 19%,33%) were positive for NRAS mutations, with 33 with Q61, two G12 and three G13 mutations. With the exception of one tumor sample, the BRAF and NRAS mutations were negatively correlated with each other, consistent with prior observations. We also observed a number of rare mutations in our samples including one in AKT1, one in AKT3, four in CDK4, five in beta-catenin, two in GNAQ and three in KIT. The results of this study demonstrated no difference between the two treatment arms in OS and PFS and no association with treatment outcome and BRAF and NRAS mutations. Consequently, the treatment arms were collapsed, and we examined the relationship of BRAF and NRAS mutations with OS and PFS. Interestingly, our data demonstrated that the BRAF and NRAS mutations did not correlate with OS or PFS. This sample population is unique in that it provides a large data set of melanoma tumor samples, all of whom were entered on a clinical trial, unselected for pre-existing mutations. Using the Sequenom assay platform, we have identified a number of mutations in a subset of genes known to be involved in melanoma in patient tumor samples. In the future, these tumor samples will be informative as we continue to examine and identify additional genetic alterations in melanoma, providing information regarding the natural history and distribution of certain mutations. Future studies will use array comparative genomic hybridization to further investigate additional somatic mutations occurring in melanoma tumors and correlate these with clinical outcome, as well as provide insight into the pathogenesis of melanoma. These results may lead to further opportunities for clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5557. doi:1538-7445.AM2012-5557
Abstract 5561: Radiogenomic analysis indicates MR images are potentially predictive of EGFR mutation status in glioblastoma multiforme
Authors: Gevaert Olivier , Mitchell Lex , Xu Jiajing , Yu Caroline , Rubin Daniel , Zaharchuk Greg , Napel Sandy , Plevritis Sylvia
Published: August 2012

Objective: To predict mutations of key genes in glioblastoma multiforme (GBM) from MR image features. Methods: We obtained mutational and MR image data from 35 patients in the Cancer Genome Atlas (TCGA) GBM database. T1-weighted axial images pre and post gadolinium contrast MRI were processed as follows: a board certified neuro-radiologist traced a region of interest (ROI) around the enhanced part of the largest lesion in the T1 post-contrast MRI and confirmed by comparing it with the T1 pre-contrast image. This ROI was then used to compute features that characterized the intensity of the enhanced lesion, the sharpness of lesion boundaries and the boundary shape. We used the resulting data set to build a linear regression model with regularization to predict the presence of a mutation in terms of image features. We focused on predicting mutations in EGFR because several drugs target it and because EGFR mutations are prevalent in the TCGA GBM data. We evaluated the performance of predicting EGFR mutations from MR imaging data using 5-fold cross validation (5F-CV) and the area under the ROC Curve (AUC). The optimal ROC operating point was defined as the point with the maximal sum of sensitivity and specificity when the cost of false positives and false negatives are considered equivalent. Results: We found that computationally-derived MR image features can predict the presence of EGFR mutations with an AUC of 0.80. The optimal operating point had a sensitivity and specificity of 83% and 93% respectively. The top ranked features in the image-based EGFR-predictor model suggest that EGFR-mutated tumors have blurrier edges than EGFR-wild-type tumors. Conclusion: Our preliminary radiogenomic analysis of GBM suggests MR images may be used to non-invasively determine the mutation status of EGFR, an important drug target in glioblastoma. The ability to determine important genomic aberrations based on image data suggests a increasingly important role for imaging in personalized medicine and warrants further investigation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5561. doi:1538-7445.AM2012-5561
Abstract 5568: Constitutively activation of STAT3 is a prognostic factor in activated B-cell subtype of diffuse large B-cell lymphoma
Authors: Huang Xin , Cao Wenfeng , Iqbal Javeed , Meng Bin , Ding B. Belinda , Bi Chengfeng , Jiang Chunsun , Ye B. Hilda , Chan Wing C. , Fu Kai
Published: August 2012

We have previously reported aberrant activation of the signal transducer and activator of transcription 3 (STAT3) pathway promotes cell proliferation and survival in the activated B-cell-like (ABC) subtype, but not in the germinal center B-cell-like (GCB) subtype of diffuse large B-cell lymphoma (DLBCL). Herein, we investigated the effects of small molecular inhibitors of STAT3 in sensitizing DLBCL cell to chemotherapy drugs. ABC-DLBCL lines Ly3 and Pfeiffer were more resistant to vincristine-induced cell death than GCB-DLBCL cells Ly1 and Ly7 were. In ABC-DLBCL cells, constitutive activation of STAT3 provided survival signal and inhibited vincristine-induced apoptosis, while inhibition of STAT3 signaling abrogated IL-6 secretion and promoted cell apoptosis. STAT3 inhibitor and chemotherapy drugs synergistically induced cell apoptosis, with increased caspases 3/7 activity and PARP cleavage, in ABC-DLBCL cells but not in the GCB-DLBCL cells. The effect was correlated with reduced expression of MCL1 in these cells. To investigate the prognostic significance of STAT3 activation in DLBCL patients, we evaluated the levels of phospho-Tyr705-STAT3 (PY-STAT3) expression by immunohistochemistry in a cohort of 309 DLBCL patients treated with R-CHOP regimen. The ABC-DLBCL subgroup had significantly higher number of PY-STAT3 positive cases compared to the GCB subgroup (59.4% vs 36.2%; P = 0.003). Constitutive activation of STAT3 pathway has a significant impact on the overall-survival (OS) and event-free-survival (EFS) in the entire DLBCL cohort (OS, P = 0.010; EFS, P = 0.006) and in the ABC subgroup (OS, P = 0.062; EFS, P=0.016). Moreover, we constructed a subgroup of genes representing STAT3 activation from the gene expression profiling data of STAT3-siRNA treated ABC cell lines and primary DLBCL samples. Attributions of these genes include repression with the STAT3 knockdown in DLBCL cell lines, overexpression in the PY-STAT3-positive DLBCL tumors, and containing one or more STAT3 binding motif within their promoter regions. Eleven genes were obtained when trained this gene subgroup with patient survival through a semi-supervised algorithm. Averaged expression of the eleven genes predicted the OS (P < 0.001) and EFS (P < 0.001) in the entire DLBCL cohort, as well as in the ABC-DLBCL subgroup (OS, P = 0.029; EFS, P = 0.025). In conclusion, our study demonstrated that STAT3 activation is a significant prognostic factor in DLBCL and suggested that STAT3 may be a new therapeutic target in this aggressive malignancy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5568. doi:1538-7445.AM2012-5568
Abstract 5569: DNA methylation status of MLH1, p16, SOCS1, ER and CDH1 in Puerto Rican patients with acute myeloid leukemia
Authors: Tirado-Gomez Maribel , Fonseca Sharon , Munoz Cristina , Cordero Paul , Bernabe Raul , Lacourt Mercedes , Rivera Noridza , Cotto Maribel , Ortiz Eddiemar , Acosta Mirelis
Published: August 2012

Background: DNA methylation of gene promoters have been show to play a role in the pathogenesis of acute myeloid leukemia (AML). We present in this study a preliminary report of the methylation status of MLH1, p16, SOCS1, ER and CDH1 in Puerto Rican patients with AML. Methods: Peripheral blood was obtained from 24 patients with diagnosis of acute myeloid leukemia before and after their induction treatments. DNA was isolated and further modified with sodium bisulfite. Methylation-specific polymerase (MSP) chain reaction was performed to detect aberrant promoter methylation of MLH1, p16, SOCS1, ER and CDH1. Results: Samples from a total of 24 patients were analyzed. The mean age was 47 years (range 22-70) with 21/24 patients younger than 60 years (88%). Most of the patients were female (13/24, 54%). Ten patients had diploid cytogenetics (42%), 4 had low risk cytogenetics (17%) and 10 had high risk cytogenetics (42%). Nineteen patients were de Novo AML (79%). Sixteen patients (67%) presented with WBC counts higher than 10 x 109/L. Seventeen patients (71%) achieved a complete response at the end of the induction therapy. Frequencies of gene methylation before treatment were as follows: MLH: 3/24 (13%); p16: 6/24 (25%); SOCS1:13/24 (54%); ER: 16/24 (67%) and CDH1: 4/24 (16.7%). Conclusions: No correlation between methylation status of p16 and MLH1 before treatment and clinicopathological characteristics was noted. Unmethylated CDH1 was more common among the novo AML patients. Methylation of SOCS1 was more common among patients that achieve a CR after induction therapy. This sample is a small one, so any correlation must be further tested in a more ample sample. Collection of more patient's samples and correlation with clinical data and patient's ultimate outcome is ongoing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5569. doi:1538-7445.AM2012-5569
Abstract 5571: Inflammatory response as a novel prognostic parameter in patients with metastatic renal cell carcinoma treated with pazopanib
Authors: Lingerfelt Brian M. , Akbashev Mikhail Y. , Master Viraj A. , Harris Wayne B.
Published: August 2012

Over sixty thousand patients were diagnosed with metastatic renal cell carcinoma (mRCC) in 2010, yet no tumor markers have been identified. Standard prognostic systems only measure characteristics at baseline and do not assess the potential benefit of therapeutic intervention. The introduction of receptor tyrosine kinase inhibitors has significantly changed the management of mRCC. Pazopanib, a selective, multi-targeted, receptor tyrosine kinase inhibitor, was approved for mRCC in 2009. The modified Glasgow Prognostic Score (mGPS), a prognostic system based on inflammatory biomarkers (serum C-reactive protein (CRP) and albumin), has been validated in the pre-treatment setting for mRCC treated with surgery or cytokines. In this study, we present data demonstrating the correlation of serial mGPS measurements with objective responses to targeted therapy with pazopanib in patients with mRCC. We conducted a retrospective chart review of patients seen at the Winship Cancer Institute for whom serial CRP and albumin measurements as well as imaging studies were available while taking pazopanib. In these patients we assessed the correlation of pre- and post-treatment mGPS scores to objective responses by imaging. Twenty patients met inclusion criteria. Of these patients, 12 (60%) had progressive disease, and 8 (40%) had clinical benefit response (defined as stable disease, partial response, or complete response) by imaging. An elevated pre-treatment mGPS score had 75% sensitivity (95% confidence interval 35.5%-95.5%; P<0.01) and 83.3% specificity (95% confidence interval 50.9%-97.1%; P<0.01) for disease progression at the end of treatment. On the other hand, an elevated post-treatment mGPS score had 100% sensitivity (95% confidence interval 59.8%-100%; P<0.01) and 100% specificity (95% confidence interval 69.9%-100%; P<0.01) for disease progression at the end of treatment. Although these data require prospective validation, they suggest that serial measurements of serum inflammatory biomarkers may improve the sensitivity and specificity of mGPS for response to therapy. If confirmed, inflammatory response rates based on mGPS may become a valuable and cost effective tool to guide patient care and drug development for mRCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5571. doi:1538-7445.AM2012-5571
Abstract 5575: Promoter methylation of SNFP4 gene as potential diagnosis biomarker in cervical cancer: Exploratory study phase I
Authors: Peña Kathleen Saavedra , Mieville Priscilla Brebi , Gangas Carmen Ili , Llaneza Maria Furrianca , Figueroa Paula González , Campos Ángela Riffo , Strauch Juan Carlos Roa
Published: August 2012

BACKGROUND: Cervical cancer (CC) is the second most prevalent neoplasia in women and fifth cause of death by cancer in this population worldwide. Pap smear and HPV infection test have been the most widely used tools to detect cytological abnormalities and women's risk of developing cancer. However, they cannot distinguish between lesions that will progress to an invasive carcinoma and those that will not. Methylation biomarkers could be useful in early detection and progression for cervical carcinoma. Thus, the aim of this study was evaluate the promoter methylation status of a selected gene as possible biomarker for early detection and progression of cervical carcinogenesis in women in the South of Chile. METHODS: DNA isolated from 12 normal samples and 7 cervical cancers was enriched with Methylated DNA Immunoprecipitation (MeDIP) and hybridized to Nimblegen 385K CpG Islands plus Promoter arrays. Bioinformatics strategies were used for background correction, array normalization and data analysis of differentially methylated genomic regions between tumor and normal tissue. The promoter methylation status of selected gene was validated by Methylation Specific PCR (MSP) in biopsies samples of the cervix: 44 normal, 22 Low Grade Lesions (LSIL), 22 High Grade Lesions (HSIL) and 45 cervical cancers (CC). The associations between methylation status of selected gene and different stages of progression of cervical cancer were analyzed using a Chi-square test. FINDINGS: SPRP4 gene was selected from microarray as potential biomarker, because it showed a high score of methylation in cancer, according to bioinformatic analysis. The methylation frequency of SPRP4 through MSP was 0% in normal samples, 50% in LSIL, 27% in HSIL and 100% in tumor samples. Thus, promoter methylation of SPRP4 was associated to cervical cancer and related lesions (p<0.05). INTERPRETATION: The promoter methylation status of SFRP4 has potential use as a biomarker for diagnosis of cervical cancer. Therefore, the validation of this marker in a case-control cohort is warranted. FUNDING: National Cancer Institute grant U01-CA84986; Oncomethylome Sciences, SA, Comisión Nacional de Ciencia y Tecnología (CONICYT; Chile, Becas Chile; Conicyt No24090095; CORFO 07CN13PBT-222). Postdoctoral Fondecyt Research Project 3120141.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5575. doi:1538-7445.AM2012-5575
Abstract 5585: Phase I clinical trial of continuous infusion of tyroserleutide in patients with advanced hepatocellular carcinoma
Authors: Jinbin Jia , Zhen-Yu Xiao , Xiao-Ping Chen
Published: August 2012

Hepatocellular carcinoma (HCC) is usually diagnosed in advanced stage, which causes difficulty of using surgical treatment. Previous studies demonstrated that tyroserleutide (YSL), an immunologically active tripeptide compound, could suppress the proliferation and tumor formation of some liver cancer cell lines. We aimed to investigate the feasibility and toxicity of continuous administration of YSL by a portable infusion pump to patients with advanced HCC and its biologically effective but non-toxic doses used in outpatient setting. Forty patients (12 in stage 1, 28 in stage 2, total 10 treated in each dose cohort) were treated with YSL 6, 12, 18 or 24 mg/d lasting for 5 days. No treatment-related mortality was observed. The overall response rates were 25% (3/12) and 7.2% (2/28) in stages 1 and 2 respectively. The median 6-month overall survival (OS) in stage 1 were 75, 64, and 41 days for 6, 18, and 24 mg/d groups respectively; all patients survived in the 12 mg/d group. The median OS in stage 2 were 68, 72, and 60 days for 12, 18, and 24 mg/d groups respectively; all survived in the 6 mg/d group. The most common adverse reactions were abnormal liver function (59/107) and hemogram (22/107). The dose limiting toxicities of 24 mg/d group contained abdominal distention (1/10), sicchasia (1/10), hyponatremia (1/10), myocardiac ischemia (1/10), and abnormal hemogram (6/10). We conclude that the continuous administration of YSL by portable infusion pump was well tolerated. Treatment responses of doses 6 and 12 mg/d were better than other two groups. Further studies of continuous infusion of YSL to determine its efficacy are warranted. Keywords tyroserleutide • hepatocellular carcinoma • clinical trial • portable infusion pump [Supported by China Postdoctoral Science Foundation-funded project (No. 20080440077 and 200902203)]

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5585. doi:1538-7445.AM2012-5585
Abstract 5593: Mechanisms of acquired crizotinib resistance in ALK-rearranged lung cancers
Authors: Katayama Ryohei , Shaw Alice T. , Khan Tahsin M. , Mino-Kenudson Mari , Iafrate Anthony J. , Engelman Jeffrey A.
Published: August 2012

ALK fusion oncogenes represent a novel molecular target in a small subset of non-small cell lung cancers (NSCLC). The vast majority of patients with ALK-positive NSCLC are highly responsive to ALK tyrosine kinase inhibitor (TKI) therapy. However, these patients invariably relapse, typically within one year, due to the development of resistance. Herein, we report findings from the first series of patients with acquired resistance to the ALK TKI crizotinib. In approximately one-third of patients, we identified a diverse array of secondary mutations distributed throughout the ALK TK domain. We also identified aberrant activation of other kinases as alternative mechanisms of crizotinib resistance. Additionally, we generated laboratory models of acquired crizotinib resistance which closely recapitulate the diversity of resistance mechanisms observed in patients. Our results highlight the marked heterogeneity of TKI resistance mechanisms in ALK-positive NSCLC, and provide the rationale for pursuing combinatorial therapeutic strategies in patients who relapse on crizotinib.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5593. doi:1538-7445.AM2012-5593
Abstract 5594: ALK-driven lung cancer: Potential therapeutic strategies for treatment and prevention of drug resistance
Authors: Pilling Amanda B. , Le Anh T. , Tan Aik-Choon , Kim Jihye , Singleton Katherine R. , Heasley Lynn E. , Helfrich Barbara , DeGregori James , Doebele Robert C.
Published: August 2012

The identification of oncogenic molecular drivers in non-small cell lung cancer (NSCLC) has allowed biologically targeted therapies to improve clinical outcomes in the treatment of NSCLC. ALK gene rearrangements are observed in a sub-set of NSCLC patients who demonstrate high response rates to treatment with the oral kinase inhibitor, crizotinib. Unfortunately, disease progression is inevitable, either through intrinsic or acquired resistance. A recently completed series of crizotinib-resistant, ALK+ NSCLC patients by our group demonstrates two broad categories of resistance: (1) continued reliance on ALK signaling (via secondary mutations in the ALK kinase domain or ALK gene fusion copy number gain) or (2) loss of reliance on ALK signaling (e.g. via reliance on alternate oncogene signaling). Here we investigate potential therapeutic strategies when ALK signaling is retained. In an effort to minimize drug resistance to crizotinib in ALK+ NSCLC, we performed a genome-wide shRNA synthetic lethal screen to identify genes whose depletion synergizes with crizotinib in ALK+ NSCLC lines. We identified several genes involved in nucleotide synthesis and DNA metabolism, including dihydrofolate reductase (DHFR) and the trifunctional enzyme encoded by GART, which are substrates for inhibition with pemetrexed. Indeed, ALK+ NSCLC cell lines show increased sensitivity to pemetrexed in vitro. Identification of this cellular process as critical for ALK+ NSCLC is also consistent with clinical data demonstrating that patients with ALK+ NSCLC exhibit an increased clinical benefit on pemetrexed compared to other molecular subgroups of NSCLC. We also identified genes involved in chaperone function in our synthetic lethal screen. As such, we investigated whether proper folding and prevention of degradation was essential for continued reliance on ALK signaling. Both non-mutated EML4-ALK wild-type and EML4-ALK containing kinase domain mutations (including a novel mutation, G1269A) are sensitive to HSP inhibition with 17AAG. By identifying critical cellular processes in ALK+ lung cancer and employing therapies that target these pathways, improved treatment strategies can be derived to treat and prevent resistance in this molecular subtype of NSCLC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5594. doi:1538-7445.AM2012-5594
Abstract 56: Stromal fibroblasts in the microenvironment of gastric carcinomas promote tumor metastasis through upregulated TAGLN expression
Authors: Liu Bingya , Yu Beiqin , Li Jianfang , Su Liping
Published: August 2012

Fibroblasts play a significant role in tumorigenesis, tumor progression and metastasis, but their detailed molecular characteristics and clinical significance are unclear. TAGLN is an actin-binding protein played an important role in tumorigenesis. Here, we aimed to the interaction between cancer cells and their microenvironments to study how the fibroblasts isolated from human gastric carcinomas facilitate tumorigenesis through TAGLN. Gene expression profiling and real-time PCR and Western blot in gastric carcinoma samples indicated that TAGLN expression was up-regulated in gastric carcinoma. However, further investigation by IHC shown that TAGLN was expressed in tumor stoma, not in tumor cells. TAGLN expression was promoted mutually by fibroblasts and MKN-45 co-culture. Decreased expression of TAGLN suggested that carcinoma-associated fibroblasts (CAF) can enhance tumor metastasis through upregulated TAGLN by MMP-2 and the FAK pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 56. doi:1538-7445.AM2012-56
Abstract 5607: TLE4 is a key modulator of resistance to mTOR pathway specific inhibition in glioblastoma
Authors: Laks Dan R. , Phillips Tiffany , Paucar Andres A. , Orozco Nicholas , Visnyei Koppany , Mischel Paul S. , Coppola Giovanni , Trinidad Jonathan C. , Burlingame Alma L. , Bradshaw Ralph A. , Kornblum Harley I.
Published: August 2012

Rapamycin, a selective inhibitor of TOR (target of rapamycin), has been used in clinical trials for the treatment of glioma (Cloughesy et al., 2008). However, even though some patients respond temporarily to rapamycin treatment, virtually all recur. The mechanisms underlying this resistance to therapy are unknown. Here, we utilized patient-derived glioblastoma (GBM) neurosphere cultures to examine these mechanisms. GBM cell cultures exposed to chronic rapamycin yield a population of rapamycin resistant cells with an increased malignant phenotype upon removal from treatment, enhanced sphere formation in vitro and enhanced tumorigenesis. We used an unbiased, proteomics-based approach to discover phosphopeptides that were upregulated during rapamycin treatment in vitro. Amongst the peptides that we uncovered was Tle4/Groucho, a global co-repressor and known modulator of several key signaling pathways, including the Wnt and Notch pathways. In an expression microarray, Tle4 was found to be significantly upregulated within chronic rapa treated GBM cells. We show that TLE4 is a key modulator of resistance to mammalian target of rapamycin (mTOR) pathway specific inhibition. While TLE4 knockdown has no clear effect on naive cells, this knockdown promotes sensitivity under chronic rapamycin treatment as evidenced by reduced neurosphere formation. TLE4 mediate rapamycin resistance through activation of the quiescent population. Our results indicate that chronic mTOR specific inhibition imparts TLE4 mediated resistance through both ERK activation and inhibition of mTOR. The identification of this role for TLE4 in rapamycin resistance provides a novel target for mTOR pathway-associated chemotherapeutic agents. References: Cloughesy, T.F., Yoshimoto, K., Nghiemphu, P., Brown, K., Dang, J., Zhu, S., Hsueh, T., Chen, Y., Wang, W., Youngkin, D., et al. (2008). Antitumor activity of rapamycin in a Phase I trial for patients with recurrent PTEN-deficient glioblastoma. PLoS Med 5, e8.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5607. doi:1538-7445.AM2012-5607
Abstract 561: Role of nucleolin in activation of NF-κB signaling with Tip≤, a carcinogenic factor of H. pylori
Authors: Watanabe Tatsuro , Hirano Kazuya , Beppu Masatoshi , Yamaguchi Kensei , Takahashi Atsushi , Fujiki Hirota , Suganuma Masami
Published: August 2012

TNF-α inducing protein (Tip≤), a carcinogenic factor secreted from H. pylori, enters the cell through binding to nucleolin, a receptor of Tip≤ on the cell surface. Internalized Tip≤ induces NF-κB activation and TNF-α gene expression and finally induces tumor promotion in the stomach. Nucleolin is a major protein in nucleoli, but significant amounts of nucleolin localize on the cell surface of several gastric cancer cell lines. We investigate the relationship between induction of both NF-κB activation and TNF-α gene expression and the presence of nucleolin, the latter of which acts as a shuttling protein of various molecules, such as Tip≤, lactoferrin and AS1411, from cell surface to cytosol or nuclei. Lactoferrin, iron-binding glycoprotein with 80 kDa, shows anti-cancer activity and preventive activity of H. pylori infection. AS1411 is a nucleolin targeted DNA aptamer associated with a potent anti-cancer activity. First, we studied the effects of lactoferrin, AS1411, and anti-NUC295 (an antibody recognizing cell surface nucleolin) on Tip≤ incorporation into the cells and Tip≤-induced TNF-α gene expression. Pretreatment of mouse gastric cancer cells (MGT-40) with lactoferrin significantly inhibited the incorporation of Tip≤ and Tip≤-induced TNF-α gene expression. However, pretreatment with AS1411 enhanced the incorporation of Tip≤, and inhibited Tip≤-induced TNF-α gene expression. It is of interest to note that Anti-NUC295 enhanced both the incorporation of Tip≤ and Tip≤-induced TNF-α gene expression. These results suggest that both lactoferrin and AS1411 inhibit tumor promoting activity of Tip≤ mediated through inhibition of NF-κB signaling. Thus we think that nucleolin acts as not only a shuttling protein but also a direct regulator of NF-κB signaling. We discuss how lactoferrin and AS1411 inhibit NF-κB activation induced by Tip≤-nucleolin complex, and how nucleolin regulates the NF-κB signaling pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 561. doi:1538-7445.AM2012-561
Abstract 5613: A specific isoform of Gli1 binds the Gli-binding site of the c-jun and c-fos promoters
Authors: Amable Lauren , Gavin Elaine , Denny Wesley , Meng Erhong , Rocconi Rodney P. , Reed Eddie
Published: August 2012

Gli1 participates in the transcriptional regulation of c-jun, which in turn, participates in the transcriptional regulation of genes in nucleotide excision repair. The c-jun promoter and the c-fos promoter have identical Gli-binding-sites that bind Gli1. C-jun and C-fos form the transcriptionally active heterodimer Activator Protein 1, AP1. AP1, is the positive transcriptional regulator for ERCC1, and other DNA repair proteins. Gli1 is a transcription factor in the Hedgehog pathway, and there are five known isoforms of the Gli1 protein that exist in human cells. We have investigated whether there is a specific isoform of Gli1 that binds to the transcriptional regulatory sequences of c-jun and of c-fos. Detailed studies were performed in cisplatin-resistant A2780-CP70 human ovarian cancer cells. EMSA studies demonstrated the presence of a Gli protein in these cells which bind to the Gli-binding-site, as well as the consensus Gli-binding sequence. Supershift EMSA assays show that Gli1 binds the Gli binding sites of c-jun and of c-fos. Southwestern blot analyses of protein lysates from A2780-CP70 cells demonstrated that only one of the five known Gli1 protein isoforms, the 130 kDa, bind the Gli-specific binding site in the promoter of c-jun and c-fos. No Gli2 protein binds this specific binding site in the c-jun promoter, in these cells. To further confirm the 130 kDa Gli1 isoform was responsible for binding the c-jun promoter, the full-length Gli1 with a C-terminal myc tag was transfected into cells, and the protein products were assessed by immunoprecipitation and Southwestern blot analysis. The transfected full length Gli1-myc generated a 130 kDa protein that binds the Gli1-specific binding site in the promoter of c-jun. The presence of this 130 kDa Gli1 isoform was also documented in: six additional human ovarian cancer cell lines, and ten clinical ovarian tissue samples. Seven were ovarian cancer tissue and three were non-cancer ovarian tissues. The 130 kDa Gli1 isoform was present in all specimens examined, but protein levels were six-fold higher in malignant tissues. We conclude that the 130 kDa isoform of Gli1 bind the Gli-binding site in the promoters of c-jun and c-fos. Therefore, Gli1 may be the transcriptional regulator of c-jun and c-fos, and thereby regulates the transcription of ERCC1 and genes of nucleotide excision repair.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5613. doi:1538-7445.AM2012-5613
Abstract 5614: Tandutinib (MLN518/CT53518) targeted to stem-like cells by inhibiting the function of ATP-binding cassette subfamily G member 2
Authors: Deng Wen , Dai Chun-Ling , Zhao Xiao-qin , Ohnuma Shinobu , Liang Yong-ju , Xiao Zhi-Jie , Zeng Mu-Sheng , Ambudkar Suresh V. , Fu Li-wu , Chen Zhe-Sheng
Published: August 2012

Tandutinib (MLN518/CT53518) is a novel quinazoline-based inhibitor of FMS-like tyrosine kinase 3, platelet-derived growth factor receptor, and KIT. In this study, we evaluated the possible interactions of tandutinib with P-glycoprotein (P-gp/ABCB1), multidrug resistance protein 1 (MRP1/ABCC1), breast cancer resistance protein (BCRP/ABCG2) and MRP4/ABCC4. Our results showed that tandutinib reverses ABCG2-mediated drug resistance in ABCG2-transfected cell lines ABCG2-482-R2, ABCG2-482-G2 and ABCG2-482-T7 and mitoxantrone-induced ABCG2-overexpressing cell line S1-M1-80. On the other hand, tandutinib did not change the sensitivity of all tested ABCG2-overexpressing cell lines and their sensitive parental cell lines to cisplatin (non-substrate of ABCG2) and had no significant reversal effect on ABCB1-, ABCC1- and ABCC4-mediated drug resistance. Tandutinib increased doxorubicin and rhodamine 123 accumulation in S1-M1-80 cells in a dose-dependent manner, but did not change their accumulation in parental S1 cells. Similarly, tandutinib also enhanced [3H]mitoxantone accumulation in ABCG2-transfected cell lines, significantly inhibited mitoxantrone efflux in ABCG2-482-R2 cells, but not in HEK293/pcDNA 3.1 cells. However, tandutinib did not stimulate the ATPase activity of ABCG2 and significantly inhibit the photolabeling of ABCG2 with [125I]Iodoarylazidoprazosin. Tandutinib also did not affect the expression of ABCG2 at mRNA or protein levels. In addition, tandutinib did not block the phosphorylation of Akt and Erk1/2 in S1-M1-80 or S1 cells. Importantly, tandutinib targeted to A549 side population cells and enhanced their sensitivity to mitoxantrone, which was not altered in non-SP cells. Collectively, our results advocate the use of tandutinib as an ABCG2 modulator and stem cell targeted agent to increase efficiency of anticancer drugs in clinic therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5614. doi:1538-7445.AM2012-5614
Abstract 563: Chemoprevention of mammary cancer in MMTV-neu transgenic mice by dietary administration of cruciferous vegetable constituent phenethyl isothiocyanate
Authors: Arlotti Julie A. , Kim Su-Hyeong , Sehrawat Anuradha , Hahm Eun-Ryeong , Powolny Anna A. , Lee Joomin , Beumer Jan H. , Sakao Kozue , Singh Shivendra V.
Published: August 2012

We have shown previously that phenethyl isothiocyanate (PEITC), which is a highly promising cancer chemopreventive constituent of edible cruciferous vegetables (e.g., watercress), inhibits growth of cultured breast cancer cells in association with apoptosis induction. The present study was undertaken to determine the efficacy of PEITC for prevention of breast cancer using MMTV-neu mouse model. PEITC administration in the diet (3 mmol PEITC/kg AIN76A diet) for 29 weeks caused an approximate 27% decrease in incidence of mammary hyperplasia compared with control, but the difference between control and PEITC treatment groups did not reach statistical significance. On the other hand, incidence of palpable tumors was reduced by about 53% by PEITC administration (P= 0.07 by two-sided Fisher's exact test). Moreover, size of microscopic carcinoma lesions was significantly lower in mice fed PEITC-supplemented diet compared with those fed basal diet (P= 0.05 by two-sided Fisher's exact test). Average body weight of the PEITC-fed mice was modestly but significantly higher than that of control mice at earlier time points (weeks 1-7) but this differential was not observed at later time points (weeks 8-29). Diet consumption was comparable for mice placed on basal diet and PEITC-supplemented diet. Mean PEITC concentration in the plasma and tumor tissue was 901 nM (95% confidence interval- 741-1060 nM) and 87 nmol/kg (95% confidence interval- 67-107 nmol/kg), respectively. Expression of HER-2 oncoprotein as well as number of CD31-positive blood vessels in the tumor did not differ between control and PEITC treatment groups. On the other hand, tumors from PEITC treatment group exhibited about 46% decrease in Ki-67 expression in comparison with control (P= 0.01). Consistent with cellular observations, number of TUNEL-positive apoptotic bodies was about 4.4-fold higher in the tumors of PEITC-fed mice as compared with controls (P= 0.01). Pulmonary metastasis was evident in 3/35 control mice but only 1/32 mice placed on PEITC-supplemented diet exhibited microscopic evidence of metastasis. Size of pulmonary metastasis was also smaller in the PEITC treatment group compared with control. The results indicate that PEITC administration in the diet inhibits carcinoma development in MMTV-neu mice without any signs of overt toxicity. This investigation was supported in part by the US PHS grants CA101753 and CA129347 awarded by the National Cancer Institute.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 563. doi:1538-7445.AM2012-563
Abstract 5631: The effect of pomegranate extract dose on prostate cancer growth in a carbohydrate restricted mouse xenograft model
Authors: Abern Michael R. , Masko Elizabeth M. , Wu Chenwei , Gaines Alexis R. , Pizzo Salvatore , Freedland Stephen J.
Published: August 2012

Introduction: Preclinical studies have shown that dietary carbohydrate restriction slows prostate tumor growth in multiple xenograft models in part due to a reduction insulin-like growth factor (IGF) axis signaling. Pomegranate extract (PE) has been shown to slow prostate tumor growth in-vitro and in vivo similarly through this pathway, as well as decrease markers of angiogenesis and inflammation. In this study, we determined the dose-response of PE on prostate tumor growth in the setting of dietary carbohydrate restriction in a slow-growing mouse xenograft model. For this study, we chose a standardized PE (POMx, Pom Wonderful, Los Angeles, CA), at doses which are easily translatable to human equivalents. Materials and Methods: Male athymic nude mice housed 5 per cage (n = 130) were subcutaneously injected with 1 × 10^5 LAPC-4 cells after a 10 day acclimation period. 14 days post-injection, all mice were all placed on an ad-libitum no-carbohydrate ketogenic diet (NCKD: 83% fat, 0% carbohydrate, 17% protein) (day 0). Each cage was randomly assigned to 1 of 4 doses of PE (placebo = 0 mg, low dose = 2.62 mg, intermediate dose = 5.25mg, or high dose = 7.87mg) suspended in 0.2 mL of PBS delivered via oral gavage 5 days per week. These doses are equivalent to 0, 1, 2, or 3 POMx capsules/day consumed by a 70 kg human. Mice will be sacrificed when tumors reach ≥1,000 mm^3. Results: At randomization, there were no differences in median body weight among groups (p = 0.623). Currently, at day 32, overall survival is decreased in the placebo group (hazard ratio 2.26, 95% confidence interval, 0.83 - 6.10, p = 0.109) relative to the low dose PE group with no survival differences among PE groups. Median tumor volumes for the placebo group are 83.3mm^3, compared to 74.5mm^3, 76.3mm^3, and 70.1mm^3 for the low, intermediate, and high PE doses respectively, which are not statistically different (p = 0.98). Conclusions: Preliminary data show a trend toward improved survival with the combination of an NCKD and PE at the selected doses. Currently, tumor volumes are not significantly different, however completed results will be presented.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5631. doi:1538-7445.AM2012-5631
Abstract 5639: Pro-drug forms of triterpenes with anticancer activity
Authors: Dzubak Petr , Vagnerova Hana , Frydrych Ivo , Konecny Petr , Rehulka Jiri , Janostakova Anna , Galandakova Adela , Hajduch Marian
Published: August 2012

Number of patients with cancer is increasing. Therapy of cancer is still extremely demanding with side effects of cytotoxic drugs. Therefore, it is necessary to develop and test new anticancer drugs. One approach, how to eliminate side effects of chemotherapy, is by using chemotherapeutic drugs in their prodrug forms, which have more favourable pharmacological properties in comparison to common drugs. Betulinic acid is one of the triterpenoid compounds with high cytotoxicity against tumor cells but with poore pharmacological properties. Therefore, our goal was to develop derivates of betulinic acid with improved pharmacological properties - better bioavailability and therapeutic effects. We have synthesised 30 pro-drugs of the betulinic acid and optimised methods to study the metabolization and possible activation of the pro-drugs by mice hepatocytes. Subsequently, we determined cytotoxicity of activated derivatives to the tumor cell line CEM (human lymphoblastic leukaemia) by MTT assay. Finally, we have identified, that some of the metabolised pro-drugs of betulinic acid were more (290 x) effective in sub -micromolar concentrations than non-metabolised. Parts of the work on this project were supported by grants from the Czech Ministry of Education (MSM 6198959216, LC07017); and The Grant Agency of Czech Republic (GACR 301/09/P433, 305/09/1216). Infrastructural part of this project (Institute of Molecular and Translational Medicine) was supported from the Operational Programme Research and Development for Innovations (project CZ.1.05/2.1.00/01.0030).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5639. doi:1538-7445.AM2012-5639
Abstract 5651: p53-mediated apoptotic signaling overcomes the resistance to oncolytic adenovirus in human osteosarcoma cells
Authors: Hasei Joe , Sasaki Tsuyoshi , Tazawa Hiroshi , Hashimoto Yuuri , Kunisada Toshiyuki , Urata Yasuo , Ozaki Toshifumi , Fujiwara Toshiyoshi
Published: August 2012

Despite major advances in the treatment of bone and soft tissue sarcomas, about one fourth of the patients show a poor response to conventional therapy, resulting in subsequent recurrence and leading to a poor prognosis. Therefore, the development of a novel therapeutic strategy is required to cure patients with bone and soft tissue sarcomas. We previously developed an oncolytic adenovirus, OBP-301, in which human telomerase reverse transcriptase (hTERT) gene promoter drives viral E1 gene for replication. OBP-301 induces tumor-specific oncolytic cell death in a telomerase-dependent manner. Recently, we revealed that OBP-301 shows cytopathic activity in human bone and soft tissue sarcoma cells. However, some human osteosarcoma cells were less sensitive to cytopathic activity of OBP-301. To enhance the cytopathic activity of OBP-301, we recently developed a novel telomerase-specific oncolytic adenovirus, OBP-702, which expresses tumor suppressor p53 gene. In this study, we investigated the antitumor effect of OBP-702 in OBP-301-sensitive (OST, HOS and U2OS) and OBP-301-resistant (MNNG/HOS and SaOS-2) human osteosarcoma cells. We compared the antitumor effects of OBP-702 and OBP-301 using XTT assay. The 50% inhibiting dose (ID50) value of OBP-702 and OBP-301 for each cell was calculated using cell viability data obtained on day 5 after virus infection. We further evaluated the expression of p53, p21 and cleaved poly (ADP-ribose) polymerase (PARP) proteins using western blot analysis in both OBP-301-sensitive and OBP-301-resistant osteosarcoma cells after infection with OBP-702 or Ad-p53, which is a p53-expressing replication-defective adenovirus. OBP-702 showed more cytopathic activity compared to OBP-301 in both OBP-301-sensitive and OBP-301-resistant osteosarcoma cells. The ID50 value of OBP-702 was lower than that of OBP-301 in all cell lines. OBP-702 induced not only oncolysis, but also apoptotic cell death with the cleavage of PARP. OBP-702 infection induced more profound p53 expression than Ad-p53. However, p53-downstream target p21 proteins were not activated by OBP-702. Taken together, we demonstrated that the p53-expressing oncolytic adenovirus OBP-702 has a much stronger antitumor effect than OBP-301 in human osteosarcoma cells. Oncolytic adenovirus-mediated p53 gene transduction would induce profound apoptosis through p53 upregulation without p21 activation, resulting in the enhancement of antitumor effect by OBP-301. OBP-702 would be a promising antitumor reagent for the treatment of OBP-301-resistant human osteosarcoma cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5651. doi:1538-7445.AM2012-5651