bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3619: The absence of cleaved caspase-3 in circulating tumor cells detected using a non-enrichment-based assay
Authors: Metzner Thomas J. , Bethel Kelly , Cho Edward H. , Luttgen Madelyn S. , Lazar Daniel C. , Uson Maria L. , Nieva Jorge J. , Bazhenova Lyudmila , Kolatkar Anand , Kuhn Peter
Published: August 2012

Background: The role of cleaved caspase-3 as an integral player in the apoptotic process of mammalian cells has prompted the investigation of an assay to detect cleaved caspase-3 in circulating tumor cells (CTCs) as one way to assess CTC viability. Methods: Using a previously developed non-enrichment based assay, nucleated blood cells and CTCs are adhered to a single slide. An antibody for cleaved caspase-3 was incorporated into the assay in addition to the existing antibodies for cytokeratin (to identify tumor cells) and CD45 (to identify white blood cells), as well as a nuclear stain (DAPI). The modified assay was initially tested on cultured MCF 10a cells spiked into normal donor blood. Before spiking, apoptosis was induced in these cells using staurosporine. Following validation of the assay in cell lines, the assay was performed using blood samples from 9 different breast cancer patients, as well as 3 different normal donor samples. Results: CTCs were detected in the patient samples as single cells, as well as clusters. The results of the assay showed that only 1 out of 896 detected CTCs was positive for cleaved caspase-3 (0.11%). Of these CTCs, 134 of them were found to be in clusters, and all 134 were negative for cleaved caspase-3, indicating that whether in clusters or as single cells, the vast majority of CTC's detected with this assay are not undergoing irreversible apoptotic processes via cleaved caspase-3. The results also showed that 0.40% of white blood cells in healthy donors expressed cleaved caspase-3, compared to 0.51% of the white cells in cancer patients, indicating that healthy donors and cancer patients express cleaved caspase-3 in their white blood cells at nearly the same rate. Conclusions: The morphologic characterization used in this platform to select healthy-appearing cells identifies a population of circulating tumor cells with virtually no evidence of caspase-related ongoing apoptosis, a finding that supports the morphologic impression of viability.

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 3619. doi:1538-7445.AM2012-3619
Abstract 362: [18F]-FLT Positron Emission Tomography can be used to image the response of sensitive tumours to PI3-Kinase inhibition with the novel agent GDC-0941.
Authors: Cawthorne Christopher , Wilson Ian A. , Danikas Antonios , Burrows Natalie , Radigois Mark , Gregory Jamil , Smigova Alison , Babar Muhammed , Williams Kaye J.
Published: August 2012

Emerging targeted therapeutics are characterised by high efficacy rates in selected patient sub-populations as well as potentially high running costs. This calls for companion diagnostic tools that contribute to the better management of such therapies, in terms of patient selection and monitoring therapeutic response. The Phosphatidylinositide 3-kinase pathway is deregulated in a range of cancers, and several targeted inhibitors are entering the clinic. This study aimed to investigate whether the PET tracer [18F]-FLT is suitable to mark the effect of the novel PI-3K inhibitor GDC-0941 which has entered Phase II clinical trial. Methods Nude mice bearing U87 and HCT116 xenografts were imaged at baseline with [18F]-FLT (10-15MBq per mouse produced using a GE Tracerlab). GDC-0941 or vehicle was then administered at a concentration of 50mg/kg twice daily by gavage and animals were rescanned at acute (∼18h) and chronic (∼186h) timepoints post-therapy. Standard uptake value (SUV)s for tumour uptake were calculated, and tissue was analysed at sacrifice for phospho-AKT. To generate normalised uptake values (NUV)s the SUV [tumour] was divided by the SUV for heart. NUV figures are given for the 100-105min post-injection time interval. Results Growth of U87 xenografts was significantly inhibited by GDC-0941 treatment throughout the course of the study (average growth rate of treated = –1.0±9.3 mm3, control = 69.2±22.1mm3 per day). Growth of HCT116 xenografts was not significantly different between groups (average growth rate of treated = 34.6±15.9 mm3, control =54.8±11.1 mm3 per day). Tumour uptake of [18F]-FLT was significantly reduced in treated animals bearing U87 xenografts at the acute timepoint compared to baseline (NUVmax 2.17 ± 0.38 vs 1.59 ± 0.29, p<0.01), whereas uptake in HCT116 tumours was unchanged. Analysis of the downstream marker phospho-AKT showed that this was decreased in U87, but not HCT116, tumours. Conclusion [18F]-FLT is a strong candidate for the non-invasive measurement of GDC-0941 action. In addition to FLT, we are assessing the performance of a range of other imaging agents, including proprietary tracers, that can potentially bring value in monitoring of PI3-kinase targeting therapies.

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 362. doi:1538-7445.AM2012-362
Abstract 3624: Circulating sEGFR arises through a fibronectin/integrin-regulated proteolytic cleavage event: Analogy to HER2 shedding
Authors: Wilken Jason A. , Perez-Torres Marianela , Nieves-Alicea Rene , Cora Elsa M. , Christensen Trace A. , Baron Andre T. , Maihle Nita J.
Published: August 2012

A circulating isoform of EGFR has been identified as an emerging serum biomarker in cancer patients. Serum concentrations of EGFR have been evaluated as screening, diagnostic, prognostic, and theragnostic markers for diverse human malignancies, including breast, ovarian, endometrial, colorectal, and lung cancer. Yet full-length EGFR is rarely shed from normal or malignant cells. Here we demonstrate that an alternately spliced EGFR isoform, termed ‘sEGFR,’ gives rise to the circulating sEGFR protein through proteolytic cleavage of a cell surface precursor. This cleavage event is inhibited by fibronectin and stimulated by both an anti-Δ5/α1 integrin antibody and the metalloprotease activator 4-aminophenylmercuric acetate. Preliminary results identify ADAM17/TACE as at least one mediator of sEGFR shedding. Shed sEGFR binds to EGF with high affinity. Two FDA-approved therapeutic anti-EGFR antibodies, cetuximab (Erbitux) and panitumumab (Vectibix), inhibit shedding of sEGFR, implicating sEGFR as a potential alternative target for these antibodies, consistent with previous studies indicating that baseline serum sEGFR concentrations may predict responsiveness to anti-EGFR-targeted therapy. These observations bear striking similarity to aspects of trastuzumab regulation of HER2, shedding, and, by analogy, may have implications for the design of future clinical trials using EGFR-directed therapeutics, including cetuximab and panitumimab.

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 3624. doi:1538-7445.AM2012-3624
Abstract 3643: Early detection of cancer in the general population - a blinded case control study of p53 auto-antibodies in colorectal cancer in UK Collaborative Trial of Ovarian Cancer Screening (UKCTOCS)
Authors: Pedersen Johannnes W. , Gentry-Maharaj Aleksandra , Fourkala Rania , Dawnay Anne , Burnell Matthew , Zaikin Alexey , Pedersen Anders , Jacobs Ian , Menon Usha , Wandall Hans H.
Published: August 2012

Purpose: Recent reports from cancer screening trials in high-risk populations suggest that autoantibodies are present prior to clinical diagnosis. However there are few data on the role of autoantibody signatures in cancer screening in the general population. Methods: Informative p53 peptides were identified on an autoantibody microarray with 15-mer overlapping peptides covering the complete p53 sequence using sera from patients with colorectal cancer. The selected peptides were evaluated in a blinded case control study using stored serum from the multimodal arm of UKCTOCS where women gave annual blood samples. Cases were postmenopausal women who developed colorectal cancer following recruitment, with 2 or more serum samples preceding diagnosis. Controls were age-matched women with no history of cancer. Results: The 50,640 women randomised to the multimodal group were followed up for a median of 6.8 (inter-quartile range 5.9-8.4) years. Colorectal cancer notification was received in 101 women with serial samples of whom 97 (277 samples) had given consent for secondary studies. They were matched 1:1 with 97 controls (276 serial samples). The four most informative peptides identified 25.8% of colorectal cancer patients with a specificity of 95%. The median lead time was 1.4 (range 0.12-3.8) years prior to clinical diagnosis. Conclusion: Our findings suggest that in the general population, autoantibody signatures are present during preclinical disease and may be of value in cancer screening. In colorectal cancer screening in particular, where the current need is to improve compliance, it suggests that p53 autoantibodies may contribute towards risk stratification.

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 3643. doi:1538-7445.AM2012-3643
Abstract 3650: Serous ovarian carcinoma patients with high alpha-folate receptor had poorer outcome by reducing cytotoxic chemo-response through anti-apoptotic pathway
Authors: Huang Ching-Ting , Chen Yu-Li , Chang Ming-Cheng , Huang Chia-Yen , Chiang Ying-Cheng , Cheng Wen-Fang
Published: August 2012

The alpha-folate receptor (α-FR) is highly expressed in various non-mucinous tumors of epithelial origin, including ovarian carcinoma. Semi-quantitative reverse-transcription polymerase chain reactions for α-FR expression were performed in 91 specimens of serous ovarian carcinomas. Furthermore, in vitro apoptotic experiments were tested in the original OVCAR-3 tumor cells and various OVCAR-3 α-FR-transfectants. In our study, the serous ovarian cancer patients with advanced stages (stage I: 0.451, stage II: 0.415, stage III: 0.652, stage IV: 0.768, p=0.004) had a significantly higher α-FR expression compared to those with early stages. Patients with an increased α-FR expression level had poorer responses to chemotherapy (odds ratio (OR): 8.97 (95% confidence interval (CI): 1.40-57.36), p=0.021). An increased α-FR expression level was an independently poor prognostic factor for DFI (Hazard ratio (HR): 2.45 (95% CI: 1.16-5.18), p=0.02) and had a negative impact on OS of these serous ovarian cancer patients (HR: 3.6 (95% CI: 0.93-13.29), p =0.03) by multivariate analyses. α-FR inhibited cytotoxic drug-induced apoptosis in our in vitro apoptotic assays. α-FR could induce chemo-resistance via regulating the expression of apoptosis-related molecules, Bcl-2 and Bax. Therefore, α-FR can be a potential biomarker for the prediction of chemotherapeutic responses and clinical prognosis. It also could be the target of treating ovarian cancer.

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 3650. doi:1538-7445.AM2012-3650
Abstract 3651: Foxp3 expression correlates with survival in urothelial carcinoma of the bladder
Authors: Zhang Hanwei , Wang Xiaoyan , Seligson David , Pantuck Allan , Chin Arnold I.
Published: August 2012

Introduction and Objective The transcription factor Forkhead box p3 (Foxp3), originally described during the development of CD4+CD25+ T regulatory cells, has recently been implicated in divergent roles in human prostate, breast, pancreatic cancer, and melanoma. Here, we investigate the inducibility of Foxp3 in bladder cancer models and assess the prognostic role of Foxp3 in patients with bladder cancer. Methods Foxp3 expression in the human bladder cancer cell line SW780 was examined in in vitro cultures by flow cytometry and in in vivo subcutaneous implanted tumor models in NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice by immunohistochemistry. Proliferation and invasion of SW780 cells in the absence of Foxp3 was performed by transfection of siRNA in vitro. Correlation of Foxp3 expression to urothelial carcinoma grade, stage, recurrence-free survival, and overall survival was examined in a human tumor microarray from a cohort of 280 patients by Cox proportional hazards regression and Kaplan-Meier analysis. Results Foxp3 expression correlates to higher cellular proliferation in vitro and in vivo. Knockdown of Foxp3 in SW780 cells resulted in decreased proliferation as well as invasion in vitro. Foxp3 expression increases with higher T stage, but not grade, as expression in adjacent normal bladder, Tis, Ta-1, T2-4, and lymph node metastasis was 1.2%, 3.8%, 15.2%, 26.5%, and 23.4% respectively, suggesting higher expression in muscle invasive and lymph node metastatic disease, but not carcinoma in situ. Using a cutoff of 10%, Foxp3 expression negatively correlates with survival following radical cystectomy with a median survival of 9.3 vs 2.8 years and a 10-year survival probability of 47% vs 27%. Conclusions This work implicates Foxp3 as novel transcription factor driving proliferation and tumor invasion in urothelial carcinoma. Expression of Foxp3 correlates with T stage and is a novel predictor of overall survival following radical cystectomy. Future studies will need to understand the mechanistic regulation of Foxp3 expression, as well examine Foxp3-mediated gene expression in urothelial carcinoma. Foxp3 expression may improve selection of patients to 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 3651. doi:1538-7445.AM2012-3651
Abstract 3661: Notch2 and Six1 are upregulated during progression of early-stage lung adenocarcinoma and define its unfavorable subset at advanced stages
Authors: Mimae Takahiro , Okada Morihito , Hagiyama Man , Miyata Yoshihiro , Tsutani Yasuhiro , Inoue Takao , Aogi Kenjiro , Murakami Yoshinori , Ito Akihiko
Published: August 2012

Lung adenocarcinoma (LA) often manifests as tumors with mainly lepidic growth (LG) pattern in which cancer cells replace alveolar cells, and is diagnosed, depending on the size of invasive foci, as in situ, minimally invasive, or invasive LA. This suggests that a subset of LAs undergoes malignant progression in this order. The present study examines how transcriptional aberrations during the early-stage of LA cells define the clinical phenotypes of advanced tumors. First, we comprehensively searched differentially expressed genes between preinvasive and invasive cancer cells in a single minimally invasive LA using laser capture microdissection and DNA microarrays, and successfully identified two transcription factors Notch2 and Six1 to be up-regulated in the invasive cancer cells from all of five minimally invasive LA tumors. These two molecules appeared to coordinately contribute to LA progression by inducing epithelial-mesenchymal transition and nuclear atypia, because exogenous Notch2 in NCI-H441 lung epithelial cells caused nuclei to enlarge and transactivated Six1, vimentin, and the TGF-β signal transducers Smad3 and Smad4. We then immunohistochemically stained 64 advanced LA tumors, and found that a third of them were double positive for Notch2 and Six1 in the invasive, but not in the LG component. The invasive potential was higher and 2-year disease-free survival rates were lower in tumors that were double-positive, than double negative in both components. These results suggested that paired up-regulation of Notch2 and Six1 during early LA progression persists in a considerable subset of advanced LAs and confers a more malignant phenotype on the subset.

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 3661. doi:1538-7445.AM2012-3661
Abstract 3668: Inhibition of HSP90 with NVP-AUY922 induces a synergistic effect in HER2-amplified, trastuzumab-resistant breast and gastric cancer
Authors: Wainberg Zev A. , Rogers Amy , O'Brien Neil , Anghel Adrian , Desai Amrita , Kalous Ondrej , Conklin Dylan , Akimov Mikhail , Finn Richard S. , Slamon Dennis J.
Published: August 2012

Background: Heat Shock Protein 90 (HSP90) enables the activation of a large number of client proteins of which the most clinically validated is HER2. NVP-AUY922 is a a highly potent, non-geldamycin HSP90 inhibitor that has shown preliminary activity in HER2 positive metastatic breast cancer. To explore its potential clinical utility specifically in HER2 amplified gastric and breast cancers, we evaluated the effect of AUY922 alone and in combination with trastuzumab in both trastuzumab-sensitive and resistant in vitro and in vivo models. Methods: A panel of 16 human gastric and 45 breast cancer cell lines, with 3 and 13 HER2-amplified lines respectively, were exposed in vitro to AUY922 over various concentrations to generate dose response curves. In both breast and gastric cancer, we used cell lines and xenograft models with conditioned trastuzumab-resistance to investigate the efficacy of AUY922 alongside trastuzumab. In all in vivo experiments, trastuzumab was dosed at 10 mg/kg (IP; twice/week) and AUY922 at 50 mg/kg, (IP; 5 days on, 2 days off). Effects of this combination on downstream markers (HER2, AKT, ERK) were analyzed via western blot. Results: AUY922 was found to have potent anti-proliferative activity in the low nanomolar range (< 40 nM) for 59 of the 61 gastric and breast cancer cell lines tested. Additionally, in both gastric and breast cancer, HER2-amplified cells expressed greater sensitivity to the compound when compared to the HER2-negative cells. In conditioned trastuzumab-resistant cell line models, AUY922 continued to have potent activity and exhibited a synergistic effect with trastuzumab. The in vitro combination also induced greater decreases in HER2, induced a G0/G1 cell cycle arrest, and increased the rates of apoptosis. In a conditioned trastuzumab-resistant gastric cancer in vivo model (N87-Res), the combination of AUY922 and trastuzumab showed greater anti-tumor efficacy than either drug alone. Conclusions: These data suggest that AUY922 in combination with trastuzumab has unique efficacy in trastuzumab-resistant models. The combination of HSP90 inhibitors and trastuzumab represents a novel approach to the treatment of HER2 amplified cancers. Clinical trials combining AUY922 and trastuzumab in HER2 amplified breast and gastric cancers are currently 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 3668. doi:1538-7445.AM2012-3668
Abstract 3673: Molecular analysis of pheochromocytoma after maternal transmission of SDHD mutation elucidates mechanism of parent-of-origin effect.
Authors: Tobias Edward S. , Yeap Phey M. , Mavraki Eleni , Fletcher Alexander , Freel Marie E. , Cooke Alexander , Bradshaw Nicola , Murday Victoria A. , Davidson Rosemarie , Perry Colin , Lindsay Robert S.
Published: August 2012

In SDHD mutation families, paragangliomas and pheochromocytomas usually occur only after paternal transmission of the mutation. This important but unexplained parent-of-origin effect is not due to imprinting of SDHD itself, as was initially suspected, since SDHD is biallelically expressed in several tissues. In clinically affected individuals who possess a paternally inherited SDHD mutation, there is loss of the entire maternal chromosome 11 in tumour DNA, implying that tumorigenesis requires loss of not only maternal (wild type) SDHD but also a further, imprinted, tumor suppressor gene (TSG). We report the second case of an SDHD-related tumor (a pheochromocytoma in a 33 year old woman possessing the common pathogenic mutation, p.Pro81Leu) occurring after maternal transmission. It is the first reported investigation of tumor DNA in this situation. Tumor DNA revealed loss of heterozygosity (LOH) at paternal 11q23 causing loss of the wild-type SDHD allele and also LOH affecting maternal 11p15, including H19. These two LOH regions were separated by a region exhibiting clearly retained heterozygosity, containing SDHAF2 (a recently reported paraganglioma TSG), which therefore appears uninvolved here. This case provides strong molecular evidence that the tumorigenic requirement for maternal 11p15 loss (in addition to inactivation of both SDHD alleles) drives the observed parent-of-origin effect. Thus, SDHD-related tumorigenesis most likely involves a “three-hit” mechanism that includes (as one of the hits) loss of an imprinted (paternally silenced and maternally active) TSG from chromosome 11, such as H19). Tumor formation more commonly results from paternal inheritance of SDHD mutations, as the necessary loss of both the wild type SDHD allele and maternal 11p15 can then occur by a single event (loss of maternal chromosome 11). These findings have important implications regarding the clinical management of carriers of maternally inherited SDHD mutations, who we confirm can develop pheochromocytomas, and the understanding of the parent-of-origin effect.

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 3673. doi:1538-7445.AM2012-3673
Abstract 3676: The hypoxia-activated pro-drug tirapazamine suppresses outgrowth of colorectal liver metastases following radiofrequency ablation
Authors: Govaert Klaas M. , Nijkamp Maarten W. , Emmink Benjamin L. , Steller Ernst J.A. , Minchinton Andrew I. , Kranenburg Onno , Rinkes Inne H.M. Borel
Published: August 2012

Background Radiofrequency ablation (RFA) is a common procedure for the management of colorectal liver metastases. RFA-generated lesions are surrounded by a rim of hypoxia that is associated with aggressive outgrowth of intrahepatic micrometastases. Hypoxia-activated pro-drugs such as tirapazamine (TPZ) are designed to selectively kill tumour cells under hypoxic conditions. Therefore we hypothesised that TPZ may have therapeutic value in limiting hypoxia-associated tumour outgrowth following RFA. Methods Murine C26 and MC38 colorectal cancer cells were grown under hypoxia and normoxia in vitro and were treated with different concentrations of TPZ. Apoptosis and cell cycle distribution were assessed by western blot and FACS analysis. Proliferative capacity was tested by colony formation assays. Mice harbouring microscopic colorectal liver metastases were treated with RFA, followed by a single injection of TPZ (60 mg/kg) or saline. Tumour load was assessed morphometrically 7 days later. Results TPZ induced apoptosis of colorectal tumour cells under hypoxia in vitro, which completely abolished clone-forming capacity. Under normoxia, TPZ caused a G2 cell cycle arrest from which cells recovered partly. This reduced, but did not abolish, colony-forming capacity. A single dose of TPZ largely prevented accelerated outgrowth of hypoxic micrometastases following RFA. TPZ administration was associated with minimal toxicity. Conclusions TPZ kills colorectal cancer cells in a hypoxia-dependent manner and potently suppresses hypoxia-associated outgrowth of liver metastases with limited toxicity. This warrants further studies to assess the potential value of TPZ, or other hypoxia-activated pro-drugs, as adjuvant therapeutics following RFA treatment of colorectal liver metastases.

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 3676. doi:1538-7445.AM2012-3676
Abstract 3680: Drug screening for glioblastoma patients
Authors: Jiang Pengfei , Mukthavaram Rajesh , Summitt IlaSri , Chao Ying , Gallagher Matthew , Kim Ryan , Pastorino Sandra , Kesari Santosh
Published: August 2012

The modest benefits of chemotherapy for Glioblastoma (GBM) patients’ makes it urgent to find potent drugs or combinations that can inhibitor tumor cell growth and proliferation. We have devised drug screening based on the NIH clinical collection compounds. This collection included 446 compounds that all have history of use in human clinical trials. The positive compounds can be rapidly entered into clinical trials for GBM. After found 22 drugs/compounds that can inhibit U87 cell proliferation and we tested 15 available positive drugs with A172, LN443, U118 cell line. After confirming in established serum-grown cell lines, we tested 8 FDA approved drugs in GBM primary cells with stem cell like features isolated from fresh brain tumor tissue samples and cultured in stem cell media. The various drug responses amongst the different patients’ primary cells suggested that we may be able to personalize drugs to optimize outcomes. In addition, we found that certain combinations of FDA approved drug had significantly enhanced activity against GBM cells in vitro when compared to the same agents used alone. Our work reveals that screening drugs and combinations for GBM patient may have clinical implications.

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 3680. doi:1538-7445.AM2012-3680
Abstract 3682: Overexpression and cell proliferation-regulating function of CRKL protein in gastric cancer
Authors: Natsume Hiroko , Shinmura Kazuya , Tao Hong , Suzuki Masaya , Goto Masanori , Yamada Hidetaka , Maeda Matsuyoshi , Konno Hiroyuki , Nakamura Satoki , Sugimura Haruhiko
Published: August 2012

Genomic DNA amplification is a genetic factor involved in cancer, and some oncogenes, such as ERBB2, are highly amplified in gastric cancer. To identify previously undiscovered amplified genes in gastric cancer, a genome-wide single nucleotide polymorphism microarray analysis was performed using three cell lines of differentiated gastric cancers. Twenty-two genes (including ERBB2) in five highly amplified chromosome regions (with a copy number of more than 6) were identified. Particular attention was paid to the CRKL gene, which has not been previously characterized in gastric cancer and the product of which is an adaptor protein containing Src homology 2 and 3 (SH2/SH3) domains. A relatively high CRKL copy number was confirmed in the MKN74 gastric cancer cell line using fluorescence in situ hybridization, and a high level of CRKL expression was also observed in the cells. The RNA-interference-mediated knockdown of CRKL in MKN74 disclosed the ability of CRKL to upregulate gastric cell proliferation. An immunohistochemical analysis revealed that CRKL protein was overexpressed in 24.4% (88/360) of the primary gastric cancers that were analyzed. Finally, we showed that MKN74 cells with CRKL amplification were responsive to the dual Src/BCR-ABL kinase inhibitor BMS354825, likely via the inhibition of CRKL phosphorylation, and that the proliferation of MKN74 cells was suppressed by treatment with a CRKL-targeting peptide. These results suggested that CRKL protein is overexpressed in a subset of gastric cancers, that it regulates gastric cell proliferation, and that it has the potential to be a molecular therapy target for gastric cancer.

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 3682. doi:1538-7445.AM2012-3682
Abstract 3694: Role of the inflammatory response in pancreatic cancer treatment
Authors: Cantoria Mary Jo , Dragovich Tomislav , Meuillet Emmanuelle
Published: August 2012

Pancreatic adenocarcinoma is an aggressive malignancy, often diagnosed late in the course of the disease, and very difficult to diagnose and treat. To date, systemic treatment for pancreatic cancer, either a single agent or combinations of agents, has shown only modest benefits; however, growing understanding of the molecular pathogenesis of pancreatic cancers, and the complex heterogeneous signals associated with them, may help to target these pathways specifically and improve patient outcome. There is evidence for a significant role of cyclooxygenase (COX)-2 and COX-2-generated prostaglandin (PG) E2 (PGE2) during pancreatic tumorigenesis. COX-2 has therefore become a potential chemotherapeutic target for pancreatic cancer. Clinical trials combining Gemcitabine and Celecoxib [1] or Gemcitabine+ Irinotecan and Celecoxib [2] have not shown a significant benefit from celecoxib but there were relatively few patients treated and they were not selected or stratified based on biomarkers. We have measured the levels of 4 cytokines, including Interleukin (IL)-6, IL-1α, Tumor Necrosis Factor (TNF)-α and Interferon (IFN)-α in patient blood samples at day 0, day 29 and day 50 following the combination Gemcitabine - Celecoxib treatment. For all samples analyzed and averaged, we show an increase in plasma TNF-α (by 1.37 fold) and IL-1α (by 2 fold) concomitantly with a dramatic increase in IFN-γ (by 7 fold on day 29, by 3 fold on day 50) and stable IL-6 using the Meso Scale Discovery multiplex platform. One patient had a stable profile of all 4 cytokines and was shown to have partial response to the treatment. All others exhibited a strong variation in their cytokines during the treatment and did exhibit progression of the disease. Although the number of patients enrolled in the trial [1] was low, these results showed an interesting trend. These findings provide a basis for investigating further the inflammation-linked pathways along the COX2/mPGE-S1/EP4/IL-6 axis in pancreatic cancer cells and animal models of pancreatic cancers which will be subjected to the comparative treatment. [1]. Dragovich et al., Am. J. Clin. Oncol. 31:157-162 (2008) [2]. Lipton et al., J. Clin. Gastroenterol. 44:286-288 (2010)

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 3694. doi:1538-7445.AM2012-3694
Abstract 3698: ODSH, a heparin derivative, enhances the efficacy of gemcitabine in a refractory human pancreatic tumor xenograft model
Authors: Marcus Stephen G. , Quintana-Diez Pedro M. , Gately Stephen , Gonzales Paul , Chavira Bernardo , Von Hoff Daniel D.
Published: August 2012

ODSH, a new chemical entity, is 2-0, 3-0 desulfated heparin, a desulfated heparin analog that has been shown to bind to the receptor for advanced glycation endproducts (RAGE). This interaction prevents RAGE from binding to a principal ligand, HMGB1, as well as to other ligands including S-100 calgranulin, which attenuates the activation of RAGE. RAGE activation is known to induce resistance to chemotherapy and promote pancreatic tumor cell survival. We have shown, in vivo, an increase in response with gemcitabine when combined with ODSH. ODSH was tested alone and in combination with standard of care agent, gemcitabine, against BxPC-3 human pancreatic tumor xenograft model. Single agent ODSH and gemcitabine treatment showed an 18.8% and 16.8% decrease in tumor weight, respectively. The combination regimen of ODSH and gemcitabine increased tumor growth inhibition to 37.9%. This decrease was statistically significant (P<0.05) compared to ODSH and gemcitabine treatments alone according to Student's t-test. We believe this is a novel approach to treat metastatic pancreatic cancer. ODSH has already shown a favorable safety profile in Phase I and II studies. A clinical trial with ODSH treatment in combination with gemcitabine+nab-paclitaxel in newly diagnosed patients with metastatic pancreatic cancer is underway.

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 3698. doi:1538-7445.AM2012-3698
Abstract 3699: Adnab-9: A prospective stool biomarker for pancreatic ductal adenocarcinoma
Authors: Weinstein Douglas , Rambus MaryAnn , Levi Edi , Yuan Mei J. , Lawson Michael J. , Tobi Martin
Published: August 2012

Sporadic pancreatic ductal adenocarcinoma (PDA) is a highly lethal cancer and with no proven screening strategies. Adnab-9, a monoclonal antibody to constituents of human colonic adenomas has been shown to be prognostic in IPMN and diagnostic in gastric and colorectal cancer or adenomas (CRN). In this study, we retrospectively and prospectively evaluated the diagnostic potential of Adnab-9 in the stool of two distinct PDA populations. Methods: As proof of principle, 249 Chinese patients submitted stool for screening. Retrospectively, 15 patients had PDA and 80 patients without cancer served as controls. Adnab-9 stool reactivity was determined by ELISA. Prospectively, 1132 American CRN screening program patients submitted stool. Six patients developed PDA over five years of follow-up. 11 Patients free of any significant CRN or other cancer but deceased due to other causes, served as controls. An additional four patients with a known family history of PDA also prospectively submitted stool for Adnab-9. Results: The prospective study results are summarized in the Table. Table: Prospective Study Data for Adnab-9 stool antibody as a biomarker of PDA. The pilot study in Chinese PDA patients found that 12 of 15 patients had stool Adnab-9 ELISA positivity compared to 10 of 80 controls (OR 28, CI 6.7-116.8, P<0.0001). 4 of 6 prospective American study PDA patients had either stool ELISA or Western blot positivity for Adnab-9 compared to 1 of 11 controls (OR=20, CI 1.4-287.6, P<0.03). 2 of 4 patients with family history of PDA had stool positive by Adnab-9 ELISA or Western blot. Conclusions: Our study suggests that Adnab-9 stool tests may be an effective PDA diagnostic marker. Based on previous data in CRC and IPMN, Adnab-9 appears to bind Paneth-like cells, which we and others have shown to be cancer-associated and the likely source of the stool antigen. Future prospective studies are needed to confirm the diagnostic efficacy of Adnab-9 stool testing in PDA.

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 3699. doi:1538-7445.AM2012-3699
Abstract 3704: Clinical efficacy and safety of bevacizumab monotherapy in patients with metastatic melanoma: predictive importance of induced early hypertension in a single-arm Phase II study
Authors: Straume Oddbjorn , Schuster Cornelia , Puntervoll Hanne , Eikesdal Hans P. , Heinrich Daniel , Geisler Stephanie B. , Geisler Jurgen , Molven Anders , Lønning Per E. , Akslen Lars
Published: August 2012

Purpose VEGF driven angiogenesis plays a key role in tumor growth. We determined clinical efficacy of bevacizumab monotherapy in unselected patients with metastatic melanoma. Patients and methods Thirty-five patients with metastatic melanoma in progression were enrolled in this phase II, single arm clinical trial. Each patient received bevacizumab monotherapy 10 mg/kg q14 d until intolerable toxicity or disease progression occurred. Clinical efficacy was evaluated by measuring objective response (OR), disease control (DC), and survival. We also evaluated potential predictive value of clinical parameters and BRAF/NRAS mutational status. Results We observed one complete (CR, 3%) and 5 partial (PR, 14%) responses. In addition, 5 patients experienced stable disease > 6 months (SD, 14%), while 24 patients had progressive disease (PD, 69%), corresponding to a total disease control (DC) at 6 months in 11 out of 35 patients (31 %). Median progression free survival (PFS) was 2.14 months and median overall survival (OS) was 9 months (1.12-49). Seven of the 11 patients experiencing DC developed early hypertension (< 2 months) after commencing bevacizumab compared to 3/24 of patients with PD (P= 0.001), and hypertension was associated with improved progression free survival (P= 0.005) and overall survival (P= 0.013). No correlation between BRAF and NRAS mutation status and response was recorded. Conclusion Bevacizumab monotherapy demonstrated promising clinical efficacy in patients with metastatic melanoma with disease control in 31% of the patients. The activity of the drug seemed particularly pronounced in patients developing by hypertension early during.

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 3704. doi:1538-7445.AM2012-3704
Abstract 3705: Multicenter phase II clinical trial of gemcitabine and liposomal paclitaxel combination chemotherapy in patients with advanced biliary cancer
Authors: Park Keon Uk , Kim Jin Young , Do Young Rok , Song Hong Suk , Ryoo Hun-Mo
Published: August 2012

OBJECTIVE: Advanced biliary tract cancer is associated with a poor prognosis due to wide resistance to chemotherapeutic agents. This phase II clinical trial was conducted to determine the safety and efficacy of weekly gemcitabine and liposomal paclitaxel (Genexol-PM®) in patients with unrectable or metastatic biliary cancer. METHODS: The eligibility criteria were patients 1) with pathologically proven unresectable or metastatic biliary cancer, 2) with an ECOG performance status 0 to 2, 3) aged more than 18, 4) with measurable lesions, 5) with adequate hematologic, renal and liver functions, and 6) who provided written informed consent. Each treatment cycle was consisted of gemcitabine 1000 mg/m*2 and liposomal paclitaxel 100 mg/m*2 on days 1, 8 followed by rest perioid of 14 days. It was repeated until the appearance of disease progression or unacceptable toxicity up to maximal 9 cycles. The primary end point of this study was reponse rate, and secondary end points included toxicity, progression free survival and overall survival. RESULTS: Thirty five patients were enrolled; median age was 63 years; male (n=30) and female (n-10). The median number of cycles administered was 3.5 (range, 1-10). Thirty patients were assessable for efficacy. Nine partial responses and 16 stable diseases were confirmed. Giving an overall response rate was 35.6% and disease control rate was 55.6% in per-protocol population. Median overall survival was 12.3 months (95% CI: 5.79-18.74) and median progression free survival was 4.07 months(95% CI: 3.12-5.01). The most frequent adverse events (AE) of all grades were anemia and neutropenia (n=17) followed by leucopenia (13). Grade 3/4 hematologic AEs: anemia (n=6), neutropenia (n=11), thrombocytopenia (n=1). Grade 3/4 non-hematologic AEs: elevated liver function tests (n=3), hyperbilirubinemia (n=1) hyponatremia (n=1). CONCLUSION: Weekly gemcitabine combined with liposomal paclitaxel appears to be effective against advanced biliary cancers. Further randomized trials are needed to confirm this finding.

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 3705. doi:1538-7445.AM2012-3705
Abstract 3713: Comparison of endpoints and data analysis methods for exposure of human tumor cells to dasatinib (NSC732517) and 6-MP (NSC755) in culture
Authors: Kaur Gurmeet , Kondapaka Sudhir B. , Teicher Beverly A.
Published: August 2012

Previous research using the NCI60 cell line combination screen suggested that dasatinib (NSC732517) and 6-mercaptopurine (6-MP; NSC755) may have greater than additive cytotoxicity in selected cell lines. We have conducted a detailed combination study using 6-MP and dasatinib in p53 wild type (MCF-7, H460, A498) and p53 mutant (MDA-MB-468, H23, 786-O) breast, lung and renal cell lines. These studies were performed measuring ATP as the endpoint using Cell Titer Glo after 72h exposure to 6-MP and dasatinib alone or in combination and measuring survival using colony formation after 72hr exposure to the compounds alone or in combination. As a single agent, the lung and renal mutant cell lines (H23, 786-O) were 2-10 fold more sensitive to 6-MP than wild type (H460, A498) with the exception of MDA-MD-468 (mutant), which was more resistant to 6-MP than MCF-7 (wild type). On the other hand all cell lines were sensitive to dasatinib with the IC50 ranging from 0.2-2 μM. For the combination studies, additivity/synergy was calculated using CompuSyn Software (Chou and Martin) or MacSynergy (Prichard and Shipman). Combination index (CI) analysis of the single agent and combination data for 6-MP and dasatinib was additive or antagonist in most of the cell lines. In A498, 786-0, MCF-7 and H460 cell lines a synergistic effect was observed at >10 fold the clinical plasma levels of 6-MP (0.59 μM) or dasatinib (0.19 μM). NCI-H23 cell line demonstrated an additive effect at 0.3 μM of 6-MP and 0.2 μM of dasatinib (CI = 0.94). MDA-MB-468 cell lines demonstrated a strong synergy (CI = 0.2-0.4) for 6-MP (0.03-1 μM) and dasatinib (0.05 μM) at concentrations less than the clinical plasma levels. In a single agent colony formation assay, the H460 cells were sensitive to 6-MP (IC50= 0.21 μM) and resistant to dasatinib (IC50= >10 μM). In a combination study, a synergistic effect (CI = 0.59-0.8) was observed in H460 cell line at concentrations below the clinical plasma level of 6-MP (0.2 μM-0.6 μM) and dasatinib (0.1 μM). These drug effects will be further evaluated in all cell lines by the determination of survival using colony formation assay.

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 3713. doi:1538-7445.AM2012-3713
Abstract 3724: Wogonin enhances tumor necrosis factor-related apoptosis-inducing ligand's antitumor activity in vivo through H2O2-mediated downregulation of X-linked inhibitor of apoptosis protein and cellular inhibitor of apoptosis protein 1 and 2
Authors: Yang Lan , Zheng Xuelian , Lin Yong , Wang Xia
Published: August 2012

Numerous malignant cells are resistant to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced cytotoxicity, limiting value of TRAIL as an antitumor therapeutics. Combination of TRAIL with other agents is a promising strategy to overcome TRAIL resistance. Wogonin (5, 7-dihydroxy-8-methoxyflavone), a flavonoid originated from the root of the medicinal herb Scutellaria baicalensis Georgi, has been shown to enhance TRAIL-induced apoptosis in malignant cells in in vitro studies. However, whether wogonin enhances TRAIL's antitumor activity in vivo has never been studied. In this study, we first found that wogonin sensitized a variety of cancer cells including A549, HeLa and SKOV3 to TRAIL-induced apoptosis, as indicated by increased annexin V staining and enhanced activation of caspase-8 and -3 in TRAIL and wogonin cotreated cells, whereas having marginal effect on non-transformed bronchial epithelial cells and cervical epithelial cells. Then the effect of combination of TRAIL and wogonin was tested in a xenografted tumor model with A549 cells in nude mice. The results show that wogonin greatly enhanced TRAIL-induced suppression of tumor growth, which was accompanied with increased apoptosis in tumor tissues determined by a terminal deoxynucleotidyl-transferase -mediated dUTP nick-end labeling (TUNEL) assay. The protein levels of X-linked inhibitor of apoptosis protein (XIAP) and cellular inhibitor of apoptosis protein 1 and 2 (cIAP-1 and cIAP-2) were markedly reduced in both cultured cells and xenografted tumor tissues with co-treatment of wogonin and TRAIL. The down-regulation of these antiapoptotic proteins was likely mediated through proteasomal degradation and involved intracellular H2O2, as wogonin robustly induced H2O2 accumulation in A549 cells and the reactive oxygen species scavengers butylated hydroxyanisole (BHA) and N-acetyl-L-cysteine (NAC) and the proteasome inhibitor MG132 restored the expression of these antiapoptotic proteins and suppressed cytotoxicity in cells co-treated with wogonin and TRAIL. These results provide important in vivo evidence supporting the application of wogonin as TRAIL sensitizer for cancer 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 3724. doi:1538-7445.AM2012-3724