bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3239: The Notch pathway as a new target in gastrointestinal stromal tumors
Authors: Dumont Amaury G. , Hughes Dennis P. , Trent Jonathan
Published: August 2012

Small molecule kinase inhibitors have dramatically improved the survival of patients with Gastrointestinal Stromal Tumors (GIST), yet most patients develop resistance, urging us to understand better the molecular mechanisms underlying this disease and identify new therapeutic strategies. Notch signaling is a conserved developmental pathway known to play a critical role in the development of several cancers, functioning as a tumor promoter or a tumor suppressor. Given that the normal progenitor cell for GIST, the Interstitial Cell of Cajal, has characteristics similar to those of cells of neuroendocrine origin, we hypothesized that Notch pathway impacts the biology of GIST cells. In a previous study, we demonstrated that constitutively active intracellular domain of Notch1 (ICN-1) induced growth arrest in GIST cell lines. Here we report that ICN-1 downregulated KIT protein by a flow cytometric analysis in the three cell lines analyzed (GIST-T1, GIST882, GIST48IM). In parallel, inhibitions of Hes1, a major target gene of ICN-1, with a dominant negative construct, upregulated KIT protein expression in vitro (60% increase on western blot). This anti-tumor effect of Notch1 in GIST leads to a therapeutic opportunity as we showed in the present study that treatment with the histone deacetylase inhibitor, SAHA, caused a dose-dependent upregulation of Notch1 (6 fold increase of the mRNA with SAHA 2 µm/L on GIST-T1, P<0.05), downregulation of KIT and pKIT (9 fold decrease of pKIT on western blot with SAHA 2 µm/L on GIST-T1), increase in apoptosis (25-fold increase in the sub-G1 population with SAHA 2 µmol/L on GIST-T1, P<0.05), decrease in viability of the GIST cells (IC50 = 2 µmol/L on GIST-T1).To confirm the clinical relevance of the Notch pathway on GIST, we performed a retrospective analysis on 15 pre-treated patient tumors and showed that GIST patients with high Hes1 mRNA levels have a significantly longer relapse-free survival than the one with low Hes1 mRNA levels (median of 37 months for the low group versus median not reached at >80 months for the high group; P=0.005). These results identify a novel anti-tumor effect of Notch1 via a crosstalk between the Notch1 and KIT pathways in GIST cells. This finding may lead to new therapeutic strategies for GIST patients, in particular after failure of tyrosine kinase inhibitors.

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 3239. doi:1538-7445.AM2012-3239
Abstract 3251: Linking prostaglandin E2 signaling, lipid rafts, and DNA protein kinase
Authors: Hawk Ernest T. , Menter David G. , Patterson Sherri , Swank Michael W. , DuBois Raymond N.
Published: August 2012

Colorectal cancer (CRC) is the second-leading cause of death from cancer in the USA. Chronic inflammation in the gastrointestinal tract is a significant risk factor for developing CRC. Generally, CRC involves the upregulation of inflammation-induced cyclooxygenase-2 (COX-2) that is blocked by non-steroidal anti-inflammatory drugs (NSAIDS). Among the downstream prostaglandin synthases that produce bioactive prostaglandins (PGs), prostaglandin E2 (PGE2) synthase products are proinflammatory. After export from cells, PGE2 can bind to G-protein coupled PG cell surface receptors EP2/4 and activate adenylate cyclase and elevate cAMP. In turn, cAMP activates downstream effectors such as protein kinase A (PKA) and/or exchange protein directly activated by cAMP (EPAC). We studied effects of PGE2 on PKA activation in CRC cells and their relationship to lipid raft function. After isolating lipid rafts from PGE2 stimulated cells we performed shotgun proteomics. Among the lipid raft proteins identified were DNA-protein kinase (DNA-PK), and heterodimer complex proteins Ku70 and Ku80, which we verified by immunoprecipitation and immunofluorescence. Of these lipid raft proteins, Ku80 contained a PKA consensus phosphorylation sites and phosphorylation increased after stimulation of CRC cells by PGE2. Typically found in the nucleus, Ku70, Ku80 and DNA-PK complexes bind to DNA double-strand breaks and initiate non-homologous end joining DNA repair. These data are the first to demonstrate the Ku70, Ku80 and DNA-PK complexes in a new non-nuclear action, which involves lipid rafts and the DNA repair complex. Furthermore, PGE2 stimulated LS174T CRC cells exhibit an increase in non-nuclear pDNA-PK (S2056). Additionally, treatment of a number of colon cancer cell lines with the selective DNA-PK inhibitor Nu7026 significantly decreased cell viability in the absence of any overt DNA damaging stimuli, suggesting that non-nuclear, and in particular, lipid raft localized DNA-PK complex may play an important role in cell growth. Finally, treatment with both PGE2 and Nu7026 causes a profound change in cell morphology involving numerous spindle-like processes. These data suggest there are important downstream effects of proinflammatory PGE2 on CRC cells that influence lipid raft associated Ku70, Ku80 and DNA-PK complexes as well as cell proliferation. Support: T. Boone Pickens Distinguished Chair for Early Prevention of Cancer to E.T.H., R37 DK47297, NCI P01 CA77839, CPRIT-RP100960, Ellen F. Knisely Distinguished Chair in Colon Cancer Research, The National Colorectal Cancer Research Alliance to R.N.D. and 5U54CA151668-02 to D.G.M.

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 3251. doi:1538-7445.AM2012-3251
Abstract 3414: Role of FGFR2 in invasion and metastasis through different activation of downstream signaling in gastric cancer
Authors: Lee Won Suk , Park Kyu Hyun , Kim Jeong Min , Lee Myung Eun , Kim Tae Soo , Kim Hyo Song , Jeung Hei Cheul , Chung Hyun Cheol , Rha Sun Young
Published: August 2012

Activation of fibroblast growth factor receptors (FGFRs), the family of receptor tyrosine kinase (RTK), initiates intracellular signaling networks for key cellular processes. Previous reports show that FGFR2 amplification induces activation of downstream signaling in gastric cancer influencing cell proliferation and survival. In this study, we evaluated activation of FGFRs and related downstream pathways, and also determine the effect of FGFRs in tumor invasion and metastasis in gastric cancer. First, we performed phospho-RTK array in 25 gastric cancer cell lines and found that FGFR2 and FGFR3 were more frequently phosphorylated than FGFR1 or FGFR4 in gastric cancer. Next, we observed genomic amplification status and phosphorylation level of FGFR2 using real-time PCR and Western blot. KATO III and SNU-16 with higher expression of FGFR2 showed high correlation between DNA amplification and phosphorylation status. Then, using invasion assay, cells with higher FGFR3 showed higher invasiveness compare to cells with FGFR2 overexpression. When we observed the activation status of downstream signaling molecules, cells with higher FGFR2 were more dependent on PI3K-AKT-mTOR pathway, while cells with higher FGFR3 were more dependent on ERK pathway. In conclusion, FGFRs have various biological roles through different signaling pathways in gastric cancer, suggesting that FGFRs are the potential molecular targets in 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 3414. doi:1538-7445.AM2012-3414
Abstract 3499: CD56 targeted chimeric antigen receptors for immunotherapy of multiple myeloma
Authors: Benjamin Reuben , Condomines Maud , Gunset Gertrude , Sadelain Michel
Published: August 2012

We have developed an adoptive T cell therapy strategy for treating multiple myeloma using chimeric antigen receptors targeting CD56. CD56 is strongly expressed by malignant plasma cells in 70% of patients with myeloma and represents a potential immunotherapy target. CD56 is also expressed at lower levels on normal tissue types including neuronal cells, NK cells and a subset of activated T cells. A second generation CAR was constructed containing the scFv of the murine monoclonal antibody against human CD56 (N901) as well as the CD28 transmembrane and cytoplasmic signaling domains. CD56 CAR cells were generated by retroviral transduction of human T cells and showed antigen dependent proliferation and cytokine secretion in vitro when stimulated with CD56 positive myeloma cells. In vitro cytotoxicity assays showed significant lysis (40-50% lysis at effector to target ratios > 5:1) of CD56 positive myeloma cell lines compared with a control prostate specific membrane antigen (PSMA) targeted CAR. To further assess the antitumor activity of CD56 CARs in vivo we developed a systemic xenograft model of myeloma by injecting the OPM2 myeloma cell line, modified to express the firefly luciferase gene, intravenously into NOD/SCID Il2rαnull mice. Bioluminescence imaging showed tumor progression predominantly within the bone marrow recapitulating the human disease phenotype. If untreated hind limb paralysis occurred at approximately 35 days following injection of 3x106 tumor cells. Cohorts of mice were then treated by intravenous injection of either 1 or 5 x106 CD56 CAR cells or control PSMA CAR cells at 7 days following tumor injection, when disease was firmly established. At the 1x106 T cell dose tumor development was significantly delayed compared to controls (median survival of 49 days compared with 34 days respectively, p=0.02) but 7/8 mice eventually progressed and had to be euthanised. In contrast all mice receiving 5x106 cells (n=8) showed complete tumor regression and remained tumor free at 3 months. Interferon-α secreting CD56 CAR cells were detected in the peripheral blood of these mice and correlated with tumor bulk with numbers eventually declining to low levels that persisted even at 3 months. These results demonstrate for the first time the impressive anti-tumor efficacy of a CD56 targeted chimeric antigen receptor in a systemic xenograft model of myeloma. CD56 CAR therapy therefore represents an attractive immunotherapy option and its use in patients with relapsed refractory myeloma should be considered.

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 3499. doi:1538-7445.AM2012-3499
Abstract 3501: Binary control of CAR-CTL by transgenic IL-7 and IL-7R expression
Authors: Bajgain Pradip , Mucharla Roopa , Heslop Helen E. , Vera Juan F.
Published: August 2012

While adoptive transfer of cytotoxic T lymphocytes (CTLs) engrafted with chimeric antigen receptors (CAR) has produced objective clinical responses in vivo, the infused cells usually fail to persist long-term, limiting benefit. We have recently demonstrated that CTL persistence can be improved by engineering cells to express the IL-7 receptor alpha chain (IL-7R) which is physiologically absent on CTLs. However, this approach requires access to clinical grade cytokine and biodistribution to T lymphocytes at tumor sites may be insufficient. To circumvent these problems we have prepared two CTL products: one expressing a tumor-specific CAR in combination with IL-7R (product #1), and the second engineered to co-express the same CAR and produce IL-7 cytokine (product #2). In this way, both products have anti-tumor activity mediated through the CAR, while cytokine produced from CTL#2 should support the survival and persistence of the IL-7R-expressing CTL#1. A binary system such as this should be intrinsically safer than incorporating a positive feedback loop of both cytokine and receptor in a single cell. As a proof of this principle, we used the SFG-CAR that targets the ≤ light chain expressed on B cell malignancies: SFG-CAR/IL-7R-GFP(#1) and SFG-CAR/IL-7cyto-mOrange(#2). EBV-CTLs from 3 donors were transduced with each vector. FACS analysis of transgene expression indicated that all were expressed at approximately equivalent levels: CTL#1 (CAR, IL-7Rα, and GFP; 58%±15, 53%±18, 57.8%±12) and CTL#2 (CAR and mOrange; 54%±18 and 52%±20). The modified CTL were functional, and cells transduced with either vector were able to kill the α+ B cell tumors Daudi as evaluated by Cr51 assay (72%±13 and 69%±25, respectively) at an R:S of 40:1. Preliminary data indicates that CTLs#1 were able to proliferate in response to exogenous IL-7 administration, and IL-7 cytokine production from CTLs#2 was directly proportional to the antigenic stimulation provided to the cells. In addition, when CTLs#1 and #2 were cultured together we observed that CTLs#2 were capable of promoting the expansion of their CTL counterpart in a more efficient way than that compared to exogenous administration of IL-7 or IL-2. These results indicate that binary control of CAR modified CTLs is possible through transgenic expression of IL-7R and IL-7 cytokine in two different CTL populations.

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 3501. doi:1538-7445.AM2012-3501
Abstract 3513: The anti-tumor effect of activated natural killer cells in xenograft mouse models with epithelial ovarian cancer
Authors: Young Mo Jin , Kyoung-jin Lee , Yoo-Kyeong Whang , Shin-Wha Lee , Yong-Man Kim , Dae-Yeon Kim , Jong-Hyeok Kim , Joo-Hyun Nam , Young-Tak Kim
Published: August 2012

Objectives: Ovarian cancer is the most common cause of death from gynecologic cancer in women. Because of the lack of effective screening and warning sign, 60-70% of patients were diagnosed at advanced stage. At present the treatment of advanced ovarian cancer is cytoreductive surgery followed by chemotherapy, and the overall response rate is 70-80%. But 50-75% of responders will relapse within about 18 -24 months. Therefore, new strategies of treatment, for examples, immunotherapy or targeted therapy, are necessary. This study is to evaluate the anti-tumor effect of activated natural killer (NK) cells in xenograft mouse model with epithelial ovarian cancer(EOC). Methods: We have made the OVCAR3-Luc stable cell line by transient transfection of pGL3-Luciferase (G418) gene. The xenograft mouse model of EOC was constructed by the intra-peritoneal injection of OVCAR3-Luc ovarian cancer cell line to the NOD/scid mice. After we confirm the tumor formation with IVIS spectrum, activated NK cells were injected with intra-peritoneal route. There was no treatment to the negative control and paclitaxel (10mg/kg) was used to the positive control. The tumor growth was evaluated by serial IVIS spectrum and biopsy at 8 weeks after any treatment. Results: Activated NK cells were injected at concentration 1 × 107 cells per mouse with intra-peritoneal route. This procedure was repeated 5 times. The NK cells significantly inhibited the tumor growth of OVCAR3-Luc-induced NOD/scid mice, up to the 57.1% comparing the negative control. Although paclitaxel decrease 69.3% of the tumor growth, it is not different to that of NK cell treating mice. Furthermore, side effects involving hair loss, abnormal behavior and weight low were not shown in NK cell-injected mouse models. Conclusion: We confirmed that activated NK cells are able to inhibit ovarian tumor growth without side effect in xenograft models. Therefore, this research demonstrates that NK cell-based immunotherapy might be desirable to the patients with ovarian cancer as an adoptive immunotherapy.

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 3513. doi:1538-7445.AM2012-3513
Abstract 3585: Second primary tumors after breast cancer: Results from the European Prospective Investigation into Cancer and Nutrition (EPIC) study
Authors: Ricceri Fulvio , Sacerdote Carlotta , Critelli Rossana ,   EPIC investigators, Vineis Paolo , Riboli Elio
Published: August 2012

During the last decades, improvements in medical and surgical treatments have substantially increased the chances of surviving from a cancer. Cancer survivors now amount to more than 3.5% of population in the US, and about 3% in Western Europe. Cancer survivors face the problem of subsequent cancers possibly related to the late effects of treatments or to a common aetiology for the first and subsequent cancers. Cancer patients have a 20% higher risk of new primary cancer compared with the general population. Approximately one third of cancer survivors aged >60 years were diagnosed more than once with another cancer. Women with breast cancer as first primary cancer were the first largest group of multiple cancer prevalence reported in the United States in 2002, while the second and third groups were men and women with a diagnosis of primary colorectal cancer and men with prostate cancer, respectively. Multiple cancers arise in the same individual due to several causes: host factors, such as hormonal and/or genetic factors, lifestyle and environment, and treatments due to the first cancer (for example, radiotherapy) The aim of this study is to assess the incidence of second primary tumors in a cohort of women with a first breast cancer and to investigate the role of shared risk factors. The study was performed in the cohort of women with a primary breast cancer from the European Prospective Investigation into Cancer and Nutrition (EPIC) study. To correctly assess the incidence of second primary tumors, we applied a Markov model studying the transition intensities from first to second tumor with the Aalen-Johansen (AJ) estimators, as usually done in competing risk models. In the cohort of 11045 women, we observed 492 second primary tumors (not metachronous) after a median follow-up of 4.92 years. About 30% (140) of them were second primary breast cancers, 65 were colorectal cancers and 39 were endometrial cancers. In this cohort, there was a slight, not statistically significant increase of cancer incidence in women with breast cancer, compared to the general population; this was for all tumors together, except non-melanoma skin cancers. Analysis by subgroups identified a significant increase for second breast cancers and for endometrial cancers. Our findings suggest that second primaries may be an important problem for cancer survivors. We will now investigate the role played by lifestyle factors, genetics and possibly therapies in second primary occurrence.

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 3585. doi:1538-7445.AM2012-3585
Abstract 3620: A multiplex quantitative immunofluorescence assay for DNA damage repair in response to cytotoxic treatment
Authors: Marrero Allison M. , Lawrence Scott M. , Balasubramanian Priya , Pommier Yves G. , Tomaszewski Joseph E. , Parchment Ralph E. , Doroshow James H. , Kinders Robert J.
Published: August 2012

The use of DNA damaging agents remains one of the most common approaches to cancer therapy. The type of damage caused by the agent will determine which of the five main DNA damage repair (DDR) processes will respond. Initially we are developing multiplex quantitative immunoflourescence assays (qIFAs) for biomarkers of active Homologous Recombination (HR) and Nucleotide Excision Repair (NER) pathway and plan to develop mutiplex assays for the other repair pathways in the near future. The first multiplex panel to quantify DDR response has been developed to include Nbs1 phosphorylated at Serine343 and γH2AX, both early markers of DNA double strand breaks as well as ERCC1, which plays a key role in NER. Quantitation of repair pathway activation after drug treatment builds upon our validated qIFA for γH2AX, which scores individual nuclei as positive or negative for the biomarker and quantifies the γH2AX response as the fraction of nuclear area that is occupied by γH2AX foci. In contrast, the multiplex qIFA was designed to quantify the global DNA repair protein response to a variety of cytotoxic, genotoxic and targeted agents administered singly and in combination. Initial evaluations were performed in vitro on a panel of cell lines, characterized for repair pathway defects: the BRCA1-null human ovarian cancer line UWB1.289 (ATCC# CRL-2945), its isogenic line with restored BRCA1 named UWB1.289+BRCA1 (ATCC# CRL-2946), and SKOV3 (BRCA1 WT). Both gemcitabine and cisplatin increased pNbs1 both in UWB1.289 and UWB1.289+BRCA1, but not in SKOV3. Co-expression of pNbs1 with γH2AX was observed but was not 100% concordant. Combination treatment with a topoisomerase 1 inhibitor and PARP1/2 inhibitor (the indenoisoquinoline NSC 724998 with ABT-888) increased nuclear ERCC1 only in UWB1.289, but not in the isogenic UWB1.289+BRCA1 line, or in the ovarian lines SKOV3 and IGROV1, or the human fibroblast line GM00637. Assessment of drug-induced DDR response in a panel of cell lines with known repair defects is planned with the expectation that this will provide a scientific foundation for interpreting qIFA results when the assay is used to analyze clinical biopsies, and will assist in the evaluation of combination therapy strategies, such as synthetic lethality, designed to take advantage of DNA repair defects. Funded by NCI Contract No. HHSN261200800001E.

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 3620. doi:1538-7445.AM2012-3620
Abstract 3664: Breast cancer companion diagnostic platform based on objectively defined tumor co-expression patterns stratifies multiple clinical and therapeutic endpoints comparison to existing molecular subtyping definitions
Authors: Rhodes Daniel R. , Tomlins Scott , Thomas Dafydd G. , Williams Paul , Wyngaard Peter , Sadis Seth , Oades Kahuku , Vo Lien , Chattopadhyay Sukla , Wang Yipeng , Lee Byung-In , Monforte Joseph
Published: August 2012

Gene expression profiles of human breast tumors have greatly expanded our understanding of the genes and pathways that underlie breast cancer. Profiling studies have also supported a molecular classification of breast cancer. The resulting molecular subtypes Luminal, Basal-like, ERBB2+, and Normal-like were shown to have different prognostic and predictive characteristics. Related studies have led to a proliferation of multigene prognostic and predictive diagnostic tests. Two independent multigene tests, OncoType Dx and MammaPrint, have been shown to be helpful in predicting the risk of recurrence of patients with early stage breast cancer. Current multigene tests consistently prioritize the proliferation, estrogen receptor (ER), and ERBB2 pathways. An alternative approach to identifying key molecular variables within breast cancer is based on a definition of objectively defined tumor co-expression patterns. To this end, we defined co-expression patterns within 56 independent breast cancer molecular profiling datasets representing >5,000 unique patients. We then performed a meta-analysis across datasets to define the most robust, consistently occurring co-expression patterns. These patterns, termed modules, recapitulate the proliferation, ER, and ERBB2 pathways, but also monitor expression of other important variables including core cancer cell growth pathways, immune signaling and microenvironment, and hallmark genomic aberrations. An important feature of co-expression patterns is that a small number of genes serve as an effective surrogate for each module. Thus, we created a single multigene qPCR test that measures the expression of 18 distinct breast cancer modules and validated the test for use with formalin-fixed paraffin-embedded (FFPE) tumor samples. In retrospective microarray scoring analyses with key clinical datasets, and with analysis of FFPE specimens from breast cancer cohorts, we demonstrate that breast cancer modules can be used to recapitulate the molecular subtypes of breast cancer and to have prognostic and predictive properties similar to the current multigene tests. Because they recapitulate existing molecular tests, while also reading out many additional axes of molecular variability, breast cancer modules provide a universal assay with broad application to companion diagnostics development.

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 3664. doi:1538-7445.AM2012-3664
Abstract 3745: The dual PI3K/mTOR inhibitor NVP-BEZ235 is a potent inhibitor of ATM- and DNA-PKcs-mediated DNA damage responses
Authors: Mukherjee Bipasha , Tomimatsu Nozomi , Amancherla Kaushik , Camacho Cristel , Burma Sandeep
Published: August 2012

Inhibitors of PI3K/Akt signaling are being actively developed for tumor therapy due to the frequent mutational activation of the PI3K-Akt-mTORC1 pathway in many cancers, including glioblastomas (GBMs). NVP-BEZ235 is a novel and potent inhibitor of PI3K/Akt signaling that is currently in phase I/II clinical trials. BEZ235 is reported to specifically inhibit PI3Ks and mTOR with little effect on other kinases. However, we find that BEZ235 also potently inhibits the two major kinases responding to DNA double-strand breaks (DSBs), ATM and DNA-PKcs, both in vitro and in vivo. Consequently, BEZ235 blocks both non-homologous end joining (NHEJ) and homologous recombination (HR) DNA repair pathways, resulting in significant attenuation of DSB repair. Additionally, phosphorylation of ATM targets and implementation of the G2/M cell cycle checkpoint are also attenuated by this drug. The consequence is profound radiosensitization at very low concentrations (100 nM) of BEZ235. This radiosensitizing effect is significantly more potent than that seen with much higher concentrations (10 μm) of inhibitors of DNA-PKcs or ATM that are currently being optimized for clinical testing. Since radiotherapy is the standard of care for GBM, we investigated the potential utility of BEZ235 as a radiosensitizing agent in a panel of GBM lines. We find that BEZ235 treatment confers an extreme degree of radiosensitization and significantly attenuates DSB repair irrespective of Akt activation status in these lines. Finally, we show that BEZ235 also significantly impairs DSB repair in a mouse tumor model thereby validating the efficacy of this drug in vivo. Our results, showing that BEZ235 is a potent and novel inhibitor of ATM and DNA-PKcs, have important implications for the informed and rational design of clinical trials involving this drug and also reveal the potential clinical utility of BEZ235 as an effective radiosensitizer.

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 3745. doi:1538-7445.AM2012-3745
Abstract 3752: Nanoparticles delivery of a novel AKT/PDK1 inhibitor inhibits pancreatic cancer tumor growth
Authors: Meuillet Emmanuelle J. , Moses Sylvestor A. , Lucero-Acuna Armando , Guzman Roberto , Jeffrey Justin , Pagel Mary
Published: August 2012

AKT (protein kinase B), a pleckstrin homology (PH) lipid binding domain and a serine/threonine kinase containing protein is a key component of the phophatidylinositol-3-kinase (PtdIns3-K) cell survival signaling pathway which is activated in pancreatic cancers. In this study, we describe the effects of a novel inhibitor of AKT/PDK1 (PH-427) that binds to the PH domain of AKT (Ki = 2.67 ± 0.37 μM) and PDK1 (Ki = 5.20 ± 0.45 μM) [1] thus preventing its binding to PtIns-(3,4,5)P3 at the plasma membrane and subsequent activation of both kinases. PH-427 inhibits AKT/PDK1 activities at low micromolar concentrations in BxPC-3 (wt-KRAS) and not in MiaPaCa2 (mt-KRAS) human pancreatic cancer cell lines. Accordingly, we have shown that in vivo, PH-427 is poorly efficient in MiaPaCa2 xenografts as compared to BxPC-3 xenografts. In order to increase the potency and delivery of PH-427 to the tumor, we have encapsulated the compound (>10% of the compound) into (poly-d, l-lactide-co-glycolide) (PLGA) nanoparticles (PNP). We demonstrate that PH-427 incorporates well in the nanoparticles (PH-427-PNP) (<200 nm in size) and is slowly released from the polymer with a 90% released over a period of 30 days. We show that PH-427-PNP were very efficient in reducing MiaPaCa-2-Luc orthotopic pancreatic tumor growth as evaluated by luminescence using an AMI-1000 scanner. MiaPaCa2-Luc pancreas tumor volume was significantly reduced in animals treated with PH-427-PNP as compared to animal treated with PH-427 alone or the PNP alone (41.4±29.9 versus 2051±387 and 631±45 mm3). In conclusion, PH-427 alone did not alter tumor growth in MiaPaCa-2-Luc but was able to reduce tumor growth significantly when encapsulated in PNP. Finally and interestingly, metastasis to the liver and other organs was also significantly inhibited by PH-427-PNP as compared to PNP or PH-427 alone. [1]. Meuillet EJ, Ihle N, Lemos R, Moses SA, Song Z, Zhang S, Mash EA, Powis G (2010). Mol Cancer Ther 9(3):706-717.

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 3752. doi:1538-7445.AM2012-3752
Abstract 3762: Second generation PIM inhibitors exhibit improved activity in solid tumor models
Authors: Foulks Jason M. , Xu Yong , Saunders Michael , Liu Xiao-Hui , Brenning Benjamin , Clifford Adrianne , Wilkes Marcus , Luo Bai , Lai Shuping , Merx Shannon , Chan Ashley , Huang Liwen , Vollmer David , Senina Anna , Liu Jihua , Ho Koc-Kan , McCullar Michael V. , Kanner Steven B.
Published: August 2012

The proto-oncogene PIM kinase family (PIM-1, -2 and -3) includes constitutively active serine/threonine kinases upregulated in multiple cancer indications, including lymphomas, leukemias, multiple myeloma, prostate and bladder cancers. Overexpression of one or more PIM family members in patient tumors frequently correlates with poor prognosis. The PIM kinases function by inhibiting apoptosis in MYC-driven tumors, and promoting tumor cell survival and proliferation. In the HEK-293T cell line, enhanced PIM kinase substrate BAD phosphorylation (pBAD) was observed following PIM and BAD overexpression. Enhancement of pBAD was inhibited by SGI-1776, a well-described PIM inhibitor, and more effectively by second generation PIM inhibitors exhibiting 4-10 fold improved potency against the PIM kinase family. The current PIM inhibitors display sub-µM activity in pharmacodynamic marker, proliferation and 2D colony formation assays using the UM-UC-3 human bladder cancer cell line. PIM1 and PIM2 overexpression models were established in the human prostate cancer cell line 22RV-1 and the non-tumorigenic mouse NIH-3T3 cell background. Overexpression of PIM kinases led to enhanced cell growth and tumorigenicity in both NIH-3T3 and 22RV-1 cell lines. In vivo xenograft studies using both PIM overexpression models and a clinically relevant solid tumor model facilitated identification of a lead candidate with demonstrated efficacy and favorable toxicity. IND-enabling studies with a lead candidate are 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 3762. doi:1538-7445.AM2012-3762
Abstract 3764: SC-1, a sorafenib derivative, induces apoptosis in epidermal growth factor receptor wild type non-small cell lung cancer through the inhibition of signal transducers and activators of transcription 3
Authors: Wang Cheng-Yi , Chao Ting-Ting , Shiau Chung-Wai , Yuan Ang , Yu Chong-Jen , Chen Kuen-Feng
Published: August 2012

Background: Lung cancer is the leading cause of cancer-related death worldwide, but currently there are no available target therapies for EGFR wild type NSCLC patients. Sorafenib, a multiple kinase inhibitors, shows therapeutic effects in renal carcinoma (RCC) and hepatocellular carcinoma (HCC). Our data shows that the inhibition of signal transducers and activators of transcription 3 (STAT3), a transcription factor involving in cell proliferation and survival, represents a major mechanism whereby sorafenib induces apoptosis in HCC cells (Clin Cancer Res 2010). Therefore, we design and synthesize a sorafenib derivative, SC-1, which is devoid of kinase activity (J Hepatol 2011). SC-1 shows higher potency in inducing apoptosis than sorafenib. Here, we tested efficacy of SC-1 to EGFR wild type NSCLC and examined the drug mechanism. Methods: EGFR wild type NSCLC cell lines, A549 H292 H322 and H460, were used for in vitro studies. Apoptosis was examined by both flow cytometry and western blot. Signal transduction pathways in cells were assessed by western blot. Results: SC-1 induced apoptosis in association with reducing the phosphorylation of STAT3 (p-STAT3) in a dose- and time-dependent manner in four NSCLC cells lines (A549 H292 H322 and H460). SC-1 demonstrates dose-dependent anti-proliferation effects on cell viability in NSCLC cell lines on MTT assay. Increased amount of SC-1 represented improvement cell apoptosis by FACS in NSCLC cells. Western blot analysis showed SC-1 exhibit dose-dependent and time-dependent effect on cell apoptosis through inhibition of STAT3 with reducing p-STAT3 and the STAT3-regulating protein, Mcl-1, survivin and cyclinD1. Conclusions: SC-1 provides a proof of concept that targeting STAT3 signaling pathway is a novel approach for the treatment of EGFR wild type 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 3764. doi:1538-7445.AM2012-3764
Abstract 3794: Inhibition of prostate cancer bone metastases by the dickkopf-1 neutralizing antibody BHQ880
Authors: Sottnik Joseph L. , Hall Christopher , Karatsinides Andreas , Keller Andrew L. , Ettenberg Seth A. , Keller Evan T.
Published: August 2012

Advanced prostate carcinoma (PCa) is usually complicated by the presence of bone metastases. Current therapies targeting bone metastases are suboptimal as the metastases progress in spite of therapy. Accordingly, novel therapeutics are needed to target these lesions. The Wnt inhibitor Dickkopf-1 (DKK-1) appears to play an important role in progression of PCa bone metastases. Overexpression of DKK-1 by PCa promotes tumor growth and bone lysis in laboratory studies and DKK1 is present in clinical PCa bone metastases. Development of a fully humanized anti-DKK-1 neutralizing antibody, BHQ880 (Novartis), provides a potential therapeutic strategy to target PCa bone metastases. The goal of this study was to test if BHQ880 could impact PCa bone metastases. PC-3-luc cells (2.5x105) were injected intratibially into mice. Treatment with anti-DKK-1 or isotype control antibody (n=12 mice per group) was initiated 0, 7, or 14 days post-tumor injection. Mice were treated three times weekly by intraperitoneal injection with 200 μg/mouse (approximately 10 mg/kg) Tumor growth and bone lysis were assessed by weekly bioluminescence and bi-weekly radiography, respectively. All mice were euthanized 31 days post-tumor injection, tumor bearing limbs were formalin fixed and serum was obtained. Tumor bearing limbs were analyzed by dual energy x-ray absorptiometry (DEXA) and histology. Serum was assayed for the bone turnover biomarkers alkaline phosphatase (ALP), osteocalcin (OC), and tartate-resistant acid phosphatase 5b (TRAP5b). BHQ880 was well tolerated with no significant side effects observed. Significant inhibition of tumor growth was observed in mice where treatment was initiated 7 or 14 days post-tumor challenge when established tumors were present. Two mice from the 14-day treatment group had tumor regressions after treatment with BHQ880. Treatment with BHQ880 resulted in decreased bone lysis as determined using DEXA. Furthermore, increased serum concentrations of ALP and OC in mice treated with BHQ880 suggest increased bone production; no significant difference in TRAP5b concentration between treated and untreated mice was found. Treatment with the DKK-1 neutralizing antibody BHQ880 was found to inhibit tumor growth, tumor-induced osteolysis and increase serum bone remodeling markers associated with increased bone production in a murine model of PCa bone metastasis. These data suggest that anti-DKK-1 treatment is a plausible mechanism for inhibition of progression of PCa bone metastases. These data suggest that inhibition of DKK-1 may be an effective strategy to inhibit PCa growth in bone.

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 3794. doi:1538-7445.AM2012-3794
Abstract 3973: Developing a public resource for structural biology and cancer pathway plasmids
Authors: Cormier Catherine Y. , Park Jin , Fiacco Michael , Steel Jason , Kramer Jason , LaBaer Joshua
Published: August 2012

Understanding the structure and function of the genes involved in cancer is crucial to dissecting the underlying mechanisms of cancer development, progression and treatment. The Protein Structure Initiative:Biology-Materials Repository (MR; http://psimr.asu.edu) connects researchers with both the genetic raw materials and the structural knowledge to better understand the roles of individual genes in cancer pathways. As a centralized plasmid repository, the MR is responsible for sequence-verification, archival, and distribution of plasmids created by the high throughput structural genomics efforts of the Protein Structure Initiative (PSI). Plasmid annotations such as the full length sequence, vector information, and associated publications are stored in a freely available, searchable database called DNASU (http://dnasu.asu.edu). Each plasmid also links to external resources, including the PSI Structural Biology Knowledgebase (http://sbkb.org) facilitating cross-referencing of plasmids to additional protein annotations and experimental data. In addition to the 50,000 protein expression plasmids and empty vectors from the PSI, DNASU distributes over 151,000 plasmids including collections of human kinases, 1,000 breast cancer-related genes, over 10,000 human genes in several cloning and expression vectors, and over 800 glyco-enzyme plasmids that were created by or in collaboration with the MR, the ORFeome Collaboration, or individual laboratories. Furthermore, the MR strives to expedite and simplify plasmid requests through its expedited material transfer agreement (EP-MTA) network, where researchers from network institutions can order and receive PSI plasmids without institutional delays. By providing these plasmids to the research community, the MR is able to facilitate and accelerate the study of the biological function of proteins involved in cancer and whose structures have been solved by the PSI.

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 3973. doi:1538-7445.AM2012-3973
Abstract 3981: Evaluation of mutation detection sensitivity in exome sequencing and other genotyping platforms
Authors: Wulur Isabella H. , Wong Swee Seong , Yu Kun , Barber Thomas D. , Yue Yong Gang
Published: August 2012

An evaluation was performed to compare mutation calls from exome capture sequencing data to publicly available data sets derived from Sequenom MassARRAY and Sanger sequencing methods. We compared hot-spots KRAS and BRAF mutations using Sanger sequencing method in patient-derived tumor grafts. Mutation data of the tumor grafts were also generated with Sequenom MassARRAY using the OncoCarta panel. We also evaluated the sensitivity of mutation detection for a panel of cancer genes in >100 cancer cell lines against the data from the Cancer Cell Line Encyclopedia Project (OncoMap mutation panel) as well as the Sanger's Cancer Genome Project. Overall, we observed high sensitivity and concordance of the mutation calls from the exome sequencing with the other platforms and technologies. The above comparisons support the use of exome sequencing as a sensitive mutation detection method for patient-derived tumor grafts and cancer cell lines.

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 3981. doi:1538-7445.AM2012-3981
Abstract 3986: Science Exchange: An integrated platform for experiment outsourcing
Authors: Connolly Jay , Abbott Ryan , Knox Dan , Iorns Elizabeth J.
Published: August 2012

Introduction: Scientific research is becoming increasingly specialized, necessitating greater use of collaborations and outsourcing to draw on experimental expertise. To facilitate this, many research institutions have established core facilities to help reduce the inefficiencies associated with the historical system of ‘bartering’ to gain access to specialized equipment and expertise. However, the current core facility system is fragmented: researchers have varying degrees of access to services, quality is hard to evaluate and pricing is not transparent. Researchers who need to collaborate outside of their institution lack an easy way to find providers, evaluate them, coordinate logistics and pay for work. Similarly, core facilities lack a robust mechanism for promoting their expertise and services to relevant researchers outside their institution. Methods: Science Exchange, an online marketplace accessible at www.ScienceExchange.com, was launched in August 2011 to address these issues. Science Exchange works by signing up core facilities as “providers” of different experiment types. Researchers can search for an experiment type they wish to outsource and choose a facility to perform the work, or post an open project and receive bids from qualified facilities. Science Exchange acts as a centralized hub for provider information and reviews, and assists with project management, including billing and payment. The goal of Science Exchange is to create an integrated and accessible marketplace for experimental services. Results: As of October 2011 more than 3,000 scientists from over 500 research institutions have registered with Science Exchange. The top 10 most searched for experiment types are, in descending order: DNA sequencing, Bioinformatics, Microarray, DNA analysis, electron microscopy, protein extraction/ purification, confocal microscopy, realtime qPCR, microRNA microarray, and NMR. On average, experiments outsourced through the platform have saved users 54% of the value of their experiment ($4,664 per experiment). Conclusions: Science Exchange has begun to address the barriers to effective experiment outsourcing. As with any marketplace, awareness and participation by key players is essential to increase utility. Experience also provides data necessary to make outsourcing as frictionless as possible for the particulars of each experiment type. Our team has an enduring commitment to making the platform as useful for researchers and providers as possible.

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 3986. doi:1538-7445.AM2012-3986
Abstract 4010: New AXIN2 germline variant in a Spanish family with an attenuated form of FAP without oligondontia or ectodermal dysplasia signs
Authors: Rivera Barbara , Perea Jose , Sanchez Eloy , Villapun Manuel , Sanchez-Tome Eva , Mercadillo Fatima , Robledo Mercedes , Benitez Javier , Urioste Miguel
Published: August 2012

Familial Adenomatous Polyposis (FAP) is a colorectal cancer syndrome caused mainly by mutations in the APC gene (50-80% of the cases), and some mutations in MUTYH, usually in recessive and milder cases. Additionally, the genetic cause of 20% of classic forms and 50% of attenuated forms of the disease remain unclear. AXIN2 is a partner of the APC protein in the b-catenin destruction complex. Mutations in AXIN2 have been described in patients with oligodontia and other ectodermal dysplasia features in association with colorectal neoplasia or adenomatous polyps. We studied the AXIN2 gene in a series of APC/MUTYH mutation negative families and identified a novel variant, c.1387C>T;p.R463P, as the possible genetic cause of an attenuated FAP phenotype (AFAP). The autosomal dominant AFAP phenotype was not associated with any extracholonic manifestations or ectodermal dysplasia features. However, hyperplastic polyps were observed in some AXIN2-variant carriers. Thus, from a clinical point of view this family displayed some features that differ from previously described families with AXIN2 mutations. The variant c.1387C>T;p.R463P is located in the b-catenin binding domain, an area highly conserved throughout evolution. The variant segregates with the affected family members and was not present in 800 healthy Spanish controls. We were able to confirm loss of heterozygosity in 3 out of 5 polyps from the proband and his affected sister. AXIN2 and b-catenin expression patterns were tested by inmunofluorescence staining: both proteins are overexpressed in adenomatous polyps compared with normal colorectal ephitelium from the proband. Moreover, no colocalization of the two proteins could be observed by confocal microscopy. These results support a possible pathogenic role for this AXIN2 variant in our family and highlight the implication of AXIN2 in some cases of FAP syndrome. We argue comprehensives and functional studies will be necessary to clarify the role of AXIN2 in familial colorectal cancer predisposition.

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 4010. doi:1538-7445.AM2012-4010