bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5370: PTEN loss confers BRAF inhibitor resistance to melanoma cells through the suppression of BIM expression
Authors: Paraiso Kim H. , Rebecca Vito , Fedorenko Inna V. , Xiang Yun , Wood Elizabeth , Anderson Alexander R. , Sondak Vernon K. , Abel Ethan V. , Koomen John M. , Messina Jane L. , Smalley Keiran S.M
Published: June 2012

The current study has addressed the role of PTEN loss in intrinsic resistance to the BRAF inhibitor PLX4720. Immunohistochemical staining of a tissue array covering all stages of melanocytic neoplasia (n=147) revealed PTEN expression to be lost in >10% of all melanoma cases. Although PTEN status did not predict for sensitivity to the growth inhibitory effects of PLX4720, it was predictive for apoptosis, with little cell death observed in melanomas lacking PTEN expression (PTEN-). Mechanistically, PLX4720 was found to stimulate AKT signaling in the PTEN- but not the PTEN+ cell lines. Liquid chromatography multiple reaction monitoring mass spectrometry (LC-MRM) was performed to identify differences in apoptosis signaling between the two cell line groups. PLX4720 treatment significantly increased BIM expression in the PTEN+ (>14-fold) compared to the PTEN- cell lines (4-fold). A role for PTEN in the regulation of PLX4720-mediated BIM expression was confirmed by siRNA knockdown of PTEN and through re-introduction of PTEN into cells that were PTEN-. Further studies showed that siRNA knockdown of BIM significantly blunted the apoptotic response in PTEN+ melanoma cells. Dual treatment of PTEN- cells with PLX4720 and a PI3K inhibitor enhanced both BIM expression and the level of apoptosis. The importance of rebound PI3K/AKT signaling in BRAF inhibitor resistance was confirmed in 3D organotypic cell culture and long-term colony formation assays. In conclusion, we have shown for the first time that loss of PTEN contributes to intrinsic BRAF inhibitor resistance via the suppression of BIM-mediated apoptosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5370. doi:10.1158/1538-7445.AM2011-5370
Abstract 5455: Integrative analysis of the Cancer Cell Line Encyclopedia reveals genetic and transcriptional predictors of compound sensitivity
Authors: Barretina Jordi , Stransky Nicolas , Caponigro Giordano , Kim Sungjoon , Margolin Adam , Venkhatesan Kavitha , Kryukov Gregory , Berger Michael , Monahan John , Morais Paula , Meltzer Jodi , Mahan Scott , Sonkin Dmitriy , Raman Pichai , Jones Michael , Meyer Vic , Wilson Christopher , Thibault Joseph , Murray Lauren , Callahan Adam , Che John , Shipway Aaron , Li Nanxin , Engels Ingo , Su Andrew , Pinchback Reid , Nedzel Jared , Liefeld Ted , Niu Lili , Hatton Charles , Palescandolo Emanuele , Gupta Supriya , Onofrio Robert , Sougnez Carrie , MacConaill Laura , Winckler Wendy , Reich Michael , Mesirov Jill , Ardlie Kristin , Getz Gad , Morrissey Michael , Warmuth Markus , Meyerson Matthew , Finan Peter , Golub Todd , Weber Barbara , Harris Jennifer , Sellers William , Schlegel Robert , Garraway Levi
Published: June 2012

Comprehensive genomic characterization of cancer is proceeding at a rapidly accelerating pace, mainly due to the expanded use of massively parallel sequencing. Despite the promise of cancer genomics, many cancer drugs still fail in the clinic due to nonresponsive patients and this translates into a significant unmet medical need. Accurate predictions of which patients are more likely to respond to drugs in development could speed clinical trials and personalize treatments.

Here we propose the use of a compendium of experimentally tractable cancer model systems, ∼1000 human genomically-annotated cancer cell lines (at the level of gene expression, DNA copy number alterations and mutations), coupled with pharmacological profiling, to systematically link genetic and transcriptional features to drug response. This resource, the Cancer Cell Line Encyclopedia (CCLE), is available online at www.broadinstitute.org/ccle.

Through computational predictive modeling we have both rediscovered molecular features that predict response to several drugs and also uncovered a number of novel potential biomarkers of sensitivity and resistance to targeted agents and chemotherapy drugs. For instance, we have found that response to topoisomerase 1 inhibitors seem to be driven by expression of a single gene. We have also observed that tissue lineage is a key predictor for sensitivity to certain compounds, providing rationale for clinical trials of these drugs in particular cancer types.

Our cell line-based platform provides a valuable tool for the development of personalized cancer medicine, revealing critical tumor dependencies and helping to stratify patients for clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5455. doi:10.1158/1538-7445.AM2011-5455
Abstract 5588: Szygium aromaticum (clove): A novel natural breast cancer inhibitor
Authors: Li Hong , Tait Larry , Raz Avraham , Nangia-Makker Pratima
Published: June 2012

The dried flower buds of clove (Syzygium aromaticum syn Eugenia aromaticum or caryophyllata) belonging to family Myrtaceae are used as a spice worldwide. Clove has been reported to exhibit antimicrobial, anti-viral, anesthetic, insecticidal, anti-convulsant, anti-oxidative and free radical scavenging activities. Eugenol, an active component of essential oil isolated from clove has anti-mutagenic, anti-genotoxic and anti-inflammatory properties. It also restricts DMBA induced skin carcinogenesis in mice. Aqueous extract of clove was effective in reducing the incidence of hyperplasia, dysplasia, and carcinoma in situ in benzo[a]pyrene induced lung carcinoma in mice. However, there are no reports on the effects of clove on human cancers.

In this study we have analyzed the inhibitory effects of the aqueous extract of clove (CAE) on breast cancer using in vitro studies like cell growth, cell migration, three-dimensional capillary tube formation / morphogenesis and apoptosis; and in vivo studies like breast tumor growth in nude mice. Our results indicate that CAE inhibited the growth and proliferation of breast cancer cell lines MDA-MB-231, MCF10ADCIS.com, and MCF10AEIII 8 cells in a dose dependent manner. In addition, it also reduced cell migration towards Matrigel. CAE also inhibited the enzymatic activity of the Matrix Metalloproteinase-2 (MMP-2), which was analyzed by using two substrates, namely galectin-3 and a fluorescence-quenched substrate MOCAcPLGLA2pr(Dnp)AR-NH2. Cell cycle analysis of MCF10ADCIS.com cells using flow cytometry after propidium iodide (PI) staining indicated an arrest mainly in S-phase. We also observed an inhibition of DNA synthesis in CAE-treated MCF10ADCIS.com cells by bromodeoxyuridine (BrdU) incorporation. Expressions of apoptosis-related proteins, poly(ADP-ribose)polymerase (PARP), caspase-3 and caspase-8 remained unchanged after CAE treatment, but CDK2, CDK4, cyclin A and cyclin D2 were down-regulated in treated cells indicating that CAE might affect tumor growth by inducing cell cycle arrest but not apoptosis. There was a 50% reduction in tumor volume in nude mice injected with MCF10ADCIS.com cells that received CAE extract in their drinking water compared to the control mice. The expression of proliferation, apoptosis, angiogenesis and tumor progression markers (PCNA, TUNEL, p63, CD31, smooth muscle actin, collagen IV, MMP-2 are being investigated in the xenografts. The results suggest that CAE may be explored further as a novel inhibitor of breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5588. doi:10.1158/1538-7445.AM2011-5588
Abstract 676: Dinitroazetidines are a novel class of anticancer agents and hypoxia-activated radiation sensitizers developed from highly energetic materials
Authors: Ning Shoucheng , Bednarski Mark , Oronsky Bryan , Scicinski Jan , Saul Gordon , Knox Susan J.
Published: June 2012

Here we report the first biologic studies of a unique class of potentially clinically useful compounds, derived from highly energetic materials. In an effort to develop unique targeted cancer therapies which maximize cytotoxicity to cancer cells while minimizing unwanted side effects to normal tissue, we have studied a series of novel, non-explosive compounds, based on the highly energetic dinitroazetidine scaffold. ABDNAZ, 1-acetylbromo-3,3-dinitroazetidine is a lead compound for clinical development both as a stand-alone chemotherapeutic drug and as a radiation sensitizer. In vitro studies demonstrated that ABDNAZ generated reactive oxygen species (ROS) in a concentration- and time-dependent manner. Modulation of intracellular redox status by buthionine-(S,R)-sulfoxime or N-acetylcysteine significantly affected ABDNAZ-induced apoptosis of tumor cells. ABDNAZ administered as a single agent was as cytotoxic to tumor cells as cisplatin with IC50s of 2.6 ± 1.6uM and 4.4 ± 2.2uM for ABDNAZ and cisplatin, respectively. ABDNAZ was more cytotoxic to tumor cells than cisplatin or tirapazamine under hypoxic conditions. ABDNAZ and cisplatin had similar effects on well established SCC VII tumors in mice, but unlike cisplatin, ABDNAZ at well-tolerated sub-maximum tolerated doses (MTD) significantly inhibited tumor growth without systemic toxicity. When combined with local tumor radiation therapy, ABDNAZ significantly enhanced the efficacy of the radiation and improved the therapeutic index of radiation therapy. Toxicity studies demonstrated that ABDNAZ was not myelosuppressive and had no apparent dose limiting toxicity when administered at sub-MTD dose daily for 14 days. These data demonstrate that ABDNAZ is a promising novel anticancer drug with a very favorable toxicity profile at therapeutic doses with activity in preclinical tumor models as both a stand-alone chemotherapeutic agent and as a radiosensitizer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 676. doi:10.1158/1538-7445.AM2011-676
Abstract 770: Lorvotuzumab mertansine (IMGN901) immune effector activity and its effect on human NK cells
Authors: Skaletskaya Anna , Setiady Yulius Y. , Park Peter U. , Lutz Robert J.
Published: June 2012

Lorvotuzumab mertansine, or IMGN901, is a conjugate consisting of the cytotoxic maytansinoid, DM1, covalently linked to the monoclonal antibody, lorvotuzumab, which selectively targets CD56 (also called neural cell adhesion molecule or NCAM). This conjugate is currently being evaluated in clinical trials for the treatment of CD56+ solid tumors and multiple myeloma. It is able to inhibit tumor cell growth by disruption of microtubule dynamics through delivery of its cytotoxic DM1 payload; however, other potential cell killing mechanisms, such as the immune effector activity of its antibody component, have not been fully investigated.

Herein, we report on the immune effector activity of the lorvotuzumab antibody and the lorvotuzumab mertansine conjugate and their effect on the function of CD56+ human primary NK cells in vitro. To investigate the effector activity of the antibody, lorvotuzumab was incubated with CD56+ tumor target cells in the presence of rabbit complement or purified human primary NK cells. The results show that this antibody fails to mediate complement dependent cytotoxicity (CDC) but induces antibody-dependent NK cell-mediated cytotoxity (ADCC) in dose-dependent manner. The complete conjugate showed ADCC activity comparable to lorvotuzumab alone, suggesting that DM1 conjugation did not alter the interaction between the Fc domain of the antibody and NK cell's Fc receptor. Since human primary NK cells express CD56, albeit at a low level, we tested whether lorvotuzumab mertansine or its antibody component could negatively impact the function of NK cells. The results demonstrate that incubation with this antibody or with this conjugate has no effect on rituximab-mediated ADCC activity of the NK cells. Furthermore, overnight exposure of human primary NK cells with lorvotuzumab mertansine did not change rituximab-mediated ADCC activity.

In conclusion, the lorvotuzumab antibody has ADCC activity that is retained after DM1 conjugation. Furthermore, neither lorvotuzumab nor its DM1 conjugate, appear to affect the ability of NK cells (which are CD56+) to mediate ADCC through another antibody.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 770. doi:10.1158/1538-7445.AM2011-770
Abstract 910: High dietary fat promotes inflammation and intestinal neoplasia, independently of diet-induced obesity, in B6.ApcMin/+ congenic-consomic mouse strains
Authors: Doerner Stephanie K. , Leung Elaine S. , Berger Nathan A. , Nadeau Joseph H.
Published: June 2012

Obesity and high fat (HF) diet are associated with increased risk of colon cancer. However, assessing their independent contributions to intestinal carcinogenesis is difficult due to their close association in humans. We utilized Chromosome Substitution Strains (CSSs) in combination with the B6.ApcMin/+ mouse model of intestinal cancer to generate Congenic-Consomic Strains (C-CSs). Using C-CSs, we tested whether a HF diet promotes the development of intestinal cancer, independent of obesity, and found that dietary fats are strong and independent determinants of polyp formation.

CSSs, which have contrasting responses to a HF diet, were used to construct C-CSs that were susceptible to intestinal cancer, but were either resistant or susceptible to diet-induced obesity, depending on the substituted A/J chromosome. A2.ApcMin/+ (obese), A7.ApcMin/+ (lean), and A17.ApcMin/+ (lean) mouse strains were maintained on diets high or low in coconut oil (HFCoco and LFCoco, respectively). If obesity is crucial in polyp formation, all obese strains should develop more polyps than the lean strains. In contrast, if diet is important, we expected to find an increase in polyp number in all strains fed the HFCoco diet, independent of their susceptibility to diet-induced obesity.

Polyp numbers were significantly increased in all strains after 60 days exposure to the HFCoco diet, regardless of obese or lean status, suggesting that a HF diet promotes polyp development. Polyp numbers were also significantly higher in B6.ApcMin/+ after similar exposure to a high fat corn oil (HFCorn) diet. We measured levels of circulating inflammatory factors and showed that adiponectin is decreased and leptin, interleukin (IL)-1beta and IL-6 are increased in B6.ApcMin/+ fed either of the HF diets compared to mice fed the corresponding LF control diets, implicating a role of inflammation in diet-induced intestinal neoplasia.

With the novel C-CS systems, we were able to elucidate complex diet-gene interactions involved in intestinal carcinogenesis and provide evidence for a crucial role of a HF diet in the development of intestinal polyps. These dietary studies have implications for both prevention and non-invasive treatment of intestinal cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 910. doi:10.1158/1538-7445.AM2011-910
Abstract 934: Chromokinesin KIF4 facilitates chromatin organization critical for immunoglobulin class switch recombination and prevents lymphoma formation
Authors: Mazumdar Manjari , Dasari Siddharth , Keen Jennifer
Published: June 2012

Un-repaired DNA damage and chromosomal translocations are the major causes of genomic instability found in various human cancers. We have observed spontaneous B-cell Follicular lymphomas (FL) in a knock-out (KO) mouse model generated by gene trapping of KIF4, a unique DNA-binding, nuclear and chromatin-associated molecular motor. We have previously shown that KIF4 is an important modulator of higher-order chromatin structure and is required for maintaining genomic stability. Since FL often results from altered levels of DNA recombination of the immune system caused by chromosome structural changes, we investigated the contributions of KIF4 in regulating chromatin structure for B cell function. We show that there is an abnormal proliferation of B cells in mice in both bone marrow and spleen of young adult mice lacking KIF4 function. Chromosome spread from young adult knockout and neoplastic splenocytes show abnormal chromosome numbers (aneuploidy) and structure in the absence of KIF4 indicating that KIF4 is critical in maintaining genome integrity in vivo. Maintaining and establishing a stable but dynamic chromatin organization is critical for successful class switch recombination during B cell development. We observed that immunoglobulin (Ig) class switch recombination (CSR) in activated B cells is severely impaired in the absence of KIF4. Moreover, our biochemical and proteomic analysis reveals that repressive chromatin marks such as trimethylation of histone H3 lysine 9 or 27 that regulate global gene expression patterns, are significantly altered in maturing B cells in the absence of KIF4 compared to wild type control cells. This data demonstrates that KIF4 is necessary for modulating dynamic chromatin structure via a remodeling activity that allows or restricts access to the Igh locus for CSR and antibody diversity.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 934. doi:10.1158/1538-7445.AM2011-934
Abstract LB-1: An investigator-initiated, phase II randomized, double-blind, placebo-controlled trial of GDC-0449 for prevention of BCCs in basal cell nevus syndrome (BCNS) patients.gg
Authors: Tang Jean Y. , Mackay-Wiggan Julian M. , Aszterbaum Michelle , Lindgren Joselyn , Chang Kris , Coppola Carol , Campbell Angela , Chanana Anita , Marji Jackleen , Callahan Chris , Yauch Robert , Bickers David R. , Epstein Ervin H.
Published: June 2012

Purpose: GDC-0449 (Genentech) is a new hedgehog pathway inhibitor that has been shown in a Phase I trial to reduce locally advanced/metastatic BCCs. We tested the efficacy of GDC-0449 for BCC prevention in a randomized, double-blind, placebo-controlled trial in patients with Basal Cell Nevus Syndrome (BCNS).

Methods: We enrolled 41 BCNS patients from 3 clinical centers from September 2009 to January 2011. Subjects were randomized 2:1 to receive once daily oral GDC-0449 (150 mg) or placebo. The primary endpoint was to assess the number of new surgically eligible BCCs per month after 3 months; secondary endpoints included assessment of the change in size of existing BCCs and safety/tolerability.

Results: 36 subjects (24 GDC-0449, 12 placebo) have been enrolled for greater than 1 month. Upon review of results at the interim analysis, the DSMB ended the placebo arm of the trial due to statistically significant differences between the 2 groups. Subjects treated with GDC-0449 developed 0.07 new BCCs per month vs. 1.74 BCCs per month for those receiving placebo (p<0.0001). The change from baseline in the aggregate size of existing BCCs was −24 cm vs. −3 cm for GDC-0449 and placebo, respectively (p=0.006). Some subjects achieved near complete remission with no BCC developing resistance. Common grade 1–2 adverse events (GDC-0449 vs. placebo) included taste loss (83% vs. 8%), muscle cramps (67% vs. 8%), and weight loss (50% vs. 8%). Two GDC-0449 subjects experienced grade 3–4 adverse events (muscle cramp, suicide attempt). Overall 20% of GDC-0449 subjects discontinued drug participation due to adverse events. BCC samples from 23 subjects treated with GDC-0449 or placebo were available for molecular analysis. The anti-BCC efficacy of GDC-0449 was associated with on-target reduction of the Hedgehog pathway as Gli1 mRNA levels were decreased by 200-fold (p<0.001) after 1month in BCCs. Gli1 mRNA levels were not changed in BCCs from placebo subjects. GDC-0449 also decreased tumor proliferation (Ki67) but had no effect on apoptosis (CC3). Biopsies of lesions taken after 3 months of GDC-0449 showed histologic clearance in 7 out of 11 samples.

Conclusion: GDC-0449 has anti-BCC efficacy in BCNS patients and may be a potential new drug for BCC prevention and treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-1. doi:10.1158/1538-7445.AM2011-LB-1
Abstract LB-156: Improved T cell survival, tumor localization and tumor eradication by folate receptor-alpha redirected T cells is achieved by provision of CD137 costimulatory signaling
Authors: Song De-Gang , Canevari Silvana , June Carl , Coukos George , Powell Daniel J.
Published: June 2012

Human T cells engineered to express a chimeric immune receptor (CIR) specific for folate receptor-alpha (FRα) have shown robust anti-tumor activity against epithelial cancers in vitro but not in the clinic due to their inability to persist and home to tumor in vivo. To address these issues, we constructed CIRs containing an FRα-specific scFv (MOv19) coupled to the T cell receptor CD3ζ chain signaling module alone or in combination with the CD137 (4–1BB) costimulatory motif in tandem. Primary human T cells transduced to express conventional MOv19-ζ or costimulated MOv19-BBζ CIRs secreted various proinflammatory cytokines and exerted cytotoxic function when cocultured with FRα+ tumor cells in vitro. However, only transfer of human T cells expressing the costimulated MOv19-BBζ CIR mediated tumor regression in immunodeficient mice bearing large, established FRα+ human cancer. MOv19-BBζ CIR T cell infusion mediated tumor regression in models of metastatic intraperitoneal, subcutaneous, and lung-involved human ovarian cancer. Importantly, tumor response was associated with the selective survival and tumor localization of human T cells in vivo and was only observed in mice receiving costimulated MOv19-BBζ CIR T cells. T cell persistence and anti-tumor activity were primarily antigen-driven, however, antigen-independent CD137 signaling by CIR did improve T cell persistence but not anti-tumor activity in vivo. Our results show that anti-FRα CIR outfitted with CD137 costimulatory signaling in tandem overcome issues of T cell persistence and tumor localization that limit the conventional FRα T cell targeting strategy to provide potent antitumor activity in vivo. Based upon these encouraging findings, we are preparing to open an anti-FRα CIR protocol for ovarian cancer in mid-2011, pending FDA approval.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-156. doi:10.1158/1538-7445.AM2011-LB-156
Abstract LB-230: Enhancing anti-angiogenic therapy by blocking focal adhesion kinase
Authors: Bottsford-Miller Justin , Sanguino Angela , Thanapprapasr Duangmani , Pecot Chad V. , Stone Rebecca L. , Auger Kurt , Nick Alpa M. , Sood Anil K.
Published: June 2012

Focal adhesion kinase (FAK) represents a central point of convergence for many signaling pathways implicated in cancer progression and metastasis. Pazopanib is a pan-VEGFR and PDGFR inhibitor. In the present study, we examined whether pazopanib treatment would result in greater anti-tumor activity in combination with the novel FAK-inhibitor, GSK2256098. The in vitro effects of GSK2256098 on invasion and migration were examined using the HeyA8 and SKOV3-IP1 human ovarian cancer cell lines. In vivo effects of pazopanib with and without GSK2256098 were then assessed using an orthotopic mouse model of human ovarian cancer. GSK2256098 resulted in reduced levels of FAK phosphorylation at Y397 (pFAKY397) at 1μM concentration in SKOV3-IP1 cells. GSK2256098 resulted in a reduced invasion (p<0.001) and decreased migration (p<0.001) in SKOV3-ip1 cells. Dose-finding studies performed in vivo demonstrated that a 75 mg/kg dose resulted in a significant reduction in pFAKY397. Monotherapy with GSK2256098 resulted in a 58% decrease in mean tumor weight compared to control (p = 0.038). The combination of GSK2256098 with pazopanib resulted in a 71% decrease in mean tumor weight compared to pazopanib monotherapy (p = 0.04). We also tested treatment combinations including traditional cytotoxic chemotherapy. The combination of GSK2256098 with docetaxel resulted in a 44% decrease in mean tumor weight compared to docetaxel monotherapy (p = 0.17). The triplet combination of GSK2256098 with pazopanib and docetaxel resulted in the greatest overall decrease mean tumor weight, 99% compared to control and 92% compared to the doublet alone (p = 0.001). Similar trends were noted with mean number of tumor nodules and ascites volume. On the basis of our recent findings that FAK is highly phosphorylated in platelets in cancer-bearing mice, we also examined FAK phosphorylation in the platelets of treated and untreated mice. Phosphorylation of platelet FAK (pFAKY397) was inhibited by 68% at 4 hours after treatment compared to control (p = 0.07). Treatment with pazopanib decreased MVD by 49% (p < 0.01), which was further enhanced in combination with GSK2256098 (p < 0.01). In summary, FAK inhibition results in substantial anti-angiogenic and anti-tumor effects in combination with pazopanib and represents a viable strategy for further development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-230. doi:10.1158/1538-7445.AM2011-LB-230
Abstract LB-248: IRF4 as a critical regulator and potential therapeutic target in ABC type diffuse large B-cell lymphoma
Authors: Tolga Emre N. C. , Shaffer Arthur L. , Wright George W. , Kohlhammer Holger , Staudt Louis M.
Published: June 2012

Interferon regulatory factor 4 (IRF4) is a transcriptional regulator with crucial roles in the development of lymphocytes and implicated in malignant transformation. Recently, we have demonstrated that cell lines derived from cancers of plasma cell origin (i.e., multiple myeloma) are addicted to an anomalous gene expression program driven by the wild-type IRF4. Here, we show that an aggressive malignancy of mature B-cells, the activated B-cell-like type of diffuse large B-cell lymphoma (ABC-DLBCL), also requires IRF4 for survival. With an integrative analysis of genome-wide IRF4 localization (ChIP-Seq) assays and gene expression profiling, we identify IRF4 target genes in ABC-DLBCL as members of diverse pathways related to B-cell biology and oncogenesis, largely distinct from the IRF4 targets in multiple myeloma cells. For instance, we identify CARD11, an adaptor in the NF-κB pathway, as a target of IRF4 in ABC-DLBCL, and confirm this by cell-based reporter assays. This novel regulatory mechanism complements the established aspects of ABC-DLBCL pathobiology, where NF-κB pathway is constitutively active and indispensable, and drives IRF4 expression. Furthermore, we find enrichment for a composite ETS-IRF DNA motif in IRF4 binding regions in ABC-DLBCL, suggesting cooperative activity between IRF4 and ETS transcription factor(s). Supporting this, we demonstrate through complementation assays that critical amino acids on IRF4 and the ETS factor SPIB, notably those residues important for their interaction, are essential for the survival of ABC-DLBCL, but not multiple myeloma cells. Taken together we show that ABC-DLBCL cells are addicted to the transcription factor IRF4, in part through a positive feedback mechanism involving CARD11, NF-κB, and IRF4. In addition, our results suggest therapeutic potential for targeting the IRF4-ETS interaction interface in ABC-DLBCL.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-248. doi:10.1158/1538-7445.AM2011-LB-248
Abstract LB-27: Dissecting signaling transduction network to infer master regulators of non small cell lung cancer
Authors: Bansal Mukesh ,   Dissecting signaling transduction network to infer master regulators of non small cell lung cancer, Bansal Mukesh , Kustagi Manjunath , Lefebvre Celine , Rieckhof Gabrielle , Girard Luc , Merberg David , Rikova Klarisa , Minna John , Comb Michael J. , White Michael , Califano Andrea
Published: June 2012

Abstract

The dissection of signal transduction networks is lagging significantly with respect to their transcriptional and protein-complex counterparts. Innovative use of multiplex and microarray-based approaches, where multiple antibodies can be used to probe an ensemble of phosphoproteins, is starting to become sufficiently mature to allow characterization of small pathways. Yet, this is still far from providing an unbiased, genome-wide view of signal-transduction processes.

Using data from the first genome-wide profiling of phosphopeptide abundance in normal and tumor related lung tissue1, we have designed a novel algorithm, pARACNe, to generate an unbiased, genome-wide network representing tyrosine kinase signal transduction pathways. Specifically, pARACNe was used to infer tyrosine kinase substrates in non-small cell lung cancer (NSCLC) from tandem mass-spec abundance profiling of phosphotyrosine enriched peptides from ∼250 samples derived from primary tumors, cell lines, and normal lung tissue. SILAC validation of EGFR, MET and ALK substrates, confirmed >80% of the pARACNe inferred interactions. Further analysis of large-scale NSCLC microarray expression profile data, using the MINDy algorithm2, showed significant convergence between the signal transduction pathways inferred by proteomics and gene expression based methods.

The availability of an accurate NSCLC Tyrosine Kinase Interaction Network (TKIN) has significant implications for our ability to elucidate master regulators of oncogenesis and tumor progression and for the identification and prioritization of druggable targets. We used the MARINa algorithm and the TKIN to infer a complete repertoire of master regulator (MRs) kinases of 46 cell lines. Initial validation shows that the method could recapitulate both cancer specific MRs, such as EGFR, as well as cell line specific ones, such as ALK and PTK2 in H2228 cells. Follow up experimental assays have shown that the PTK2, which is active in tumor-related but not in normal cells, is required for H2228 cell viability and may constitute a novel oncogene addiction in NSCLC and a candidate therapeutic target. The TKI suggests that current pathway models are incomplete and sets the stage for quantitative, network-based methods to identify key regulators of physiologic and pathologic cell phenotypes.

References

1. Rikova K et al. Cell 2007;131:1190–203.

2. Wang K et al. Nat Biotechnol 2009;27:829–39.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-27. doi:10.1158/1538-7445.AM2011-LB-27
Abstract C61: The role of prostanoid signaling in lung cancer
Authors: Pamarthi Pavan Kumar
Published: June 2012

Rationale: An increasingly large body of evidence indicates that PGE2 promotes tumor growth by stimulating EP4 receptor signalling with subsequent enhancement of cellular proliferation, metastasis and suppression of immune responses. However, the role of other EP receptors is not known.

Objective: We hypothesized that prostanoid receptors other than the EP4 receptor are involved in progression of Lung cancer.

Methods: We analyzed the expression profile of EP receptors in lung adenocarcinoma cell lines A549 and H1299 by using Real Time PCR, western blotting and immunofluorescence. All four EP receptors are getting expressed in both the cell lines. Both L-161982 (EP4 receptor specific antagonist) and SC-51322 (EP1 receptor specific antagonist) inhibited proliferation of both cell lines as assessed by thymidine incorporation assay. EP2 and EP3 receptors have not shown inhibitory affects in proliferation assays. SC-51322 has not decreased cAMP levels in A549 and decreased intracellular calcium levels. SC-51322 has inhibited pERK levels as shown in the western blots. In addition, SC-51322 inhibited migration of A549 cells in transwell migration assay.

Conclusion: We conclude that EP1 receptor may play a role in lung cancer progression and metastasis. Finding out downstream molecules to EP1 receptor will give us a better understanding of this pathway in lung cancer and can provide therapeutic opportunities.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr C61.
Abstract C9: Quantitative modeling of androgen receptor-mediated transcriptional complexes in human prostate cancer cells
Authors: Hsiao Jordy J. , Martinez Hairyl D. , Wright Michael E.
Published: June 2012

The androgen receptor (AR) is a member of the nuclear receptor superfamily and the key transcription factor of androgen-mediated signaling pathways. AR translocates to the nucleus in response to ligand binding, where it selectively binds canonical androgen response elements (AREs) in DNA to assemble transcriptional complexes at target genes. Aberrant AR signaling resulting from unbalanced AR transcriptional responses at genes involved in cellular proliferation and differentiation drives the development and progression of human prostate cancers. In addition, the AR coregulator proteins, members of a large family that bind AR and modulate AR transcriptional activity, are known to elicit aberrant AR activity in prostate cancers. Although numerous AR coregulators have been identified, in most cases this was through static, binary protein-protein interaction assays (e.g., GST-fusion, yeast two-hybrid assays). Thus, to what degree androgen-sensitive coregulators mediate pathological AR actions in the context of human prostate cancer remains poorly defined. We have devised a biochemical strategy to identify and quantify androgen-sensitive coregulators (coactivators and corepressors) of AR-mediated transcription, based on their in vitro affinity for a model androgen-regulated gene promoter. Specifically, we have used DNA-affinity purification techniques in combination with label-free quantitative mass spectrometry (qMS) to do so. Our preliminary findings show that androgen-sensitive coregulators represent a functionally diverse group of protein involved in a number of signal transduction pathways. Furthermore, future studies utilizing selected reaction monitoring (SRM), a quantitative proteomics method, will be exploited to study the molecular functions of androgen-sensitive coregulators and facilitate the construction of quantitative models of AR-mediated transcription that will make it possible to predict androgenic responses at disease-associated, androgen-regulated genes. Our findings will broadly benefit the biomedical research community in that they define a novel class of androgen-sensitive coregulators and determine how they influence AR-mediated transcriptional processes underlying the development and progression of human prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr C9.
Abstract IA11: New insights into the microenvironment and metastasis
Authors: Werb Zena
Published: June 2012

Both extrinsic and intrinsic mechanisms regulate the development of the aberrant tumor organ and regulate its progression. We have used genetic and in vivo imaging techniques to study the interaction between epithelial cancer cells and the tumor microenvironment during tumor progression. Transcriptional regulation of epithelial differentiation, cell-cell interaction and motility is a key intrinsic event in tumor progression. We have investigated GATA3, a master regulator for luminal differentiation, which is lost upon malignant conversion, and found that it allows the malignant tumor cells to disseminate throughout the body. The progressing tumor cells develop in an extrinsic microenvironment rich in inflammatory cells, growth factors, angiogenesis and activated stroma that promotes neoplastic risk. All of these components communicate with each other to contribute to the aberrant tumor organ. The tumor microenvironment regulates access to therapeutics and may alter expression of drug resistance genes. To explore the relationship between microenvironment and cancer cells in drug response, we treated mouse models of mammary carcinoma with doxorubicin. Primary cancer cells isolated from different tumor sizes had the same sensitivity in 2D and 3D cultures. Live imaging confirmed that individual cancer cells in early carcinoma stages were more sensitive to doxorubicin than in earlier or later stages. Imaging intact tumors during treatment with chemotherapy revealed marked differences in drug penetration and sensitivity of cancer cells in different tumor microenvironments. The inflammatory cells modulate the tumor ecology, altering vascular permeability and responses to chemotherapy. These experiments establish that chemosensitivity is determined by cooperation between cancer cell intrinsic and extrinsic factors.

Supported by grants from the National Cancer Institute

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr IA11.
Abstract A103: The synthetic triterpenoid CDDO-methyl ester targets tumor-associated macrophages to delay carcinogenesis in the PyMT model of estrogen receptor negative breast cancer
Authors: Tran Kim M. , Risingsong Renee , Royce Darlene , Williams Charlotte R. , Sporn Michael B. , Liby Karen
Published: June 2012

Breast cancer is the most common cancer among women in the United States. The incidence rates of breast cancer are no longer significantly declining, and approximately 40,000 women die from the disease each year. Hence, novel drugs are needed for the prevention and treatment of the disease. Synthetic triterpenoids are a promising new class of compounds with chemopreventive activity in a variety of preclinical cancer models. We tested the methyl ester derivative of the synthetic triterpenoid, 2-cyano-3, 12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO-Me), in a relevant model of ER-negative breast cancer. In this mouse model, the polyoma-middle T (PyMT) oncoprotein drives carcinogenesis in the mammary gland. The developing tumors recapitulate key features of the human disease including significant infiltration of tumor-associated macrophages (TAM). Depletion of TAMs has been previously shown to delay tumor progression. PyMT mice were fed CDDO-Me (50 mg/kg diet), starting at 4 weeks of age. CDDO-Me significantly increased the age of mice at onset of first tumor by an average of 4.3 weeks (P < 0.001) and overall survival by 5.2 weeks (P < 0.001). CDDO-Me significantly (P < 0.05) inhibited the infiltration of TAMs into mammary glands of PyMT mice along with levels of the chemokines CXCL12 and CCL2 in primary PyMT mammary tumor cells. Treatment with CDDO-Me also inhibited secretion of MMP-9 in primary tumor cells from PyMT mice and decreased proliferation of these cells by inhibiting cyclin D1 and decreasing phosphorylation of EGFR and STAT3, key breast cancer biomarkers. Thus, CDDO-Me delayed breast carcinogenesis via inhibition of TAM infiltration and by inhibition of key pathways in breast carcinogenesis. Taken together, these findings implicate CDDO-Me as a promising chemopreventive agent for ER-negative breast cancer via inhibition of TAM infiltration and diverse oncogenic pathways.

Citation Information: Cancer Prev Res 2011;4(10 Suppl):A103.
Abstract A106: Induction of synthetic lethality in mutant KRAS cells for lung cancer prevention and therapy
Authors: Huang Shaoyi , Ren Xiaoyang , Wang Haizhen , Wu Xiangwei
Published: June 2012

Lung cancer is the leading cause of cancer death in both men and women in the United States, with a low five-year survival rate despite improved treatment strategies. Mutations and activation of KRAS occur frequently in non-small cell lung cancers (NSCLCs) and are associated with higher risk of drug-resistance, tumor recurrence, and poor prognosis. Currently, there are no effective therapies for targeting mutant KRAS and there is a great need to develop such a strategy. We have previously discovered a synthetic lethal interaction among TNF-related apoptosis-inducing ligand (TRAIL), the second mitochondria-derived activator of caspase Smac/DIABLO (Smac), and mutant KRAS in premalignant lung epithelial cells, which allows for chemoprevention of KRAS-driven lung cancer by the combination of TRAIL and Smac mimic [Cancer Prev Res (Phila). 2011 May;4(5):666–73.]. Here we demonstrated that this synthetic lethal interaction also exists in malignant lung cancer cells. We showed that 3 out of 4 lung cancer cell lines harboring mutant KRAS displayed high sensitivity to the combination of TRAIL and Smac mimic and KRAS wild type lung cancer cell lines were resistant to the treatment. Expression of mutant KRAS in KRAS wild type lung cancer cells sensitized these cells to TRAIL and Smac mimic and knocking down of mutant KRAS in mutant KRAS expressing cells resulted in resistance to the treatment. Furthermore, we showed that TRAIL and Smac mimic treatment inhibited xenograft tumor growth in mutant KRAS dependent manner. Mechanistically, we found that mutant KRAS repressed cellular FADD-like interleukin 1β–converting enzyme (FLICE)-like inhibitory protein (c-FLIP) expression through the activation of Erk/mitogen-activated protein kinase (MAPK)-mediated activation of c-Myc. Mutant KRAS also enhanced the secretion of Prostate Apoptosis Response-4 (Par4) to promote TRAIL-induced apoptosis. Our study provided a potential new approach for lung cancer chemoprevention and therapy based on synthetic lethality that targets mutant KRAS for apoptosis using the combination of and Smac mimic.

Citation Information: Cancer Prev Res 2011;4(10 Suppl):A106.
Abstract A108: Reproductive and hormonal factors in relation to risk of fatal versus non-fatal invasive epithelial ovarian cancer
Authors: Merritt Melissa A. , Poole Elizabeth M. , Tworoger Shelley S. , De Pari Mary , Cramer Daniel W. , Terry Kathryn L.
Published: June 2012

Ovarian cancer is the fifth leading cause of cancer death for women in the U.S. and the 7th most fatal worldwide. Reproductive and hormonal exposures are known to influence risk of ovarian cancer. We therefore investigated whether these exposures also influence ovarian cancer survival.

We studied survival in relation to hormonal and reproductive history in invasive epithelial ovarian cancer cases and controls from the New England based Case-Control Studies (NECC, 1,642 cases and 2,100 controls). Cases and controls were identified between 1992 and 2008 in New Hampshire and eastern Massachusetts. At the time of enrollment, participants were interviewed in person on a wide range of known and suspected risk factors, including hormone use, childbearing, lifestyle habits and anthropometric characteristics. Polytomous logistic regression was applied to evaluate whether a risk factor was associated differently with fatal (those that died within 3 years of diagnosis) versus non-fatal cases while controlling for other covariates and P-values for heterogeneity (comparing the Odds Ratios (ORs) for fatal ovarian cancer to the ORs for non-fatal ovarian cancer) were calculated based on the maximum likelihood estimates.

A total of 360 of the 1642 women with ovarian cancer (22%) died within 3 years. We found that fatal ovarian cancer was associated with increasing age (multivariate OR 1.05 per 1 year increase in age, 95% CI 1.04 − 1.07, P-value < 0.0001), post-menopausal status (multivariate OR 1.31 vs pre-/dubious menopause, 95% CI 0.97 − 1.77, P-value < 0.0001) and oral contraceptive pill use was consistently more protective across all duration of use categories (P-value = 0.01) as compared with the non-fatal cases. Associations with tubal ligation, total number of ovulatory years, body mass index and family history of breast/ ovarian cancer did not differ significantly between fatal and non-fatal ovarian cancer cases.

These results suggest that the most aggressive subset of invasive epithelial ovarian cancer exhibits important differences in its reproductive and hormonal risk factor profile as compared with non-fatal cases, and that selected protective factors such as oral contraceptive pill use may be especially important for the prevention of the most aggressive subgroup of disease.

Citation Information: Cancer Prev Res 2011;4(10 Suppl):A108.
Abstract A114: Dietary magnesium and calcium intake and risk of prostate cancer in Caucasian and African American men
Authors: Rico Stephanie , Forman Michele , Etzel Carol , Strom Sara S. , Hoque Ashraful
Published: June 2012

Dietary magnesium intake is significantly less in African Americans than Caucasians. Deficiency of magnesium influences immune-inflammatory response and induces oxidative stress that leads to increased cell proliferation and DNA damage. Low magnesium level also increases the physiologic effects of calcium. Therefore, we hypothesize that low dietary intake of magnesium and high calcium intake will increase the risk of developing prostate cancer among African American men. To test this hypothesis, we conducted a pilot case-control study to determine whether deficiency of dietary magnesium and a high calcium intake are associated with an increased risk of prostate cancer. This study included 209 cases with histologically confirmed adenocarcinoma of the prostate (110 African American and 99 Caucasian men). 199 population-based controls were frequency matched by age (±5 years) and race. Risk factor questionnaire and nutrition data were collected via telephone interview. Dietary Assessment was performed using Modified Block-NCI Food Frequency Questionnaire with 129 food items, questions about restaurant intake and vitamin mineral supplementation. Dietary intake was calculated using DIETSYS + Plus version 5.9 dietary analysis program. Nutrition data for 166 cases and 155 controls were available for this analysis. Mean magnesium and calcium intake did not vary significantly between cases and controls. The odds ratios for magnesium was 0.79 (95% CI 0.39–1.61) and for calcium 1.35 (95% CI 0.87–2.09). However, among African Americans, the odds ratio for magnesium was 0.28 (95% CI 0.08–1.05) and for calcium 1.9 (95% CI 1.05–3.40). These preliminary data suggest that high dietary magnesium intake is inversely associated with prostate cancer, while higher calcium intake is associated with increased risk among African American men. These data may have significant public health implications since the identification of African American men deficient of magnesium can be targeted for preventive interventions. Results of this study needs to be confirmed in future studies with larger sample size.

Citation Information: Cancer Prev Res 2011;4(10 Suppl):A114.