bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2381: ApoStreamtm, an antibody independent platform, captures circulating tumor cells in patients with hepatocellular carcinoma
Authors: Poklepovic Andrew , Roberts John D. , Quigley Maria , Melnikova Vladislava , Woo Jacky , Wu Weigou , Davis Darren
Published: August 2012

Biopsies of HCC can be technically difficult, given the vascular nature of the liver and underlying liver disease. Many HCC patients do not get biopsies, and molecular characterization of these tumors is not possible. Capture of CTCs from blood allows for analysis of cancer cells in metastatic dissemination. The use of EpCAM-based enrichment platforms limits the type of tumor cells that can be recovered, as it selects only for cells which express the antigen of interest. Nonselective methods of CTC analysis are needed. An ongoing study is evaluating the recovery of CTCs in HCC patients with elevated serum alpha-fetoprotein (AFP) using the novel antibody-independent ApoStream™ platform, which utilizes the principle of dielectrophoretic field-flow fractionation to position cells in a hydrodynamic flow profile for sorting. Paired blood samples from HCC patients were analyzed by CellSearch and ApoStream™. Collected cells were immunophenotyped using antibodies against Cytokeratin (CK), CD45, DAPI, and AFP, and analyzed by quantitative laser scanning cytometry. In comparison with the CellSearch platform, ApoStream™ isolated higher number of CK+/CD45-/DAPI+ CTCs in HCC cancer patients. A majority of the CK+/CD45-/DAPI+ CTCs enriched by the ApoStream™ platform (62-98%) were AFP+. ApoStreamTM also isolated AFP+, putative CTCs with the CK-/CD45-/DAPI+ phenotype (9-85 cells), and cells with CK+/CD45+/DAPI+ phenotype (1-978 cells). Conclusion: The ApoStreamTM CTC enrichment platform successfully isolated a greater number of CTCs from HCC patients. ApoStream™ CTC analysis is also being used for pharmacodynamic analysis in an ongoing phase I study of the combination of sorafenib and vorinostat in patients with advanced HCC. In preclinical models, the combination synergistically induces tumor cell death through activation of CD95. In this study, CTC analysis will compare the expression of CD95 and other markers prior to and after drug combination exposure.

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 2381. doi:1538-7445.AM2012-2381
Abstract 2528: 3C23K: an anti-human Müllerian inhibiting substance type II receptor humanized monoclonal antibody for ovarian cancer targeted therapy
Authors: Jacquet Alexandra , Kersual Nathalie , Garambois Véronique , Fontayne Alexandre , Pujols Fanny , Fournier Nathalie , Pugnière Martine , Chavanieu Alain , de Romeuf Christophe , Prost Jean-François , Urbain Rémi , Pèlegrin André , Gaucher Christine , Teulon-Navarro Isabelle
Published: August 2012

Asymptomatic in early stages, ovarian cancer is a “silent killer” representing the fifth leading cause of female deaths in western countries. Every year 56,967 women in Europe and USA die as a consequence of this disease. The incidence of ovarian cancer is forecast to undergo a 13 % increase in the next eight years in the seven major developed countries to reach about 72,000 annual cases in 2019. Due to the limitations of the current therapeutic approaches, there is a strong need for novel, more efficient, therapies. For this reason, we have produced a humanized monoclonal antibody 3C23K targeting the human Müllerian Inhibiting Substance type II Receptor (MISRII), expressed on most ovarian cancer subtypes, including epithelial ovarian cancer (EOC) representing more than 90% of ovarian cancers. This monoclonal antibody derives from the murine monoclonal antibody 12G4 and displays a particular glycosylation profile known to favor effector recruitment (EMABling®) as previously demonstrated in vitro. In vivo, we also showed that 3C23K exhibited a significant effect on tumor growth against several ovarian tumor xenografted models derived from patient primary EOC tumors. In this study, we first confirmed by peptide microarray that the epitope of 3C23K antibody was strictly identical to that of 12G4 antibody and a 3D-model of the full MISRII molecule was generated in order to better localize the 3C23K epitope. Furthermore, SPR studies demonstrated cross-reactivity of 3C23K with MISRII of rabbit, dog, pig, cow and primate. In vivo, antitumor activity of 3C23K against xenografted EOC tumor models was confirmed with various concentrations and treatment schedules. Moreover, interestingly, we constructed a mutant form of 3C23K harboring two mutations in the Fc region (G236R/L328R) in order to prevent binding to both murine and human FcαRs, and demonstrated that such a modification abolished antitumor activity. This data confirmed in vivo that recruitment of effectors is essential for 3C23K efficacy. Finally, in order to anticipate clinical treatment of ovarian cancers, 3C23K was tested in vivo in combination with carboplatin. Combination, when compared with treatment of each product alone, resulted in more than additive antitumor activity. Altogether these data showed that humanized monoclonal antibody 3C23K represents a promising candidate for ovarian cancer 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 2528. doi:1538-7445.AM2012-2528
Abstract 2531: Reduction of In Vivo Tumor Growth and Angiogenesis by a Humanized IgG4 Monoclonal Antibody to SEMA4D
Authors: Jonason Alan S. , Fisher Terrence L. , Torno Sebold , Bussler Holm , Doherty Michael A. , Winter Laurie A. , Reilly Christine , Kirk Renee , Howell Alan , Huang He , Caplan Jeff , Kenney Patrick , Scrivens Maria , Pandina Tracy , Croy Leslie , Evans Elizabeth E. , Paris Mark , Watkins Raymond , Zauderer Maurice , Leonard John E. , Smith Ernest S.
Published: August 2012

Semaphorin 4D (SEMA4D; CD100) has been implicated in several key mechanisms of tumor progression, including metastasis, tumor invasion, and neovascularization. SEMA4D binding to its receptor plexin-B1 (PLXNB1) on endothelial cells activates RhoA and AKT signaling pathways, which promotes formation of new blood vessels and tumor growth in vivo. SEMA4D is over-expressed in a wide array of tumor types, and is also produced by inflammatory cells recruited to the tumor microenvironment, such as tumor-associated macrophages. In addition to its effects on endothelial cells, the interaction of PLXNB1 with MET and ERBB2 can lead to SEMA4D-mediated transactivation of these membrane receptor kinases with a direct effect on tumor cell migration and invasive growth. Collectively, these results suggest that expression of SEMA4D, either by tumor cells or by tumor associated inflammatory cells, functions as a crucial factor in tumor metastatic potential, and that expression of SEMA4D and/ or its high affinity receptor in tumors induces neovascularization and increases overall tumor aggressiveness. Antibody neutralization of SEMA4D represents a new therapeutic strategy for cancer treatment. We selected a humanized IgG4 antibody, VX15/2503, that binds with roughly 3 nM affinity to rat, mouse, primate, and human SEMA4D. We utilized cellular collapse, flow cytometric and other in vitro functional assays to demonstrate that this antibody blocks SEMA4D interaction with PLXNB1. Using both this antibody and its mouse IgG1 equivalent, MAb 67-2, we have demonstrated that antibody-mediated SEMA4D neutralization blocks tumor growth in CT26 tumor grafts with a tumor growth delay (TGD) of 42% and BCA34 (TGD 18%) tumor grafts, as well as RIP-Tag2, with a tumor growth inhibition (TGI) of 56%, and Tyr:NRAS (TGD 52%) spontaneous tumor models. In summary, we demonstrate that antibody mediated neutralization of SEMA4D in vivo inhibits tumor growth and tumor angiogenesis in a variety of tumor models. The humanized antibody, VX15/2503, has successfully completed IND-enabling toxicology testing and a Phase I trial is currently being conducted in adult patients with advanced solid tumors.

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 2531. doi:1538-7445.AM2012-2531
Abstract 2600: SLX4 mutation in hereditary breast cancer
Authors: Shah Sohela , Lach Frank , Kim Yonghwan , Sarrel Kara , Rau-Murthy Rohini , Hansen Nichole , Rendleman Justin , Kirchhoff Tomas , Joseph Vijai , Offit Kenneth , Smogorzewska Agata
Published: August 2012

SLX4 is a DNA repair protein that serves as a docking platform for three structure-specific endonucleases. Recent studies have reported mutations in SLX4 in a new subtype of Fanconi Anemia (FA-P). Monoallelic defects in several Fanconi Anemia genes are known to confer susceptibility to breast and ovarian cancer. Therefore, we resequenced the entire SLX4 coding region in 739 (288 Ashkenazi Jewish and 480 Non-Ashkenazi Jewish) breast cancer patients with 3 or more family members affected by breast cancer and no known BRCA1/BRCA2 mutations, to determine if SLX4 is involved in breast cancer susceptibility. We found a novel nonsense (p.W823*) mutation in one patient. We are sequencing the gene in the breast tumor from the patient to determine the status of the wild-type allele. In addition we also found 8 novel, 33 rare (MAF<1%), and 3 common (MAF> 0.5) missense variants, of which 12 (5 novel and 10 rare) are predicted as damaging by Polyphen2. Functional analysis of these missense mutations is being performed to determine their role in the pathogenicity of the SLX4 gene. Acknowlegements- This project was funded by The Starr Cancer Consotium (Agata Smorgorzewska and Kenneth Offit), The Rita Allen Foundation Scholars Program Grant (Agata Smorgorzewska) and Burroughs Wellcome Fund Career Award for Medical Scientists (Agata Smorgorzewska). We would like to thank the Geoffrey Beene Translational Oncology and Diagnostic Molecular Genetics laboratories for their help with the study.

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 2600. doi:1538-7445.AM2012-2600
Abstract 2648: Gene-asbestos interaction and lung cancer risk from a genome-wide association study
Authors: Liu Chen-Yu
Published: August 2012

Occupational asbestos exposure has been found to increase lung cancer risk based on epidemiologic studies. In order to clarify the respective roles of genetic factors and asbestos exposure on lung cancer risk, we conducted the asbestos exposure-gene interaction analyses. The first discovery stage included 745 Caucasian cases and 839 Caucasian controls using Illumina Human 610-Quad BeadChips. Several independent populations were included in the second replication stage. Cumulative lifetime asbestos exposure score (AES), which was previously developed and validated in our study population, was calculated from self-reported duration and intensity of occupational and nonoccupational exposures. We have found several suggested interesting / significant regions that might be associated with lung cancer risk in our SNP-asbestos exposure interaction tests, especially on 2p25.3, which has been replicated in another independent population. Further replication is still in process.

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 2648. doi:1538-7445.AM2012-2648
Abstract 2698: Improved therapeutic activity of Isoflavone-G2535 and Docetaxel combination in hormone refractory prostate cancer
Authors: Banerjee Sanjeev , Ali Shadan , Azmi Asfar , Kong Dejuan , Ahmad Aamir , Bao Bin , Sarkar Fazlul H.
Published: August 2012

The emergence of Castrate Resistant Prostate Cancer (CRPC) and its subsequent metastasis to the bone is in part driven by Androgen Receptor (AR) signaling, and contributes to overall poor survival of prostate cancer patients. Here, we present in vitro and in vivo preclinical evidence of chemo-sensitization of CRPC cells to the most clinically useful therapeutic agent, Docetaxel (Taxotere) by G2535, an isoflavone that attenuates AR signaling. In vitro studies were done using AR+ prostate cancer (PCa) cell lines (LNCaP and C4-2B) to understand the molecular basis of combination therapy. We found that the combination was significantly more effective in inhibiting cell growth and induction of apoptosis (p<0.01), down-regulation of AR and its target, prostate specific antigen (PSA) (p<0.01) with suboptimal concentrations of Docetaxel (1nmole/L) compared to monotherapy. It is known that resistance to chemotherapy in CRPC is partly associated with constitutively activated NF-κB/Akt signaling pathways, which becomes further activated by chemotherapeutic drugs, and these molecular targets were found to be down-regulated by G2535 treatment. Of greater significance were the results from in vivo tumor growth inhibition studies conducted in a SCID-hu model of experimental bone metastasis induced by C4-2B cells. Tumor volume, AR and serum PSA was used as response markers to evaluate therapeutic activity of the combination therapy relative to a single regimen and untreated control. At termination, we found 75% reduction in tumor volume elicited by the combination treatment relative to the untreated control group. Most importantly, the anti-tumor activity was associated with the down-regulation of molecular markers in tumor tissue remnants that were similar to those observed in vitro (down regulation of NF-κB, Bcl-2 and survivin) accompanied by a significant reduction in serum PSA. Using qPCR, we found that the level of miR-34a, which target AR, was increased by 1.5 fold after G2535 treatment alone whereas this increase was found to be 3-fold in the combination group. Increased expression of miR-34a resulted in the down regulation of AR and PSA in this in vivo model. Based on our findings, we conclude that the combination of G2535 and Taxotere could be an alternative promising regimen for the treatment of CRPC and its metastases for which curative treatment is urgently needed.

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 2698. doi:1538-7445.AM2012-2698
Abstract 2700: The use of a novel receptor targeted (CD206) Tc99m-labeled intraoperative node mapping agent for preoperative SPECT/CT in imaging head/neck cancer, breast cancer and melanoma Phase 3 trials: Localization and concordance in lymph nodes
Authors: Metz Wendy Rich , Hall Nathan , Povoski Stephen , Agrawal Amit , Shuping Jo , Abbruzzese Bonnie , Blackburn Simon , Potter Beth , Cope Frederick O.
Published: August 2012

The intraoperative lymphaticmapping (ILM) concept, based on evidence that colored agents injected into tissues leads to tracing of the agents into lymphatic structures, has carried over from observations dating back to the 1920s. The modern concept of using the pathological status of the lymph nodes to decide whether regional lymphatic clearance should be done was clearly articulated by Cabanas in 1977. Refinement of the concept as it would be reduced to a practical methodology of lymph node identification both in Europe and the United States began in earnest in the 1990s. Development of the application of sentinel node mapping has been built upon the concept of an anatomic/biologic nexus between the solid tumor bed and the immediate versus the distal lymphatic structures. It is the “biological proximity” that defines the probability of found disease in the adnexa, between the tumor bed and the lymphatic structures. It is inferred that the probability of disseminated disease in any lymphatic anatomy is highest when anatomic or biologic nexuses actually exist, as opposed to the lack of any anatomic nexuses where disease dissemination probabilities in lymphatic structures are lowest. To date, agents that have been employed in this procedure bear only simple designs that utilize simple physiologic pulse/draining of the lymphatics and provide no specificity for targeting the lymphatic tissue of interest. We present here patient results of three Phase III studies in head/neck squamous cell carcinoma, breast cancer, and melanoma where a novel 99mTc-labeled receptor targeted agent (99mTc-tilmanocept) not only out performed standard of care agents, but lent itself to remarkable concordance of intraoperative nodal localization with preoperative SPECT/CT imaging. The conclusion of these studies is that 99mTc-tilmanocept provides higher localization rates, degree of localization, lower failed detection rates, and reliable concordance between imaging and intraoperative findings. This work was supported by Neoprobe Corporation, Dublin, OH 43017

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 2700. doi:1538-7445.AM2012-2700
Abstract 2707: Long-term significance (15 years) of pathological complete response after dose-dense neo-adjuvant chemotherapy in breast cancer
Authors: Wang-Lopez Qian , Abrial Catherine , Planchat Eloise , Mouret-Reynier Marie-Ange , Cure Hervé , Gimbergues Pierre , Dubray-Longeras Pascale , Gallon-Bernard Dominique J. , Kwiatkowski Fabrice , Chollet Philippe J. , Durando Xavier
Published: August 2012

Purpose: As a contribution to the discussion of pathological complete response (pCR) as surrogate end-point, after neo-adjuvant chemotherapy (NACT), we studied disease-free survival (DFS) and overall survival (OS) at 15 years with THP-doxorubicin, vinorelbine, 5-Fluorouracil and cyclophosphamide (TNCF regimen). Patients and methods: This retrospective cohort study has been done in 61 patients treated with neo-adjuvant semi-intensive chemotherapy. PCR rate was analysed separately in breast (no histological evidence of invasive tumor cells and / or presence of in situ carcinoma tumor cells) and in nodes (no metastasis). Univariate and multivariate analysis (COX regression) have been used to assess the potential prognostic factors with a median follow up of 13.8 years. Results: Thirty two patients relapsed and 21 died. At 15 years, DFS and OS were 53% and 88% respectively for the patients who achieved pCR compared to 33% and 53% respectively for those who did not achieve pCR. Multivariate analysis with COX model showed that pCR in breast and the number of involved nodes at surgery were 2 factors who can significant predict patients’ DFS and these two factors were significantly related. Conclusion: In this current study, pCR in breast can be used as surrogate marker to predict survival outcome, even with a long-term follow-up of 15 years. When pathological response was incomplete in primary tumor, the number of involved nodes (n > 4 vs n </= 4) significantly predicted survival.

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 2707. doi:1538-7445.AM2012-2707
Abstract 271: Validation of Polo-like Kinase 1 as a novel therapeutic target in alveolar rhabdomyosarcoma
Authors: Thalhammer Verena , Herrero-Martin David , Hecker Regina , Niggli Felix K. , Schäfer Beat W.
Published: August 2012

Oncogenic transcription factors are the second most frequently mutated class of proteins. They represent ideal therapeutic targets because oncogene addiction is common to a broad spectrum of tumors and their inhibition may help to develop further alternative treatment strategies to improve therapy and to minimize treatment related side effects. However, thus far it has not been possible to target these oncogenic transcription factors as they usually do not respond to small-molecule inhibition. As a model system we use alveolar rhabdomyosarcoma (aRMS), an aggressive childhood tumor, which often displays resistance to conventional chemo-and radiotherapy. aRMS is characterized by the expression of the tumor-specific chimeric transcription factor PAX3/FKHR that is essential for survival of tumor cells. This transcription factor is post-translationally modified by phosphorylation which is required for efficient transcriptional activity. We believe that identification of upstream protein kinases mediating PAX3/FKHR phosphorylation will be an efficient strategy to impair the activity of the oncogenic fusion protein and subsequently induce tumor cell specific apoptosis. Having performed both siRNA and kinase-inhibitor library screens against the human kinome, we claim that Polo-like Kinase 1 (PLK1) might represent a very attractive and specific potential therapeutic target in aRMS. Its inhibition by RNAi and the small-molecule inhibitor BI 2536 reduces PAX3/FKHR activity and consequently aRMS cell viability. Moreover drug treatment causes tumor regression in xenograft mouse models. Therefore we are elucidating mechanisms causing these antitumorigenic effects and especially focus on the interaction of PLK1 and PAX3/FKHR. Our data suggest that PLK1 warrants further investigations as a novel drug target for the treatment of aRMS and may lead to a general strategy for treatment of tumors addicted to post-translationally modified oncogenic transcription factors.

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 271. doi:1538-7445.AM2012-271
Abstract 2786: Potent inhibition of human liposarcoma cell growth and survival by novel modulators of the MDM2-p53 interaction
Authors: Zhang Yixiang , Remillard Stephen P. , Demetri George D. , Wang Shaomeng , Debussche Laurent , Wagner Andrew J.
Published: August 2012

The tumor suppressor p53 is commonly inactivated in human malignancies. One mechanism is by genomic amplification and consequent overexpression of the ubiquitin E3 ligase MDM2, which targets p53 for proteosome-mediated degradation. The MDM2 gene is amplified in >95% of human liposarcomas, a disease for which the efficacy of systemic therapies is limited. Here, we compared the effects of two novel highly potent and selective inhibitors of the MDM2-p53 interaction, with the classic MDM2 inhibitor Nutlin-3 on the expression and activity of p53 and the resulting changes in human liposarcoma cell proliferation and survival. We determined MDM2 copy number, p53 mutational status, and relative expression levels of MDM2 and p53 proteins in 8 human liposarcoma cell lines and normal human preadipocytes/adipocytes. TP53 status was wild type in 7 of 8 liposarcoma cell lines, which also had genomic amplification of MDM2. MDM2 protein expression correlated with the degree of amplification and was highly expressed in comparison to normal preadipocytes/adipocytes. The two spiro-oxindole derivatives - Compounds A and B- (1) and Nutlin-3 were applied to liposarcoma cell lines with or without p53 siRNA and the biological consequences were determined by immunoblot, cell viability, cell cycle and apoptosis assays. Treatment with MDM2 inhibitors led to increased p53 protein levels and induction of its transcriptional targets MDM2 and p21. Cell viability was markedly decreased in a dose-dependent manner in liposarcoma cells with wild-type p53, with IC50 values of 0.40-4.23 μM, 0.37-1.88 μM and 2.72-9.83 μM for Compound A, Compound B, and Nutlin-3, respectively. These effects were markedly abrogated by siRNA-mediated p53 knockdown and in a cell line harboring a de novo p53 mutation. Compound A and Compound B also induced cell cycle arrest and apoptosis at approximately 5 fold lower concentrations than Nutlin-3. At low concentrations, G0/G1 cell arrest occurred, whereas higher concentrations resulted in G2/M arrest and induction of apoptosis. Cell line and tumor xenograft experiments are ongoing. These data demonstrate that Compounds A and B are highly potent and specific inhibitors of MDM2 with activity in genetically characterized liposarcoma cells. Further study and clinical development in this disease is warranted. 1. Wang S. et al., AACR, Orlando FL, 2011, Abstract LB-204

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 2786. doi:1538-7445.AM2012-2786
Abstract 2795: Safety profile and therapeutic potential in breast cancer of STX140, an original tubulin binding agent
Authors: Meyer-Losic Florence , Fiore Ann , Carlson Mark , Lauer Jeffrey , Roubert Pierre , Bernetière Sonia , Oligo Charleen , Vignaux Laurent , Gaudry Hervé , Foster Paul A. , Newman Simon P. , Purohit Atul , Leese Matthew P. , Kasprzyk Philip G.
Published: August 2012

STX-140 is an anti-tubulin binding agent which has demonstrated a good anti-tumor efficacy profile. Although the binding site on tubulin differs from other classical tubulin binders, one could anticipate the same type of safety issues with such an agent, namely neurotoxicity. In order to differentiate this compound, we evaluated its efficacy in a transgenic model of mammary cancer, its neurotoxicity potential, and its therapeutic index in a breast cancer model. STX140 efficacy was evaluated in C3(1) Tag transgenic mice which develop mammary carcinomas. In those transgenic animals, the disease progresses in a similar fashion to human breast cancer, with the development of tumors which show the same histological characteristics as the human pathology. In these mice, STX140 was able to significantly increase the survival of the animals and to suppress the emergence of metastases, whereas paclitaxel was completely inactive on both parameters. Neurotoxicity potential was assessed in a thermal hyperalgesia model in comparison with paclitaxel. Continuous oral treatment of mice with STX-140 did not induce an increase sensibility to heat as measured by the latency of paw withdrawal after a heat stimulus. However, paclitaxel increased this heat sensitivity starting after the second i.v. administration, underlying the neurotoxicity of this compound, which is observed in the clinic. Finally, in MDA-MB-231 breast cancer xenografts, two different schedules of oral administration were evaluated in order to determine the therapeutic index and PK parameters were measured in parallel to the anti-tumor efficacy. Continuous (QD) and intermittent (Monday, Wednesday, Friday; MWF) treatments were evaluated and a large panel of doses were tested. The therapeutic index was determined as the ratio of the exposure observed between the first toxic dose and first active dose. The TI for the QD schedule was 2.1 and was 1.7 for the MWF schedule, showing the potential good safety index of this compound. Taking into account the previous data reported elsewhere and new data presented here, which underline the great efficacy and safety profile of STX140, this compound is considered for further preclinical and clinical 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 2795. doi:1538-7445.AM2012-2795
Abstract 2796: Efficacy and biomarker modulation by AZD1208, a novel, potent and selective pan-Pim kinase inhibitor, in models of acute myeloid leukemia
Authors: Keeton Erika , McEachern Kristen , Alimzhanov Marat , Wang Suping , Cao Yichen , Bao Larry , Palakurthi Sangeetha , Grondine Michael , Chen Yuching , Dillman Keith , Chinnappan Dharmaraj , Shen Minhui , Dakin Les , Zheng Xiaolan , Lamb Michelle , Wu Allan , Chen Huawei , Lyne Paul , Huszar Dennis
Published: August 2012

The Pim serine/threonine kinase family is composed of three highly homologous members; Pim-1, Pim-2 and Pim-3, identified by the ability of the prototype member Pim-1 to drive lymphomagenesis in mice. Upregulation of Pim-1 and Pim-2 is observed in leukemias and lymphomas, including AML, NHL and CLL, highlighting the potential of these kinases as therapeutic targets in these indications. Overexpression of Pim-1 or Pim-3 has also been observed in prostate, pancreatic, gastric, bladder and hepatocellular cancers. Pim kinases are downstream effectors of many cytokine and growth factor signaling pathways and are direct transcriptional targets of STAT transcription factors activated by these pathways. Pims can phosphorylate multiple substrates to mediate cell proliferation and survival. Here we describe the activity of AZD1208, an orally available, potent and highly selective Pim inhibitor that effectively inhibits all three isoforms. AZD1208 inhibits the growth of several AML cell lines and sensitivity correlates with the level of Pim-1 expression, STAT5 activation and presence of protein tyrosine kinase mutation. AZD1208 causes cell cycle arrest and apoptosis in MOLM-16 cells in culture. This is accompanied by a dose-dependent reduction in phosphorylation of BAD, 4EBP1 and p70S6K. AZD1208 suppresses the growth of MOLM-16 and KG-1a xenograft tumors in vivo in a dose proportional manner. In addition, AZD1208 leads to potent inhibition of colony growth of primary AML cells from bone marrow aspirates and downregulates phosphorylation of Pim targets. These results underscore the therapeutic potential of Pim kinase inhibition by AZD1208 for the treatment of AML. They also further support investigation of this inhibitor in other hematological and solid tumor malignancies where PIM signaling may play a role in tumorigenesis and survival.

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 2796. doi:1538-7445.AM2012-2796
Abstract 28: The Netrin-1 receptor UNC5C is a tumor suppressor in colorectal malignancies
Authors: Coissieux Marie-May , Tomsic Jerneja , Lasset Christine , Hampel Heather , Scoazec Jean-Yves , Wang Qing , Tanner Stephan M. , Bernet Agnès , de la Chapelle Albert , Mehlen Patrick
Published: August 2012

The Netrin-1 receptors UNC5A, B and C belong to the functional family of dependence receptors that share the ability to induce apoptosis in the absence of their ligands. Such a trait has been hypothesized to confer a tumor suppressor activity to these receptors. Expression of one of these dependence receptors at the surface of a tumor cell is thus speculated to render this cell dependent on ligand for its survival, hence inhibiting cell proliferation or migration outside the ligand availability zone. Here we show that the expression of the UNC5C gene is down-regulated in colorectal cancers, mainly because of tumor-associated specific promoter methylation. In addition, inactivation of UNC5C in mice is associated with increased intestinal tumor progression, demonstrating its tumor suppressor role. Moreover, mutations in UNC5C that turn down the pro-apoptotic activity of the receptor are found in patients with familial form of colorectal cancer. This germline coding mutation in UNC5C has been detected in five different families from France and USA and co-segregates with the disease. We further, determine that this anti-apoptotic mutation arose from a common founder. In summary, our study strongly indicates that the genetic inactivation of the tumor suppressor UNC5C may be a major promoter of familial predisposition to colorectal tumorigenesis.

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 28. doi:1538-7445.AM2012-28
Abstract 280: Genetics and functions of Kaposi's sarcoma-associated herpesvirus (KSHV) glycoproteins in virion egress, infectivity and tumorigenesis
Authors: D'Auvergne Oswald , Subramanian Ramesh
Published: August 2012

Kaposi's sarcoma-associated herpes virus (KSHV), or human herpes virus 8, is a lymphotropic oncogenic virus that has been implicated in the pathogenesis of Kaposi's sarcoma; body cavity-based B-cell lymphoma (BCBL), or primary lymphoma; and some forms of Multicentric Castleman's disease. We have developed a conditional silencing system for partial inhibition of KSHV viral glycoprotein synthesized genes that are not susceptible to siRNA inhibition. We have successfully used this system to demonstrate that KHSV glycoprotein B (gB) is necessary for virion egress from BCBL-1 cells and virus infectivity (Subramanian, D'Auvergne, et al. J. Virol. 2008, 82:7144-54). To investigate the potential role of gB in tumorigenesis, BCBL-1 cells were transiently transfected with anti-gB siRNAs and codon optimized gB were mixed with matrigel and injected subcutaneously in nude mice. Direct measurement of tumors revealed that BCBL-1 cells transfected with anti-gB siRNAs produced tumors significantly smaller than mock-transfected BCBL-1 cells. Co-transfection of codon optimized gB appeared to abrogate the inhibition of tumor formation by siRNAs. These results show that gB is important for infectivity, virion egress and pleural effusion lymphoma (PEL) formation in mice.

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 280. doi:1538-7445.AM2012-280
Abstract 2802: Selective inhibition of signal transducers and activators of transcription 3 overexpressing endometrial cancer cells, using HO-3867, a novel STAT3 inhibitor
Authors: McCann Georgia , Tierney Brent J. , Cohn David , Sudhakar Meryl , Naidu Shan , Eisenhauer Eric , Karuppaiyah Selvendiran
Published: August 2012

OBJECTIVE: Signal transducer and activator of transcription 3 (STAT3) proteins have important roles in cancer cell survival and proliferation. Recent studies show constitutive activation of STAT3 is associated with tumor growth and oncogenic transformation in several human cancers; however, little is known about STAT3 expression in endometrial cancer. The objectives of this study were to examine the expression of the STAT family of proteins in endometrial cancer cell lines and to evaluate the efficacy of HO-3867, a novel STAT3 inhibitor. METHODS: Expression of STAT family proteins, cell cycle and apoptotic proteins were evaluated via western blot on Ishikawa, AN3-CA, RL95-2, HEC1B, HEC-1A, and SK-UT1B human endometrial cancer cell lines. Cell viability and proliferation of these cell lines were analyzed with MTT and nucleo counter assay after treatment with increasing concentrations of HO-3867. Cell cycle analysis and apoptosis were performed using flow cytometry and annexin V staining. An ubiquitination assay was used to examine the inhibition of CDK5 degradation. Finally, STAT3 over-expression or suppression experiments were performed using wild-type STAT3 cDNA and STAT3 siRNA. RESULTS: Expression of activated STAT1, 2, 5, and 6 was inconsistent among cell lines tested, while pSTAT3 Ser727 had consistently high expression. HO-3867 decreased cell viability through induced G2/M arrest and apoptosis within 24h of treatment. HO-3867 inhibited pSTAT3 Ser727 with no change in total STAT3. This was associated with an increase in expression of cell cycle related proteins p53 and p21, a decrease in anti-apoptotic proteins Bcl2 and Bcl-XL, as well as increased cleavage of caspases. After 24h of treatment with HO-3867, 55-88% of cells were apoptotic. The downregulation of Ser727 was, at least in part, due to an accelerated inhibition of ubiquitin-dependent CDK5 degradation, as a positive regulator of STAT3 Ser727. Suppression of STAT3 expression with siRNA significantly reduced cell survival. In addition, we confirmed that over-expression of STAT3 cDNA provides resistance to HO-3867. CONCLUSION: Endometrial cancer cells express high levels of pSTAT3 Ser727. HO-3867 induces cell cycle arrest and apoptosis by targeting pSTAT3 Ser727. Our findings suggest a specific role of serine-phosphorylated STAT3 that is independent of tyrosine phosphorylation. HO-3867 may be a potential adjunct to chemotherapy and radiation in the treatment of endometrial 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 2802. doi:1538-7445.AM2012-2802
Abstract 2804: Radiosensitization by the Dual PI3K/mTOR inhibitor NVP-BEZ235
Authors: Cerniglia George J. , Karar Jayashree , Koumenis Constantinos , Maity Amit
Published: August 2012

The PI3K/Akt pathway has been implicated in cancer progression and in the survival of cancer cells following DNA damage induced by radiation. Hence, we chose to study the radiosensitizing ability of the Novartis compound, NVP-BEZ235, a dual PI3K/mTOR inhibitor. Clonogenic cell survival assays following ionizing radiation, cell cycle analysis, immunofluorescence for γ-H2AX foci, and Western blotting were performed using standard protocols.Treatment of SQ20B head and neck squamous cell carcinoma cells and U251 glioblastoma cells with NVP-BEZ235 led to a decrease in phosphorylation of Akt at serine 473 and of S6 and 4E-BP1, both of which are downstream markers of mTOR activation. Pre-treatment with the drug (25-50 nM) led to radiosensitization of these cell lines with a dose enhancement ratio (DER) ranging from 1.32 - 1.72. Knockdown of Akt1, p110alpha, or mTOR in these cell lines, also resulted in radiosensitization, but not to the same extent as with NVP-BEZ235. Of note, treatment with rapamycin led to minimal radiosensitization of these cells. Reduced survival with NVP-BEZ235 does not result from cell cycle redistribution during the treatment intervals tested, although combining NVP-BEZ235 with radiation enhances the G2/M delay seen after radiation. We examined how cells died after radiation and found no suggestion of increased apoptosis or necrosis. We did find evidence of increased conversion of microtubule-associated protein light chain 3 (LC3)-I to LC3-II, suggestive of autophagy. In split course experiments, in which cells were either irradiated with a single dose of radiation or with two half-doses separated by time, as the interval between doses increased, so did clonogenic survival. This increase in survival was abolished by pre-treatment with NVP-BEZ235, suggesting that the drug interfered with DNA damage repair. This was confirmed by assessing levels of gamma-H2AX after ionizing radiation. Initial post-radiation levels of gamma-H2AX foci were similar in NVP-BEZ235-pretreated and control cells; however, the resolution of these foci was significantly delayed in the BEZ235-treated cells. We then examined DNA-PKcs, a protein important for DNA damage repair, and found that the drug abrogated the increased phosphorylation normally seen after radiation. These results indicate that NVP-BEZ235 is a potent radiosensitizer of HNC cells in vitro. As siRNA against either p110alpha or mTOR leads to radiosensization, both of these targets of NVP-BEZ235 may be critical mediators of its to radiosensitizing ability; however, a second mechanism involved, may be via delayed repair of DNA damage following radiation, via interference of DNA-PKcs phosphorylation.

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 2804. doi:1538-7445.AM2012-2804
Abstract 2809: BMS-777607, a small molecule RON/MET kinase inhibitor induces polyploidy in breast cancer cells: molecular mechanism and therapeutic implication
Authors: Sharma Sharad , Zhuang Chun-Mei , Zeng Junying , Wang Minghai
Published: August 2012

Aberrant RON expression contributes significantly to breast cancer tumorigenic progression. This observation has provided the basis for targeting RON in potential breast cancer treatment. In the study of breast cancer tumorigenesis, we discovered that inhibition of RON by small molecule inhibitor BMS-777607 causes polyploidy followed by apoptosis in various breast cancer cells including T-47D and ZR-75-1. BMS-777607 is a tyrosine kinase inhibitor specific to RON and MET receptor tyrosine kinases and blocks cancer cell proliferation in vitro and tumor growth in xenograft model in vivo. Using breast cancer cells as the model, we observed that BMS-777607 at experimental doses induces extensive polyploidy and apoptosis in breast cancer cells. The effect of BMS-777607 has the following features. First, BMS-777607 treatment dramatically altered cell morphology, increased cell size, and created multinucleate structures. Chromosome spreading at metaphase and flow cytometric analysis confirmed polyploid cells with 6N to 16N chromosome content, indicating that abortive mitosis and endoreduplication are responsible for the observed polyploidy. Second, the effect of BMS-777607 was highly specific and dependent on RON or MET expression. MCF-7 cells that do not express RON/MET showed only low levels of polyploidy, suggesting that RON and MET expression are required for the observed polyploid activity. Third, the effect of BMS-777607 is time and dose-dependent. A substantial polyploid population of cells accumulated within 24 h of drug treatment. Continuous cycle of abortive mitosis further increased cell ploidy with increased 16N cell fractions. Moreover, an experimental dose of 500 nM is sufficient to induce polyploidy. The maximal effect occurred when BMS-777607 was used at 5.0 μM for 72 h. Fourth, polyploidy induction promoted apoptosis and senescence of breast cancer cells, which led to significant changes in cellular sensitivity towards cytotoxic activity of chemoagents. Finally, we demonstrated that Aurora B kinase is the major signaling molecule responsible for BMS-777607-induced cancer cell polyploidy. BMS-777607 inhibited Aurora B kinase activity, which affected mitosis leading to polyploidy. In summary, BMS-777607 is a potent inducer of polyploidy in breast cancer cells. By blocking RON/MET-mediated signaling events, BMS-777607 impaired Aurora B kinase activity leading to polyploidy and apoptosis. Such observations provide mechanistic and therapeutic insight in BMS-777607 targeted cancer therapy (*supported in part by NIH grant R01 CA91980 and funds from Amarillo Area Foundation).

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 2809. doi:1538-7445.AM2012-2809
Abstract 2811: Preclinical evaluation of combined TKI258 and RAD001 in hepatocellular carcinoma
Authors: Wong Chi Hang , Yeo Winnie , Lui Vivian , Lau Cecilia P.Y. , Hui Connie W.C. , Chan Anthony T.C. , Chan Stephen L.
Published: August 2012

Hepatocellular carcinoma (HCC) is one of the most lethal cancers. Signaling pathways including AKT-mTOR axis is often dysregulated in HCC. RAD001 principally targets at the mTOR complex 1. mTOR inhibition is reportedly associated with activation of feedback loop and parallel pathways. TKI258 is a potent tyrosine kinase inhibitor targeting the FGFRs and VEGFRs, which blocks subsequent PI3K/AKT signaling pathways in cancer cells. In the current study, we aim to study the in-vitro and in-vivo effects of the drug combination with reference to the parallel and upstream pathway of AKT-mTOR axis. We have treated the HCC cell line Hep3B with TKI258 and RAD001. RAD001 could suppress the phosphorylation of down stream signal mediators of mTOR pathway including p70S6 kinase 1 (S6K) and eukaryotic initiation factor binding protein 1 (4E-BP1) and led to G1 phase arrest. The results demonstrated a significant increase in suppression of cell proliferation in vitro with the combination of TKI258 (300nM) and RAD001 (either 200nM or 200pM) compared with either drug alone. Although the addition of the TKI258 only slightly suppressed the AKT positive feedback loop induced by RAD001, the combination of drugs could significantly suppress the phosphorylation of mTOR (Ser2448), ERK1/2 (Ser217/221) and p38 (Thr180/Tyr182) as well as their endogenous protein expression levels. The expression levels of FGFR1 and FGFR2 were also suppressed. The overall protein reduction was due to the synergistic inhibition of p70S6 (Ser240/244) and 4E-BP1 (Ser65) phosphorylation, which indicated a strong suppression in translation and protein synthesis. Pro-caspases 3 and PARP cleavage were not observed at 24 hours and only slightly detected at 48 hours post-treatment, suggesting that the drug combination did not further increase cytotoxic effects but mainly increased in cytostatic arrest ability. In Hep3B derived xenograft model, TKI258 (15mg/ml) and RAD001 (2.5mg/kg) had shown a synergistic inhibition of tumor growth in volume and weight. In addition, there was a significant reduction in microvessel density (MVD) in the xenograft, which indicated an improved effect in angiogenesis inhibition. Our data showed that the combination of RAD001 and TKI258 were active in controlling HCC proliferation via double inhibition of both AKT-mTOR axis and its parallel pathway. Acknowledge: The works are supported by Novartis Oncology.

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 2811. doi:1538-7445.AM2012-2811