bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract C20: Reversal of mesenchymal phenotype of triple-negative breast cancer and glioblastoma cells and its therapeutic implication
Authors: Yoshikawa Kiyotsugu , Sakai Hiroaki , Shimazaki Masahiro , Okada Nobuhiro , Matsumoto Yoshiaki , Reinherz Ellis , Toi Masakazu
Published: May 2016

Mesenchymal transition is common to many aggressive cancers including breast, glioblastoma (GBM), lung and pancreatic cancers characterized by chemo/radiotherapy-resistance and cancer stem cell-like property. Claudin-low msenchymal subtype in triple-negative breast cancer (TNBC) is a major problem due to the lack of molecularly-targeting drugs. So far, we have identified an shRNA (shP1) which can induce mesenchymal to epithelial transition (MET) in shRNA library screening using positive selection reporter system in MDA-MB-231 mesenchymal TNBC cell line. Here, we have analyzed the effect of shP1 on morphology, transcription and cell proliferation in various mesenchymal cancer cell lines derived from breast, glioblastoma (GBM), lung and pancreatic cancers.

The shP1 increased E-cadherin promoter-driven GFP in a wide range of mesenchymal cancers including brain, breast, pancreatic, lung origin. Drastic morphology change introduced by shP1 was observed both in 2D and 3D cultures (matrigel, collagen), reducing their invasive and branching morphogenesis, especially in U251 GBM and MDA-MB-231 TNBC cell lines. The shP1-introduced cells are more sensitive to EGFR-inhibition and paclitaxel in TNBC cell line, reminiscent of reversal of therapy-resistant phenotype. Gene expression analysis reveals alteration of pathways related to cell adhesion, ERRB signal pathway, apoptosis and cytokines signaling. Of note, shP1-introduced U251 cells increased gene expression characteristic of proneural subtype, indicating shP1 reverted the subtype from mesenchymal to proneural. These results suggest that although U251 and MDA-MB-231 cell lines are of different tissue origins, their mesenchymal property can be reverted by the same program targeted by single shRNA.

These results provide a rationale of MET as therapeutic for a wide range of cancers with mesenchymal property.

Citation Format: Kiyotsugu Yoshikawa, Hiroaki Sakai, Masahiro Shimazaki, Nobuhiro Okada, Yoshiaki Matsumoto, Ellis Reinherz, Masakazu Toi. Reversal of mesenchymal phenotype of triple-negative breast cancer and glioblastoma cells and its therapeutic implication. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C20.
Abstract C35: Low-dose imatinib mesylate suppressed PDGF-BB mediated motility and invasion of gastric cancer cells
Authors: Kim Hong Jun , Lee Suk Young , Oh Sang Cheul , Kim Jun Suk , Kim Jung Lim , Kim Bo Ram , Na Yoo Jin
Published: May 2016

Purpose

Imatinib mesylate is a powerful tyrosine kinase inhibitor that specifically targets BCR-ABL, KIT, and PDGFR kinases and is used in treatment of chronic myelogenous leukemia (CML), gastrointestinal stromal tumors (GIST), and other cancers. Although imatinib has not yet been used in gastric cancer, we verified that high-dose imatinib leads gastric cancer cell lines to apoptosis via down regulation of Akt phosphorylation in previous study. This study was undertaken to evaluate the in vitro effect of low-dose imatinib in inhibition of invasion and migration of gastric cancer cells and to define molecular mechanism underlying these effects.

Methods

This study was to determine the expression of PDGFR molecules in gastric cancer cells (four human gastric cancer cell lines; AGS, MKN28, MKN45, and SNU638) by semi-quantitative polimerase chain reaction (PCR). The effects of low-dose imatinib (≤20uM.ml) on cell viability were examined using MTT assay. The effects of imatinib and PDGF-BB on cell motility and cell invasion were studied using wound healing assay and matrigel invasion assay respectively.

Results

The results showed expression of PDGFR in all four gastric cancer cells. We measured the sensitivity of gastric cancer cell line (AGS, MKN28, MKN45, SNU638) to imatinib using MTT assay. Cell viability of low-dose imatinib treated cell was not decreased. However, cell migration and invasion was decreased with low-dose imatinib. When PDGF-BB was added, cell motility and invasion were activated. However, when both PDGF-BB and imatinib were added, that activation was not shown.

Conclusion

In conclusion, low-dose imatinib mesylate inhibits PDGF-BB mediated cell migration and invasion not decreasing cell viability. The results suggest that imatinib mesylate may be useful in the treatment of gastric cancer.

Citation Format: Hong Jun Kim, Suk Young Lee, Sang Cheul Oh, Jun Suk Kim, Jung Lim Kim, Bo Ram Kim, Yoo Jin Na. Low-dose imatinib mesylate suppressed PDGF-BB mediated motility and invasion of gastric cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C35.
Abstract C51: Chemosensitisation to cisplatin by STAT3 inhibitors is mediated through the inhibition of DNA interstrand crosslink unhooking
Authors: Valentine Helen , Dixon Emily , Hartley John A. , Kiakos Konstantinos
Published: May 2016

Signal Transducer and Activator of Transcription 3 (STAT3) is constitutively activated in many cancer cell lines, leading to survival, proliferation, angiogenesis and metastasis. The inhibition of STAT3 by natural or synthetic compounds has been shown to sensitise cancer cell lines to the chemotherapy agent cisplatin, however the molecular mechanisms contributing to this chemosensitisation have not yet been fully elucidated. Therefore we investigated the effect of STAT3 inhibition on cisplatin-induced DNA damage and key DNA repair factors.

STAT3 inhibitors stattic and curcumin were investigated in combination with cisplatin using the Suphorhodamine B cell growth inhibition assay in the DU145 prostate cancer and A549 non-small cell lung cancer cell lines. Combination treatments result in a significant (P<0.0001) reduction in cisplatin GI50 values. These combinations were synergistic as determined by combination index value analysis using the software Calcusyn, confirming that inhibition of STAT3 enhances the cytotoxic effect of cisplatin. Using a modification of the alkaline comet assay we observed that STAT3 inhibitors significantly prevent the unhooking step of repair of cisplatin induced interstrand crosslinks (ICLs). Treatment with cisplatin alone induces formation of ICLs which peak at 9 hours and unhook by 24 hours post cisplatin exposure in both cell lines. Pre-treating cells with 12μM stattic or 48μM curcumin followed by cisplatin results in an equivalent peak formation of ICLs at 9 hours, however 24 and 48 hours post cisplatin no unhooking is observed. Therefore both STAT3 inhibitors used in this study block the unhooking stage of ICL repair. Using RT-PCR and immunoblotting we show that treatment with STAT3 inhibitors dose-dependently down-regulates EME1, BRCA1 and FANCD2 mRNA and protein levels. These three DNA repair proteins have been shown to be involved in the early stages of ICL repair and therefore down-regulation of these genes may be responsible for the impairment of ICL unhooking seen with STAT3 inhibition.

As the effects observed here have been replicated with two STAT3 inhibitors of differing chemical structure, this suggests that STAT3 is directly involved in the regulation of cisplatin-induced ICL repair through modulation of key ICL repair genes. We propose this as a contributing mechanism to the synergy reported between STAT3 inhibitors and cisplatin. The data presented here will inform the design of future drug combination strategies.

Citation Format: Helen Valentine, Emily Dixon, John A. Hartley, Konstantinos Kiakos. Chemosensitisation to cisplatin by STAT3 inhibitors is mediated through the inhibition of DNA interstrand crosslink unhooking. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C51.
Abstract C78: ATM/G6PD-dependent metabolic pathways promote mitochondrial redox homeostasis and resistance to FLT3 inhibition in acute myeloid leukemia
Authors: Gregory Mark A. , D'Alessandro Angelo , Alvarez-Calderon Francesca , Kim Jihye , Nemkov Travis , Tan Aik Choon , Hansen Kirk C. , DeGregori James
Published: May 2016

Acute myeloid leukemia (AML) is the most common adult acute leukemia and accounts for approximately 20% of childhood leukemias. Although frontline treatment of AML with cytotoxic chemotherapy is capable of achieving high remission rates, 75-80% of patients will either not respond to or will relapse after initial therapy, and most patients will die of their disease. More effective and better-tolerated therapies for AML are required. Activating mutations in fms-like tyrosine kinase 3 (FLT3) are the most frequently observed genetic defect in AML and drive leukemic cell growth and survival. FLT3 tyrosine kinase inhibitors have shown impressive anti-leukemic activity in clinical trials, however, sustained remissions using these inhibitors as monotherapy have not been achieved. In order to identify genetic targets that can sensitize AML cells to killing by FLT3 inhibitors, we performed a large-scale RNA interference-based screen. The screen identified several genes involved in metabolic regulation, including ataxia telangiectasia mutated (ATM), as being synthetic lethal with FLT3 inhibition in FLT3 mutated AML. Genetic or pharmacological inactivation of ATM or its downstream effector glucose-6-phosphate dehydrogenase (G6PD) sensitized AML cells to FLT3 inhibition through enhancing apoptosis. Whole metabolome profiling revealed that FLT3 inhibition causes severe and widespread metabolic deficiencies, including depletion of the antioxidant factor glutathione. Inactivation of either ATM or G6PD exacerbated glutathione depletion upon FLT3 inhibition. Subsequent analyses revealed that FLT3 inhibition elicits severe mitochondrial oxidative stress that is causative in apoptosis and accentuated by ATM or G6PD inhibition. The use of a drug that promotes the production of mitochondrial reactive oxygen species (ROS) in combination with a FLT3 inhibitor augmented elimination of AML cells both in vitro and in vivo. Our data support the hypothesis that FLT3 mutated AML cells are highly dependent on FLT3 activity to maintain glutathione levels and sufficient mitochondrial antioxidant capacity to sustain cell survival. Moreover, these data support the novel strategy of employing mitochondrial ROS inducing agents as adjuvant to FLT3 inhibitor therapy to more effectively treat FLT3 mutated AML.

Citation Format: Mark A. Gregory, Angelo D'Alessandro, Francesca Alvarez-Calderon, Jihye Kim, Travis Nemkov, Aik Choon Tan, Kirk C. Hansen, James DeGregori. ATM/G6PD-dependent metabolic pathways promote mitochondrial redox homeostasis and resistance to FLT3 inhibition in acute myeloid leukemia. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C78.
Abstract CN05-03: Personalized cancer diagnostics and therapeutics based on the computational modeling of signal transduction networks
Authors: Kolch Walter , Fey Dirk , Halasz Melinda , Rauch Nora , Munoz Amaya Garcia , Pilkington Ruth , Kholodenko Boris N. , Croucher David R. , Kennedy Sean P. , Hastings Jordan F. , Westermann Frank , Dreidax Daniel , Fischer Matthias , Duffy David , Krstic Aleksandar , Schwarzl Thomas
Published: May 2016

Signalling pathways exert finely tuned control over cell fate decisions that ultimately determine the behaviour of cancer cells. It could therefore be expected that the dynamics of key pathway activation may contain prognostically relevant information over and above that which is contained in the static nature of traditional biomarkers. To investigate this hypothesis we have characterised the network architecture regulating JNK stress signalling in neuroblastoma cells and applied an experimentally calibrated and validated computational model of this network to extract prognostic information from neuroblastoma patient-specific simulations of JNK activation. Survival analysis based upon the dynamics of these simulations revealed that an inability to initiate switch-like JNK signalling in silico was significantly associated with poor overall survival for both MYCN amplified and non-amplified neuroblastoma patients. Furthermore, our analysis demonstrated that in order to extract prognostic information from a signalling pathway, deciphering the extant network structure is a vital consideration in model development. We also show that network based analysis can lead to the discovery of new therapeutic targets.

Citation Format: Walter Kolch, Dirk Fey, Melinda Halasz, Nora Rauch, Amaya Garcia Munoz, Ruth Pilkington, Boris N. Kholodenko, David R. Croucher, Sean P. Kennedy, Jordan F. Hastings, Frank Westermann, Daniel Dreidax, Matthias Fischer, David Duffy, Aleksandar Krstic, Thomas Schwarzl. Personalized cancer diagnostics and therapeutics based on the computational modeling of signal transduction networks. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr CN05-03.
Abstract CN06-04: Combination strategies
Authors: Bono Johann S. De
Published: May 2016

Targeting the endocrine axis remains a mainstay of the treatment of many breast and most prostate cancers. Recent studies indicate that these tumors commonly remain nuclear steroid hormone receptor driven once endocrine resistance ensues. Multiple novel, rationally designed, agents are in development that are showing promise in the treatment of these diseases including drugs targeting p110beta, AKT, TORC1/2, as well as drugs targeting PARP, bromodomain proteins BRD2-4 and heat shock proteins including HSP90 and HSP27. Furthermore novel agents targeting the amino terminus of the androgen receptor and degrading these receptors are being evaluated. This presentation will describe challenges and solutions to their successful drug combination development.

Citation Format: Johann S. De Bono. Combination strategies. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr CN06-04.
Abstract CN08-02: Metabolic adaptations and liabilities of TCA cycle-truncated tumors
Authors: Gottlieb Eyal
Published: May 2016

Last century, Otto Warburg observed that cancer tissues have high rates of glycolysis in the presence of oxygen. This was attributed to defects in mitochondrial respiration, and is now considered a key hallmark of cancer. Nevertheless, convincing evidence that mitochondrial dysfunction causes metabolic switch and tumorigenesis, were scarce. Recently, the identification of mutations in key metabolic enzymes with a causal link to tumorigenesis awakened interest in Warburg's hypothesis. Cancer-associated mutations have been identified in genes coding for tricarboxylic acid (TCA) cycle and closely related enzymes. Succinate dehydrogenase (SDH) is a TCA cycle enzyme and complex II of the mitochondrial respiratory chain. Loss of function in any of the SDH subunits is associated with susceptibility to cancers. Loss of SDH causes succinate accumulation, which activates hypoxia-inducible factors and inhibits α-ketoglutarate-dependent histone and DNA demethylases, establishing pseudohypoxic and hypermethylator phenotypes. However, so far no mechanism enabling survival and proliferation of SDH-defective cells has been provided. We generated genetically modified kidney mouse cells in which Sdhb has been deleted and identified metabolic pathways dispensable in normal, but essential in SDH-null cells. We demonstrated that SDH deficient cells depend on extracellular pyruvate to sustain glycolytic flux and to channel glucose-derived carbons to aspartate biosynthesis through pyruvate carboxylase. Our study provides a new potential therapeutic target for treating SDH defective neoplasms.

Citation Format: Eyal Gottlieb. Metabolic adaptations and liabilities of TCA cycle-truncated tumors. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr CN08-02.
Abstract LB-A07: Using Impedance-Based Approaches for Measuring Cell-Mediated Cytotoxicity and Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)
Authors: Zhao Leyna , Abassi Yama
Published: May 2016

The most common method for measuring cell mediated cytotoxicity is the release assay based on the loss of target cell membrane integrity. Within up to 4 hours following effector cell addition resulting in target cell lysis, either the radioactive release from target cells pre-labeled with Chromium-51 or Indium-111 is measured, or the release of naturally occurring substances, such as lactate dehydrogenase (LDH), into the culture medium is assayed. Release of these substances thus serves as an indirect measure of the extent of cell damage due to effector cell-mediated target cell lysis. Alternative endpoint methods also include flow cytometry, enzyme-linked immunosorbent assay-based granzyme measurement, and morphometric analyses by microscopy.

Here we describe an impedance-based real-time label-free method that can automated capture the kinetics of the cell mediated or antibody-dependent cell mediated cytolysis (ADCC) of the target cancer cells. To determine if cell-mediated cytotoxicity, and specifically ADCC, can be investigated using an impedance-based xCELLigence system, the response of tumor cells (as target cells) to natural killer (NK) cell activity (as effector cells), in the presence or absence of immunoglobulin G isotype-specific antibody, was measured. Importantly, it is shown that the addition of NK cells in suspension, over a monolayer of adherent tumor cells, does not produce impedance changes, because the NK cells do not come in contact with the electronic sensor. However, the secretion of perforins and granzymes by these non-adherent NK cells does activate caspases inducing tumor cell apoptosis. Dysfunctional and dying tumor cells detach from the sensor electrode, reducing the number of viable and adhering cells on the electrode surface.

Overall, our results show that the impedance-base technology is a label-free alternative to the traditional end-point assays. The automated readouts provide direct, sensitive and specific measurement of target cell changes both short-term (hrs) and long-term (days). It allows the easy quantification of the cell-mediated cytotoxicity and evaluation of the potencies for specific antibodies.

Citation Format: Leyna Zhao, Yama Abassi. Using Impedance-Based Approaches for Measuring Cell-Mediated Cytotoxicity and Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC). [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A07.
Abstract LB-A10: The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway
Authors: Kwon Yeo-Jung , Shin Sangyun , Ye Dong-Jin , Baek Hyoung-Seok , Chun Young-Jin
Published: May 2016

The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway

Yeo-Jung Kwon*, Sangyun Shin, Dong-Jin Ye, Hyoung-Seok Baek, and Young-Jin Chun

Department of Pharmaceutical Biochemistry, College of Pharmacy, Chung-Ang University, Seoul, Korea

Cytochrome P450 1B1 (CYP1B1) belongs to the CYP1 family and shares the feature as an enzyme for oxidative metabolisms. It has been reported that CYP1B1 expression is higher in the tumor tissues than the normal ones, especially in hormone-related cancers such as breast, ovarian, and prostate cancer cells. To explore the role of CYP1B1 on cancer cell proliferation, we studied the effects of CYP1B1 inhibition and activation in MCF-7 and MCF-10A cells. Here, we confirmed that expression of CYP1B1 affects the level of cell proliferation using CCK assay. To elucidate whether the cell proliferation induced by CYP1B1 is related to Wnt/β-catenin signaling, we examined the alteration of β-catenin and expression using western blot, RT-PCR, immunostaining, and qPCR followed by over-expression of CYP1B1 or treatment with DMBA (7,12-dimethylbenz[α]anthracene), a well known CYP1B1 inducer and these studies also performed when CYP1B1 gene was knockdown or cells were treated with TMS (tetramethoxystilbene), a specific CYP1B1 inhibitor. These investigations all showed that CYP1B1 promotes β-catenin expression in mRNA and protein levels and induces β-catenin nuclear localization. To identify the effect of CYP1B1 on Wnt/β-catenin signaling further, the promoter activity of β-catenin/TCF/LEF or expression level of Wnt/β-catenin signaling target genes such as c-myc, cyclin D1 were examined and all factors also showed to be up-regulated by CYP1B1. To elucidate whether CYP1B1 induces cell metastasis, cell migration and invasion assay were conducted and it has been identified that CYP1B1 enhances cell motility by induction of mesenchymal-like cell morphological changes. Moreover, the uPAR-uPA pathway has been identified to be related to the CYP1B1-induced cell metastasis by detection of uPAR-uPA pathway factors using western blot and qPCR in MCF-7 and HeLa cells. Taken together, these data suggest that CYP1B1 promotes cell proliferation and metastasis via inducing Wnt/β-catenin signaling and uPAR pathway.

Citation Format: Yeo-Jung Kwon, Sangyun Shin, Dong-Jin Ye, Hyoung-Seok Baek, Young-Jin Chun. The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A10.
Abstract LB-A17: Hospital volume determines mortality amongst neutropenic cancer patients within the United States
Authors: Lawrence Yaacov Richard , Urban Damian , Sella Tal , Bar Jair , Berger Raanan , Goldstein Jeffrey , Shacham-Shmueli Einat , Symon Zvi , Golan Talia
Published: May 2016

Introduction:

Hospital Volume is associated with improved outcomes in cancer surgery, and some aspects of cancer care. The influence of hospital volume on outcomes in neutropenic patients is unknown. Neutropenia secondary to bone-marrow suppression is a common, and dangerous complication of cancer treatment. Known factors associated with increased mortality include age, race and co-morbidities. We hypothesized that large-volume hospitals would have reduced mortality rates for neutropenic patients compared with small-volume institutions.

Methods:

We utilized the NIS database of the Healthcare Cost and Utilization Project, focusing on the years 2000-2006. Only inpatient episodes with a diagnosis of both neutropenia and cancer were included in the study. Hospital volume was defined as the number of neutropenic cancer episodes listed for each institution per year. Mortality was defined as death during admission.

Results:

155,512 hospitalizations were included in the study, from 3049 different institutions. Median age was 60 years. The overall inpatient mortality was 7.1%. Mortality increased with patient age, for instance amongst those aged 30-39 years it was 4.3% compared to 10.7% for those aged 70-79 years. The average number of “neutropenic” inpatient episodes in each institution per year ranged from less than one to 225 (median 22.7). The population was split into four equal groups based upon hospital volume. Mortality was 8.5%, 7.7%, 7.0% and 5.2% for each group (from lowest to highest volume), likewise% discharged home was 77.3%, 82.4%, 85.2% and 90.1% (for both p<0.001). In a logistic regression model incorporating age, race and sex, ‘hospital volume’ remained statistically associated with reduced mortality (p<0.001).

Conclusion:

Neutropenic patients hospitalized in large-volume institutions have a substantially lower mortality and a higher likelihood of returning home compared to those hospitalized at low-volume institutions. Our findings have implications for the setting in which phase I/II clinical trials of combination treatments with new biological agents and cytotoxic therapies be performed.

Citation Format: Yaacov Richard Lawrence, Damian Urban, Tal Sella, Jair Bar, Raanan Berger, Jeffrey Goldstein, Einat Shacham-Shmueli, Zvi Symon, Talia Golan. Hospital volume determines mortality amongst neutropenic cancer patients within the United States. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A17.
Abstract LB-A23: Galectin-8 favors growth of endothelial cells and induces production of extracellular matrix by cancer associated fibroblasts
Authors: Gal Peter , Varinska Lenka , Kello Martin , Mojzis Jan , Dvorankova Barbora , Smetana Karel
Published: May 2016

Galectins (Gal) belong to a family of endogenous lectins specifically recognizing sugar motives and playing an important role in the biology of tissue repair and cancer. It has been shown that the tandem-repeat lectin Gal-8 plays an essential role in tumor angiogenesis by improving endothelial cell migration and capillary tube formation. Interestingly by a more prominent way than the proangiogenic growth factor VEGF. Thus, new question arises, i.e. does Gal-8 improves endothelial cell proliferation? To provide answer to that question the MTT and BrDu proliferation assays using HUVEC (human umbilical vein endothelial cells) as a model were used. We found that Gal-8 induces cell proliferation in combination with VEGF, but Gal-8 alone does not exhibit any significant effect. Furthermore, we studied whether human Gal-8 induces production of a bioactive extracellular matrix (ECM) scaffold by cancer associated fibroblasts (CAF). In parallel, the effect of Gal-8 on ECM production by human dermal fibroblasts (HDF) was studied as well. Adding Gal-8 to the culture medium led the CAF to generate a very fine structured 3D meshwork especially rich in fibronectin. Contrary, the ECM production by HDF was not that apparent. Since ECM-cell interactions play an important role in the ability of tumors to spread and form metastasis further research focused on roles of the tandem-repeat-type Gal-8 in other steps of tumorigenesis is warranted by present in vitro study.

This study was supported in part by the Grant Agency of Ministry of Education, Science, Research, and Sport of the Slovak Republic (VEGA-1/0404/15 and VEGA-1/0048/15) and the Agency for Science and Research under the contract no. APVV-0408-12 and APVV-14-0731.

Citation Format: Peter Gal, Lenka Varinska, Martin Kello, Jan Mojzis, Barbora Dvorankova, Karel Smetana Jr.. Galectin-8 favors growth of endothelial cells and induces production of extracellular matrix by cancer associated fibroblasts. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A23.
Abstract LB-A24: Molecular alteration of SMAD4 in hindgut-derived colorectal tumors identifies a distinct subset of patients and is associated with worse recurrence-free survival
Authors: Smith Jesse Joshua , Lee Lik Hang , Chen Xi , Wu Chao , Pelossof Raphael , Nash Garrett M. , Temple Larissa R. , Guillem Jose G. , Weiser Martin R. , Paty Philip B. , Shia Jinru , Garcia-Aguilar Julio , Sawyers Charles L.
Published: May 2016

This abstract has been withheld from publication due to its inclusion in the AACR-NCI-EORTC Molecular Targets Conference 2015 Official Press Program. It will be posted online at the time of its presentation in a press conference or in a session: 10:00 AM ET Friday, November 6.

Citation Format: Jesse Joshua Smith, Lik Hang Lee, Xi Chen, Chao Wu, Raphael Pelossof, Garrett M. Nash, Larissa R. Temple, Jose G. Guillem, Martin R. Weiser, Philip B. Paty, Jinru Shia, Julio Garcia-Aguilar, Charles L. Sawyers. Molecular alteration of SMAD4 in hindgut-derived colorectal tumors identifies a distinct subset of patients and is associated with worse recurrence-free survival. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A24.
Abstract LB-B09: Somatic mutation detection via sequencing using circulating tumor cell samples from patients with renal cell and prostate cancer
Authors: Amato Robert J. , Brobey Reynolds , Dehghani Mehdi , Rosenblatt Kevin , Souza Glauco , Tseng Hubert , Jensen Jeff , Ionescu-Zanetti Cristian
Published: May 2016

Introduction: Tumor genotyping using fluid samples such as blood can potentially allow tracking of dynamic changes in mutational profiles over time and allow better access than biopsies. We present a method to detect somatic mutations from a blood draw, where circulating tumor cell (CTC) enrichment above 10% of total cell numbers allows the use of standard gene panels typically used to analyze tissue-based biopsies.

Methods: Clinical samples were obtained from 9 prostate cancer (PC) patients and 6 renal cell cancer (RCC) patients, followed by CTC enrichment using the IsoFluxTM System. Cells were lysed and DNA amplified by whole genome amplification (WGA) using the NGS Kit (Fluxion Biosciences) and quantified via qPCR. CTCs defined as CK+, CD45- nucleated cells (DAPI+) for cell enumeration. Analytical samples were prepare by spiking tumor derived cell lines into whole blood and parallel analysis. Next-generation sequencing was performed using 3 targeted cancer panels on the Ion torrentTM PGM platform: the Ion ampliseqTM cancer hot spot panel (50 genes; 6 PC samples), Oncomine (143 genes; 3 PC samples), and a 29-gene panel of actionable mutations in RCC (6 samples). Data was analyzed using a customized variant calling/filtering pipeline. Variant filtering and functional interpretation was performed using VarSeqTM. All data was analyzed in a blinded manner.

Results: Our method was able to isolate CTCs from all patient samples. WGA DNA concentrations were at a range of 25-164 ng/μL (median, 69) in PC and 29-180 ng/μL (median, 69) in RCC. CTC purity after the first enrichment step was in 2.9-33.7% (median, 10.5%) of PC samples and 1.9-33% (median, 14.5%) of RCC; final CTC purity is estimated at 5-40%. We found 1 variant/sample using hotpot, 12/sample using Oncomine, and 3/sample using the RCC panel.

Conclusions: Our assay consistently detected somatic variants from blood draw using standard gene panels in both PC and RCC. Obtaining repeat tumor biopsies from patients during treatment and/or at time of progression is both challenging and impractical from a clinical perspective. Our assay provides molecular characterization using standard blood draws and will be prospectively validated in clinical trials.

Citation Format: Robert J. Amato, Reynolds Brobey, Mehdi Dehghani, Kevin Rosenblatt, Glauco Souza, Hubert Tseng, Jeff Jensen, Cristian Ionescu-Zanetti. Somatic mutation detection via sequencing using circulating tumor cell samples from patients with renal cell and prostate cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-B09.
Abstract LB-B15: A novel PI3K gamma isoform selective small molecule kinase inhibitor demonstrates single agent anti-tumor activity and enhanced combination activity with checkpoint blockade in syngeneic mouse models of cancer
Authors: Saha Saurabh , Zhang Linping , Hoock Thomas , Aronov Alex , Mahajan Sudipta , Boyd Michael , Come Jon , Damagnez Veronique , Dworakowski Wojciech , Khare-Pandit Suvarna , LeTiran Arnaud , Moody Cameron , O'Dowd Harwin , Prezioso Joseph , Roday Setu , Zhang Xiaoyan M.
Published: May 2016

Immune checkpoint inhibitors have generated impressive clinical responses, however, a number of patients and cancer types remain resistant to checkpoint blockade. Immunosuppressive cells such as Tregs and MDSCs in the tumor microenvironment are hypothesized to mediate this resistance. The gamma (γ) isoform of PI3K has long been known to modulate myeloid and lymphocyte cell migration and function, including trafficking of monocytic and granulocytic cells into inflamed tissues such as tumors. Here we report the identification and characterization of BVD-723, a PI3Kγ selective small molecule kinase inhibitor which demonstrated single agent and enhanced combination anti-tumor activity[xx]with either an anti-PD-1 or anti-CTLA-4 antibody. In a mouse syngeneic colon cancer model,[xx]BVD-723 in combination with anti-PD-1 or anti-CTLA-4 resulted in complete regression[xx]of tumors in 35% (7/20) and 60% (12/20) of animals treated with each antibody respectively [versus 0% (0/20) and 5% (1/20) with either antibody alone]. Interestingly, the anti-tumor activity was coupled with a decrease in tumor infiltrating Tregs, granulocytic MDSCs, CD4+ T cells and an increase in the CD8+/Treg ratio, suggesting that inhibition of PI3Kγ by BVD-723 promotes immune-reactivation in the tumor microenvironment. Additionally, BVD-723 demonstrated potent synergy with an anti-PD-1 antibody in a syngeneic mouse model of lymphoma. Single agent BVD-723 activity was also observed in syngeneic mouse models of pancreas, colon, lymphoma and bladder cancers. Results from completed safety pharmacology, toxicology and in vivo efficacy studies support clinical evaluation of BVD-723[xx]as a monotherapy or in combination with the checkpoint inhibitors in cancer.[xx]

Citation Format: Saurabh Saha, Linping Zhang, Thomas Hoock, Alex Aronov, Sudipta Mahajan, Michael Boyd, Jon Come, Veronique Damagnez, Wojciech Dworakowski, Suvarna Khare-Pandit, Arnaud LeTiran, Cameron Moody, Harwin O'Dowd, Joseph Prezioso, Setu Roday, Xiaoyan M. Zhang. A novel PI3K gamma isoform selective small molecule kinase inhibitor demonstrates single agent anti-tumor activity and enhanced combination activity with checkpoint blockade in syngeneic mouse models of cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-B15.
Abstract LB-B22: CCNY2 promotes lung cancer cell migration and invasion via regulating F-actin expression
Authors: zhao xiaoting , Jiang Mei , Yue wentao
Published: May 2016

Background: Cyclin Y (CCNY) is a novel cyclin and is highly conserved in metazoan species. Cyclin Y mRNA has several transcripts and only ccny1 and ccny2 has been documented. A potential CDK partner of Cyclin Y is PFTAIRE kinase (PFTK1). In hepatocellular carcinoma, cell motility and invasion was enhanced by PFTK1 expression. But the function of CCNY1 and CCNY2 on cell migration and invasiveness has not been reported yet. Methods: Recombined plasmids carrying CCNY1 and CCNY2 were constructed and transfected to H1299 cells to obtain CCNY up-regulation cells. A lentivirus-based RNAi delivery system was used to inhibit CCNY mRNA expression. The role of CCNY in cell motility and invasion was investigated using wound healing and transwell assay. The protein levels in lung cancer cells were determined by western-blot, immunofluorescence technique and high-content cell analysis. Mouse xenograft experiments were carried out to study the metastasis ability of CCNY1 and CCNY2 in vivo. Immunohistochemistry was used to detect the CCNY protein level of lung cancer tissues. Results: cell motility and invasion activity were inhibited and MET (Mesenchymal - Epithelial Transition) was caused by down-regulating of CCNY in 95D and H1299 cells. CCNY2 could enhance cell migration and invasion activity in vivo and vitro. The F-actin level was regulated by CCNY2 expression. In non-small cell lung cancer tissues, CCNY2 was highly expressed and the CCNY2 expression was associated with histological grade. Conclusion: CCNY2 was firstly detected in lung cancer cells and non-small cell lung cancer tissues. Our findings demonstrated CCNY2 not CCNY1 promoted cell motility and invasion by regulating the expression of F-actin and modulating intracellular cytoskeletal components.

Citation Format: xiaoting zhao, Mei Jiang, wentao Yue. CCNY2 promotes lung cancer cell migration and invasion via regulating F-actin expression. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-B22.
Abstract LB-B23: Medulloblastoma regulatory circuitries reveal subgroup-specific cellular origins
Authors: Lin Charles Y. , Erkek Serap , Tong Yiai , Yang Linlin , Federation Alexander J. , Zapatka Marc , Haldipur Parthiv , Kawauchi Daisuke , Risch Thomas , Warnatz Hans-Jörg , Worst Barbara , Ju Bensheng , Orr Brent A. , Zeid Rhamy , Polaski Donald R. , Segura-Wang Maia , Waszak Sebastian M. , Jones David TW , Kool Marcel , Hovestadt Volker , Buchhalter Ivo , Sieber Laura , Johann Pascal , Gröschel Stefan , Ryzhova Marina , Korshunov Andrey , Chen Wenbiao , Chizhikov Victor V. , Millen Kathleen J. , Amstislavskiy Vyacheslav , Lehrach Hans , Yaspo Marie-Laure , Eils Roland , Lichter Peter , Korbel Jan O. , Pfister Stefan , Bradner James E. , Northcott Paul A.
Published: May 2016

Medulloblastoma is a highly malignant paediatric brain tumour, often inflicting devastating consequences on the developing child. Genomic studies have revealed four distinct molecular subgroups with divergent biology and clinical behaviour. An understanding of the regulatory circuitry governing the transcriptional landscapes of medulloblastoma subgroups, and how this relates to their respective developmental origins, is currently lacking. Using H3K27ac and BRD4 ChIP-Seq, coupled with tissue-matched DNA methylation and transcriptome data, we describe the active cis-regulatory landscape across 28 primary medulloblastoma specimens. Analysis of differentially regulated enhancers and super-enhancers reinforced inter-subgroup heterogeneity and revealed novel, clinically relevant insights into medulloblastoma biology. Computational reconstruction of core regulatory circuitry identified a master set of transcription factors responsible for subgroup divergence that validated by ChIP-Seq and implicated candidate cells-of-origin for Group 4. Our integrated analysis of cis-regulatory elements in a large series of primary tumour samples reveals insights into cis-regulatory architecture, unrecognized dependencies, and cellular origins.

Citation Format: Charles Y. Lin, Serap Erkek, Yiai Tong, Linlin Yang, Alexander J. Federation, Marc Zapatka, Parthiv Haldipur, Daisuke Kawauchi, Thomas Risch, Hans-Jörg Warnatz, Barbara Worst, Bensheng Ju, Brent A. Orr, Rhamy Zeid, Donald R. Polaski, Maia Segura-Wang, Sebastian M. Waszak, David TW Jones, Marcel Kool, Volker Hovestadt, Ivo Buchhalter, Laura Sieber, Pascal Johann, Stefan Gröschel, Marina Ryzhova, Andrey Korshunov, Wenbiao Chen, Victor V. Chizhikov, Kathleen J. Millen, Vyacheslav Amstislavskiy, Hans Lehrach, Marie-Laure Yaspo, Roland Eils, Peter Lichter, Jan O. Korbel, Stefan Pfister, James E. Bradner, Paul A. Northcott. Medulloblastoma regulatory circuitries reveal subgroup-specific cellular origins. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-B23.
Abstract LB-B25: Consensus tissue-independent epithelial tumour subtypes and their differing responses to therapy
Authors: Sadanandam Anguraj
Published: May 2016

Background: Epithelial cancers are highly heterogeneous, and each epithelial tumour is composed of a number of subtypes. It is becoming clear that the same dysregulated pathways are common to different tumours, regardless of their tissue of origin. Therefore, the goal of the current study was to identify intrinsic tissue-independent epithelial tumour subtypes by analysing a large cohort of different epithelial tumours.

Methodology: Different gene expression profiles from various epithelial tumours (n = ∼5000) and cell lines (n = ∼200) from 12 different organs were merged using distance weight discrimination analysis, followed by consensus clustering using non-negative matrix factorization (NMF). The association of subtypes with response to various therapeutic compounds was identified using publicly available cell line drug response data.

Results: Three to seven consensus gene expression subtypes were identified and validated in the epithelial tumours and cell lines, irrespective of the tissue of origin. Certain epithelial subtypes were not present in melanoma or haematological cancers. The consensus subtypes had gene expression and biological profiles similar to normal epithelial and stem cells, and could be readily compared to the transcriptomic subtypes previously reported for individual epithelial tumour types. Moreover, specific gene signatures were identified that would be suitable as clinical assays in future applications of this classification system. There were significant differential responses between subtypes to different therapeutic compounds.

Conclusion: Several consensus transcriptomic subtypes were identified that are applicable to different organ-specific tumours. This study provides new avenues for precision therapy irrespective of the tissue of origin.

Late-Breaking Poster Session C:[xx]Animal Models

Citation Format: Anguraj Sadanandam. Consensus tissue-independent epithelial tumour subtypes and their differing responses to therapy. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-B25.
Abstract LB-C13: Selective GPR55 antagonism attenuates EGFR-PKM2-β-catenin signaling and reduces chemoresistance in PANC-1 pancreatic cancer cells
Authors: Singh Nagendra , Bernier Michel , Wainer Irving W.
Published: May 2016

G protein-coupled receptor GPR55 possesses pro-oncogenic activity, but the nature of GPR55 signaling pathways in mammalian tumor cells remain unclear. The effect of (R,R')-4'-methoxy-1-naphthylfenoterol (MNF), a potent GPR55 competitive inhibitor, on the expression of cancer biomarkers, such as EGFR, PKM2, β-catenin and HIF-1A, was investigated in human pancreatic cancer cell line, PANC-1. We also examined the modulation in expression of P-glycoprotein (Pgp) and other multidrug resistance drug exporters (MDR) in response to MNF. Incubation of PANC1 cells with MNF (1 μM) for 24h significantly decreased EGFR (∼2-fold), PKM2 (∼2.5-fold) and β-catenin (∼2-fold) protein levels and was accompanied by significant reduction in nuclear accumulation of HIF-1A(∼1.5-fold) and phospho-active forms of PKM2 (∼3-fold) and B-catenin (∼2-fold). MNF and the GPR55 antagonist CID16020046 consistently reduced the expression of Pgp (∼2-fold), BCRP (∼2-fold), MRP1 (∼1.5-fold) and MRP5 (∼1.5-fold) in total cellular extracts while diminishing the nuclear expression of Pgp (∼2.5-fold) and BCRP (∼4.5-fold). Further, a significant increase in the nuclear accumulation of doxorubicin was observed in MNF-treated PANC-1 cells using laser scanning confocal microscopy. Importantly, a 24-h preincubation with MNF increased the cytotoxicity of doxorubicin (IC50 reduced from 170nM to 24nM) and gemcitabine (IC50 reduced from 486nM to 157nM) in PANC-1 cells. In the mouse xenograft model, daily MNF treatment (10mg.kg-1, i.p.) for 21 days led to a ∼2-fold reduction in the expression of EGFR, PKM2, β-catenin and Pgp in PANC-1 tumor tissue relative to vehicle-treated controls (P < 0.001). These data suggest that MNF exerts its antitumor effects by inhibiting GPR55-mediated activation of select cancer biomarkers, and suppressing multidrug resistance to chemotherapeutic agents in pancreatic cancer cells.

Citation Format: Nagendra Singh, Michel Bernier, Irving W. Wainer. Selective GPR55 antagonism attenuates EGFR-PKM2-β-catenin signaling and reduces chemoresistance in PANC-1 pancreatic cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-C13.
Abstract LB-C16: Safety and tumor responses of the anti-Trop-2 antibody drug conjugate, sacituzumab govitecan (IMMU-132), in refractory, metastatic, triple-negative breast cancer (TNBC): An ongoing Phase II trial
Authors: Bardia Aditya , Mayer Ingrid , Diamond Jennifer , Starodub Alexander , Moroose Rebecca , Isakoff Steven , Ocean Allyson , Guarino Michael , Kalinsky Kevin , O'Shaugnessy Joyce , Wilhelm Francois , Maliakal Pius , Sharkey Robert , Goldenberg David , Vahdat Linda
Published: May 2016

This abstract has been withheld from publication due to its inclusion in the AACR-NCI-EORTC Molecular Targets Conference 2015 Official Press Program. It will be posted online at the time of its presentation in a press conference or in a session: 12:30 PM ET Sunday, November 8.

Citation Format: Aditya Bardia, Ingrid Mayer, Jennifer Diamond, Alexander Starodub, Rebecca Moroose, Steven Isakoff, Allyson Ocean, Michael Guarino, Kevin Kalinsky, Joyce O'Shaugnessy, Francois Wilhelm, Pius Maliakal, Robert Sharkey, David Goldenberg, Linda Vahdat. Safety and tumor responses of the anti-Trop-2 antibody drug conjugate, sacituzumab govitecan (IMMU-132), in refractory, metastatic, triple-negative breast cancer (TNBC): An ongoing Phase II trial. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-C16.