bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3717: Therapeutic strategies to overcome the crizotinib resistance in ROS1-rearranged lung cancers
Authors: Katayama Ryohei , Kobayashi Yuka , Koike Sumie , Fujita Naoya 
Published: May 2015

Abstract
ROS1 fusion oncogene represent a novel molecular target in a small subset of non-small cell lung cancers (NSCLC).The ROS1 fusion oncogene, such as CD74-ROS1, leads to constitutive ROS1 activation with potent transforming activity. In the ongoing phase 1 clinical trial, the response rate of crizotinib, ALK/MET/ROS1 tyrosine kinase inhibitor (TKI), in the patients with NSCLC harboring ROS1 fusions was around 60%. However, despite crizotinib has shown remarkable initial responses, cancers eventually develop resistance to crizotinib. In a recent report of a clinical resistance to crizotinib, G2032R mutation in ROS1 kinase domain was identified, which is analogous to the G1202R mutation found in crizotinib-resistant ALK-rearranged lung cancers. To further identify the mechanism of resistance to crizotinib in ROS1 fusion positive NSCLC, we established a model of acquired resistance by ENU mutagenesis screening using CD74-ROS1 addicted Ba/F3 cell line models. As the results, we identified several novel crizotinib resistance mutations including G2032R mutation. To overcome the resistance, we performed the drug screening with small molecular inhibitors, and identified several inhibitors including Foretinib, cMET/VEGFR inhibitor, effectively inhibit the survival of CD74-ROS1 expressing cells. Furthermore, we also identified that some of those inhibitors can overcome the resistance caused by G2032R mutation. Thus, we have developed a comprehensive model of acquired resistance to crizotinib in ROS1 fusion harboring NSCLC and identified therapeutic strategies to overcome the resistance.
Citation Format: Ryohei Katayama, Yuka Kobayashi, Sumie Koike, Naoya Fujita. Therapeutic strategies to overcome the crizotinib resistance in ROS1-rearranged lung cancers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3717. doi:10.1158/1538-7445.AM2014-3717
Abstract 3720: Intrinsic and acquired resistance mechanisms of alectinib in ALK rearranged cells
Authors: Hibino Yukiko , Takaaki Sasaki , Minami Yoshinori , Janne Pasi A. , Ohsaki Yoshinobu 
Published: May 2015

Abstract
Background
However potency of Alectinib, a selective ALK inhibitor, for ALK rearranged NSCLC and for that with secondary mutation have been reported, intrinsic and acquired resistance are likely to emerge. Recent study showed tumor microenvironment triggered resistance to Crizotinib and TAE684. In this study, we examine whether emerging of intrinsic and acquired resistance are affected by tumor microenvironment in H3122, EML4-ALK positive lung cancer cell line, and in Alectinib-resistant clone of H3122 (AFR) which is generated in vitro. Elucidating intrinsic resistant mechanisms and preceding development of acquired resistance to understand its mechanisms will provide beneficial information for the development of novel therapeutic strategies in clinical practice.
Methods
We examined the effect of EGF produced by endothelial cells on Alectinib sensitivity of H3122 cell line, and EGF and HGF produced by fibroblasts on Alectinib sensitivity of H3122AFR by clonogenic assay. Downstream pathways of ALK in both cells were analyzed by western blotting. Combination treatments of Alectinib and other TKI were tested both in vitro and in vivo.
Results
H3122 cells which had been cultured with EGF acquired resistance to Alectinib treatment. Combination treatment of Alectinib and Erlotinib overcome EGF induced resistance and showed down-regulation in phosphorylation of AKT and ERK1/2. H3122AFR cells were also reduced their sensitivity to Alectinib at 100 uM or higher concentration when the media contained EGF or HGF. Erlotinib or Crizotinib combined with Alectinib canceled the EGF-induced or HGF-induced resistance by down-regulating phosphorylation of AKT and ERK1/2. In vivo, tumor xenografts of nude mice which had been orally administered Alectinib and Erlotinib showed tumor regression in both H3122 and H3122AFR, and those received Alectinib and Crizotinib showed regression in H3122AFR.
Conclusion
In our study, we showed ligands produced by tumor microenvironment, such as EGF and HGF, contributed the emerging of intrinsic resistance and acquired resistance by activating survival pathway. Further, the efficacy of combination treatment of Alectinib with Erlotinib or that with Crizotinib was shown. Collectively, Alectinib combined with EGFR-TKI would be an effective strategy for Alectinib-resistant lung cancer, and Alectinib with crizotinib is another considerable strategy.
Citation Format: Yukiko Hibino, Sasaki Takaaki, Yoshinori Minami, Pasi A. Janne, Yoshinobu Ohsaki. Intrinsic and acquired resistance mechanisms of alectinib in ALK rearranged cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3720. doi:10.1158/1538-7445.AM2014-3720
Abstract 3728: A discovery platform for allosteric modulators of c-Abl kinase function
Authors: Grover Prerna , Smithgall Thomas E. 
Published: May 2015

Abstract
The c-Abl protein-tyrosine kinase regulates intracellular signaling pathways controlling cell growth, adhesion, and responses to genotoxic stress. The kinase activity of c-Abl is repressed by intramolecular interactions involving its non-catalytic SH3 and SH2 domains. The SH3 domain interacts with a polyproline type II helix formed by the SH2-kinase linker, while the SH2 domain interacts with the C-lobe of kinase domain away from the active site. In addition, the myristoylated N-terminal cap region (Ncap) binds a hydrophobic pocket in the C-lobe of the kinase domain, clamping the SH3 and SH2 domains against the back of the kinase domain. Small molecules that allosterically regulate c-Abl kinase activity through its non-catalytic domains may represent selective probes of c-Abl function in diverse tumor types. In this study, we developed a chemical library screening assay designed to identify chemical modulators of c-Abl kinase activity that either disrupt or stabilize the regulatory interaction of the SH3 domain with the SH2-kinase linker. This fluorescence polarization (FP) assay is based on a recombinant purified protein consisting of the c-Abl Ncap, SH3 and SH2 domains plus the linker (N32L protein) and a short fluorescein-labeled probe peptide that binds to the SH3 domain. In assay development experiments, we found that the probe peptide binds to the recombinant c-Abl N32L protein in vitro, producing a robust FP signal that can be competed with an excess of unlabeled peptide. The FP signal is not observed with control N32L proteins bearing either an inactivating mutation in the SH3 domain or enhanced SH3:linker interaction. The assay was optimized with respect to probe peptide sequence, probe and N32L protein concentrations, DMSO tolerance, and FP signal stability over time. Pilot library screens were then performed with the NCI Diversity Set III and an FDA-approved drug library (2800 compounds total). Seven compounds were identified that specifically reduced the FP signal by at least three standard deviations from the untreated controls. Secondary assays showed that one of these hit compounds enhances c-Abl kinase activity in vitro, suggesting that it disrupts regulation of kinase activity through the non-catalytic region. These results show that screening assays based on the non-catalytic domains of c-Abl can identify small molecule regulators of kinase function.
Citation Format: Prerna Grover, Thomas E. Smithgall. A discovery platform for allosteric modulators of c-Abl kinase function. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3728. doi:10.1158/1538-7445.AM2014-3728
Abstract 373: An siRNA screen identifies CHD4 as a target for epigenetic therapy
Authors: Okamoto Yasuyuki , Yamazaki Jumpei , Sato Takahiro , Cesaroni Matteo , Chung Woonbok , Garriga Judith , Jelinek Jaroslav , Katz Richard A , Issa Jean-Pierre 
Published: May 2015

Abstract
Background and aims: In the past decade, epigenetic mechanisms in the regulation of tumor-specific gene expression have been elucidated partially, prompting the development of therapeutic approaches in cancer cells. In this study, we used an unbiased screen to investigate therapeutic targets which might be effective combination with DNMT inhibitors in reactivating hypermethylated genes. Methods: We screened an siRNA library targeting 188 gene predicted as epigenetic regulators using colon cancer cells that harbor a GFP locus stably integrated and silenced by a hypermethylated cytomegalovirus (CMV) promoter. GFP-positive cell percentages were measured using Guava EasyCyte Plus flow analyzer software. For genome wide gene expression analysis, affymetrix microarrays were performed. DNA methylation status was evaluated by pyrosequencing analysis and Digital Restriction Enzyme Analysis of Methylation (DREAM), based on next generation sequencing analysis. To access biological effect of the combination therapy, we evaluated cell growth using several cell lines. Results: We identified the chromodomain helicase DNA-binding protein 4 (CHD4) as a target; CHD4 depletion in combination with DNMT inhibition synergistically reactivated expression of GFP and endogenously methylated genes. Synergy was most pronounced for genes with very low basal expression. DREAM analysis revealed that methylated genes with promoter CpG islands were enriched among reactivated genes compared to background genes (57% vs 27%, P&lt;0.0001). DNA methylation levels of reactivating genes after combination therapy were identical with DNMT inhibitor monotherapy, suggesting that siCHD4 as a supplement to DNMT inhibition works by a parallel, synergistic mechanism. Combination treatment inhibited cell growth of colon cancer cell lines better than treatment with a single DNMT inhibitor with minimal effects on the cell growth of non-cancerous cell lines. Interestingly, depletion of CHD4 alone also inhibited cell growth of colon cancer cell line but not of non-cancerous cell lines. Conclusions: Combination treatment of siCHD4 and DNMT inhibitor strongly reactivated hypermethylated and silenced genes in cancer cell lines. CHD4 inhibition is a promising new approach to epigenetic targeted therapy.
Citation Format: Yasuyuki Okamoto, Jumpei Yamazaki, Takahiro Sato, Matteo Cesaroni, Woonbok Chung, Judith Garriga, Jaroslav Jelinek, Richard A Katz, Jean-Pierre Issa. An siRNA screen identifies CHD4 as a target for epigenetic therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 373. doi:10.1158/1538-7445.AM2014-373
Abstract 3732: Utilizing Bimolecular Fluorescent Complementation to Identify Inhibitors of RAL and KRAS
Authors: Ifediba Marytheresa , Burnett David , Hovda Nicholas , Burnett William , VanBrocklin Matthew 
Published: May 2015

Abstract
We have identified RAL activity to be vital for NSCLC survival independent of KRAS or EGFR mutation status in a large panel of adenocarcinomas and have developed a novel high throughput screening assay based on Bimolecular Fluorescent Complementation (BiFC) to identify small molecules that impair RAL function. Coupling RALA and RALB to the C-terminus of the Venus protein (VC) and coexpressing either with direct binding effector RALBP1 fused to the N-terminus of the Venus protein (VN) result in a fluorescent signal when RAL-RALBP1 binding occurs in cells. This allows for the interrogation of small molecules that disrupt RAL-RALBP1 binding interactions (resulting in loss of fluorescence) in a real time, high throughput manner. Our preliminary screen with a natural compound library identified 14 small molecules that demonstrated high activity in disrupting both RALA- and RALB-RALBP1 binding and fluorescence. We are currently investigating these compounds' mode of action and have recently expanded our assay to screen for molecules that impair mutant KRAS activity (VC-KRAS / RAF1-VN). BiFC provides a rapid, more physiologic method to identify molecules that can disrupt protein-protein interactions of targets considered ‘undruggable’, including membrane bound small GTPases such as RAL and KRAS.
Citation Format: Marytheresa Ifediba, David Burnett, Nicholas Hovda, William Burnett, Matthew VanBrocklin. Utilizing Bimolecular Fluorescent Complementation to Identify Inhibitors of RAL and KRAS. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3732. doi:10.1158/1538-7445.AM2014-3732
Abstract 3743: Bioluminescent, non-lytic, real-time cell viability assay
Authors: Duellman Sarah J. , Vidugiriene Jolanta , Zhou Wenhui , Osterman Jean , Arbit Ruslan , Bernad Laurent , Meisenheimer Poncho , Cali James J. 
Published: May 2015

Abstract
Cell-based assay development for cancer drug discovery continues to advance toward more innovative methods that provide efficient workflow and meaningful data. Rapid, sensitive techniques that are non-lytic and therefore facilitate multiplexing with a wide range of assays allow more data to be gained per well and conserve precious samples. Real time measurement of cell viability also allows numerous measurements over time without the need for separate plates at each time point. Real time measurements allow unique analyses including interrogation of the timing of drug killing, identification of static vs toxic drug effects, and determination of the ideal time to multiplex an additional assay. Cancer drug discovery would benefit from novel assay technologies that allow real time measurements with multiplexing capability.
We have developed a homogeneous, non-lytic, and bioluminescent method to analyze cell viability in real time through measurement of the reducing potential of the cell. The signal correlates with the number of viable cells making it well-suited for cytotoxicity studies. This Real Time Cell Viability Assay utilizes the NanoLuc luciferase enzyme, which is 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase, and the novel substrate, furimazine. Together these reagents produce high intensity luminescence through an ATP-independent reaction.
The assay can be performed in two formats, live cell endpoint or continuous read. The live cell endpoint method is sensitive, rapid, and enables extensive multiplexing opportunities due to the non-lytic measurement and signal depletion upon cell lysis. When the assay is set up in the continuous read format, the cell viability reagents can be added at the same time as cell plating, drug dosing, or at whatever point in the assay the researcher would like to start obtaining viability readings. The luminescent signal can be continually monitored from the same wells over an extended period of time to analyze cell viability in real time.
This real time cell viability assay enables non-lytic cell viability measurements, extensive multiplexing options including both fluorescent and luminescent assays with no special filter requirements, straightforward normalization studies, conservation of precious samples and reagents, and a sensitive, real time measurement of cell viability.
Citation Format: Sarah J. Duellman, Jolanta Vidugiriene, Wenhui Zhou, Jean Osterman, Ruslan Arbit, Laurent Bernad, Poncho Meisenheimer, James J. Cali. Bioluminescent, non-lytic, real-time cell viability assay. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3743. doi:10.1158/1538-7445.AM2014-3743
Abstract 3762: Downregulation of TRPC3 enhances sensitivity to cisplatin and inhibits motility in epithelial ovarian cancer
Authors: Tao Xiang , Jin Hongyan , Zhang Zhenbo , Bast Robert C. , Feng Youji , Yu Yinhua 
Published: May 2015

Abstract
Epithelial ovarian cancer (EOC) remains the most lethel gynecological malignancy. In the absence of an effective screening strategy, two thirds of patients are diagnosed in advanced clinical stage. Despite chemotherapy with cisplatin and paclitxel, primary or acquired chemoresistance faciltates survival of cancer cells, resulting in disease recurrence and death. Our previous studies indicate that ovarian cancer patients whose tumors overexpress transient receptor potential channel C3 (TRPC3) protein tend to relapse early and have shorter overall survival. In vitro studies indicated that follicle-stimulating hormone (FSH) induced resistance to cisplatin-induced apoptosis could be partially reversed by blocking expression of TRPC3 in EOC cell lines. To enhance sensitivity to platinum compounds, we have investigated the relationship between resistance to cisplatin and TRPC3. We have now found that expression of TRPC3 in recurrent cancer following chemotherapy is higher than in specimens from primary cytoreductive surgery. The expression of TRPC3 in the cisplatin-resistant EOC line A2780cp is higher than in the sensitive parental cell line A2780. An in vitro study showed that transient treatment of the EOC cell line HEY with cisplatin upregulated TRPC3 mRNA and membrane localized protein. Induced cisplatin-resistant cell line HEYcp augmented the expression of TRPC3 and enhanced invasive capacity in cell migration assays. Knockdown of TRPC3 expression in EOC cell lines with specific siRNA or miRNA decreased cell migratory ability and enhanced cisplatin-induced apoptosis when compared to EOC cells treated with scrambled control RNA. Our data indicate that TRPC3 may play an important role in cisplatin-resistance of EOC and could serve as a therapeutic target to resensitize cancer cells to cisplatin in culture and in xenografts.
Citation Format: Xiang Tao, Hongyan Jin, Zhenbo Zhang, Robert C. Bast, Youji Feng, Yinhua Yu. Downregulation of TRPC3 enhances sensitivity to cisplatin and inhibits motility in epithelial ovarian cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3762. doi:10.1158/1538-7445.AM2014-3762
Abstract 3795: Cabozantinib affects multiple signaling pathways in glioblastoma and is effective in a subset of xenograft tumors
Authors: deCarvalho Ana C. , Arnold Kimberly , Mueller Claudius , Petricoin Emanuel F. , Poisson Laila M. , Mikkelsen Tom 
Published: May 2015

Abstract
Background: Glioblastoma (GBM) is the most common and aggressive form of brain tumor characterized by poor prognosis and high recurrence rate. Several new targeted agents are being considered for the adjuvant treatment of GBMs. Cabozantinib, a multi-kinase currently in clinical trials for several malignancies, including glioblastoma, targets c-Met and VEGFR2 receptors, as well as Ret, Kit, Flt-1/3/4, Tie2, and AXL. Our goal is to first evaluate sensitivity and molecular response of patient-derived GBM cancer stem cells (CSCs) to XL184 in vitro. Anti-tumor efficacy of cabozantinib as a single agent and in combination with the DNA-alkylating agent temozolomide (TMZ) in orthotopic xenografts was then evaluated.
Methods: Neurospheres enriched in CSCs were cultured from resected GBM tumors. Sensitivity to cabozantinib was determined in vitro. Cells were treated (IC40) in triplicate, and cell lysates were analyzed by reverse phase protein microarrays. GBM CSCs were implanted intracranially into nude mice. Cabozantinib was administered by oral gavage at a dose of 60mg/kg for 4 weeks (5 days/week) as a single agent or in combination with 40mg/kg TMZ. Tumor growth and response to treatment were monitored by non-invasive in vivo biolumiescent imaging (BLI) using the Xenogen IVIS System (Caliper Life Sciences), and overall survival.
Results: Sensitivity to cabozantinib treatment varied for the different GBM CSCs. From 70 proteins and phosphoproteins measured, 29 distributed among several signaling pathways were significantly altered after treatment in both resistant and sensitive GBM CSCs. Cabozantinib single agent treatment reduced GBM tumor growth and increased mouse survival in two xenograft lines. Cabozantinib monotherapy reduced tumor size, as measured by BLI, but had no significant effect on overall survival for another xenograft line, however, the combination treatment resulted in sensitization of these xenografts to TMZ treatment.
Conclusion: The use of multi-kinase inhibitors is a promising strategy for GBMs, and given the range of responses, identifying predictive biomarkers, though challenging, will be invaluable for patient stratification.
Citation Format: Ana C. deCarvalho, Kimberly Arnold, Claudius Mueller, Emanuel F. Petricoin, Laila M. Poisson, Tom Mikkelsen. Cabozantinib affects multiple signaling pathways in glioblastoma and is effective in a subset of xenograft tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3795. doi:10.1158/1538-7445.AM2014-3795
Abstract 3799: Urokinase kringle-derived peptide UP-7 potently inhibits angiogenesis and tumor growth
Authors: Kim Hyun-Kyung , Naidansuren Purevjargal , Lee Seung Woo , Joe Young Ae 
Published: May 2015

Abstract
The recombinant kringle domain of urokinase-type plasminogen activator (UK1) has been shown to inhibit angiogenesis in vitro and in vivo and suppress brain tumor in vivo. UK1 consists of 83 amino acids and has 2 short helices, rudimentary ß-sheets and one extended antiparallel ß-sheets regions. In order to identify functional core sequence of the UK1 for angiogenesis inhibition, we examined the anti-angiogenic and anti-tumor activities for the seven UK1-derived peptides. Among the tested peptides, UP-7 potently inhibited the proliferation and migration of endothelial cells in vitro and suppressed vessel formation in Matrigel plugs in vivo. However, such anti-angiogenic activities were not exerted by the scrambled peptide UP7-SC7 or UP7-derived shorter peptides. UP-7 also suppressed VEGF-induced phosphorylation of FAK and ERK1/2 in endothelial cells. In a NCI-H460 lung cancer xenografted animal model, UP-7 effectively inhibited tumor growth. In addition, treatment of mice bearing MDA-MB231 tumor with UP-7 peptide resulted in a dose-dependent inhibition of tumor growth. Taken together, these results suggest that UK1- derived novel peptide UP-7 can be used for treatment of cancers. (NRF-2012R1A1A2007175).
Citation Format: Hyun-Kyung Kim, Purevjargal Naidansuren, Seung Woo Lee, Young Ae Joe. Urokinase kringle-derived peptide UP-7 potently inhibits angiogenesis and tumor growth. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3799. doi:10.1158/1538-7445.AM2014-3799