bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2912: CRISPR-Cas9: A tool for rapid target discovery and validation
Authors: Wiggins Ceri M. , Walter David , Russell Paul , Sheridan Clare , Hudson Chantelle , Yarker Joanne , Sage Carlos le , McCarthy Nicola , Moore Jonathan D. 
Published: August 2018

Abstract
Genetic perturbations mediated by CRISPR-Cas9 are transforming the drug discovery process. In the hunt for novel drug targets, researchers finally have a screening tool with sufficient penetrance and power to identify new interactions, followed by a rapid gene editing technology to streamline validation approaches. At Horizon Discovery, we have now initiated more than 250 CRISPR-Cas9 knockout (KO) screens, including a significant effort to identify novel synthetic lethal interactions in colon and lung cancer genotypes. These screens have identified a reassuring number of known essential interactions, such as MDM2 and PPMID in TP53 wild type cancer cell lines. These results provide confidence in the ability of these screens to also discover novel targets that should make it through the validation process. Indeed, we have discovered novel targets for selective ablation of TP53 mutant colon cancers, several of which are strongly supported by the literature. In FBXW7 mutant colon cancers, CRISPR-Cas9 KO screens identified a set of potential novel druggable targets in addition to MCL1 - a known substrate of FBXW7, which showed a strong dependency in FBXW7 mutant lines. An important step in triaging primary hits ahead of extensive validation work has been secondary screens that target functional domains with high density sgRNA tiling. They allow more targets to be examined than would be feasible by any arrayed approach. Data from such screens, which test tens to hundreds of guides in unison, not only increases hit calling power, but can also yield information regarding important domains and mechanism of action. This was evident in the FBXW7 secondary screen data, where guides targeting the key functional BH domains of MCL1 proved to be the most effective. This information can be used to prioritise individual targets for further validation, using techniques that include an inducible CRISPR-Cas9 system to assess the effect of individual guides in an arrayed format on cell proliferation and survival, and CRISPRi. These data exemplify how CRISPR-Cas9 can enable the discovery of a new, more robust generation of drug and antibody targets.
Citation Format: Ceri M. Wiggins, David Walter, Paul Russell, Clare Sheridan, Chantelle Hudson, Joanne Yarker, Carlos le Sage, Nicola McCarthy, Jonathan D. Moore. CRISPR-Cas9: A tool for rapid target discovery and validation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2912.
Abstract 2924: Preclinical analysis of synergistic antitumor effect of eribulin and HDAC inhibitor for triple-negative breast cancer cells
Authors: Oba Takaaki , Izumi Hiroto , Ito Ken-ichi 
Published: August 2018

Abstract
Eribulin (ERI), an inhibitor of microtubule dynamics, is a key drug for metastatic breast cancer. While HDAC inhibitors have been shown to presents antitumor effect for various malignancies. Among HDAC family, HDAC6 is known as a deacetylase of tubulin. To develop a novel therapeutic strategy for triple negative breast cancer (TNBC), we tested the combinational effect of ERI and HDAC inhibitor for TNBC cells in vitro. Two TNBC cell lines (MDA-MB-231, Hs578T) and their ERI-resistant cells (MDA-MB-231/E, Hs578T/E) established in our laboratory were used in the study. Vorinostat (VOR), a pan-HDAC inhibitor, and ricolinostat (RICO), a selective HDAC6 inhibitor were used. The sensitivity to VOR or RICO was not different between the parental and the ERI-resistant cells. In the protein expression analysis, HDAC6 expression was not altered between parental and ERI-resistant cells. Prior treatment with VOR or RICO increased the expression of acetylatedα-tubulin in both parental and ERI-resistant TNBC cell lines. When low concentration of VOR (0.5 μM) or RICO (0.5 μM) was added simultaneously with ERI, the sensitivity to ERI was enhanced significantly in both parental MDA-MB-231 and Hs578T cell lines. Furthermore, when both parental cell lines were treated with VOR (5 μM) or RICO (5 μM) for 48 hrs prior to addition of ERI, the sensitivity to ERI was significantly enhanced. The enhancement of sensitivity to ERI both by the simultaneous addition with low concentration of VOR or RICO and by the prior treatment with VOR or RICO was observed in the ERI-resistant cells as well. Moreover, inhibition of HDAC6 by small interfering RNA partially enhanced the sensitivity to ERI in the both parental cell lines. Our study demonstrated the possibility that the combination of ERI with HDAC inhibitor might enhance the antitumor effect of ERI and overcome ERI resistance through acetylation of α-tubulin by HDAC6 inhibition in TNBC cells.
Citation Format: Takaaki Oba, Hiroto Izumi, Ken-ichi Ito. Preclinical analysis of synergistic antitumor effect of eribulin and HDAC inhibitor for triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2924.
Abstract 2926: The glycogen synthase kinase-3 inhibitor, 9-ING-41, synergizes with chemotherapy to inhibit pancreatic tumor growth in vivo
Authors: Ding Li , Zhang Jin-san , Dube John R. , Madamsetty Vijay S. , Schmitt Daniel M. , Mukhopadhyay Debabrata , Billadeau Daniel D. 
Published: August 2018

Abstract
Glycogen synthase kinase-3 (GSK3) is a ubiquitously expressed serine-threonine protein kinase involved in multiple cellular functions ranging from the control of glycogen metabolism to transcriptional regulation. We have previously demonstrated that GSK-3β is overexpressed in human pancreatic ductal adenocarcinoma (PDAC) and aberrant nuclear accumulation of GSK-3β serves as a hallmark of poorly differentiated PDAC. Moreover, genetic ablation of GSK-3β or GSK-3 inhibition led to PDAC tumor cell killing, identifying GSK-3β as a therapeutic target in PDAC. Herein, we describe a novel GSK-3 inhibitor, 9-ING-41. The effect of 9-ING-41 on tumor cell growth was tested using several PDAC and patient-derived xenograft (PDX) cell lines in vitro and in vivo using an orthotopic pancreatic cancer animal model. Interestingly, 9-ING-41 treatment resulted in the stabilization of cyclin B levels as well as enhanced serine-10 phosphorylated histone H3 (pS10HH3) and consequently M phase arrest and blocked mitotic exit. The prolonged M phase arrest induced mitotic catastrophe in pancreatic cancer cell lines and inhibited in vitro colony formation. An in vivo orthotopic pancreatic cancer animal model study using PDX-derived cell lines demonstrated that 9-ING-41 hindered tumor growth in combination with several standard chemotherapeutic agents. Significantly, we found that 9-ING-41 treatment impaired the DNA damage response pathway activated by Gemcitabine in pancreatic cancer cell lines. In conclusion, our study indicates that GSK-3 plays an important role in pancreatic cancer progression and 9-ING-41 in combination with existing gemcitabine or irinotecan could be a new strategy for treating patients with PDAC.
Citation Format: Li Ding, Jin-san Zhang, John R. Dube, Vijay S. Madamsetty, Daniel M. Schmitt, Debabrata Mukhopadhyay, Daniel D. Billadeau. The glycogen synthase kinase-3 inhibitor, 9-ING-41, synergizes with chemotherapy to inhibit pancreatic tumor growth in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2926.
Abstract 293: Heterogeneous nucleocytoplasmic regulation of an incompletely penetrant ErbB2 onco-phenotype
Authors: Wang Lixin , Janes Kevin A. 
Published: August 2018

Abstract
Perturbation of cancer cells often leads to heterogeneous outcomes, in that most cells exhibit a dominant phenotype, but the rest appear resistant or hypersensitive to the perturbation. If the penetrance of such a phenotype is heritably incomplete, then it becomes extremely difficult to decipher the upstream molecular events that heterogenize the population and cause response variability. By combining quantitative measurements with dynamical models, systems approaches should be useful if provided with a core network of important biomolecules. The daunting hurdle lies in identifying phenotype-relevant regulatory heterogeneities that define the network for penetrance at the single-cell level. Here, we exploit a new approach, called stochastic frequency matching (SFM), which elaborates the molecular networks upstream of incompletely penetrant phenotypes. SFM identifies and parameterizes single-cell heterogeneities—which emerge after a uniform perturbation but before the appearance of a variable phenotype—to hone in on regulatory states corresponding to future penetrance. For a multi-acinar onco-phenotype incompletely triggered by ErbB receptor tyrosine kinase signaling in 3D cultured breast epithelia, we implemented SFM using microarrays to uncover a network of critical nucleocytoplasmic regulators, including the inner-ring nucleoporin NUP37 and the exportin CSE1L. Gain- and loss-of-function perturbations of NUP37 and CSE1L alter the frequency of ErbB2-triggered multiacinus formation, supporting that SFM can identify mechanisms of incompletely penetrant phenotypes. In addition, population-level transcriptomics revealed other ErbB2-induced changes in nucleocytoplasmic shuttling proteins: KPNA2, RANBP1, and XPO1. The net result of these alterations on shuttling is complicated, and the specific cargo relocalized by ErbB2 signaling is not known. We hypothesize that ErbB signaling heterogeneously reconfigures the nucleocytoplasmic shuttling state of cells to determine incomplete penetrance of the onco-phenotype. Using proximity labeling, we seek to identify novel ErbB2-induced NUP37 and CSE1L interactors that may serve as effectors for the multiacinar phenotype. Systems models of the nucleocytoplasmic shuttling network will evaluate the impact of observed abundance changes and identify characteristics of cargo predicted to be most sensitive to network reconfiguration. The importance of key effectors will ultimately be tested in vivo with murine cells engineered for spontaneous Erbb2 amplification and in patient samples with HER2 amplification.
Citation Format: Lixin Wang, Kevin A. Janes. Heterogeneous nucleocytoplasmic regulation of an incompletely penetrant ErbB2 onco-phenotype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 293.
Abstract 2933: Synergistic antitumor effect of ONC201 in combination with radiation therapy
Authors: Zhou Lanlan , Wagner Jessica , El-Deiry Wafik S. 
Published: August 2018

Abstract
Radiation therapy is used as the main treatment when surgery is not feasible, before/after surgery, and to palliate symptoms in advanced stage. About 50% of cancer patients receive radiation therapy. 40% of the cure is achieved by radiation therapy. First-in-class small-molecule imipridone ONC201 can cause dual inactivation of ERK and AKT and induce an integrated stress response, proapoptotic TRAIL receptor DR5 activation, cancer stem cell depletion, and cell cycle arrest. ONC201 could synergize with radiation therapy because of its proapoptotic and antimetastasis effects. We tested this hypothesis in colorectal cancer, non-small cell lung cancer and glioblastoma cell lines (HCT116, RKO, LS513, H460, A549 and SNB19). Firstly, CellTiter-Glo® luminescent cell viability assays were performed with single treatment or combination of radiation therapy (0 to 4 Gy) and ONC201 (0 to 20 μM). For combination treatment, cancer cells were pretreated with ONC201 for 24 hours followed by radiation therapy. Next, colony formation assays with single treatment or combination were used to evaluate the effective dose range from cell viability assays. Flow cytometry and Western blot analysis were used for determination of apoptosis. We observed synergism between ONC201 and radiation therapy in several colorectal cancer, non-small cell lung cancer and glioblastoma cell lines. Combination indices are 0.53, 0.59 and 0.68 when non-small cell lung cancer cell line H460 was treated with ONC201 at 5 μM and radiation therapy at 1, 2 and 4 Gy. We are currently pursuing this promising combinational therapy in vivo, using the H460 xenograft model and potential syngeneic lung tumor models with wild-type p53. We plan to pursue the mechanism of this combination from in vivo samples. Our results indicate that the combination of radiation therapy and ONC201 could be an important approach in the application of ONC201 in clinical practice.
Citation Format: Lanlan Zhou, Jessica Wagner, Wafik S. El-Deiry. Synergistic antitumor effect of ONC201 in combination with radiation therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2933.
Abstract 2942: Enhanced cytotoxic activity of BRAF, MEK and PI3K inhibition with combination CDK4/6 inhibition in melanoma cells
Authors: Zhang Yanping , Peng Guangyong , Hsueh Eddy C. 
Published: August 2018

Abstract
Introduction: We have previously reported that CDK4/6 inhibition enhanced the anti-proliferation and anti-migration effects of BRAF, MEK and PI3K inhibitors in melanoma cells. (Zhang Y et al, AACR 2017) In this study, we investigated the effect of combination CDK4/6 inhibition and BRAF, MEK and PI3K inhibition on induction of cell cycle arrest and apoptosis in melanoma.
Methods: Five human melanoma cell lines were used. CDK4/6 inhibitor (PD0332991; PD), BRAF inhibitor (PLX4032; PLX), MEK inhibitor (AZD6244; AZD), and PI3K/AKT inhibitor (XL765; XL) were used alone or in combination. Cell apoptosis analysis was performed using the FITC Annexin V Apoptosis Detection Kit. Autophagy was evaluated by immunofluorescence using anti-LC3A/B and NECN-1 antibodies. Lysosomal morphology was evaluated using lysosomal associated membrane protein 1-green fluorescent protein fusion construct. Western blotting was performed for apoptosis markers (PARP and caspase-9) and autophagy markers (LC3A/B and Beclin1) using GAPDH as internal control. Data were presented as means ± SD for triplicate experiments. For comparison between groups, the student's t test was used and p&lt; 0.05 was considered to be statically significant.
Results: Apoptosis was evaluated by Annexin V assay. Various levels of apoptosis were observed with single agent PD, PLX, XL and AZD in all 5 melanoma cell lines. Combination of PD enhanced the apoptosis effects of PLX, XL, and AZD in all 5 melanoma cell lines. Increased LC3 and BECN1 detection and increased density of autophagosomes on immunofluorescence was observed after combining CDK4 inhibition with BRAF, MEK, and PI3K/AKT inhibition. Inhibition of PARP and Beclin-1 expression and increase in caspase-9 expression were observed with combination of PD compared with single agent PLX, XL, and AZD.
Conclusion: CDK4 inhibition enhanced anti-melanoma activity of BRAF , PI3K/AKT and MEK inhibitions through increased induction of early autophagy and caspase-mediated apoptosis. This suggests that inhibition of CDK4/6 plus BRAF , MEK and PI3K/AKT inhibition may help understanding the interplay between autophagy and apoptosis and inform the development of future targeted agents.
Citation Format: Yanping Zhang, Guangyong Peng, Eddy C. Hsueh. Enhanced cytotoxic activity of BRAF, MEK and PI3K inhibition with combination CDK4/6 inhibition in melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2942.
Abstract 2949: Synergistic effects of tyrosine kinase inhibitors and conventional chemotherapeutics on canine and human osteosarcoma cells
Authors: Yang Ya-Ting , Yuzbasiyan-Gurkan Vilma 
Published: August 2018

Abstract
Osteosarcoma (OS) is the most common bone tumor in both humans and dogs, and has a nearly ten fold higher incidence in dogs than humans. Despite advances in treatment of other cancers, the overall survival rates for OS have stagnated for the past decades. Therefore, there is a great need to identify novel and effective treatments. In the past few years, several classes of novel small molecules with anti-tumor activities have become available. Among them, we selected tyrosine kinase inhibitors (TKIs) with potential anti-tumor medications to evaluate the effects of treating OS. In our previous studies, two TKIs (sunitinib and sorafenib) showed good growth inhibition effects at low concentrations on three canine OS cell lines (D17, Abrams, and Gracie) and two human OS cell lines (SAOS2 and U2OS) in a series of drug screening assay. In addition to viability, migration, and invasion ability of OS cells were decreased by these two TKIs. Furthermore, we investigated the effect of combing one of the TKIs with current chemotherapeutics for OS. We determined the combination index to assess whether sorafenib and sunitinib have synergistic effects with conventional chemotherapeutics including cisplatin, carboplatin, or doxorubicin. Data from our cell lines point to synergistic effects of sorafenib combined with doxorubicin, but not when combined with cisplatin or carboplatin in both human and canine OS. Combination studies with sunitinib are ongoing. Based on current findings, clinical trials using combination of doxorubicin and sorafenib in proof of concept studies in dogs are warranted. These studies can be carried out relatively quickly in dogs where case load is high, and in turn, provide useful data for initiation of clinical trials in humans.
Citation Format: Ya-Ting Yang, Vilma Yuzbasiyan-Gurkan. Synergistic effects of tyrosine kinase inhibitors and conventional chemotherapeutics on canine and human osteosarcoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2949.
Abstract 295: Efficacy of MCL1 inhibitor S63845 in small cell lung cancer
Authors: Yasuda Yuto , Ozasa Hiroaki , Tsuji Takahiro , Nomizo Takashi , Funazo Tomoko , Yoshida Hironori , Sakamori Yuichi , Hirai Toyohiro , Kim Young Hak 
Published: August 2018

Abstract
Lung cancer is the leading cause of cancer death. Small cell lung cancer (SCLC) is a histologic subtype of lung cancer and the proportion of SCLC is approximately 15%. New molecular targeted drugs and immune checkpoint inhibitors are successfully effective for non-small cell lung cancer. However, treatment of SCLC has not been improved over recent decades. It is important to explore new treatment strategies of SCLC. MCL1 is a member of the BCL-2 family, which regulates apoptosis. Targeting MCL1 represents a potential breakthrough of cancer treatment. We tested S63845, a MCL1 inhibitor, in four SCLC cell lines (DMS114, DMS53, SW1271, and NCI-H69) and, in addition, one patient derived SCLC cell (KTOR201). S63845 had greater efficacy in two of five SCLC cells (DMS114 and KTOR201). These two SCLC cells had higher expression of MCL1 and lower expression of BCL-XL, which is another member of the BCL-2 family. The other three SCLC cells (DMS53, SW1271 and NCI-H69) were resistant to S63845 and had a higher expression of BCL-XL or lower expression of MCL1. These data suggested that S63845 might be a powerful treatment of SCLC as a new therapeutic strategy. It is possible that the expressions of MCL1 and other members of the BCL-2 family predict the sensitivity of S63845.
Citation Format: Yuto Yasuda, Hiroaki Ozasa, Takahiro Tsuji, Takashi Nomizo, Tomoko Funazo, Hironori Yoshida, Yuichi Sakamori, Toyohiro Hirai, Young Hak Kim. Efficacy of MCL1 inhibitor S63845 in small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 295.
Abstract 2950: Development of a 4-aminopyrazolo[3,4-d]pyrimidine-based dual IGF1R/Src inhibitor as a novel anticancer agent with minimal toxicity
Authors: Lee Ho-Jin , Min Hye-Young , Pham Phuong Chi , Baek Byungyeob , Kim Byungjin , Kim Yunha , Lee Jeeyeon , Lee Ho-Young 
Published: August 2018

Abstract
Both the type I insulin-like growth factor receptor (IGF1R) and Src pathways are associated with the development and progression of numerous types of human cancer, and Src activation confers resistance to anti-IGF1R therapies. Hence, targeting both IGF1R and Src concurrently is one of the main challenges in combating resistance to the currently available anti-IGF1R-based anticancer therapies. However, the enhanced toxicity from this combinatorial treatment could be one of the main hurdle for this strategy, suggesting the necessity of developing a novel strategy for co-targeting IGF1R and Src to meet an urgent clinical need. In the present study, We synthesized a series of 4-aminopyrazolo[3,4-d]pyrimidine-based dual IGF1R/Src inhibitors, selected LL28 as an active compound, and evaluated its potential antitumor effects in vitro and in vivo. LL28 markedly suppressed the activation of IGF1R and Src and significantly inhibited the viability of several NSCLC cell lines in vitro by inducing apoptosis. Administration of mice with LL28 significantly suppressed the growth of H1299 NSCLC xenograft tumors without overt toxicity and substantially reduced the multiplicity, volume, and load of lung tumors in the KrasG12D/+-driven lung tumorigenesis model. These results suggest the potential of LL28 as a novel anticancer drug candidate targeting both IGF1R and Src, providing a new avenue to efficient anticancer therapies. Further investigation is warranted in advanced preclinical and clinical settings.
Citation Format: Ho-Jin Lee, Hye-Young Min, Phuong Chi Pham, Byungyeob Baek, Byungjin Kim, Yunha Kim, Jeeyeon Lee, Ho-Young Lee. Development of a 4-aminopyrazolo[3,4-d]pyrimidine-based dual IGF1R/Src inhibitor as a novel anticancer agent with minimal toxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2950.
Abstract 296: Loss of Romo1 enhances TRAIL-induced apoptosis through Bax upregulation in colon cancer
Authors: Jo Minjee , Kim Jung Lim , Jeong So Yeon , Kim Bo Ram , Na Yoo Jin , Park Seong Hye , Jeong Yoon A , Lee Dae-Hee , Oh Sang Cheul 
Published: August 2018

Abstract
Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a protein which is induces cell apoptosis in various cancer. To identify novel sensitizer of TRAIL, we knocked down Reactive Oxygen Species modulator 1 (Romo1). Romo1 is a mitochondrial membrane protein that is in charge of increasing ROS. Romo1 was highly upregulated in various cancers including colorectal cancer. Inhibition of Romo1 significantly increased TRAIL-induced apoptosis in colorectal cancer cells, not colon normal cells. Silencing of Romo1 has resulted in upregulation of Bax protein expression. It has been reported that Bax degraded by ubiquitination. We identify interaction between Bax and Parkin decreased in knockdown of Romo1 condition. Also, suppression of Romo1 lead to mitochondrial dysfunction. Our study suggests that inhibition of Romo1 enhanced TRAIL-mediated apoptosis by regulating Bax ubiquitination.
Citation Format: Minjee Jo, Jung Lim Kim, So Yeon Jeong, Bo Ram Kim, Yoo Jin Na, Seong Hye Park, Yoon A Jeong, Dae-Hee Lee, Sang Cheul Oh. Loss of Romo1 enhances TRAIL-induced apoptosis through Bax upregulation in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 296.