bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3504: Arginylated BiP/oxidized DJ-1 complex mediates crosstalk between the endoplasmic reticulum and mitochondria
Authors: Lee. Dae-Hee 
Published: July 2016

Abstract
In cell death induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), although much is known about signal pathways involving external signals through cell surface receptors or internal signals through mitochondrial dysfunction, little is known about signal pathways activated by endoplasmic reticulum (ER) stress during TRAIL treatment. Here, we report that TRAIL-induced caspase activation is a prerequisite for TRAIL-induced ER stress response. During TRAIL treatment, ER chaperon BiP dissociates from ER stress sensors such as PERK and IRE1α and the activation of the PERK-eIF2α-ATF4-CHOP apoptotic signal transduction pathway occurs. Meanwhile ER-stress induces BiP release to the cytosol and ATE1-encoded Arg-tRNA transferase arginylates BiP. TRAIL also induces loss of ΔΨm and redox sensor DJ-1 detects mitochondria-derived ROS. Oxidized DJ-1 binds to arginylated BiP (R-BiP) in the cytosol. DJ-1 binding activity was abrogated in C106A and C53A but not C46A mutant. R-BiP/oxidized DJ-1 complex may mediate crosstalk between the ER and mitochondria.
Key words: TRAIL; endoplasmic reticulum; apoptosis; colorectal cancer
Citation Format: Dae-Hee Lee. Arginylated BiP/oxidized DJ-1 complex mediates crosstalk between the endoplasmic reticulum and mitochondria. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3504.
Abstract 3510: Tolfenamic acid induces apoptosis and growth inhibition in anaplastic thyroid cancer: involvement of nonsteroidal anti-inflammatory drug-activated gene-1 expression and intracellular reactive oxygen species generation
Authors: Kim Yeon Soo , Chang Jae Won , Kang SungUn , Choi JaeWon , Shin YooSeob , Baek Seung Joon , Lee Seong-Ho , Oh Kyoung Ho , Kim Chul-Ho 
Published: July 2016

Abstract
Nonsteroidal anti-inflammatory drugs (NSAIDs) are usually used for the treatment of inflammatory diseases. However, certain NSAIDs also have antitumor activities in various cancers, including head and neck cancer, through cyclooxygenase-dependent or independent pathways. Nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1), a TGF-β superfamily protein, is induced by NSAIDs and has been shown to be induced by several antitumorigenic compounds and to exhibit proapoptotic and antitumorigenic activities. In this report, we demonstrate for the first time that tolfenamic acid (TA) transcriptionally induced the expression of NAG-1 during TA-induced apoptosis of anaplastic thyroid cancer (ATC) cells. TA reduced the viability of ATC cells in a dose-dependent manner and induced apoptosis, findings that were coincident with NAG-1 expression. Overexpression of the NAG-1 gene using cDNA enhanced the apoptotic effect of TA, whereas suppression of NAG-1 expression by small interfering RNA attenuated TA-induced apoptosis. Subsequently, we found that intracellular ROS generation plays an important role in activating the proapoptotic protein NAG-1. Then, we confirmed antitumorigenic effects of TA in a nude mouse orthotopic ATC model, and this result accompanied the augmentation of NAG-1 expression and ROS generation in tumor tissue. Taken together, these results demonstrate that TA induces apoptosis via NAG-1 expression and ROS generation in in vitro and in vivo ATC models, providing a novel mechanistic explanation and indicating a potential chemotherapeutic approach for treatment of ATC.
Citation Format: Yeon Soo Kim, Jae Won Chang, SungUn Kang, JaeWon Choi, YooSeob Shin, Seung Joon Baek, Seong-Ho Lee, Kyoung Ho Oh, Chul-Ho Kim. Tolfenamic acid induces apoptosis and growth inhibition in anaplastic thyroid cancer: involvement of nonsteroidal anti-inflammatory drug-activated gene-1 expression and intracellular reactive oxygen species generation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3510.
Abstract 3514: Inhibition of PI-3 kinase signaling potentiates the apoptotic response to mitotic arrest
Authors: Kassim Roaa M. , Shuster Charles B. 
Published: July 2016

Abstract
Anti-mitotic drugs represent a common strategy for cancer chemotherapy by inducing a prolonged prometaphase delay and eventual cell death by apoptosis. Kinesin Spindle Protein (KSP) inhibitors arrest cells in mitosis by blocking bipolar spindle assembly, and these drugs have demonstrated limited side effects in clinical trials. However, they are not as effective as Taxol, whose efficacy is limited by the frequent development of resistance as well as the deleterious effects on non-dividing cells. Recent studies suggest that the ability of cells to maintain energy levels during mitosis could account for the differential responses of cancer cells to mitotic arrest. To determine if manipulating cellular metabolism could potentiate the efficacy of KSP inhibitors, we arrested cells in mitosis in the absence or presence of the PI3-kinase LY294002, and probed for the induction of apoptosis. Because tumor cells respond differently to mitotic arrest, we employed two cell lines that either survive mitotic arrest (MCF7) or undergo apoptosis shortly following mitotic slippage (HeLa). In both cases, simultaneous inhibition of KSP and PI3-kinase induced apoptosis more effectively than mitotic arrest or PI3-K inhibition alone. In the case of HeLa cells, there also appeared to be an acceleration of cell death such that arrested cells underwent apoptosis prior to mitotic slippage. Similar effects were observed using Akt inhibitors, providing additional evidence that PI3-kinase signaling was critical for surviving mitotic arrest. Lastly, we found that increased glucose had a protective effect, further supporting the notion that the ability to survive mitotic arrest depended on the ability to maintain energy levels. If confirmed, this may represent a novel approach for potentiating the anti-tumor activity of KSP inhibitors in the clinic.
Citation Format: Roaa M. Kassim, Charles B. Shuster. Inhibition of PI-3 kinase signaling potentiates the apoptotic response to mitotic arrest. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3514.
Abstract 3517: Expression studies of RBBP6 in breast and cervical cancer suggests a role in apoptosis activation
Authors: Choene Mpho , Moela Pontsho , Motadi Lesetja 
Published: July 2016

Abstract
Ubiquitin-like DWNN domain and RING finger-like domain presence on Retinoblastoma Binding protein 6 (RBBP6), is linked to its function as cancer promoting gene through p53 degradation via its p53-binding domain. RBBP6 bind and regulate the expression of p53 and pRB in the cell machinery implicates RBBP6 in cell proliferation during cancer development (Moela et al 2014). Studies indicate that overexpression of the mouse spliced variants led to apoptosis and cell cycle arrest whilst deregulation in mice led to embryonic lethality followed by p53 accumulation and a widespread apoptosis (Li et al., 2007). In this study, focuses on determining the expression pattern of RBBP6 in human tissue models of breast and cervical cancer. The study further aims to analyze apoptosis and cell cycle following either knockdown or overexpression of RBBP6 coupled to treatment with potential anticancer compounds, camptothecin and GABA. Quantitative analysis by qPCR in HeLa, SiHa, MCF-7and MDA-MB-231 cell lines shows that RBBP6 expression is fairly high in cancer cells when compared to normal lung fibroblast. Following co-treatment with GABA and cDNA or siRNA, in cDNA treated there was a reduction in both apoptosis as measured by flow cytometer and TP53 expression while in siRNA there was increased expression of TP53 and those of Bax and Bak1 which seemed to account for increased apoptosis. Cell cycle presented increase in G0/G1 and the G2/M following treatment with siRNA and GABA. GABA alone could not induce a significant increase in apoptotic cells whereas when combined with siRNA it was able to induce over 30% apoptosis. These results suggest that RBBP6 might be a cancer promoting gene.
Citation Format: Mpho Choene, Pontsho Moela, Lesetja Motadi. Expression studies of RBBP6 in breast and cervical cancer suggests a role in apoptosis activation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3517.
Abstract 3518: Flow cytometry as a single platform tool to evaluate multiple mechanisms of actions of therapeutic antibodies
Authors: Chadderton Antony R. , Harvey Shane , Strake Brandy , Giles-Komar Jill , Capocasale Renold J. , Raju T. Shantha 
Published: July 2016

Abstract
The unique ability of flow cytometry to simultaneously examine intricate details of multiple cell subsets is unparalleled and the platform is now a dominant tool for measuring antibody mediated effector functions on cancer cells. The objective of this study is to illustrate the utility of flow cytometry as a single platform for concurrently measuring the multiple mechanisms of actions of therapeutic antibodies such as ADCC, ADCP, Apoptosis, CDC and Trogocytosis on human cancer cells and immune functioning cells. Rituximab® was tested with the Burkett's Lymphoma cell line (Daudi) for all assays between the ranges of 0.01 and 100ug/mL. Apoptosis was evaluated by measuring Caspase 3/7 staining. ADCC assays were established with Rituximab®/CFSE labeled Daudi cells and human PBMC’s. ADCP was assessed with Rituximab®/ CFSE labeled Daudi cells and CD14+/CD11b+ in vitro generated phagocytes using a 4 hour exposure. CDC assays were established with Daudi cells/ 10% pooled human serum using 7-AAD as the cytotoxic indicator in a 4 hour assay. Trogocytosis was measured up to 4 hours with Rituximab®/ Daudi and PBMC's and CD19/20 and CD14. All assays were analyzed using a FACS ARIA III flow cytometer and replicates of 3 were evaluated for statistical relevance. Results demonstrated that Rituximab® induced effects were observed in all 5 assays using flow cytometry. Apoptosis was induced when 0.01ug/mL Rituximab® was present. ADCP occurred at levels as low as 0.01ug/mL Rituximab®. ADCC and CDC occurred at levels as low as 0.1ug/mL Rituximab®. Trogocytosis, indicated by the transfer of CD19+ B cells to CD14+ monocytes, occurred at 0.1ug/ml Rituximab®. In summary, we demonstrated the clinical applicability of flow cytometry as a single platform to simultaneously evaluate five different mechanisms of actions that can occur when therapeutic antibodies are used to treat cancer cells.
Citation Format: Antony R. Chadderton, Shane Harvey, Brandy Strake, Jill Giles-Komar, Renold J. Capocasale, T. Shantha Raju. Flow cytometry as a single platform tool to evaluate multiple mechanisms of actions of therapeutic antibodies. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3518.
Abstract 3531: Autophagy facilitates leukemogenesis in a murine model of MLL-AF9-driven AML
Authors: Liu Qiang , Chen Longgui , Wang Hong-Gang 
Published: July 2016

Abstract
Acute myeloid leukemia (AML) is a hematopoietic malignancy characterized by the clonal expansion of immature myeloid cells. There is an urgent and unmet need for the development of novel therapeutics against AML. Autophagy is a lysosomal catabolic process that has been implicated in leukemia cell survival and chemotherapy response. However, the biological role of autophagy in AML leukemogenesis remains poorly defined. In this study, we genetically ablated ATG5, a gene that is essential for autophagy, in AML cells during leukemogenesis in a murine model of MLL-AF9-driven AML. The homozygous deletion of ATG5 in AML cells significantly delayed MLL-AF9-induced leukemogenesis and progression in vivo. Likewise, the in vitro deletion of ATG5 in MLL-AF9 driven AML cells demonstrated decreased growth as well as reduced colony forming units. Interestingly, ATG5 deficiency demonstrated an altered differentiation hierarchy of MLL-AF9-induced AML cells in vivo that could be linked to decreased proliferation and suppressed leukemogenesis. Overall, this study provides insight at how autophagy can facilitate AML progression. A more comprehensive understanding on autophagy in AML is needed prior to the deployment of autophagy modulators for the treatment of AML.
Citation Format: Qiang Liu, Longgui Chen, Hong-Gang Wang. Autophagy facilitates leukemogenesis in a murine model of MLL-AF9-driven AML. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3531.
Abstract 3532: Compound Tc inhibits ATG4 to attenuate autophagic flux and sensitize cancer cells to chemotherapy
Authors: Shu. Chih-Wen 
Published: July 2016

Abstract
Autophagy is induced to protect cancer cells against stresses, including starvation, hypoxia and chemotherapy. Antimalarial drugs chloroquine and its derivative hydroxychloroquine are used to inhibit autophagy and being investigated with clinical trials for certain types of cancer. However, the tumor suppressive effects of the drugs might be autophagy independent, which require more specific inhibitors target autophagy components for clinical uses. Here, we screened FDA-approved drug library with both biochemical and cellular ATG4 reporter assays and found compound Tc blocked proteolysis activity of ATG4, essential proteases in autophagy machinery. Docking and MD simulation results appeared compound Tc was directly associated with active site of ATG4. Further, compound Tc obstructed the binding between ATG4 and MAP1LC3 and accumulated autophagosomes, which in turn reduce autophagic flux in cells. Moreover, compound Tc suppressed tumor viability and sensitized chemotherapy efficacy in two-dimensional and sphere culture model. Silencing ATG4 or overexpression of ATG4 catalytic mutant had no additional effects on tumor suppression compared to compound Tc alone, indicating compound Tc inhibited ATG4s activity to diminish tumor viability. In addition, compound Tc enhanced genotoxic stress-induced apoptosis to synergize chemotherapy efficacy in vitro and in vivo. Thus, we report that the clinical drug compound Tc is a potent ATG4 inhibitor, which could interrogate the role of autophagy in clinical setting and provide compound Tc as an anticancer drug or chemosensitizer.
Citation Format: Chih-Wen Shu. Compound Tc inhibits ATG4 to attenuate autophagic flux and sensitize cancer cells to chemotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3532.
Abstract 3534: Inhibition of melanoma cell survival through p62/LC3B autophagic signaling
Authors: Hambright Heather G. , Kumar Addanki P. , Ghosh Rita 
Published: July 2016

Abstract
The primary objectives of this study were i) systematic evaluation of the differences in basal autophagic activity between melanocytes and malignant melanoma cells to determine whether these differences may be selectively targeted using the ROS inducer, Nexrutine (NX); ii) determination of NX selectivity in inhibiting melanoma cell survival and iii) specific involvement of autophagy signaling protein, SQSTM1/p62 in NX-mediated autophagic cell survival. NX is obtained from the bark of the cork tree, Phellodendron amurense. A panel of melanocyte and melanoma cells was evaluated for basal autophagy level and effect of NX treatment, including protein levels and turnover of p62 and LC3B (+/- CQ), and number of autophagic puncta per cell. Trypan blue, MTT, and Annexin V-APC were used to evaluate the effect of NX on cell death, proliferation and apoptosis. Genetic approach using siRNA for p62 and overexpression using HA-p62 followed by survival, proliferation, and autophagy measurements were used to further evaluate the possibility of p62 as a molecular target for inhibition of autophagy seen using NX. We found that i) melanoma cells displayed high autophagic flux compared with melanocytes, ii) NX treatment a) inhibited autophagy in melanoma cells, b) inhibited melanoma cell survival, and c) induced apoptosis in melanoma cells, iii) knockdown of p62 resulted in a lesser inhibition of cell viability and autophagy after NX treatment in high p62-expressing melanoma cells, and iv) overexpression of p62 sensitized low p62-expressing normal melanocytes and melanoma cells to NX-mediated cell death and autophagy inhibition, suggesting that p62 is a bonafide molecular target of NX. Overall, we show that Nexrutine is able to inhibit the inherent high level of pro-survival autophagy in melanoma cells, leading to loss of cell viability and induction of apoptosis, while melanocytes remain unaffected by NX at the same doses. Further, SQSTM1/p62 was found to be a critical mediator of the NX-mediated inhibition of autophagy and cell viability. This study demonstrates that NX maybe a promising novel, non-toxic agent for melanoma.
Supported by R21 CA125719 & ACRCF (RG); NIDCR T32 DE14318/COSTAR (HGH)
Citation Format: Heather G. Hambright, Addanki P. Kumar, Rita Ghosh. Inhibition of melanoma cell survival through p62/LC3B autophagic signaling. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3534.
Abstract 354: Physiologically relevant target engagement using CETSA
Authors: Karén Jakob , Sioberg Catrine , Niekrasz Pawel , Neumuller Magnus , Martinez Molina Daniel 
Published: July 2016

Abstract
Targeted therapies are dependent on adequate occupancy and target engagement to reach their efficacy measure. Yet, more than half of novel candidate drugs entering clinical trial stage fail before proof of concept is achieved. Inadequate target engagement is often at the root cause of these failures. Therefore we have developed a versatile label-free method to quantify the target engagement of drugs on their native physiological relevant targets. The method is validated in multiple sample matrices from bench to bedside: in living cells and tissue from rodents and man.
CETSA is a patented biophysical method measuring the thermal stability of drug target proteins and the shift induced by their ligands. By quantifying the melting temperature and shift induced by the ligand we can quantify the potency of target engagement. This potency determination then allows filtering of candidate drugs by their ability to engage the target in its physiologically relevant form, in relevant sample formats such as intact cells or tissue.
We present a number of applications and readouts of the CETSA method quantifying target engagement of drugs used in the clinic and in development. Furthermore, we show that the target engagement of cyclin dependent kinase inhibitors correlated both to viability and to a dose dependent decrease of a cell cycle activity marker in cellular sample matrices. The biomarker, thymidine kinase activity, was analyzed with the commercially available test DiviTum®. By combining CETSA with DiviTum® in the same samples, it was thus possible to correlate CETSA outcomes to pharmacodynamic effects of a drug.
The utility and value of the CETSA method enabling preclinical development of novel drugs is substantiated by the examples included here. Combining quantifications of efficacy with the molecular target engagement of the drug yields an orthogonal dataset, which has a potential to validate the target-mediated efficacy. Such datasets can be generated from early in vitro systems to clinical studies and may reduce the high failure rate in drug development by facilitating the discovery, development and evaluation of novel drugs.
Citation Format: Jakob Karén, Catrine Sioberg, Pawel Niekrasz, Magnus Neumuller, Daniel Martinez Molina. Physiologically relevant target engagement using CETSA. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 354.
Abstract 3540: Exploring novel treatment agent in GIST by using connectivity map
Authors: Yen Chueh-Chuan , Chen Li-Tzong , Li Chien-Feng 
Published: July 2016

Abstract
Gastrointestinal stromal tumors (GIST) is gastrointestinal tract sarcoma which commonly contains a mutation in the KIT or PDGFRA. Imatinib mesylate, a specific tyrosine kinase inhibitor of the KIT and PDGFRA, causes regression or stabilization of disease in most of the patients with metastatic GIST. However, acquired resistance to imatinib is inevitable. Recently, E26 variant 1 (ETV1) pathway was found to be a key downstream effector of KIT. In this study, we explored the role of ETVI and its downstream effectors pathway in GIST by bioinformatics analysis, and identified potential novel agents that target ETV1 pathway of GISTs by using expression profile and a pattern-matching software “Connectivity Map” (CMAP), and confirmed the efficacy of novel agents by using GIST cell line in vitro assay. By using CMAP, four drugs with potential ETV1-inhibition effect were identified: suberanilohydroxamic acid (SAHA,Vorinostat) and trichostatin, two histone deacetylase inhibitors (HDACI), and trifluoperazine (TFP) and thioridazine (TDZ), two drugs of phenothiazine class. All four drugs were found to have ETV1-down regulating effect by immunoblotting. Phenothiazine could induce apoptosis and autophagy in GIST. Treatment of phenothiazine was found paradoxically up-regulate MEK activity. Combination of MEK inhibitor and phenothiazine showed great synergistic effect in exerting cytotoxicity against GIST cell lines. Our study established phenothiazine as a new class of agent with therapeutic effect in GIST, and combination of phenothiazine and MEK inhibitor showed great potential in GIST treatment.
Citation Format: Chueh-Chuan Yen, Li-Tzong Chen, Chien-Feng Li. Exploring novel treatment agent in GIST by using connectivity map. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3540.