bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract C142: Mortalin/HSPA9 regulates p21CIP1 expression in Raf/MEK/ERK-activated cancer cells.
Authors: Wu Pui Kei , Hong Seung-Keun , Veeranki Sudhakar , Karkhanis Mansi , Starenki Dmytro , Plaza Jose A. , Park Jong-In 
Published: August 2014

Abstract
Dysregulated Raf/MEK/ERK signaling is a common hallmark of tumorigenesis. However intriguingly, aberrant Raf/MEK/ERK activation triggers senescence-like growth arrest as a primary response in many cell types. This response is recognized as an innate tumor suppressive mechanism, which should be bypassed for Raf/MEK/ERK to drive carcinogenesis. Therefore, understanding of the bypass mechanisms is likely to provide an opportunity to design a novel therapeutic strategy. In this study, we report that mortalin, a member of the heat shock protein 70 (HSP70) family, has a role in bypassing the Raf/MEK/ERK-induced growth inhibition in cancer. Using tandem affinity purification and proteomic mass spectrometry, we found that mortalin is present in the MEK1/2 proteome. A specific physical interaction between mortalin and MEK was validated by co-immunoprecipitation analyses and in vitro binding assays of recombinant proteins. Immunohistochemical analysis revealed that mortalin is upregulated in human melanoma biopsies in correlation with tumor malignancy. Of note, mortalin levels were inversely correlated with p21CIP1 levels in a number of Raf/MEK/ERK-activated cancer cell lines, suggesting a role for mortalin in p21CIP1 regulation. Indeed, mortalin depletion elicited increased p21CIP1 transcription and MEK/ERK activation which were accompanied by G2/M phase cell cycle arrest and cell death in different B-Raf or K-Ras mutated cancer cell lines. Remarkably, blocking MEK/ERK activation, using the MEK1/2 inhibitor, AZD6244, dominant negative mutants of MEK or ERK, or RNA interference, abrogated p21CIP1 induction by mortalin depletion in cancer cells harboring B-RafV600E regardless of their p53 status. In contrast, mortalin overexpression suppressed B-RafV600E or ΔRaf-1:ER-induced MEK/ERK activation, p21CIP1 expression, and cell cycle arrest in cell types exhibiting normal MEK/ERK status. Other HSP70 family chaperones could not effectively replace mortalin for the observed effects on p21CIP1 regulation, suggesting a unique role for mortalin. These findings suggest that mortalin upregulation is a mechanism that underlies p21CIP1 silencing in Raf/MEK/ERK-activated cancer, identifying mortalin as a novel negative regulator of the pathway signaling. Mortalin may provide a target to reactivate the tumor suppressive signaling of Raf/MEK/ERK in cancer.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C142.
Citation Format: Pui Kei Wu, Seung-Keun Hong, Sudhakar Veeranki, Mansi Karkhanis, Dmytro Starenki, Jose A. Plaza, Jong-In Park. Mortalin/HSPA9 regulates p21CIP1 expression in Raf/MEK/ERK-activated cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C142.
Abstract C143: Overcoming melanoma dual resistance to Vemurafenib and adoptive immunotherapy by chromatin remodeling drug.
Authors: Jazirehi Ali R. , Nazarian Ramin , Toress-Collado Antonio Javier , Economou James S. 
Published: August 2014

Abstract
BRAFV600E-inhibitors (BRAFi) and modern immune-based therapies have significantly improved the care of melanoma patients. We developed an in vitro model to determine whether tumor resistance mechanisms to this small molecule inhibitor of a driver oncogene and to cytotoxic T lymphocyte (CTL)-delivered apoptotic death signals were exclusive or intersecting. We generated melanoma sublines resistant to BRAFi Vemurafenib and to CTL recognizing the MART-1 melanoma antigen. Vemurafenib-resistant (VemR) sublines were cross-resistant to MART CTL indicating a common apoptotic pathway governing response to both modalities was disrupted. Pretreatment of VemR melanomas with a histone deacetylase inhibitor (HDACi) restored sensitivity to MART CTL apoptosis. These findings suggest that aberrant apoptotic pathways, amenable by a chromatin remodeling drug, regulate tumor resistance mechanisms and identify potential obstacles and solutions for combining these therapies clinically. TModulation of apoptotic machinery by inclusion of HDACi in BRAFV600E targeted therapy protocols as adjuvant will sensitize VemR (also CTL cross-resistant) human melanomas to immunotherapy.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C143.
Citation Format: Ali R. Jazirehi, Ramin Nazarian, Antonio Javier Toress-Collado, James S. Economou. Overcoming melanoma dual resistance to Vemurafenib and adoptive immunotherapy by chromatin remodeling drug. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C143.
Abstract C144: MAPK1 mutation E322K modulates ERK pathway output and feedback.
Authors: Guan Nin , Delpuech Oona , Greenawalt Danielle , Jacques Kelly , Gharahdaghi Farzin , Zinda Michael , Smith Paul J. , Guichard Sylvie M. 
Published: August 2014

Abstract
Mutations of MAPK1 E322K are observed in ∼8% of cervical ∼1% and head and neck cancer but there is currently little evidence of their impact on the ERK pathway. This study evaluated the impact of the E322K mutation on the ERK pathway output and feedback using an inducible system pTRIPZ in MCF10A immortalised cells. Expression of E322K MAPK1 following doxycycline induction reduced RSK phosphorylation on T359/S363 but increased pELK1 S383. DUSP6 protein levels were also increased in MCF10A expressing E322K mutant compared to WT. Pathway output was determined using the mRNA MEK signature (Dry et al. 2010). Following Dox induction, MCF10A E322K showed a 4-fold increase in DUSP2 mRNA and a 2-fold increase in DUSP6 mRNA while levels were unchanged in MCF10A expressing WT MAPK1. Conversely, &gt;2-fold decrease in ETV5 levels was observed in E322K MCF10A compared to WT cells. Exposure of WT and E322K MCF10A cells to an ATP competitive ERK inhibitor showed a greater reduction in pRSK T359/S363 levels in E322K cells compared to WT cells (likely due to lower baseline levels) but abrogation of pELK1 required higher concentrations of the inhibitor in mutant compared to WT cells. Consistent with its mechanism of action, the ERK inhibitor increased pERK levels. Interestingly, the combination of the MEK inhibitor selumetinib with the ERK inhibitor completely abrogated pERK in WT but not in E322K mutant MCF10 cells. Overall, the results suggest that the E322K mutation increases ERK-regulated genes such as DUSP2 which may only partially de-phosphorylate ERK due to the E322K mutation affecting substrates’ binding to the D site. The impact of E322K on the sensitivity to ERK and MEK inhibition is currently under evaluation.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C144.
Citation Format: Nin Guan, Oona Delpuech, Danielle Greenawalt, Kelly Jacques, Farzin Gharahdaghi, Michael Zinda, Paul J. Smith, Sylvie M. Guichard. MAPK1 mutation E322K modulates ERK pathway output and feedback. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C144.
Abstract C147: Mechanism of resistance to a semisynthetic anticancer andrographolide derivative: Possible involvement of Ras-MAPK signaling pathway.
Authors: Stanslas Johnson , Wong Charng Choon , Sagineedu Sreenivasa Rao , Sidik Shiran , Sumon Shariful Hasan , Phillips Roger , Lajis Nordin Haji 
Published: August 2014

Abstract
SRJ09 (3,19-(2-bromobenzylidene)andrographolide) has been identified as a lead anticancer agent on the basis of its selectivity towards breast and colon cancers in the USA NCI 60 cancer cell lines screen (Jada et al (2008) Br J Pharmacol, 155(5):641-54). In our recent seminal publication (Hocker et al (2013), PNAS, 110(25):10201-6), we revealed that the compound is a binder of oncogenic mutant K-Ras. In the present investigation, HCT116 colon cancer cells were made resistant to SRJ09 to elucidate the mechanism of resistance. MTT cell viability assay was utilized to screen for cross-resistance to other analogues of andrographolide (AGP) and standard anticancer drugs. Flow cytometry technique was utilized for cell cycle study. Western blot was performed to determine the expression of cell cycle, apoptosis regulatory and cell signaling proteins. Additionally, reverse transcriptase (RT)-PCR was carried out to determine the cell cycle regulatory genes. SRJ09-resistant HCT116 displayed approximately 40-fold resistance towards SRJ09 compared with the parental cells. However, the cells were not resistant towards AGP analogues and standard anticancer drugs. The cells failed to arrest in G1 phase upon treatment with SRJ09, unlike the parental cells. RT-PCR confirmed the findings that protein expression of p21 was not up-regulated and CDK4 was not down-regulated, contrary to the parental cells. The resistance of the cells to undergo apoptosis upon treatment with SRJ09 was mainly attributed to the unchanged levels of Bax, Bid, caspase 8 and 9, otherwise activated in the parental cells. The inability of the cells to experience cell cycle arrest and apoptosis might be attributed in part to the marginal changes in phosphorylated c-Raf and ERK1/2 protein levels, which are in complete contrast to the parental cells. In conclusion, the mechanism of resistance is very likely due to failure of c-Raf and ERK1/2 to undergo dephosphorylation, implicating Ras-MAPK pathway as a target of SRJ09. This study was funded by Universiti Putra Malaysia Research University Grant Scheme (04-01-09-0713RU; 04-02-12-2017RU).
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C147.
Citation Format: Johnson Stanslas, Charng Choon Wong, Sreenivasa Rao Sagineedu, Shiran Sidik, Shariful Hasan Sumon, Roger Phillips, Nordin Haji Lajis. Mechanism of resistance to a semisynthetic anticancer andrographolide derivative: Possible involvement of Ras-MAPK signaling pathway. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C147.
Abstract C154: Kinetic analysis of the glutaminase activity of L-asparaginases derived from Erwinia chrysanthemi (Erwinase®) and Eshericia coli (Kidrolase).
Authors: Phillips Roger M. , Saleem Mohammed U. , Williams Adrian , Morgan Jon 
Published: August 2014

Abstract
Glutamine is an essential nutrient for cancer cells and increased glutaminolysis is widely recognised as a key feature of the metabolic reprogramming that occurs during malignant transformation. As a consequence of the role glutamine plays in cancer biology, targeting glutamine metabolism is of considerable interest with inhibition of the mitochondrial enzyme glutaminase showing promise in experimental models. An alternative or adjunct to the strategy of inhibiting glutamine metabolism intracellularly is to deplete extracellular pools of glutamine. L-asparaginases are known to possess glutaminase activity and the purpose of this study is to characterise the kinetics of the glutaminase activity of two clinically approved L-asparaginases, Kidrolase (derived from E. coli) and Erwinase (derived from Erwinia chrysanthemi). Glutaminase activity was measured using a two step reaction: initial reduction of glutamine to glutamate by Kidrolase/Erwinase followed by the NAD dependent conversion of glutamate to ammonia by L-glutamate dehydrogenase (GDH). Each reaction consisted of NAD (2mM), K2PO4 (50mM), GDH (3U), Erwinase or Kidrolase (10U), L-glutamine (concentrations ranged from 5mM to 1.25 μM) in a final volume of 1 ml Tris Acetate buffer (50mM, pH8.6). The reaction was started by the addition of Erwinase/Kidrolase and the formation of NADH monitored for 30 seconds at 340nm using a Cary UV/Vis spectrophotometer and a NADH molar extinction coefficient of 6,220 M-1cm-1. The reaction was linear between glutamine concentrations 100 and 1.25 μM. Using 10U in each reaction, Hanes Woolf kinetic plots generated Vmax parameters of 46.29 and 7.57 μM/min for Erwinase and Kidrolase respectively. Km values were 96.05 and 33.06 μM for Erwinase and Kidrolase respectively. Preliminary studies demonstrate that glutamine depletion induced a dose dependent reduction in cell proliferation in HCT116 p53+/+, HCT116 p53-/-, MDA-MD-231, MDA-MB-453, MCF7, TK10, A549 and T47D cell lines in vitro. In conclusion, these studies demonstrate that Erwinase has significant glutaminase activity and is superior to Kidrolase in terms of glutaminase activity. Erwinase is a good candidate for exploring the efficacy of glutamine depletion strategies and further studies are required to determine whether Erwinase can potentiate the activity of other approaches aimed at restricting the glutamine dependence of tumors.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C154.
Citation Format: Roger M. Phillips, Mohammed U. Saleem, Adrian Williams, Jon Morgan. Kinetic analysis of the glutaminase activity of L-asparaginases derived from Erwinia chrysanthemi (Erwinase®) and Eshericia coli (Kidrolase). [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C154.
Abstract C155: Targeting glucose metabolism to suppress Ph+B-ALL progression.
Authors: Liu Tingyu , Xiang Handan , Nichols Amanda , Gerriets Valerie , Kishton Rigel , Rizzieri David , Rathmell Jeffrey C. 
Published: August 2014

Abstract
Philadelphia chromosome positive B cell derived acute lymphoblastic leukemia (Ph+B-ALL) is a type of aggressive leukemia that lacks effective treatment. The high glucose metabolism observed in many other cancers suggests that targeting glucose metabolism may provide a novel therapeutic approach for Ph+B-ALL. However, metabolic features of Ph+B-ALL and metabolic stress responses have not been described. In this study, we examined human primary B-ALL samples, B-ALL cell lines and primary murine Ph+B-ALL to identify metabolic program and metabolic stress responses of Ph+B-ALL cells. Metabolic measurements of extracellular flux and glucose consumption showed that B-ALL cell lines are highly glycolytic and preferentially utilize glucose. Consistent with a dependence on elevated glycolysis, primary human B-ALL cells were more sensitive than normal B cells to inhibition of glycolysis with 2-deoxyglucose (2-DG). This metabolic stress induced cell death was likely to be mediated through p53 and Bcl-2 family proteins. 2-DG treatment induced expression of pro-apoptotic Bcl-2 family protein Bim and primary murine Ph+B-ALL cells generated from p53-/- or Bim-/- background remained viable even when treated with 2-DG. To specifically investigate how inhibition of glucose metabolism impacts B-ALL survival and disease progression in vivo with genetic tools, we generated murine Ph+ B-ALL cells on a Glut1fl/fl Ubi-CreER background that allow specific deletion of Glut1 in cancer cells upon treatment of tamoxifen. In vitro deletion of Glut1 reduced glucose uptake and glucose metabolism, although glycolysis was not wholly suppressed. This partial reduction of glucose metabolism led to greatly reduced cell proliferation and some cell death in vitro. Importantly, in vivo deletion of Glut1 after transfer of B-ALL cells into congenic recipients suppressed B-ALL progression and prolonged animal survival. These data show that B-ALL cells exhibit high glucose metabolism similar to other types of cancer and are sensitive to glucose metabolism inhibition. Reduction of glucose metabolism can impede B-ALL proliferation or cause cell death through p53 and Bim and thus suppress B-ALL progression in vivo.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C155.
Citation Format: Tingyu Liu, Handan Xiang, Amanda Nichols, Valerie Gerriets, Rigel Kishton, David Rizzieri, Jeffrey C. Rathmell. Targeting glucose metabolism to suppress Ph+B-ALL progression. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C155.
Abstract C165: Discovery of a lectin-like receptor family protein as a novel target for monoclonal antibody therapy.
Authors: Campagnoli Susanna , Parri Matteo , Grandi Alberto , De Camilli Elisa , Terracciano Luigi , Jin Boquan , Viale Giuseppe , Sarmientos Paolo , Grandi Guido , Pileri Piero , Grifantini Renata Maria 
Published: August 2014

Abstract
In our recent research activities, we identified a panel of novel candidate markers for prevalent cancers by a systematic tissue microarray analysis (TMA) of a large collection of polyclonal antibodies (approximately 1600) raised against membrane-associated and secreted human proteins currently marginally characterized. Of major interest were those proteins specifically detected in one or more of the five tumors with significant frequencies (data from 50 patients per each of the five cancers) and showing surface exposure on cancer cells. Among them, here we describe the molecular characterization of novel surface-associated protein belonging to the lectin-binding family proteins (involved in the innate immune response) for which very little knowledge exists. The protein was mainly over-expressed in breast, lung and ovary cancers (approximately 30-40% of tested cases) and it is involved in cell proliferation, migration and and invasiveness, as demonstrated by gene silencing using specific siRNA technology. Highly specific murine monoclonal antibodies (mAbs) were generated toward it are able to recognize the protein on the surface of breast and ovary cell lines. Of particular interest are two mAbs, which, upon binding, are efficiently internalized by breast and ovary cancer cells, indicating that they are amenable for the generation of antibody-drug conjugates. These two mAbs influence viability of breast and ovary cells in vitro. One of them shows antibody-dependent cell-mediated cytotoxicity in vitro on breast cancer cells. In vivo efficacy studies are ongoing to evaluate their therapeutic activity. Overall, data indicate that the lectin-binding protein and the specific mAbs here described could be developed as a novel tool for a targeted therapy alone or in combination with other treatments.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C165.
Citation Format: Susanna Campagnoli, Matteo Parri, Alberto Grandi, Elisa De Camilli, Luigi Terracciano, Boquan Jin, Giuseppe Viale, Paolo Sarmientos, Guido Grandi, Piero Pileri, Renata Maria Grifantini. Discovery of a lectin-like receptor family protein as a novel target for monoclonal antibody therapy. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C165.
Abstract C17: Genome-wide siRNA screen identifies diverse signaling pathways converging at mitochondria to regulate cell death.
Authors: Deng Jing , Letai Anthony 
Published: August 2014

Abstract
Programmed cell death (apoptosis) is regulated by the interplay of BCL2 family proteins, and by signals transduced to/from mitochondria. Recent studies in our group suggest that priming, a mitochondrial state involving interaction between anti- and pro-apoptotic proteins, can determine the sensitivity of chemotherapy. To understand how such priming status is regulated, we performed genome-wide siRNA screen using potent BCL2 inhibitor ABT737. The lead targets generated by the screen were validated and studied for potential mechanisms, revealing multiple signaling pathways that can regulate mitochondrial priming thus affect cell death. Knockdown of lead targets not only protects cells from ABT737 treatments, but also from other chemotherapeutics, such as paclitaxel and etoposide. The protection correlates with the protection of mitochondrial potential, suggesting that they may function upstream of mitochondrial outer membrane permeablization (MOMP). Targets initially identified in MDAMB231 breast cancer cell line were further validated in other cancer types, and demonstrated cell protection from chemo-treatments as well when down-regulated. Analysis of patients bearing deletions in genes or down-regulated RNA expression in some of these targets showed a trend to a poorer survival, supporting our finding that these targets may play important roles in regulating cell death and response to chemotherapy.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C17.
Citation Format: Jing Deng, Anthony Letai. Genome-wide siRNA screen identifies diverse signaling pathways converging at mitochondria to regulate cell death. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C17.
Abstract C186: Tyro3 is a potential target for the treatment of breast cancer.
Authors: Ekyalongo Roudy Chiminch , Mukohara Toru , Funakoshi Yohei , Tomioka Hideo , Kataoka Yu , Shimono Yohei , Toyoda Masanori , Kiyota Naomi , Fujiwara Yutaka , Minami Hironobu 
Published: August 2014

Abstract
Background: We previously identified the involvement of receptor tyrosine kinase Tyro3 in the cell growth mechanism of MCF-7 breast cancer cells. In the present study, we evaluated the potential of Tyro3 as a therapeutic target in various types of breast cancer cell lines.
Material and Methods: We compared outcomes following Tyro3 knockdown by siRNA in four estrogen receptor (ER)-positive/HER2-nonamplified (luminal-type; CAMA-1, T47D, MCF-7, and HCC1428), two ER-negative/HER2-amplified (HER2-type; HCC1419 and HCC1954), and two ER-negative/HER2-nonamplified (triple negative [TN]-type; MDA-MB-231 and HCC1143) breast cancer cell lines. The effect of Tyro3 knockdown on cell growth, cell cycle distribution, and cell signaling was evaluated by MTS assay, FACS, and Western blot, respectively.
Results: Tyro3 knockdown by siRNA induced the most prominent growth suppression in luminal-type cells, and to a lesser extent in HER2-type cells. In contrast, no growth inhibition following Tyro3 knockdown was observed in TN-type cells. The Tyro3 siRNA-induced growth inhibition in luminal-type cells was observed in both the estradiol (E2)-rich and -null conditions. Cellular sensitivity to Tyro3 siRNA was not correlated with baseline Tyro3 expression level or level of secretion of Tyro3 ligand Gas6. Phosphorylation of Tyro3 was detectable in five of six sensitive lines, but not in two resistant TN-type lines. The results of the growth assays significantly correlated with those of FACS analysis, with G0-G1/S cell cycle arrest observed only in Tyro3 siRNA-sensitive cells. Western blot analysis showed a decrease in phosphorylation of downstream signaling molecules, including ERK1/2, S6K, and STAT3, and in cyclin D1 in cell lines sensitive to Tyro3 siRNA only, and not in resistant TN-type cell lines.
Conclusions: Tyro3 is a potential therapeutic target in breast cancer, particularly in luminal-type cells. Phosphorylation of Tyro3 might be a predictive marker of sensitivity to Tyro3-targeted therapy.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C186.
Citation Format: Roudy Chiminch Ekyalongo, Toru Mukohara, Yohei Funakoshi, Hideo Tomioka, Yu Kataoka, Yohei Shimono, Masanori Toyoda, Naomi Kiyota, Yutaka Fujiwara, Hironobu Minami. Tyro3 is a potential target for the treatment of breast cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C186.