tbuckley/cabot-search huggingface dataset:
Abstract 3299: Radiation induced p53-dependent cytoprotective autophagy fails to reduce radiation sensitivity in H460 non-small cell lung cancer cells
Authors: Jingwen Xu, Tareq Saleh, Liliya Tyutynuk-Massey, Yingliang Wu, David A. Gewirtz
Journal: Cancer Research (2017)

Abstract Our previous studies showed that radiation-induced autophagy could be either cytoprotective or nonprotective in A549 non-small cell lung cancer cells, HN30 and HN6 head and neck cancer cells and 4T1 breast tumor cells, and that this functional difference was related to the status of p53 in the cells (i.e. cytoprotective autophagy in p53 wild type (wt) cells and nonprotective autophagy in cells with mutant or null p53) [1]. Despite the fact that inhibition of autophagy has been proposed as a strategy to overcome resistance to radiation and chemotherapy, there is little direct information in support of the premise that the induction of autophagy confers resistance. As an example, studies in HCT-116 cells that are either wt or null in p53 demonstrate essentially identical radiation sensitivity [2], although the nature of radiation induced autophagy was not determined in these cell lines. To address this question, p53 wt and p53 null H460 cell lines were utilized to evaluate whether cytoprotective autophagy actually suppressed radiation sensitivity. Acridine orange staining, flow cytometric analysis and western blotting (LC3B, p62, Atg5) showed that following exposure to 6 Gy of radiation wtp53 and p53-null H460 cells exhibited a significant induction of autophagy. p53 wt H460 cells showed sensitization to radiation upon pharmacological autophagy inhibition and Atg5 knock down. However, pharmacological inhibition of autophagy and knock down of Atg5 genes failed to influence sensitivity to radiation in p53-null H460 cells. Furthermore, clonogenic survival assays showed that while exposure to radiation induced a dose-dependent (2, 4, 6, 8 Gy) decrease in clonogenicity in both p53wt and p53-null H460 cells, radiation sensitivity was not decreased in the p53 wt cells. Taken together, these findings indicate that cytoprotective autophagy may not actually alter the nature of radiation (and possibly chemotherapeutic) sensitivity in tumor cells. [1] Chakradeo, S, et al. Mol Pharm (2015): [2] Lindgren, T, et al. Int J Rad Biol (2015). Acknowledgments We would like to thank Dr. Santiago Lima (Virginia Commonwealth University) for the H460 p53-null cells. Citation Format: Jingwen Xu, Tareq Saleh, Liliya Tyutynuk-Massey, Yingliang Wu, David A. Gewirtz. Radiation induced p53-dependent cytoprotective autophagy fails to reduce radiation sensitivity in H460 non-small cell lung cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3299. doi:10.1158/1538-7445.AM2017-3299
Abstract 395: Activation mechanisms of cancer associated MEK1 mutants
Authors: Yijun Gao, Matthew T. Chang, Daniel McKay, Rona D. Yaeger, Merna Torres, Keven Muniz, Drosten Matthias, Omar I. Abdel-Wahab, Mariano Barbacid, Giordano Caponigro, Darrin Stuart, David Solit, Barry S. Taylor, Zhan Yao, Neal Rosen
Journal: Cancer Research (2017)

Abstract Activating BRAF mutants drive human tumors by dysregulating ERK signaling despite ERK-dependent feedback suppression of RAS. These RAF mutants become RAS independent, by either of two mechanisms, and they are thus unaffected by feedback inhibition of upstream signaling. Recently, it has become clear that mutation activation of MEK1 or MEK2 occur at appreciable frequency in human tumors, but the mechanism of activation of these mutants and whether they remain dependent on upstream activation of RAS or RAF signaling remains unknown. Here we characterized the mechanism of activation of 18 recurrent MEK1 mutants identified in human cancer. Based on these data, they fall into three classes. The kinase activity of first class remains dependent on RAF mediated phosphorylation of S218 and S222; the second class of MEK mutants has basal RAF-independent activity, but can be further stimulated by RAF and the third class no longer requires phosphorylation of S218 and S222 and signals in a RAF independent manner. These features determined their sensitivity to ERK-dependent feedback regulation and the ability to drive ERK signaling output in cells. The more RAF-independent activity the mutants acquired, the better they could activate downstream ERK pathway, leading to increased transforming activities in MEF cells in the absence of RAF. This is consistent with their genetic association with RAS, RAF and NF1 mutations in human tumors. The Class 3 RAF-independent MEK1 mutants tend to be mutually exclusive with those mutants, while coexistence with RAS/RAF/NF1 mutations is frequently observed in tumors with Class 1 or 2 MEK1 mutants. Moreover, functional class correlated with sensitivities of ERK signaling driven by these mutants to MEK inhibitors that function by different mechanisms. Unlike RAF dependent or regulated MEK1 mutants, the ERK signaling driven by RAF independent MEK1 mutants is insensitive to an allosteric MEK inhibitor that functions by preventing RAF mediated MEK phosphorylation. However, signaling driven by all classes of MEK1 mutants is sensitive to an ATP competitive MEK1 inhibitor which targets MEK1 kinase activity. Citation Format: Yijun Gao, Matthew T. Chang, Daniel McKay, Rona D. Yaeger, Merna Torres, Keven Muniz, Drosten Matthias, Omar I. Abdel-Wahab, Mariano Barbacid, Giordano Caponigro, Darrin Stuart, David Solit, Barry S. Taylor, Zhan Yao, Neal Rosen. Activation mechanisms of cancer associated MEK1 mutants [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 395. doi:10.1158/1538-7445.AM2017-395
Abstract 1999: Cofactor Paragon: a novel tool to analyze the tumor microenvironment using RNAseq
Authors: Ryan Bloom, Raman Talwar, Jeff Hiken, Jon Armstrong
Journal: Cancer Research (2017)

Abstract The impact of cancer immunotherapy on clinical cancer care is growing rapidly. However, there are several mechanisms by which a tumor can avoid recognition and/or elimination by the immune system. Immune recognition, activation, and infiltration are each required for effective clearance of a tumor by the immune system. Impairment of any one of these mechanisms requires a different therapeutic strategy or combination of strategies to significantly address. The development of new multi-modal biomarker technologies that have the ability to provide insight into each of these factors in a cost-effective manner is essential to effectively guide treatment choice for patients and to discover new therapeutic targets. We have developed Paragon, which is an RNA sequencing based panel that utilizes novel informatics technologies to provide a comprehensive picture of a tumor’s transcriptomic and immune micro-environment. There are three distinct outputs of Paragon: measurement of the expression levels of ten immune checkpoint genes, quantification of the total mutational burden of the tumor, and the levels of infiltration of 24 different immune cell subtypes within the tumor microenvironment. These outputs collectively inform the various mechanisms that tumors use to evade recognition and clearance by the immune system. We show that Cofactor Paragon accurately reports on gene expression, mutational burden and infiltrating leukocytes compared to gold standard methods from just 20 ng of FFPE tumor RNA. We further analyze the data to show the correlation between mutational burden and infiltrating CD4+ and CD8+ T-cell levels. Finally, we show how Paragon can identify expression of genes involved in inhibitory tumor metabolism that correlate with resistance to PD-1 blockade therapy. These results position Cofactor Paragon as a novel tool that can help identify ideal therapeutic strategies in immuno-oncology. Citation Format: Ryan Bloom, Raman Talwar, Jeff Hiken, Jon Armstrong. Cofactor Paragon: a novel tool to analyze the tumor microenvironment using RNAseq [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1999. doi:10.1158/1538-7445.AM2017-1999
Abstract 5427: Distinctions between the metabolic changes in glioblastoma cells and glioma stem-like cells following irradiation
Authors: Elizabeth I. Spehalski, Cord Peters, Philip Tofilon, Kevin Camphausen
Journal: Cancer Research (2017)

Abstract Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor in adults and, even with aggressive treatment that includes surgical resection, radiation, and temozolomide administration, prognosis is poor due to tumor recurrence. There is evidence that within GBMs a small number of glioma stem- like cells (GSCs) exist, which are thought to be radiation resistant and may be capable of repopulating a tumor after treatment. Like most cancers, GBMs largely employ aerobic glycolysis to create ATP, a phenomenon known as the Warburg effect. Experiments have shown that the cellular metabolism of GSCs differs from that of differentiated cells, making them an attractive target for novel therapeutic approaches. Much work has been done to analyze the metabolic profiles of GSCs with the goal of identifying potential therapeutic targets, but little data exists linking metabolic changes to radiation resistance. The purpose of this study is to characterize the metabolic differences between glioma stem- like cells and traditional GBM tumor cells with and without radiation treatment (IR). To this end, we compared the metabolism of a human derived GSC line with two commonly used GBM cell lines before and after IR. At baseline, we find that glioma stem-like cells are more quiescent than GBM cells, which have higher levels of both glycolysis and oxidative phosphorylation. GBM lines show higher levels of both basal and maximal respiration, as well as basal glycolysis and glycolytic capacity than GSCs. They also express higher levels of energy, glycolysis, and TCA cycle metabolites than GSCs. Inversely, GSCs demonstrate metabolic signs of quiescence such as decreased NEAA synthesis. After IR, the radiation- sensitive GBM tumor cell line (U251) exhibits increases in all metabolic pathways, whereas the levels of glycolytic and oxidative metabolites in the GSCs remain unchanged. All cell lines show an increase of ATP and NAD production following IR. These findings indicate that the metabolism of GSCs undergoes different alterations than that of GBM tumor cells after IR, making them an attractive target for novel therapeutic approaches in conjunction with radiation therapy. Additionally, differences in metabolic signatures between GSC lines could be useful for non-invasive diagnostic modalities such as 13C MR spectroscopy. Citation Format: Elizabeth I. Spehalski, Cord Peters, Philip Tofilon, Kevin Camphausen. Distinctions between the metabolic changes in glioblastoma cells and glioma stem-like cells following irradiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5427. doi:10.1158/1538-7445.AM2017-5427
Abstract 1815: Extracellular vesicles derived from lung cancer cell may contribute to the formation of malignant pleural effusion via activating Stat3/Tissue factor/VEGF
Authors: Chien-Chung Lin, Wei-Lun Huang, Wu-Chou Su
Journal: Cancer Research (2017)

Abstract Malignant pleural effusion (MPE) is a poor prognostic sign for patients with non-small-cell lung cancer (NSCLC) and is noted around 15% at the time of initial diagnosis and 50% in their later courses. Previously, we have shown that IL-6/Stat3/tissue factor (TF)/VEGF pathway enhances lung cancer angiogenesis, metastasis and the generation of MPE. In the current study, we demonstrated that EVs derived from lung cancer cells may also contribute MPE formation. We first used ultra-filtration (UF) method as well as the classical ultra-centrifugation (UC) method to isolate EV from lung cancer cell line and demonstrated that the EVs isolated by UF method were characteristic with exosome marker in Western Blotting (Alix, CD63, and Tsg101), morphology at Transmission electron microscopy (TEM) and size in Nanosight system (30 to 150 nm). After labeling these EVs with the green fluorescent dye Cellvue, EVs uptake in AS2 cell can be imaged by the fluorescent microscopy. After the EV uptake of recipient cell, the oncogenic signal such as Stat3 and AKT were activated with increasing phosphorylation at Western blotting comparing to those adding serum free medium. Using the ELISA method, we found the cargos in these EV were enriched in IL6. We also found these cargo derived from EVs, containing high VEGF and TF, induced more prominent extravasation than serum free medium did in Miles assays. In clinical sample, we also isolated cancer cell from MPE and the purity of the cells was confirmed by immunofluorescent staining with specific TTF-1expression of lung adenocarcinoma. After short-term culture, EV from MPE cancer cell have the characteristic of exosome marker and induced more prominent extravasation than serum free medium as EV from AS2 cell did. And we also isolated EV direct from MPE and pleural effusion from congestive heart failure (CHF) patients. And we found EV derived from MPE induced more prominent extravasation than those derived from CHF in Miles assays. In conclusion, we demonstrated the UF is clinical assessable method for isolating EV from body fluid and EV derived from lung cancer cell and MPE may contribute oncogenic and angiogenic signal transmission accelerating the formation of MPE. Citation Format: Chien-Chung Lin, Wei-Lun Huang, Wu-Chou Su. Extracellular vesicles derived from lung cancer cell may contribute to the formation of malignant pleural effusion via activating Stat3/Tissue factor/VEGF [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1815. doi:10.1158/1538-7445.AM2017-1815
Removing Firearms From the Home After Attempted Suicide Can Be Life Saving
Authors: Merete Nordentoft, Annette Erlangsen, Trine Madsen
Journal: American Journal of Psychiatry (2017)

In this issue, Olfson et al. present data showing a 37-fold higher risk of suicide within the first year after nonfatal deliberate self-harm compared with the U.S. population (1).These alarming findings stem from a large national study based on Medicaid data from 45 states.The cohort consisted of 61,297 patients who were clinically diagnosed with deliberate selfharm between 2001 and 2007.Although the study is not likely to be representative of the entire U.S. population, it provides relevant observations that could help improve follow-up after deliberate self-harm.Olfson et al. found that among persons who presented with nonfatal deliberate self-harm, the suicide rate was 439 per 100,000 person-years and the rate of repeated self-harm was 26,320 per 100,000 person-years over the following 12 months.In absolute terms, this means that 0.4% died by suicide and 19.7% repeated deliberate self-harm within a year of the initial presentation.The high proportion of suicides is confirmed by recent findings from Sweden, where 4.9% of people with deliberate self-harm died by suicide within a median followup of 5.3 years (2).Nationwide Danish studies have found that between 0.9% and 1.2% of people presenting with deliberate self-harm had died by suicide within 12 months, and an international meta-analysis reported a proportion of 1.6% (95% CI51.2, 2.1) (3-5).Nonfatal repetitions were reported to range between 11.8% and 12.2% in the Danish studies, and the metaanalysis reported a repeat deliberate self-harm rate of 16.3% (95% CI515.1, 17.7) (4).In comparison, in the Olfson et al. study, a slightly lower 12-month postdischarge suicide rate was reported, but also a slightly higher rate of repeat deliberate self-harm.As in previous studies (6), elevated risks of both suicide and repeat deliberate self-harm were found across all types of mental disorders among patients after deliberate self-harm.Considering that 20% repeat deliberate self-harm during the first year, the need for effective treatment irrespective of psychiatric diagnosis is evident.Psychotherapeutic or psychosocial treatment administered to patients with deliberate selfharm may reduce the risk of suicide and repeated nonfatal deliberate self-harm (5), as indicated by a recent Cochrane review (7).The World Health Organization recommends that patients presenting with deliberate self-harm be offered mental health care (8).However, as Olfson et al. highlighted, those who did not survive their initial deliberate self-harm episode were less likely to have received treatment for a mental
Abstract 4292: Indole-3-carbinol (I3C) enhances efficacy of Gemcitabine in leiomyosarcoma
Authors: Sujit Suwal, Alexandra Moran, Ana Paz-Mejia, Lorena Flor, Omar Picado, Basem Azab, Jonathan Trent, Fiorella Pendola, Alan S. Livingstone, Danny Yakoub
Journal: Cancer Research (2017)

Abstract Introduction: Adjuvant Gemcitabine have recently been shown to improve progression free survival in some types of leiomyosarcoma. Adequate cellular uptake of the drug remains a challenge. We aimed to evaluate the role of nucleoside transporter activator (I3C) in increasing the in vitro efficacy of gemcitabine in the treatment of leimyosarcoma. Methods: Leiomyosarcoma cells (SKLMS) were incubated with DMEM+10% FBS + 1% Antibiotics. Dose response curves were generated for gemcitabine and I3C to identify the IC-50 values. Triplicates of cells were then plated in 96 well plates (3000 cells/well) and allowed to adhere. The adherent cells were further treated with gemcitabine in the presence or absence of I3C and compared with controls (DMSO treatment). MTS cell viability assays were done and absorbance was monitored in a micro-plate reader. Flow cytometry was performed with Propidium Iodide (PI) staining. Scratch assays were done to assess cell migration. Results: Viability assay showed significant increase in the cytotoxicity of gemcitabine in presence of I3C compared to controls. Flow cytometry data showed gemcitabine alone treatment has arrested the cell cycle at G0/G1 phase without increase of cell death. On the contrary, use of gemcitabine in combination with I3C increased the cell death by at least three folds compared to control as well as gemcitabine alone treated cell population. Scratch assay over a 16 hour period showed that the rate of cellular migration significantly dropped by 1.5 folds upon treatment with gemcitabine combined with I3C compared to gemcitabine alone. Western blots showed increased expression of membrane transporter hENT-1 with I3C treatment. Conclusion: The use of I3C enhanced cellular uptake, transport and cytotoxicity of gemcitabine in SKLMS cells. Further studies are warranted and underway to elaborate on intracellular mechanisms of action to optimize the use of I3C as a potential adjunct in preclinical and clinical leiomyosarcoma treatment. Note: This abstract was not presented at the meeting. Citation Format: Sujit Suwal, Alexandra Moran, Ana Paz-Mejia, Lorena Flor, Omar Picado, Basem Azab, Jonathan Trent, Fiorella Pendola, Alan S. Livingstone, Danny Yakoub. Indole-3-carbinol (I3C) enhances efficacy of Gemcitabine in leiomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4292. doi:10.1158/1538-7445.AM2017-4292
Abstract 4644: A cell-line based TP53 expression pattern control panel with interobserver variability and next-generation sequencing analysis
Authors: Yaling Zhou, Sorana Raiciulescu, Viqar Syed, Patrick Malafronte, Justin Wells, Mica Grantham, Michael Allgaeuer, Heping Gong, Joel Moncur, Deyin Xing, John D. Andersen
Journal: Cancer Research (2017)

Abstract Introduction: TP53 gene mutation status is known to be manifested through distinct p53 immunohistochemical staining patterns; diffuse, wild-type and null. This stratified staining pattern is routinely utilized in subtyping ovarian and endometrial cancer subtypes and distinguishing low- and high-grade serous carcinoma. Experimental Procedure: Three ovarian cancer cell lines were used in the construction of an immunohistochemical control panel slide that highlights each respective TP53 expression pattern. Cell lines were expanded, harvested and resuspended in agarose gel and subsequently sectioned onto a microscope slide. Procured tissue specimens were submitted along with either standard control tissue or the TP53 cell line control panel to pathologists of varying experience. Next-generation sequencing analysis was performed on cases in which there was discordance in expression pattern interpretation. Results: Interobserver variability was assessed on 44 samples between five raters of varying experience. The results were similar with the use of the TP53 expression panel (Fleiss kappa= 0.6 vs 0.53). Next-generation sequencing analysis demonstrated low-frequency variant mutations in cases in which there was reviewer discordance. Conclusion: This study suggests the addition of a TP53 cell line expression pattern control panel could potentially increase diagnostic accuracy for ovarian and endometrial cancer subtypes and distinguishing ovarian low-grade and high-grade serous carcinoma. The cell line control has refined quality assurance potential for antibody optimization.This cell line control panel has the potential to be utilized in routine immunohistochemical staining with difficult to interpret p53 expression patterns directed to submission for next-generation sequencing. Citation Format: Yaling Zhou, Sorana Raiciulescu, Viqar Syed, Patrick Malafronte, Justin Wells, Mica Grantham, Michael Allgaeuer, Heping Gong, Joel Moncur, Deyin Xing, John D. Andersen. A cell-line based TP53 expression pattern control panel with interobserver variability and next-generation sequencing analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4644. doi:10.1158/1538-7445.AM2017-4644