bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 702: Kinetic analysis identifies determinants of sensitivity to MEK inhibitor-induced cell death
Authors: Inde Zintis , Han Kyuho , Bassik Michael C. , Dixon Scott J. 
Published: August 2019

Abstract
Successful identification of the genetic determinants of drug sensitivity has been a long-standing goal of cancer research and, more recently, of precision medicine. In cancer cells, drug treatment can induce both growth arrest and death. The distinction between these outcomes is important but can be overlooked in large-scale analyses that rely on traditional metabolism-based methods for quantifying cell viability. We recently demonstrated that Scalable Time-Lapse Analysis of Cell Death Kinetics (STACK) effectively quantifies cell death in response to diverse lethal stimuli. We investigated cases in which this approach captured drug-induced lethality more effectively than traditional assays. We found that inhibitors of mitogen activated protein kinase 1 and 2 (MEK1/2) arrested proliferation in diverse cancer cell lines, but induced cell death in only a subset of these lines. We sought to identify the factors that determine whether a cancer cell will undergo growth arrest or death in response to MEK inhibition. MAP kinase pathway activation has previously been reported as a determinant of MEK inhibitor sensitivity; we determined that the dynamics of Bcl-2 family proteins in response to MEK inhibitor treatment correlate with death sensitivity independent of pathway activation. In a genome-wide CRISPR screen, we identified a lipid metabolic enzyme that is a novel modulator of MEKi sensitivity. Our results suggest new approaches for predicting and enhancing sensitivity to cancer therapeutics in tumors and demonstrate that direct measurement of cell death provides clear advantages over proliferation-based metrics.
Citation Format: Zintis Inde, Kyuho Han, Michael C. Bassik, Scott J. Dixon. Kinetic analysis identifies determinants of sensitivity to MEK inhibitor-induced cell death [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 702.
Abstract 703: An anthelmintic drug, flubendazole, exerts antitumor effects in triple-negative breast cancer via targeting cancer stem-like properties
Authors: Kim Yoon-Jae , Oh Eunhye , Cho Tae-Min , Jang Seojin , Park Jung Min , Park Soeun , Park Minsu , Kim Ji Young , Seo Jae Hong 
Published: August 2019

Abstract
Triple-negative breast cancer (TNBC) remains challenging due to the absence of established molecular targets for the phenotype. These tumors tend to be more aggressive with a higher risk of recurrence and metastasis, leading to poorer clinical outcomes. Flubendazole (FLU) is a widely used anthelmintic agent that also exhibits anticancer activity in several cancer types. The objective of the present study was to investigate the effects of FLU on cell proliferation, apoptosis and breast cancer stem cell (BCSC)-like properties in TNBC. The effect of FLU on TNBC cell lines in vitro was evaluated in terms of apoptosis, cell cycle distribution, caspase-3/-7 activity and BCSC-like characteristics. An orthotopic allograft model with 4T1 mammospheres was used to examine the effect of FLU on tumor growth and angiogenesis in vivo. FLU efficiently induced apoptosis with accompanied by activation of caspase-3 and -7, and PARP cleavage in TNBC. FLU-induced apoptosis is associated with the accumulation of G2/M phase cells by tubulin degradation, leading to mitotic catastrophe. Interestingly, FLU significantly suppressed BCSC-like properties in TNBC, as evidenced by a marked inhibition of ALDH1 activity, the CD44high/CD24low and CD49fhigh/CD24high subpopulations, and the subsequent impairment of mammosphere formation in vitro. This appears to contribute toward the suppression of BCSC-enriched tumor burden and intra-tumoral angiogenesis in TNBC allografts, together with a significant downregulation of ALDH1A1 and CD49f expression in vivo. Our findings suggest that FLU kills not only rapid proliferating tumor cells but also effectively eradicates BCSC-like cells in vitro and in vivo. These findings warrant further investigation of flubendazole as a potential treatment for triple-negative breast cancer.
Citation Format: Yoon-Jae Kim, Eunhye Oh, Tae-Min Cho, Seojin Jang, Jung Min Park, Soeun Park, Minsu Park, Ji Young Kim, Jae Hong Seo. An anthelmintic drug, flubendazole, exerts antitumor effects in triple-negative breast cancer via targeting cancer stem-like properties [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 703.
Abstract 704: RALB GTPase: A critical regulator of DR5 cell surface expression and TRAIL sensitivity in RAS mutant colorectal cancer
Authors: Khawaja Hajrah , Roberts Jamie Z. , Javadi Arman , O'Reilly Paul , McArt Darragh , Allen Wendy L. , Morrison Markus , Kennedy Richard , Vitale Nicolas , Johnston Patrick G. , Harrison Tim , Longley Daniel B. , Evergren Emma , Schaeybroeck Sandra Van 
Published: August 2019

Abstract
RAS mutant (MT) metastatic colorectal cancer (mCRC) remains a difficult-to-treat group. RALA and RALB GTPases function downstream of RAS and have been found to be key regulators of several cell functions and implicated in KRAS-driven tumourigenesis. However, their role as regulators of the apoptotic machinery remains to be elucidated. Here, we found that inhibition of RALB expression, but not RALA, resulted in caspase 8-dependent cell death in KRASMT cells, that was not further increased following MEK1/2 inhibition. Proteomics analysis and mechanistic studies revealed that RALB inhibition induces marked upregulation of the pro-apoptotic cell-surface TRAIL Death Receptor 5 (DR5) receptor in KRASMT CRC. Moreover, DR5 knockout also attenuated siRALB-induced apoptosis, confirming the role of the extrinsic apoptotic pathway as regulator of siRALB-induced cell death. Importantly, TRAIL treatment resulted in acute association of RALB with the death-inducing signalling complex (DISC) and targeting RALB using pharmacologic inhibition or RNAi approaches, resulted in a potent increase in TRAIL-induced cell death in RASMT CRC. Significantly, high RALB mRNA levels were found in the poor prognostic CRIS-B CRC subgroup. Collectively, this study provides the first evidence for a role of RALB in apoptotic priming and that RALB inhibition may be a promising strategy to improve response to TRAIL treatment in poor prognostic RASMT CRC.
Citation Format: Hajrah Khawaja, Jamie Z. Roberts, Arman Javadi, Paul O'Reilly, Darragh McArt, Wendy L. Allen, Markus Morrison, Richard Kennedy, Nicolas Vitale, Patrick G. Johnston, Tim Harrison, Daniel B. Longley, Emma Evergren, Sandra Van Schaeybroeck. RALB GTPase: A critical regulator of DR5 cell surface expression and TRAIL sensitivity in RAS mutant colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 704.
Abstract 707: Emerging role of mitochondrial pathway in crosstalk between ferroptosis and apoptosis
Authors: Lee Yong J. 
Published: August 2019

Abstract
Introduction: Several processes lead to cell death; ferroptosis, apoptosis, necrosis, and autophagy are the primary mechanisms. Each type of biological death has been believed to have a distinct biochemical and morphological fingerprint. However, emerging evidence suggests that these different types of cell death often share common pathways. In this study, we investigated whether mitochondrial signaling pathways play an important role in the intracellular communication between ferroptosis and apoptosis.
Methods: The human colorectal carcinoma HCT116 cell line was used to evaluate the interplay between cell death pathways. Cells were treated with the apoptotic agent TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) and ferroptotic agents such as artesunate and erastin. Cell death was determined by trypan blue exclusion assay. Apoptosis was measured by western blot. Ferroptosis was assessed using MDA assay. Bax oligomerization was performed by mitochondrial fractionation followed by SDS-PAGE under nondenaturing conditions.
Results: We observed that the combinatorial treatment induced synergistic apoptosis, which was mediated through an increase in caspase activation. We also observed an increase in pro-apoptotic Bax oligomerization in the mitochondrial fraction. The combined treatment-induced apoptosis was suppressed in Bax-deficient HCT116 cells. Ferroptotic agents enhanced the intracellular level of anti-apoptotic Mcl-1. However, the combined treatment decreased the level of Mcl-1. Interestingly, the combined treatment-induced apoptosis was abolished in HCT116 cells with knockin of Mcl-1 phosphorylation site mutant (S121A/E125A/S159A/T163A).
Conclusions: These data suggest a novel mechanism for the combined treatment of the apoptotic agent TRAIL and ferroptotic agent erastin/artesunate-induced synergistic apoptosis through the mitochondrial pathway.
Citation Format: Yong J. Lee. Emerging role of mitochondrial pathway in crosstalk between ferroptosis and apoptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 707.
Abstract 708: Targeting fatty acid synthase induces apoptosis and senescence
Authors: Steen Travis Van der , Kemble George , Lupu Ruth 
Published: August 2019

Abstract
Fatty acid synthase (FASN) is a novel therapeutic target for cancer as it has increased expression in many different types of cancers. There has been significant understanding that cancer cells can hijack and modify tightly regulated metabolic pathway for its own survival. FASN is the enzyme that synthesizes palmitate from malonyl-CoA for energy storage, structure and biomolecules. FASN expression is low to undetectable in normal cells with the exception of liver and adipose tissue. In this study, we investigated the role of FASN in breast cancer cells. We observed that in highly expressing FASN breast cancer cells, TVB-3166, FASN inhibitor, results in a decrease in cellular proliferation and an increase in cellular death. FASN inhibition increased reactive oxygen species (ROS) and altered NADPH/NADP+ ratio. Increase in ROS and NADPH have been linked to the intrinsic apoptosis pathway. Additionally, there is an increase in expression of the BH3-only proteins Bim, Puma and Noxa, leading to the permeabilization of the mitochondria and the release of cytochrome c. Furthermore, after prolong exposure to TVB-3166 a number of cells enter into a senescent state. This was observed the in detection of β-galactosidase and confirmed by the expression of senescent markers including ARF, INK4A, and CIP. Additionally, when the cells were treated with a CDK4/6 inhibitor in combination with the FASN inhibitor TVB-3166, an increase in stress induced premature senescence was observed. Our data demonstrates that inhibition of FASN induces intrinsic apoptosis while it stimulates cellular senescence to the apoptotic resistant cells. Therefore, the ability for TVB-3166 to both eradicate and overturn cell growth makes it potential therapeutic modality.
Citation Format: Travis Van der Steen, George Kemble, Ruth Lupu. Targeting fatty acid synthase induces apoptosis and senescence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 708.
Abstract 713: BBI-608 modulates DNA repair pathways, induces apoptosis and impacts response to 5-Fluorouracil and ionizing radiation in pancreatic cancer cells
Authors: Farran Batoul , Nagaraju Ganji P. , Shipra Sneha G. , Bethi Shipra R. , Lesinski Gregory B. , El-Rayes Bassel 
Published: August 2019

Abstract
Background: BBI-608 has been shown to modulate multiple oncogenic cellular pathways which are often constitutively active in pancreatic ductal adenocarcinoma (PDAC). Among these are key mediators of DNA repair and cellular survival pathways. We hypothesized that BBI-608 would sensitize PDAC cells to chemoradiotherapy.
Methods: The combined effects of BBI-608 and chemoradiotherapy (5-FU + IR) were evaluated in human (MIA PaCa-2) and murine (PANC-02) PDAC cell lines using a Br-dU cell proliferation assay. To elucidate the mechanism of action, effects on the expression of DNA damage and repair (pATM, γ-H2AX, p53; pATR, and Rad51) molecules were examined by Western blot. Apoptosis was assessed via Annexin V staining and flow cytometric analysis to evaluate mechanism of cell death following treatment of MIA PaCa-2 and PANC-02 cell lines. The in vivo activity of BBI-608, 5-FU + IR, alone and in combination, were evaluated in C57BL/6 mice bearing subcutaneous PANC-02 tumors.
Results: The combination of BBI-608 and chemoradiotherapy significantly decreased (p&lt;0.001) cell proliferation and induced Annexin V positive cells in both MIA PaCa-2 and PANC-02 cell lines. BBI-608 and chemoradiotherapy also decreased the activation of pATR, MDM2, and Rad51 and increased pATM, γ-H2AX and p53. Administration of BBI-608 potentiated the effects of chemoradiotherapy in vivo in mice bearing subcutaneous PANC-02 tumors as measured by tumor volume.
Conclusion: These data identify a role for BBI-608 as a modulator of DNA damage and apoptosis. These effects enhanced the efficacy of chemoradiotherapy in cell lines and in vivo models of PDAC and provide rationale for continued investigation in the pre-clinical and clinical setting.
Note: This abstract was not presented at the meeting.
Citation Format: Batoul Farran, Ganji P. Nagaraju, Sneha G. Shipra, Shipra R. Bethi, Gregory B. Lesinski, Bassel El-Rayes. BBI-608 modulates DNA repair pathways, induces apoptosis and impacts response to 5-Fluorouracil and ionizing radiation in pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 713.
Abstract 744: Clinically targetable genomic alterations in acral melanoma
Authors: Broit Natasa , Dutton-Regester Ken , Johansson Peter , Pritchard Antonia L. , Boyle Glen M. , Hayward Nicholas K. 
Published: August 2019

Abstract
Background: Acral melanoma is a rare subtype of melanoma, which has a distinct genomic profile from cutaneous melanoma. While survival outcomes for late-stage cutaneous melanoma have significantly improved in the last decade, treatment options for acral melanoma remain limited. The purpose of this study was to assess therapeutically targetable genomic alterations in acral melanoma.
Methods: The AACR project GENIE database was accessed via cBioPortal (http://cbioportal.org/genie/; version 3) to identify recurrent gene alterations in acral melanomas analyzed using the MSK-IMPACT oncopanel. These alterations were assessed through Cancer Genome Interpreter (http://cancergenomeinterpreter.org) to identify clinically actionable alterations.
Results: Thirty-six patients with acral melanoma were identified from the cBioPortal-GENIE database. These patients were sequenced using three different versions of oncopanel, which included 341 (n = 6), 410 (n = 22) and 468 (n = 8) genes. Seventy-six genes were found to be somatically mutated in this cohort in at least one sample. The most commonly mutated genes were NF1 (n = 5), BRAF (n = 5), PTPRT (n = 4), NOTCH3, HRAS and KRAS (n = 3). Copy number variations greatly contributed to aberration burden, with 132 unique genes found to either carry an amplification or deletion event. The most commonly amplified genes were CDK4 (n = 9), CCND1 (n = 9), FGF19/FGF4 (n = 8), PAK1 (n = 8), MDM2 (n = 7) and FGF3 (n = 7). The most recurrently deleted genes included CDKN2A (n = 9) and CDKN2B (n = 8), and less commonly JAK2 and PTEN (n = 2). Some genes were altered by various mechanisms; for example, some tumors had KIT activated by amplification while other tumors carried missense mutations. BRAF exhibited two fusion events: BRAF-METTL2B (with concomitant amplification) and BRAF-KIAA1549. Cancer Genome Interpreter identified several therapeutic targets, some of which have been the focus of clinical trials in cutaneous melanoma and other solid tumors, e.g. CDK4/6 inhibitors to target amplified CDK4 and CCND1, or mTOR inhibitors for tumors with NF1 mutations.
Conclusion: Several inhibitors are available which could show efficacy in a subset of acral melanomas with particular genomic alterations. Many of these inhibitors have yet to be tested in the context of acral melanoma and should be explored.
Citation Format: Natasa Broit, Ken Dutton-Regester, Peter Johansson, Antonia L. Pritchard, Glen M. Boyle, Nicholas K. Hayward. Clinically targetable genomic alterations in acral melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 744.
Abstract 75: Anti-tumor activity of a novel selenonucleoside via targeting Skp2 degradation in paclitaxel-resistant prostate cancer
Authors: Byun Woong Sub , Jin Minkyung , Yu Jinha , Kim Won Kyung , Jeong Lak Shin , Lee Sang Kook 
Published: August 2019

Abstract
Prostate cancer (PC) is the most common disease in men over age 50, and its prevalence rate has been gradually increasing since 1980. Taxane-derived anticancer agents are the primary agents used to treat metastatic prostate cancer patients; however, the side effects and acquired drug resistance limit the success of these therapies. Because there is no specific treatment for paclitaxel-resistant prostate cancer, it is necessary to develop new targets and therapeutic strategies to overcome the acquired resistance. In this study, the antitumor activity of a novel selenonucleoside (4′-selenofuranosyl-2,6-dichloropurine, LJ-2618), a third-generation nucleoside, and its plausible mechanisms of action in paclitaxel-resistant prostate cancer (PC-3-Pa) cells were investigated. The established PC-3-Pa cells exhibited over 100-fold resistance against paclitaxel compared to the paclitaxel-sensitive PC-3 cells. LJ-2618, however, effectively inhibited the proliferation of both cell lines with similar IC50values in vitro. In PC-3-Pa cells, the activated PI3K/Akt signaling pathway was suppressed by LJ-2618 treatment. In addition, Skp2 was found to be over-expressed in paclitaxel-resistant cells, and the transfection of Skp2 siRNA recovered the sensitivity of paclitaxel in PC-3-Pa cells. Furthermore, LJ-2618 significantly down-regulated Skp2 expression in PC-3-Pa cells by promoting degradation and inducing destabilization of Skp2, which triggers G2/M cell cycle arrest. In a xenograft mouse model implanted with PC-3-Pa cells, LJ-2618 (3 or 10 mg/kg) effectively inhibited tumor growth with the enhancement of Skp2 degradation and induction of p27 expression in tumor tissues. These findings suggest that LJ-2618 may have a potential for overcoming paclitaxel resistance via promoting Skp2 degradation and stabilizing p27 expression in PC-3-Pa cells. Therefore, the novel selenonucleoside LJ-2618 may lead to the development of a new treatment strategy for patients with paclitaxel-resistant, castration-resistant prostate cancer. This study was supported by a National Research Foundation of Korea (NRF) Grant funded by the Korean Government (NRF-2016M3A9B6903499).
Citation Format: Woong Sub Byun, Minkyung Jin, Jinha Yu, Won Kyung Kim, Lak Shin Jeong, Sang Kook Lee. Anti-tumor activity of a novel selenonucleoside via targeting Skp2 degradation in paclitaxel-resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 75.
Abstract 753: In situ RNA expression profiling of 1600+ immuno-oncology targets in FFPE tissue using NanoString GeoMx™Digital Spatial Profiler
Authors: Hoang Margaret , Zhou Zoey , Kriner Michelle , Sorg Kristina , Norgaard Zach , Piazza Erin , Merritt Chris , Kim Dae , Beechem Joseph 
Published: August 2019

Abstract
Clinical specimens including formalin-fixed, paraffin embedded (FFPE) tumor sections preserve spatial and molecular information of tumor cells and their surrounding microenvironment. This valuable spatial information is loss with bulk RNA-seq, the most prevalent method for gene expression profiling of archival FFPE samples. In contrast, the NanoString GeoMx™ Digital Spatial Profiler (DSP) is a high multiplexing assay that can profile thousands of RNAs from user selectable regions of interest (ROIs) in FFPE sections. Here we introduce a 1600+ gene panel of tumor, stroma, and immune cell-specific content derived from NanoString nCounter® PanCancer series. We compare RNA profiling using our 1600+ immune-oncology panel to the RNA-seq methodology in FFPE samples. DSP RNA in situ probes are photocleavable oligonucleotides tags coupled to hybridization sequences that bind to mRNA transcripts in the FFPE tissue section. We gridded 96 ROIs upon the FFPE section, photocleaved oligonucleotide tags from each ROI were collected and prepared into sequencing libraries with our NGS readout workflow. After sequencing, reads were charted back to each ROI in the tissue section, generating a map of transcript activity within the tissue. We found high concordance between “bulk” DSP RNA (counts from all 96 gridded ROIs) and RNA-seq from the same FFPE block. Individually, however, each ROI showed different expression patterns than bulk and ROI expression patterns clustered based on similar tissue morphology. We further profiled the tumor and microenvironment compartments from multiple FFPE cancer tissues, comparing our 1600+ RNA expression profile to RNA-seq. DSP was able to detect a higher number of genes with low expression within each spatial compartment compared to bulk RNA-seq. These data demonstrate that DSP offers unparalleled sensitivity for large-scale gene expression while preserving critical information about tissue architecture. GeoMx™ DSP technology is for Research Use Only and not for use in diagnostic procedures.
Citation Format: Margaret Hoang, Zoey Zhou, Michelle Kriner, Kristina Sorg, Zach Norgaard, Erin Piazza, Chris Merritt, Dae Kim, Joseph Beechem. In situ RNA expression profiling of 1600+ immuno-oncology targets in FFPE tissue using NanoString GeoMx™Digital Spatial Profiler [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 753.