bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 793: The prometastatic effects of subcytotoxic doses of doxorubicin are countered by agents inactivating mitochondrial reactive oxygen species
Authors: Porporato Paolo E. , Sonveaux Pierre 
Published: August 2019

Abstract
Mitochondrial reactive oxygen species (mtROS) possess both tumor-promoting and tumor-suppressive activities according to their net concentration resulting for production levels and antioxidant detoxification. mtROS promote cancer cell fitness at low doses (mitohormesis), are cytotoxic at high doses and promote cancer migration, invasion and metastasis at intermediate doses. These intracellular doses vary from a model to another. Yet, we recently identified that mitochondria-targeted ROS scavengers mitoTEMPO and mitoQ prevent cancer metastasis in several in vivo models (Porporato et al. Cell Reports 2014;8:754-66). Interestingly, several anticancer drugs, among which doxorubicin, are well known to trigger mtROS production in cancer cells, but whether this response participates in cell killing or in resistance is still debated. We therefore tested whether combining mitochondria-targeted antioxidants with doxorubicin could help controlling cancer cell proliferation in vitro and tumor growth in mice. Using breast cancer cells lines MCF7 and 4T1 as models, we first found that subcytotoxic doses of doxorubicin increased cell respiration and, thereby, mitochondrial superoxide production. Downstream, mtROS activated the TGFβ pathway and induced cancer cell migration, invasion, epithelial-mesenchymal transition (EMT) and metastatic take in lungs. MitoTEMPO and mitoQ almost totally blocked these phenotypical changes. Together, our data provide a strong rationale for combining doxorubicin chemotherapy with drugs lowering mtROS production in breast cancer.
Citation Format: Paolo E. Porporato, Pierre Sonveaux. The prometastatic effects of subcytotoxic doses of doxorubicin are countered by agents inactivating mitochondrial reactive oxygen species [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 793.
Abstract 796: Progestins target mitochondrial respiration to shift metabolic phenotype in luminal breast cancer
Authors: Matthews Shawna B. , Fettig Lynsey M. , Ward Ashley V. , Jackman Matthew R. , MacLean Paul S. , Sartorius Carol A. 
Published: August 2019

Abstract
The dichotomous nature of progesterone receptor (PR) in estrogen receptor (ER)-positive breast cancers has confounded its use as an endocrine target. Historically, progestins were used to treat some advanced breast cancers and ongoing clinical trials are testing several PR ligands. PR generally opposes ER activity through multiple mechanisms, most notably by altering global ER transcriptional activity to suppress breast tumor growth. However, progestins can also stimulate expansion of endocrine-resistant cancer stem cells (CSCs). Thus, better understanding of potential consequences of progestins in breast cancer is necessary to gauge therapeutic efficacy. Given that ER-driven tumor growth relies on mitochondrial respiration, we evaluated whether PR modulates cell metabolism machinery as a mechanism of ER antagonism. We used functional and morphometric assays in an ER+ breast cancer cell line and patient-derived tumor models to show that progestin treatment dramatically alters mitochondrial function. Multivariate metabolomics analysis highlights a progestin-associated metabolic signature involving accumulation of multiple TCA cycle intermediates, including fumarate, malate, and succinate. Functional assays indicate that progestins impair mitochondrial oxidative reserve capacity and oxygen consumption while ablating estradiol-induced accumulation of ATP in PR+ but not PR− cells. Morphometric assessment of mitochondria via laser-scanning confocal and transmission electron microscopy indicate a shift away from reticular filamentous mitochondria towards smaller, rounder, more perinuclear mitochondria with altered membrane potential. These results suggest that PR targets mitochondrial respiration as part of a broader strategic shift from ER-driven tumor growth to PR-driven thrifty metabolic dormancy. Whether the latter is indicative of an acquired propensity for glycolytic metabolism in indolent tumor cells, CSCs, or perhaps both, is addressed in ongoing studies.
Citation Format: Shawna B. Matthews, Lynsey M. Fettig, Ashley V. Ward, Matthew R. Jackman, Paul S. MacLean, Carol A. Sartorius. Progestins target mitochondrial respiration to shift metabolic phenotype in luminal breast 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 796.
Abstract 805: Adaptation of mitochondrial organization to aggregation in serous ovarian cancer
Authors: Grieco Joseph P. , Edwards Stephanie , Allen Mitchell , Wang Yao , Perry Justin , Song Yipei , Swami Nathan , Brown David , Schmelz Eva 
Published: August 2019

Abstract
During metastasis, ovarian cancer cells exfoliate from the primary tumor and disseminate throughout the peritoneal cavity mostly as aggregates with a higher survival rate and metastatic potential. To determine how the mitochondrial morphology and dynamic expression of fusion and fission-regulating proteins contribute to spheroid survival, in this study we investigated changes in mitochondrial morphology in order to identify phenotypic alterations that are associated with changes in mitochondrial function. We compared monolayers and spheroids of murine and human benign, late-stage, and tumor-initiating cells in hypoxia using fluorescent microscopy and western blot analysis. In addition, we investigated how the mitochondrial membrane potential (Δψm) and ROS production contributed to this phenotype using the InCell analyzer with TMRM and Amplex Red dyes respectively. The data from this study suggest that the morphology and dynamic protein expression are highly altered during tumorigenesis, aggregation, and in hypoxic conditions, indicating a direct correlation of mitochondrial morphology in regulating survival, apoptosis and proliferation. The identification of these molecular events and changes that allow for the survival of metastatic cancer cells in an environment low in oxygen and nutrients may provide specific targets for interventions that would suppress successful metastatic outgrowth of disseminating cells and could potentially be used as a biomarker for treatment efficacy.
Citation Format: Joseph P. Grieco, Stephanie Edwards, Mitchell Allen, Yao Wang, Justin Perry, Yipei Song, Nathan Swami, David Brown, Eva Schmelz. Adaptation of mitochondrial organization to aggregation in serous ovarian 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 805.
Abstract 807: HDAC1 -dependent epigenetic loss of ATP5H confers a stem cell-like phenotype with features associated with metastatic capacity through mitochondrial reprogramming
Authors: Kim Suyeon , Song Kwon-Ho , Kim Tae-Woo 
Published: August 2019

Abstract
The failure of most cancer therapies is the emergence of tumor cells that are refractory to available agent. Here we report that single-modality cancer therapy triggers broad-spectrum cross-resistance to multiple clinical interventions. This multi-modality resistance arises through a core metabolic reprogramming pathway instigated by loss of the ATP synthase subunit ATP5H, which sequentially leads to accumulation of reactive oxygen species and stabilization of HIF-1α under normoxia. Furthermore, we found that NANOG/HDAC1 axis regulates epigenetic loss of ATP5H, this pathway confers tumor cells a stem cell-like phenotype with features associated with metastatic capacity. ATP5H loss in the tumor is strongly linked to failure of therapy, and poor survival in cancer patients. Integration of these biological features provides the best basis for understanding patient outcome in this setting and suggests that loss of ATP5H shapes the tumor resistance to multiple anti-tumor therapy.
Citation Format: Suyeon Kim, Kwon-Ho Song, Tae-Woo Kim. HDAC1 -dependent epigenetic loss of ATP5H confers a stem cell-like phenotype with features associated with metastatic capacity through mitochondrial reprogramming [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 807.
Abstract 818: Changes in tumor suppressor methylation in SKBR3 cells during treatment with Olaparib or Doxorubicin
Authors: Sichmanova Zuzana , Ottesen Asta , Lonning Per Eystein , Berge Elisabet Ognedal , Knappskog Stian 
Published: August 2019

Abstract
DNA methylation is an epigenetic mechanism that can alter gene expression without modifying the DNA sequence. Reactivation of tumor suppressor genes that have been epigenetically silenced in cancer could be a potential therapeutic approach. In this study, we performed long-term treatment of the breast cancer cell line SKBR3, with Olaparib or Doxorubicin in order to assess the methylation status of tumor suppressor genes, after the cell cultures had become resistant to these drugs. Using bisulphite conversion and massive parallel sequencing we examined CpG islands in promoter regions of 283 tumor suppressor genes at several time points under the two treatment regiments. Methylation analysis was carried out for each sample using our in-house bioinformatic pipeline established in collaboration with Roche and publicly available tools. We assessed differentially methylated CpG’s for each drug, compared to control samples treated with dimethyl sulfoxide (DMSO) using the R package methylKit. Gene ontology and pathway analysis revealed hypermethylation in genes belonging to homologous recombination pathway and hypomethylation in genes responding to cellular stress and protein kinase signaling in Olaparib-treated samples. In Doxorubicin-treated samples, we identified genes involved in inter-strand cross-link repair to be hypomethylated, while other genes associated with more general cellular response to DNA damage were hypermethylated. While in Olaparib-treated samples, differential methylation seemed to be reversible and dependent on the dosage of Olaparib, methylation patterns in Doxorubicin-treated samples appeared to change in an incremental manner, as a function of exposure time to the drug.
Citation Format: Zuzana Sichmanova, Asta Ottesen, Per Eystein Lonning, Elisabet Ognedal Berge, Stian Knappskog. Changes in tumor suppressor methylation in SKBR3 cells during treatment with Olaparib or Doxorubicin [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 818.
Abstract 822: PureQuant Methylation Assay for the accurate quantification of immune cell populations
Authors: Pradhan Suman , Dargitz Carl , Guzman Jerry , Lakshmipathy Uma 
Published: August 2019

Abstract
A key challenge during the development and manufacturing of cell based immunotherapies is the reliable estimation of cell purity. Current flow cytometric methods, while effective for the detection of surface antigens such as CD8 that positively identifies Cytotoxic T lymphocytes, are not ideal for intracellular markers used to positively identify Regulatory T (Treg) cells and T Helper 17 (Th17) cells. Additionally, characterization methods that utilize live cells pose a challenge in large scale GMP manufacturing environments due to complicated logistics, limited throughput and difficulty in standardization. Therefore, there is an emerging need for alternative assay methods that address some of these challenges.
Here, we introduce our newly developed PureQuant™ Assays that specifically measure the unique DNA methylation status of specific immune cell types. Assays were developed specifically for Cytotoxic T Cells, Regulatory T cells and T helper 17 cells by detecting methylation status of CD8, FoxP3 and IL17A, respectively, via qPCR of bisulfite converted genomic DNA. These assays are robust with minimal sample requirement, utilizing fresh/frozen cells or isolated gDNA. Results are represented as a percentage of the total cell population similar to flow cytometry based methods. Comparison of the two methods for surface markers such as CD8 show comparable results. Our results demonstrate that this method which combines accuracy, low sample requirement and flexibility, is an ideal measurement system for confirmation of T cell identity and purity that is critical for cell therapy research and development.
Citation Format: Suman Pradhan, Carl Dargitz, Jerry Guzman, Uma Lakshmipathy. PureQuant Methylation Assay for the accurate quantification of immune cell populations [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 822.
Abstract 824: LKB1 loss in lung adenocarcinoma leads to global hypomethylation and altered FOXA binding
Authors: Koenig Michael J. , Amann Joseph , Agana Bernice , Kaufman Jacob , Wysocki Vicki , Oakes Christopher , Carbone David P. 
Published: August 2019

Abstract
LKB1 is functionally lost in 30% of lung adenocarcinoma cases. There are no targeted therapies available for LKB1-loss tumors. Previous studies of LKB1-loss in a mouse model of pancreatic cancer suggested that LKB1 loss drives DNA hypermethylation and serine biogenesis. DNA methyltransferases (DNMTs) maintain CpG methylation, which suppresses gene transcription. The primary substrate for DNMTs is S-adenosyl Methionine (SAMe), and the maintenance of CpG methylation has been shown to be dependent on continued regeneration of SAMe. KRAS status is associated both with LKB1 loss and with a CpG island hypermethylation phenotype (CIMP). Using a previously published 16-gene signature for LKB1 loss, we set out to confirm whether the changes that occur in the mouse model of pancreatic cancer also occur in lung cancer. We applied our signature to samples in The Cancer Genome Atlas (TCGA) lung adenocarcinoma dataset (signature-positive samples are described as “LKB1-loss”), which allowed us to compare both gene expression data as well as methylation data from Illumina 450k microarrays. We then developed a cell-line model for LKB1-loss in order to study the mechanism by which LKB1 regulates DNA methylation.
We observed that LKB1-loss is associated with widespread demethylation of the genome in TCGA samples. Approximately 14,000 CpG sites were significantly hypomethylated in LKB1-loss tumors, while only 200 were hypermethylated (adjusted p-value cutoff = 0.05). Furthermore, Student’s t-test showed that LKB1-loss samples have significantly lower average methylation (β-values), regardless of KRAS status. Using motif analysis, we found that demethylated CpG sites are significantly enriched for FOXA1/2/3 consensus binding sites. FOXA transcription factors are well known as pioneering transcription factors that mediate demethylation. We discovered in vitro that FOXA1 localization and turnover is dependent on LKB1 expression, specifically the activity of downstream kinase SIK. Rather than showing enhanced serine biogenesis, LKB1-loss tumors express less DNMT1 and LC-MS/MS analysis shows depletion of SAMe. Finally, in vitro models of LKB1-loss show an increased sensitivity to DNMT inhibitors and FOXA1 silencing.
Together, these observations suggest that LKB1-loss lung adenocarcinoma patients have a distinct hypomethylation profile. This profile may make them uniquely sensitive to DNMT-inhibition in combination with other therapies, a hypothesis which we will test in future studies.
Citation Format: Michael J. Koenig, Joseph Amann, Bernice Agana, Jacob Kaufman, Vicki Wysocki, Christopher Oakes, David P. Carbone. LKB1 loss in lung adenocarcinoma leads to global hypomethylation and altered FOXA binding [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 824.
Abstract 828: Accelerated epigenetic aging in bladder cancer patients
Authors: Chew Yap Ching , Guo Wei , Yang Xiaojing , Piatti Paolo , Jin Mingda , Booher Keith , Jara Benjamin , Jia Xi-Yu 
Published: August 2019

Abstract
Aging represents the most important risk factor for many chronic diseases including cardiovascular diseases, diabetes, and cancer, therefore understanding the mechanisms of aging is a fundamental step for designing new treatments for chronic diseases. DNA methylation is the most reliable and accurate molecular marker for aging quantification, however, genome-wide DNA methylation profiling techniques, such as reduced representative bisulfite sequencing and Illumina Bead Array that are widely used in aging research are prohibitively expensive and have poor data quality at low-read coverage sites. Here we report a robust targeted bisulfite sequencing approach, called SWARM® (Simplified Whole-panel Amplification Reaction Method), for the accurate biological age determination. SWARM™ is flexible and low cost, requires relatively low DNA starting material, allows the simultaneous amplification and sequencing of hundreds of loci, and has shown to increase sample throughput. Using the SWARM® approach, we were able to analyze the methylation level of several hundreds of age-associated loci including the published Horvath Clock sites. Gender-specific age-predictive models were built using the elastic net regression of DNA methylation levels of the loci and chronological age of urine DNA samples of over 300 healthy subjects of 18 to 88 years old. Urine samples from bladder cancer patients exhibit significant age acceleration, with an average of &gt;10 years. In brief, our gender-specific urine DNAge® analysis is a tool for the precise biological aging quantification and can be used to address questions in aging and urinary track cancers.
Citation Format: Yap Ching Chew, Wei Guo, Xiaojing Yang, Paolo Piatti, Mingda Jin, Keith Booher, Benjamin Jara, Xi-Yu Jia. Accelerated epigenetic aging in bladder cancer patients [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 828.
Abstract 845: c-MET nuclear transportation via membrane-bounded retrograde trafficking in breast cancer cells
Authors: Chen Mei-Kuang , Du Yi , Sun Linlin , Hsu Jennifer L. , Wang Yu-Han , Gao Yuan , Huang Jiaxing , Hung Mien-Chie 
Published: August 2019

Abstract
Reactive oxygen species (ROS) can be induced by anti-cancer treatments, such as ionizing irradiation and chemotherapy drugs. Hydrogen peroxide (H2O2) is one of the ROS species highly produced in cancer cells due to rapid proliferation or anti-cancer treatments. ROS serve as important intracellular messengers in cell growth and proliferation signaling pathways. However, ROS also causes cellular stress in cancer cells through oxidizing DNA, protein and lipids. Therefore, balancing ROS stress between signaling and damages at a tolerable level is important for cancer cell survival. c-MET is one of the receptor tyrosine kinases overexpressed in breast cancer. Full-length c-MET is known to be oncogenic for triggering signal transduction cascade from plasma membrane to upregulate cell proliferation and migration in cancer cells. Moreover, c-MET is reported to phosphorylate PARP1 under H2O2 stimulation, and the phosphorylated PARP1 resulting in PARP inhibitor resistance in breast cancer cells. However, it is not yet clear how c-MET travels from cell membrane to the nucleus. Here, we demonstrate that H2O2 induces retrograde transport of c-MET from plasma membrane into the nucleus and that knocking down either COPI or Sec61β attenuates H2O2-induced full-length nuclear c-MET accumulation. However, c-MET kinase inhibitor did not block the nuclear transport of c-MET, suggesting the transportation mechanisms is independent from kinase activity. Our results indicate that membrane-bound retrograde vesicles involve in membrane-to-nucleus transportation of c-MET in breast cancer cells. Moreover, nuclear c-MET interacts with both PARP1 and Ku proteins in breast cancer cells, suggesting the role of full-length nuclear c-MET in mediating repair of ROS-induced DNA damages.
Citation Format: Mei-Kuang Chen, Yi Du, Linlin Sun, Jennifer L. Hsu, Yu-Han Wang, Yuan Gao, Jiaxing Huang, Mien-Chie Hung. c-MET nuclear transportation via membrane-bounded retrograde trafficking in breast 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 845.
Abstract 852: Determining the roles of ERO1-alpha and ERO1-beta in progression of multiple myeloma and drug resistance
Authors: Chen Weichih , Hazlehurst Lori 
Published: August 2019

Abstract
Multiple myeloma (MM) is a hematological malignancy characterized by dysregulated growth and accumulation of monoclonal plasma cells within the bone marrow. It is a treatable but incurable disease. Despite improvement in progression-free and overall survival of MM patients nearly all MM patients will relapse and become resistant to current treatments. Therefore, to find an effective therapeutic target that is applicable to MM, our laboratory analyzed a retrospective study, patients with relapsed MM who had received bortezomib therapy were enrolled in phase 2 and phase 3 clinical trial. Patients who had higher ERO1-alpha expression demonstrated significant poorer outcomes as compared to patients with lower ERO1-alpha expression. Moreover, patients with relapsed myeloma had higher expression of ERO1-alpha protein levels compared to newly diagnosed MM patients, suggesting that ERO1-alpha correlated with disease progress and poor survival of MM patients. Disulfide bond formation is critical for the folding, stability and function of newly synthesized secretory and membrane proteins in endoplasmic reticulum (ER). Endoplasmic reticulum oxidoreductase 1 (ERO1) is an essential component in this process. ERO1-alpha and ERO1-beta are two flavoprotein isomers that have been identified in mammalian cells. ERO1-alpha is widely expressed and up-regulated during oxidative stress and ERO1-beta is only expressed in select tissue and induced by unfold protein response (UPR). We found that knockdown of ERO1-alpha MM cells caused cell growth inhibition and decreased colony formation and lambda secretion. Moreover, overexpression of ERO1-alpha enhanced cell growth. Additionally, reducing the expression of ERO1-alpha resulted in upregulated ERO1-beta by transcriptional and translational levels. Reducing the expression of ERO1-beta in MM cells increased cell growth and induced resistance to current treatments, including the proteasome inhibitor (bortezomib), alkylating agent (melphalan), corticosteroids (dexamethasome) and histone deacetylase inhibitor (panobinostat). We found reducing the expression of ERO1-beta resulted in decreased expression of glucocorticoid receptor (GR). Taken together, our data indicate that ERO1 -alpha and -beta are not functionally redundant as increased alpha expression correlates with MM progression while increased beta expression correlates with increased sensitivity to chemotherapeutic agents. Future studies are aimed at delineation of mechanism underlying the divergent phenotypes that result from reducing ERO1- alpha or -beta in MM cells.
Citation Format: Weichih Chen, Lori Hazlehurst. Determining the roles of ERO1-alpha and ERO1-beta in progression of multiple myeloma and drug resistance [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 852.