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      Multiple Active Compounds from Viscum album L. Synergistically Converge to Promote Apoptosis in Ewing Sarcoma

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          Abstract

          Ewing sarcoma is the second most common bone cancer in children and adolescents, with poor prognosis and outcome in ~70% of initial diagnoses and 10–15% of relapses. Hydrophobic triterpene acids and hydrophilic lectins and viscotoxins from European mistletoe ( Viscum album L.) demonstrate anticancer properties, but have not yet been investigated for Ewing sarcoma. Commercial Viscum album L. extracts are aqueous, excluding the insoluble triterpenes. We recreated a total mistletoe effect by combining an aqueous extract (viscum) and a triterpene extract (TT) solubilized with cyclodextrins. Ewing sarcoma cells were treated with viscum, TT and viscumTT in vitro, ex vivo and in vivo. In vitro and ex vivo treatment of Ewing sarcoma cells with viscum inhibited proliferation and induced apoptosis in a dose-dependent fashion, while viscumTT combination treatment generated a synergistic effect. Apoptosis occurred via intrinsic and extrinsic apoptotic pathways, evidenced by activation of both CASP8 and CASP9. We show that viscumTT treatment shifts the balance of apoptotic regulatory proteins towards apoptosis, mainly via CLSPN, MCL1, BIRC5 and XIAP downregulation. ViscumTT also demonstrated strong antitumor activity in a cell line- and patient-derived mouse model, and may be considered an adjuvant therapy option for pediatric patients with Ewing sarcoma.

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          Most cited references54

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          Natural products and drug discovery. Can thousands of years of ancient medical knowledge lead us to new and powerful drug combinations in the fight against cancer and dementia?

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            Risk of recurrence and survival after relapse in patients with Ewing sarcoma.

            The prognosis in patients with relapsed Ewing sarcoma is unfavorable. Our investigation identifies factors predicting for the outcome following relapse. We analyzed type of relapse, time to relapse and overall survival after relapse (OSr) in 714 patients with first recurrence. All patients had been treated within the Cooperative Ewing Sarcoma Studies (CESS) 81 or 86, or the European Intergroup CESS (EICESS 92). OSr time was calculated from diagnosis of first relapse to last follow-up or death. Median follow-up time from diagnosis of primary disease was 2.2 years (mean = 4.0; range: 0.2-24.9). Relapse sites were local in 15%, combined local and systemic in 12%, and systemic in 73%. Among patients with a localized primary tumor, 20% relapsed locally, while 12% showed combined and 68% systemic relapse. When the primary disease was disseminated, 82% developed systemic, 13% combined, and 5% local relapse. Five-year OSr was 0.13 (SE = 0.01). Outcome following local relapse, with a 5-year survival rate of 0.24 (P 2 years) and strictly localized relapse. Copyright © 2011 Wiley-Liss, Inc.
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              X-Linked Inhibitor of Apoptosis Protein – A Critical Death Resistance Regulator and Therapeutic Target for Personalized Cancer Therapy

              Defects in apoptosis regulation are one main cause of cancer development and may result from overexpression of anti-apoptotic proteins such as inhibitor of apoptosis proteins (IAPs). IAPs are cell death regulators that, among other functions, bind caspases, and interfere with apoptotic signaling via death receptors or intrinsic cell death pathways. All IAPs share one to three common structures, the so called baculovirus-IAP-repeat (BIR)-domains that allow them to bind caspases and other proteins. X-linked inhibitor of apoptosis protein (XIAP) is the most potent and best-defined anti-apoptotic IAP family member that directly neutralizes caspase-9 via its BIR3 domain and the effector caspases-3 and -7 via its BIR2 domain. A natural inhibitor of XIAP is SMAC/Diablo, which is released from mitochondria in apoptotic cells and displaces bound caspases from the BIR2/BIR3 domains of XIAP thereby reactivating cell death execution. The central apoptosis-inhibitory function of XIAP and its overexpression in many different types of advanced cancers have led to significant efforts to identify therapeutics that neutralize its anti-apoptotic effect. Most of these drugs are chemical derivatives of the N-terminal part of SMAC/Diablo. These “SMAC-mimetics” either specifically induce apoptosis in cancer cells or act as drug-sensitizers. Several “SMAC-mimetics” are currently tested by the pharmaceutical industry in Phase I and Phase II trials. In this review, we will discuss recent advances in understanding the function of IAPs in normal and malignant cells and focus on approaches to specifically neutralize XIAP in cancer cells.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 September 2016
                2016
                : 11
                : 9
                : e0159749
                Affiliations
                [1 ]Department of Pediatric Oncology/Hematology, Otto Heubner Centre for Pediatric and Adolescent Medicine (OHC), Charité, Universitätsmedizin Berlin, Germany
                [2 ]Institute of Pharmacy, Department of Biology, Chemistry, Pharmacy, Freie Universität, Berlin, Germany
                [3 ]EPO GmbH, Experimental Pharmacology & Oncology, Berlin, Germany
                [4 ]Birken AG, Niefern-Öschelbronn, Germany
                Johns Hopkins University, UNITED STATES
                Author notes

                Competing Interests: The authors have the following interests: Jana Rolff is employed by EPO GmbH and Sebastian Jäger by Birken AG. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

                • Conceived and designed the experiments: GS CD MT.

                • Performed the experiments: MT SK JR.

                • Analyzed the data: MT CD GS.

                • Contributed reagents/materials/analysis tools: SJ JR AE.

                • Wrote the paper: MT AE.

                Article
                PONE-D-15-48437
                10.1371/journal.pone.0159749
                5010293
                27589063
                623eff5f-3608-42bd-afaa-a9020557e644
                © 2016 Twardziok et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 24 November 2015
                : 7 July 2016
                Page count
                Figures: 7, Tables: 0, Pages: 18
                Funding
                Funded by: Software AG Stiftung
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100006360, Bundesministerium für Wirtschaft und Energie;
                Award Recipient :
                This work was supported by the Software AG-Stiftung (Darmstadt, Germany) and the Federal Ministry for Economic Affairs and Energy on the basis of a decision by the German Bundestag. EPO GmbH and Birken AG provided support in the form of salaries for authors JR and SJ respectively, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Sarcomas
                Ewing Sarcoma
                Medicine and Health Sciences
                Oncology
                Cancer Treatment
                Biology and Life Sciences
                Biochemistry
                Proteins
                Lectins
                Research and Analysis Methods
                Model Organisms
                Animal Models
                Mouse Models
                Research and Analysis Methods
                Spectrum Analysis Techniques
                Spectrophotometry
                Cytophotometry
                Flow Cytometry
                Biology and Life Sciences
                Biochemistry
                Bioenergetics
                Energy-Producing Organelles
                Mitochondria
                Mitochondrial Membrane
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Energy-Producing Organelles
                Mitochondria
                Mitochondrial Membrane
                Biology and Life Sciences
                Toxicology
                Cytotoxicity
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Cytotoxicity
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

                Uncategorized
                Uncategorized

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