DOCSLIB.ORG

The Novel Arsenical Darinaparsin Circumvents BRG1-Dependent, HO-1-Mediated Cytoprotection in Leukemic Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Novel Arsenical Darinaparsin Circumvents BRG1-Dependent, HO-1-Mediated Cytoprotection in Leukemic Cells Leukemia (2013) 27, 2220–2228 & 2013 Macmillan Publishers Limited All rights reserved 0887-6924/13 www.nature.com/leu ORIGINAL ARTICLE The novel arsenical Darinaparsin circumvents BRG1-dependent, HO-1-mediated cytoprotection in leukemic cells N Garnier1,2, LA Petruccelli1,2, MF Molina1,2, M Kourelis1,2, S Kwan1,2, Z Diaz1,2, HM Schipper2, A Gupta3, SV del Rincon1,2, KK Mann1,2 and WH Miller Jr1,2 Darinaparsin (Dar) is a more potent cytotoxic arsenical than arsenic trioxide (ATO). We hypothesized that the increased cytotoxicity of Dar may be because of a decreased cytoprotective response. We observed that, unlike ATO, Dar does not induce heme oxygenase-1 (HO-1), even though it induces expression of other nuclear factor (erythroid-derived 2)-like 2 (NRF2)-dependent detoxifying enzymes to a greater extent than ATO, in both cancer cell lines and patient-derived leukemic cells. This strengthens the emerging evidence, showing that response to reactive oxygen species (ROS) is stimuli specific. Dar treatment prevents recruitment of the transcriptional coregulator Brahma-related gene 1 (BRG1) to the HMOX1 promoter, which is required for HMOX1 expression. The inability of Dar to induce HO-1 correlates with arrest in G2/M cell cycle phase and BRG1 phosphorylation. Inhibition of HO-1 increases the toxicity of ATO, but has no effect on Dar-induced apoptosis. Accordingly, the lack of HO-1 induction is involved in Dar’s enhanced antileukemic properties. Our data highlight cytoprotective responses mediated by HO-1 and BRG1 as a novel target for enhancing the therapeutic range of arsenicals. Leukemia (2013) 27, 2220–2228; doi:10.1038/leu.2013.54 Keywords: arsenic; leukemia; Darinaparsin; HO-1; BRG1; NRF2 INTRODUCTION we reviewed.1 We postulate that within the signaling differences Darinaparsin (Dar; ZIO-101, S-dimethylarsino-glutathione) is a lies the mechanism responsible for the enhanced antitumor effect promising new arsenic-based, anticancer drug candidate, of Dar. which is currently undergoing clinical studies in both hematolo- In response to oxidative stress, cells induce an arsenal of gical malignancies and solid tumors.1 This organic arsenical, protective proteins including the nuclear factor (erythroid-derived which consists of dimethylarsenic conjugated to glutathione, 2)-like 2 (NRF2) gene battery, which includes heme oxygenase-1 shows significant activity against multiple cancers in vitro.2 (HO-1, HMOX1 gene), NAD(P)H:quinone oxidoreductase (NQO1), Another arsenical, arsenic trioxide (ATO), is a proven thioredoxin-reductase-1 (TR1, TXNRD1 gene) and glutamate- 15 chemotherapy for acute promyelocytic leukemia (APL), but has cysteine ligase regulatory subunit (gGCS). NRF2 is a basic substantially reduced antitumor effects at clinically achievable leucine zipper transcription factor that mediates the expression of doses in other malignancies.3–5 We have shown previously that these key protective enzymes by binding antioxidant response Dar is a more potent inducer of apoptosis than ATO in various elements (ARE)16–18 in the regulatory regions of these genes. malignant cell lines and is highly active against APL cells.2 Under unstressed conditions, Kelch-like ECH-associated protein 1 Importantly, Dar has a maximum tolerated dose that is 50-fold confines NRF2 in the cytoplasm, where it is constitutively higher than ATO in mice and is active against xenograft tumors.6 degraded.19–21 In contrast, ROS liberate NRF2 from Kelch-like However, the mechanisms of its increased antitumor efficacy have ECH-associated protein 1, leading to NRF2 translocation and not been elucidated. accumulation in the nucleus.15 Transcriptional regulation of some Oxidative damage is postulated to be a key mechanism by of these detoxifying enzymes involves a competition between the which arsenicals initiate the apoptotic process.7–12 Consistent with activator NRF2 and the repressor BTB, and CNC homolog 1 its increased cytotoxicity, we have reported that Dar induces (BACH1) for interaction with their AREs.22 Of the NRF2 target - significantly more superoxide (O2 ), hydrogen peroxide and genes, regulation of HMOX1 expression is unique because it is oxidative damage than ATO in arsenic-sensitive NB4 and further regulated by the binding of Brahma-related gene 1 (BRG1, arsenic-resistant AR2 cells.2 Both ATO and Dar-induced ROS SMARCA4 gene), the catalytic subunit of SWI2/SNF2-like chromatin- 23 activate the c-Jun NH2-terminal kinase/stress-activated protein remodeling complex, to the ARE. kinase pathway, leading to apoptosis.2,13,14 However, we have also Here, we show that Dar selectively fails to induce HO-1 reported that Dar triggers apoptosis by inducing cellular expression in leukemic cell lines in vitro and in APL patient blasts responses that do not completely overlap with ATO.1,2 The ex vivo, even though it induces the expression of other NRF2 characterization of similarities versus differences in triggered target genes. We provide evidence that Dar prevents BRG1 pathways and intracellular ROS modulation between ATO and recruitment to the HMOX1 promoter. We also show that specific Dar have elicited interest among the labs working on arsenic, as inhibition of HO-1 enhances the toxicity of ATO, but not Dar, in 1Department of Oncology, Lady Davis Institute for Medical Research, Segal Cancer Center, Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montre´al, Quebec, Canada; 2Faculty of Medicine, Department of Experimental Medicine, McGill University, Montre´al, Quebec, Canada and 3Osta Biotechnologies, Inc., Dollard-des-Ormeaux, Quebec, Canada. Correspondence: Dr WH Miller Jr, Department of Oncology, Lady Davis Institute for Medical Research, Segal Cancer Centre, Sir Mortimer B. Davis Jewish General Hospital, McGill University, 3755 Coˆte Ste-Catherine Road, Montreal, Quebec, Canada H3T 1E2. E-mail: [email protected] Received 9 November 2012; revised 4 February 2013; accepted 11 February 2013; accepted article preview online 21 February 2013; advance online publication, 15 March 2013 Darinaparsin prevents heme oxygenase-1 induction N Garnier et al 2221 leukemic cells. This suggests that the inability of Dar to induce have previously shown that Dar induces more ROS than ATO.2 HO-1 expression is partially responsible for the increased We hypothesized that the enhanced cytotoxicity of Dar is because pro-apoptotic potential we observe. of its inability to elicit a complete antioxidant enzyme response. Therefore, we investigated whether Dar induced significant levels of HO-1, the prototypic arsenic-induced phase II antioxidant MATERIALS AND METHODS enzyme. Figure 1b shows that Dar did not induce HO-1 protein Ethics statement expression in any of the tested cell lines, as opposed to ATO that The patient samples were obtained, collected and used in accordance to strongly induced expression in all cell lines. We were able to the ethical requirements and regulations of the Jewish General Hospital. reproduce these results in HeLa cells, suggesting that our results Informed written consent was obtained from all patients involved. are not limited to hematological malignancies (Figures 1a and b). To verify this observation, we treated both NB4 and U937 cells Cell culture with ATO or Dar and assessed HMOX1 mRNA levels by quantitative All cells were grown in a humidified chamber at 37 1C with 5% CO2. NB4, PCR (qPCR). Treatment with Dar failed to induce significant HMOX1 U937, U266, Kasumi-1, LY8 and HeLa cells were obtained from the expression in either cell line. This result remained consistent in american type culture collection (ATCC) and cultured as recommended. NB4 (Figure 1c) and U937 (Figure 1d), regardless of treatment dose Blasts from APL patients were isolated from either peripheral blood or and/or duration. In addition, we tested all the cell lines in Figure 1a bone marrow (see Supplementary Materials). and found that Dar did not induce significant HMOX1 mRNA expression in any of them (Supplementary Figure 1). Cell viability and caspase-3/7 assay Cell viability and caspase-3/7 activity were assayed using CellTiter-Glo Dar fails to induce HO-1, but not other NRF2 target genes, in NB4 Luminescent Cell Viability Assay and Caspase-Glo 3/7 Assay (Promega, cells Madison, WI, USA) according to manufacturer’s instruction (see Supplementary Materials). We investigated whether Dar would induce significant levels of other phase II antioxidant enzymes, which are induced through activation of the NRF2-signaling pathway.15,17,18 NB4 cells were Propidium iodide staining treated for 3 or 6 h with increasing concentrations of ATO or Dar. As 2 Quantification of apoptotic cells was performed as previously described expected, HO-1 protein levels increased with ATO, but not with Dar (see Supplementary Materials). (Figure 2a). In contrast, Figures 2b, c and d show that both ATO and Dar increase, in a time- and dose-dependent manner, protein levels Protein quantification of TR1, gGCS and NQO1, with Dar consistently inducing expression Cells were lysed, whole-cell extracts were run on SDS-polyacrylamide gel at lower concentrations. To expand our observations, we also tested electrophoresis followed by western blot analysis (see Supplementary the ability of ATO and Dar to induce TR1 and gGCS protein levels in Table 1). Levels of phospho-DNA-PKcs (T2604) were measured by flow Kasumi-1 and U937, which are two non-APL cell lines. As expected, cytometry (see Supplementary Materials). NRF2 protein levels were both ATO and Dar increased protein levels of TR1 and gGCS assessed by densitometry using Image J (imagej.nih.gov). The intensity of each NRF2 band is normalized with the corresponding loading control (Supplementary Figure 2A and 2B). NQO1 protein expression is band (GAPDH or H3) and compared with the corresponding nuclear or shown as a time course because of the transient nature of its cytoplasmic control-treated bands, which were given the value of 1. expression. NQO1 protein levels increase quickly following exposure to arsenicals but decrease at 6 h (Supplementary Figure 2C). Our mRNA analysis data indicate that the activation of HO-1 expression in response to NRF2 signaling is different than for the other NRF2 target genes.
Load more

Recommended publications
  • Genome-Wide CRISPR Screening Identifies BRD9 As a Druggable Component Of
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.04.429732; this version posted February 4, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Genome-Wide CRISPR Screening Identifies BRD9 as a Druggable Component of 2 Interferon-Stimulated Gene Expression and Antiviral Activity 3 Jacob Börolda,b,†, Davide Elettoa,†,#, Idoia Busnadiegoa, Nina K. Maira,b, Eva Moritza, 4 Samira Schiefera,b, Nora Schmidta,$, Philipp P. Petricc,d, W. Wei-Lynn Wonge, Martin 5 Schwemmlec & Benjamin G. Halea,* 6 7 aInstitute of Medical Virology, University of Zurich, Zurich, Switzerland. 8 bLife Science Zurich Graduate School, ETH and University of Zurich, Zurich, 9 Switzerland. 10 cInstitute of Virology, Freiburg University Medical Center, Faculty of Medicine, University 11 of Freiburg, Freiburg, Germany. 12 dSpemann Graduate School of Biology and Medicine, University of Freiburg, Freiburg, 13 Germany. 14 eInstitute of Experimental Immunology, University of Zurich, Zurich, Switzerland. 15 *Correspondence: Benjamin G. Hale, [email protected] 16 †Equal first authors listed in alphabetical order. 17 #Present address: Department of Biosystems Science and Engineering, ETH Zurich, 18 Basel, Switzerland. 19 $Present address: Helmholtz Institute for RNA-based Infection Research, Helmholtz- 20 Center for Infection Research, Wurzburg, Germany. 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.02.04.429732; this version posted February 4, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Increased ACTL6A Occupancy Within Mswi/SNF Chromatin Remodelers
    bioRxiv preprint doi: https://doi.org/10.1101/2021.03.22.435873; this version posted March 22, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Increased ACTL6A Occupancy Within mSWI/SNF Chromatin Remodelers Drives Human Squamous Cell Carcinoma Chiung-Ying Chang1,2,7, Zohar Shipony3,7, Ann Kuo1,2, Kyle M. Loh4,5, William J. Greenleaf3,6, Gerald R. Crabtree1,2,5* 1Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA. 2Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA. 3Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA. 4Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California 94305, USA. 5Department of Developmental Biology, Stanford University School of Medicine, Stanford, California 94305, USA. 6Department of Applied Physics, Stanford University, Stanford, California 94305, USA. 7These authors contributed equally. *Corresponding author: Gerald R. Crabtree, [email protected] Summary Mammalian SWI/SNF (BAF) chromatin remodelers play dosage-sensitive roles in many human malignancies and neurologic disorders. The gene encoding the BAF subunit, ACTL6A, is amplified at an early stage in the development of squamous cell carcinomas (SCCs), but its oncogenic role remains unclear. Here we demonstrate that ACTL6A overexpression leads to its stoichiometric assembly into BAF complexes and drives its interaction and engagement with specific regulatory regions in the genome. In normal epithelial cells, ACTL6A was sub-stoichiometric to other BAF subunits. However, increased ACTL6A levels by ectopic expression or in SCC cells led to near-saturation of ACTL6A within BAF complexes.
    [Show full text]
  • Ten Commandments for a Good Scientist
    Unravelling the mechanism of differential biological responses induced by food-borne xeno- and phyto-estrogenic compounds Ana María Sotoca Covaleda Wageningen 2010 Thesis committee Thesis supervisors Prof. dr. ir. Ivonne M.C.M. Rietjens Professor of Toxicology Wageningen University Prof. dr. Albertinka J. Murk Personal chair at the sub-department of Toxicology Wageningen University Thesis co-supervisor Dr. ir. Jacques J.M. Vervoort Associate professor at the Laboratory of Biochemistry Wageningen University Other members Prof. dr. Michael R. Muller, Wageningen University Prof. dr. ir. Huub F.J. Savelkoul, Wageningen University Prof. dr. Everardus J. van Zoelen, Radboud University Nijmegen Dr. ir. Toine F.H. Bovee, RIKILT, Wageningen This research was conducted under the auspices of the Graduate School VLAG Unravelling the mechanism of differential biological responses induced by food-borne xeno- and phyto-estrogenic compounds Ana María Sotoca Covaleda Thesis submitted in fulfillment of the requirements for the degree of doctor at Wageningen University by the authority of the Rector Magnificus Prof. dr. M.J. Kropff, in the presence of the Thesis Committee appointed by the Academic Board to be defended in public on Tuesday 14 September 2010 at 4 p.m. in the Aula Unravelling the mechanism of differential biological responses induced by food-borne xeno- and phyto-estrogenic compounds. Ana María Sotoca Covaleda Thesis Wageningen University, Wageningen, The Netherlands, 2010, With references, and with summary in Dutch. ISBN: 978-90-8585-707-5 “Caminante no hay camino, se hace camino al andar. Al andar se hace camino, y al volver la vista atrás se ve la senda que nunca se ha de volver a pisar” - Antonio Machado – A mi madre.
    [Show full text]
  • TP53 Mutations As a Driver of Metastasis Signaling in Advanced Cancer Patients
    cancers Article TP53 Mutations as a Driver of Metastasis Signaling in Advanced Cancer Patients Ritu Pandey 1,2,*, Nathan Johnson 3, Laurence Cooke 1, Benny Johnson 4, Yuliang Chen 1, Manjari Pandey 5, Jason Chandler 5 and Daruka Mahadevan 6,* 1 Cancer Center, University of Arizona, Tucson, AZ 85724, USA; [email protected] (L.C.); [email protected] (Y.C.) 2 Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ 85724, USA 3 School of Medicine, Vanderbilt University, Nashville, TN 37325, USA; [email protected] 4 MD Anderson Cancer Center, Houston, TX 77030, USA; [email protected] 5 West Cancer Center, 7945 Wolf River Blvd, Germantown, TN 38138, USA; [email protected] (M.P.); [email protected] (J.C.) 6 Mays Cancer Center, University of Texas Health San Antonio, San Antonio, TX 78229, USA * Correspondence: [email protected] (R.P.); [email protected] (D.M.) Simple Summary: The DNA sequencing of cancer provides information about specific genetic changes that could help with treatment decisions. Tumors from different tissues change over time and acquire new genetic changes particularly with treatment. Our study analyzed the gene changes of 171 advanced cancer patients. We predicted that tumors gain new mutations but that TP53 mutations (guardian of the human genome) are conserved as the tumor progresses from primary to metastatic sites and across tissue types. We analyzed the primary and metastatic site gene changes in 25 tissue types and conducted in-depth analysis of colon and lung cancer sites for substantial changes. TP53 site specific mutations were different across tissue types and suggest different molecular changes.
    [Show full text]
  • SMARCA4 Regulates Spatially Restricted Metabolic Plasticity in 3D Multicellular Tissue
    bioRxiv preprint doi: https://doi.org/10.1101/2021.03.21.436346; this version posted March 22, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. SMARCA4 regulates spatially restricted metabolic plasticity in 3D multicellular tissue Katerina Cermakova1,2,*, Eric A. Smith1,2,*, Yuen San Chan1,2, Mario Loeza Cabrera1,2, Courtney Chambers1,2, Maria I. Jarvis3, Lukas M. Simon4, Yuan Xu5, Abhinav Jain6, Nagireddy Putluri1, Rui Chen7, R. Taylor Ripley5, Omid Veiseh3, and H. Courtney Hodges1,2,3,8,9,‡ 1. Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA 2. Center for Precision Environmental Health, Baylor College of Medicine, Houston, TX, USA 3. Department of Bioengineering, Rice University, Houston, TX, USA 4. Therapeutic Innovation Center, Baylor College of Medicine, Houston, TX, USA 5. Department of Surgery, Division of General Thoracic Surgery, Baylor College of Medicine, Houston, TX, USA 6. Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Houston, TX, USA 7. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA 8. Center for Cancer Epigenetics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA 9. Dan L Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, USA * These authors contributed equally to this work ‡ Corresponding author: H. Courtney Hodges, Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA, [email protected]. Abstract SWI/SNF and related chromatin remodeling complexes act as tissue-specific tumor suppressors and are frequently inactivated in different cancers.
    [Show full text]
  • BRG1 Knockdown Inhibits Proliferation Through Multiple Cellular Pathways in Prostate Cancer Katherine A
    Giles et al. Clin Epigenet (2021) 13:37 https://doi.org/10.1186/s13148-021-01023-7 RESEARCH Open Access BRG1 knockdown inhibits proliferation through multiple cellular pathways in prostate cancer Katherine A. Giles1,2,3, Cathryn M. Gould1, Joanna Achinger‑Kawecka1,4, Scott G. Page2, Georgia R. Kafer2, Samuel Rogers2, Phuc‑Loi Luu1,4, Anthony J. Cesare2, Susan J. Clark1,4† and Phillippa C. Taberlay3*† Abstract Background: BRG1 (encoded by SMARCA4) is a catalytic component of the SWI/SNF chromatin remodelling com‑ plex, with key roles in modulating DNA accessibility. Dysregulation of BRG1 is observed, but functionally uncharacter‑ ised, in a wide range of malignancies. We have probed the functions of BRG1 on a background of prostate cancer to investigate how BRG1 controls gene expression programmes and cancer cell behaviour. Results: Our investigation of SMARCA4 revealed that BRG1 is over‑expressed in the majority of the 486 tumours from The Cancer Genome Atlas prostate cohort, as well as in a complementary panel of 21 prostate cell lines. Next, we utilised a temporal model of BRG1 depletion to investigate the molecular efects on global transcription programmes. Depleting BRG1 had no impact on alternative splicing and conferred only modest efect on global expression. How‑ ever, of the transcriptional changes that occurred, most manifested as down‑regulated expression. Deeper examina‑ tion found the common thread linking down‑regulated genes was involvement in proliferation, including several known to increase prostate cancer proliferation (KLK2, PCAT1 and VAV3). Interestingly, the promoters of genes driving proliferation were bound by BRG1 as well as the transcription factors, AR and FOXA1.
    [Show full text]
  • Intrinsic Disorder of the BAF Complex: Roles in Chromatin Remodeling and Disease Development
    International Journal of Molecular Sciences Article Intrinsic Disorder of the BAF Complex: Roles in Chromatin Remodeling and Disease Development Nashwa El Hadidy 1 and Vladimir N. Uversky 1,2,* 1 Department of Molecular Medicine, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd. MDC07, Tampa, FL 33612, USA; [email protected] 2 Laboratory of New Methods in Biology, Institute for Biological Instrumentation of the Russian Academy of Sciences, Federal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Pushchino, 142290 Moscow Region, Russia * Correspondence: [email protected]; Tel.: +1-813-974-5816; Fax: +1-813-974-7357 Received: 20 September 2019; Accepted: 21 October 2019; Published: 23 October 2019 Abstract: The two-meter-long DNA is compressed into chromatin in the nucleus of every cell, which serves as a significant barrier to transcription. Therefore, for processes such as replication and transcription to occur, the highly compacted chromatin must be relaxed, and the processes required for chromatin reorganization for the aim of replication or transcription are controlled by ATP-dependent nucleosome remodelers. One of the most highly studied remodelers of this kind is the BRG1- or BRM-associated factor complex (BAF complex, also known as SWItch/sucrose non-fermentable (SWI/SNF) complex), which is crucial for the regulation of gene expression and differentiation in eukaryotes. Chromatin remodeling complex BAF is characterized by a highly polymorphic structure, containing from four to 17 subunits encoded by 29 genes. The aim of this paper is to provide an overview of the role of BAF complex in chromatin remodeling and also to use literature mining and a set of computational and bioinformatics tools to analyze structural properties, intrinsic disorder predisposition, and functionalities of its subunits, along with the description of the relations of different BAF complex subunits to the pathogenesis of various human diseases.
    [Show full text]
  • SMARCA4 Regulates Gene Expression and Higher-Order Chromatin Structure in Proliferating Mammary Epithelial Cells
    Downloaded from genome.cshlp.org on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press Research SMARCA4 regulates gene expression and higher- order chromatin structure in proliferating mammary epithelial cells A. Rasim Barutcu,1 Bryan R. Lajoie,2 Andrew J. Fritz,3 Rachel P. McCord,4 Jeffrey A. Nickerson,1 Andre J. van Wijnen,5 Jane B. Lian,3 Janet L. Stein,3 Job Dekker,2,6 Gary S. Stein,3 and Anthony N. Imbalzano1 1Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA; 2Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA; 3Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont 05405, USA; 4Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee 37996, USA; 5Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA; 6Howard Hughes Medical Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA The packaging of DNA into chromatin plays an important role in transcriptional regulation and nuclear processes. Brahma- related gene-1 SMARCA4 (also known as BRG1), the essential ATPase subunit of the mammalian SWI/SNF chromatin re- modeling complex, uses the energy from ATP hydrolysis to disrupt nucleosomes at target regions. Although the transcrip- tional role of SMARCA4 at gene promoters is well-studied, less is known about its role in higher-order genome organization. SMARCA4 knockdown in human mammary epithelial MCF-10A cells resulted in 176 up-regulated genes, includ- ing many related to lipid and calcium metabolism, and 1292 down-regulated genes, some of which encode extracellular ma- trix (ECM) components that can exert mechanical forces and affect nuclear structure.
    [Show full text]
  • A Role for SMARCB1 in Synovial Sarcomagenesis Reveals That SS18-SSX Induces Canonical BAF Destruction
    Author Manuscript Published OnlineFirst on June 2, 2021; DOI: 10.1158/2159-8290.CD-20-1219 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. SMARCB1 in Synovial Sarcomagenesis 1 A role for SMARCB1 in synovial sarcomagenesis reveals that SS18-SSX induces canonical BAF destruction Jinxiu Li*1,2,3, Timothy S. Mulvihill*2,3, Li Li1,2,3, Jared J. Barrott1,2,3, Mary L. Nelson1,2,3, Lena Wagner6, Ian C. Lock1,2,3, Amir Pozner1,2,3, Sydney Lynn Lambert1,2,3, Benjamin B. Ozenberger1,2,3, Michael B. Ward3,4, Allie H. Grossmann3,4, Ting Liu3,4, Ana Banito6, Bradley R. Cairns2,3,5† and Kevin B. Jones1,2,3† 1Department of Orthopaedics, 2Department of Oncological Sciences, 3Huntsman Cancer Institute, 4Department of Pathology, 5Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah. 6Hopp Children’s Cancer Center (KiTZ), German Cancer Research Center (DFKZ), Heidelberg, Germany. *These authors contributed equally to this work. †These authors are co-corresponding authors. Please Address Correspondence to: Kevin B. Jones and Bradley R. Cairns Address: 2000 Circle of Hope Drive, Salt Lake City, UT 84112 Phone: 801-585-0300 Fax: 801-585-7084 Email: [email protected], [email protected] Running Title: SMARCB1 in Synovial Sarcomagenesis Key Words: SWI/SNF; Chromatin Remodeling; Mouse Genetic Model; Epigenetics; Biochemistry Financial Support: This work was supported by R01CA201396 (Jones and Cairns), U54CA231652 (Jones, Cairns, and Banito), and 2P30CA042014-31, from the National Cancer Institute (NCI/NIH), as well as the Paul Nabil Bustany Fund for Synovial Sarcoma Research (Jones), and the Sarcoma Foundation of America (Barrott).
    [Show full text]
  • Residual Complexes Containing SMARCA2 (BRM) Underlie the Oncogenic Drive of SMARCA4 (BRG1) Mutation
    Residual Complexes Containing SMARCA2 (BRM) Underlie the Oncogenic Drive of SMARCA4 (BRG1) Mutation The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Wilson, B. G., K. C. Helming, X. Wang, Y. Kim, F. Vazquez, Z. Jagani, W. C. Hahn, and C. W. M. Roberts. 2014. “Residual Complexes Containing SMARCA2 (BRM) Underlie the Oncogenic Drive of SMARCA4 (BRG1) Mutation.” Molecular and Cellular Biology 34 (6): 1136–44. doi:10.1128/MCB.01372-13. Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:41542741 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA Residual Complexes Containing SMARCA2 (BRM) Underlie the Oncogenic Drive of SMARCA4 (BRG1) Mutation Boris G. Wilson,a Katherine C. Helming,a Xiaofeng Wang,a Youngha Kim,a Francisca Vazquez,c,d Zainab Jagani,f William C. Hahn,c,d,e Charles W. M. Robertsa,b,d,e Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, USAa; Division of Hematology/Oncology, Boston Children’s Hospital, Boston, Massachusetts, USAb; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, USAc; Cancer Program, Broad Institute of Harvard and Massachusetts Institute of Technology, Boston, Massachusetts, USAd; Center for Cancer Genome Discovery, Dana-Farber Cancer Institute, Boston, Massachusetts, USAe; Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USAf Collectively, genes encoding subunits of the SWI/SNF (BAF) chromatin remodeling complex are mutated in 20% of all human Downloaded from cancers, with the SMARCA4 (BRG1) subunit being one of the most frequently mutated.
    [Show full text]
  • Comprehensive Analysis of SWI/SNF Inactivation in Lung Adenocarcinoma Cell Models
    cancers Article Comprehensive Analysis of SWI/SNF Inactivation in Lung Adenocarcinoma Cell Models 1,2, 1,2, 2,3,4, 2,3,4, Paola Peinado y, Alvaro Andrades y , Marta Cuadros y, Maria Isabel Rodriguez y , Isabel F. Coira 5,6 , Daniel J. Garcia 2,4 , Juan Carlos Álvarez-Perez 1,2 , Carlos Baliñas-Gavira 1,2, Alberto M. Arenas 1,2 , Juan Rodrigo Patiño-Mercau 1,2 , Juan Sanjuan-Hidalgo 2 , Octavio A. Romero 7, Luis M. Montuenga 8,9 , Julian Carretero 10, Montserrat Sanchez-Cespedes 7 and Pedro P. Medina 1,2,3,* 1 Department of Biochemistry and Molecular Biology I, University of Granada, 18071 Granada, Spain; [email protected] (P.P.); [email protected] (A.A.); [email protected] (J.C.Á.-P.); [email protected] (C.B.-G.); [email protected] (A.M.A.); [email protected] (J.R.P.-M.) 2 GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, 18016 Granada, Spain; [email protected] (M.C.); [email protected] (M.I.R.); [email protected] (D.J.G.); [email protected] (J.S.-H.) 3 Health Research Institute of Granada (ibs.Granada), 18014 Granada, Spain 4 Department of Biochemistry and Molecular Biology III and Immunology, University of Granada, 18016 Granada, Spain 5 School of Pharmaceutical Sciences, University of Geneva, 1211 Geneva, Switzerland; [email protected] 6 Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, 1211 Geneva, Switzerland 7 Genes and Cancer Group, Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research
    [Show full text]
  • PRC2-Mediated Repression of SMARCA2 Predicts EZH2 Inhibitor Activity in SWI/SNF Mutant Tumors
    PRC2-mediated repression of SMARCA2 predicts EZH2 inhibitor activity in SWI/SNF mutant tumors Thomas Januarioa,1, Xiaofen Yea,1, Russell Bainerb, Bruno Alickec, Tunde Smitha, Benjamin Haleyd, Zora Modrusand, Stephen Gouldc, and Robert L. Yaucha,2 aDepartment of Discovery Oncology, Genentech, Inc., South San Francisco, CA 94080; bDepartment of Bioinformatics, Genentech, Inc., South San Francisco, CA 94080; cDepartment of Translational Oncology, Genentech, Inc., South San Francisco, CA 94080; and dDepartment of Molecular Biology, Genentech, Inc., South San Francisco, CA 94080 Edited by Joan S. Brugge, Harvard Medical School, Boston, MA, and approved October 3, 2017 (received for review March 8, 2017) Subunits of the SWI/SNF chromatin remodeling complex are fre- of the ovary, hypercalcemic-type (SCCOHT) (3, 7–9). Although the quently mutated in human cancers leading to epigenetic dependen- mechanisms underlying tumorigenesis in these specific contexts cies that are therapeutically targetable. The dependency on the have yet to be fully elucidated, data are further supportive of a polycomb repressive complex (PRC2) and EZH2 represents one such tumor-suppressive function (10, 11). vulnerability in tumors with mutations in the SWI/SNF complex Efforts to therapeutically target SWI/SNF-defective cancers subunit, SNF5; however, whether this vulnerability extends to other have focused on identifying novel vulnerabilities that may be a SWI/SNF subunit mutations is not well understood. Here we show consequence of the altered chromatin state caused by muta- that a subset of cancers harboring mutations in the SWI/SNF ATPase, tions in BAF complex subunits. One such described vulnera- bility was based on the initial discovery in Drosophila of an SMARCA4, is sensitive to EZH2 inhibition.
    [Show full text]