DOCSLIB.ORG

BRD4 Regulates Breast Cancer Dissemination Through Jagged1/Notch1 Signaling Guillaume Andrieu1, Anna H

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
BRD4 Regulates Breast Cancer Dissemination Through Jagged1/Notch1 Signaling Guillaume Andrieu1, Anna H Published OnlineFirst September 20, 2016; DOI: 10.1158/0008-5472.CAN-16-0559 Cancer Molecular and Cellular Pathobiology Research BRD4 Regulates Breast Cancer Dissemination through Jagged1/Notch1 Signaling Guillaume Andrieu1, Anna H. Tran1, Katherine J. Strissel1, and Gerald V. Denis1,2 Abstract The bromodomain and extraterminal (BET) proteins are epige- expression and Notch1 signaling. BRD4-selective knockdown sup- netic "readers" of acetylated histones in chromatin and have been pressed Notch1 activity and impeded breast cancer migration and identified as promising therapeutic targets in diverse cancers. How- invasion. BRD4 was required for IL6-stimulated, Notch1-induced ever, it remains unclear how individual family members participate migration and invasion, coupling microenvironment inflamma- in cancer progression and small molecule inhibitors such as JQ1 tion with cancer propagation. Moreover, in patients, BRD4 and can target functionally independent BET proteins. Here, we report a Jagged1expression positivelycorrelated with the presence of distant signaling pathway involving BRD4 and the ligand/receptor pair metastases. These results identify a BRD4/Jagged1/Notch1 signal- Jagged1/Notch1 that sustains triple-negative breast cancer migra- ing pathway that is critical for dissemination of triple-negative tion and invasion. BRD4, but not BRD2 or BRD3, regulated Jagged1 breast cancer. Cancer Res; 76(22); 1–13. Ó2016 AACR. Introduction recent study reported that BRD4 interacts with the transcription factor Twist to regulate Twist-dependent transcription pro- The bromodomain and extra-terminal (BET) proteins form a grams and elicit breast cancer tumorigenesis (20). However, family of chromatin-associated proteins that recognize epige- the molecular mechanisms by which BET proteins participate in netic marks, such as N-e-acetylated lysine residues in nucleo- cancer progression are not fully established. We build on these somal histones. BET proteins interact with chromatin and insights to develop a hypothesis that BET proteins play a role in recruit members of the transcription machinery to target pro- the regulation of breast cancer aggressiveness. moters to regulate gene expression (1, 2). In mammals, the BET Triple-negative breast cancer is an aggressive subtype that family is composed of three ubiquitously expressed proteins: lacks estrogen receptor, progesterone receptor, and epidermal BRD2, BRD3, BRD4, and a testis-specificform,BRDT(3).We growth factor receptor 2. These tumors are extremely challeng- were the first to establish a link between expression and ing for therapy. Patients with triple-negative breast cancer function of a BET protein and human cancer (3). A consistent cannot be treated by hormone therapy and present a worse literature has since reported essential roles of BRD2 and BRD4 outcome after conventional chemotherapy (21). Moreover, the in cell-cycle control and cell proliferation, notably through lack of identified "druggable" molecular targets severely limits their crucial role in regulating gene transcription (4–7). Like the development of targeted therapeutic strategies. Triple-neg- numerous other chromatin-regulatory proteins, the cancers in ative breast cancer cells activate multiple transcription pro- which BET proteins have been implicated are diverse. Small- grams, such as the epithelial–mesenchymal transition (EMT), molecule BET-specific bromodomain inhibitors, e.g., (S)-JQ1 which confers high plasticity, increased migration, and inva- (8), highlight the therapeutic impact of targeting BET proteins sion capabilities that can promote tumor dissemination (22). in important malignancies, including acute myeloid leukemia The breast tumor microenvironment is composed of various (9, 10), B-cell lymphoma (11), lung (12), prostate (13), breast cell types, including adipocytes; fibroblasts; infiltrating (14), pancreatic (15), and colorectal cancers (16). Several immune cells, such as macrophages and lymphocytes; and studies have reported that BET inhibition downregulates the stem cells (23). Numerous factors secreted within this micro- proto-oncogene MYC and its associated transcription (9, 10, environment have been reported to sustain tumor progression, 17, 18). Moreover, BRD2 has been shown to associate with including growth factors, hormones, cytokines, and chemo- chromatin remodeling complexes to regulate transcription pro- kines. Amid this complexity, inflammatory signaling molecules grams associated with cell proliferation (19). Interestingly, a have focused investigators' attention, because a direct associ- ation exists between chronic inflammation and cancer devel- opment (24). Notably, multiple proinflammatory mediators 1Cancer Center, Boston University School of Medicine, Boston, Mas- sustain the acquisition of an aggressive phenotype by tumor sachusetts. 2Department of Pharmacology and Experimental Thera- cells that can lead to resistance and dissemination. Significant- peutics, Boston University School of Medicine, Boston, Massachusetts. ly, BET protein inhibitors like (S)-JQ1 have anti-inflammatory Corresponding Author: Gerald V. Denis, Boston University School of Medicine, properties (25, 26), suggesting that BET protein targeting may 72 East Concord Street, Room K520, Boston, MA 02118. Phone: 617-414-1371, disrupt inflammation-induced cancer aggressiveness. Here, we Fax: 617-638-5673; E-mail: [email protected] identify a role for BRD4 function in invasive properties of doi: 10.1158/0008-5472.CAN-16-0559 triple-negative breast cancer and reveal the underlying molec- Ó2016 American Association for Cancer Research. ular mechanisms. www.aacrjournals.org OF1 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst September 20, 2016; DOI: 10.1158/0008-5472.CAN-16-0559 Andrieu et al. Materials and Methods Samples containing 25 mg of protein were resolved by SDS- PAGEandtransferredontonitrocellulosemembranes.After Cell culture saturation of non-specific binding sites in TBS-BSA 5%, mem- Human breast cancer cell lines maintained at the NCI Office of branes were probed with primary antibodies, then visualized Physical Sciences-Oncology Centers (PS-OC) Network Biore- with HRP-conjugated secondary antibodies. After incubation in source Core Facility (PBCF) were contractually obtained through ECL, membrane-associated light emission was quantified with the ATCC, under a Material Transfer Agreement. The cell lines a gel imager. have been authenticated by the NIH Physical Sciences Oncology Consortium. MDA-MB-231 and MCF-7 were cultured in DMEM. SUM149PT cells were cultured in DMEM/F12 þ 5 mg/mL insulin qRT-PCR and 0.5 mg/mL hydrocortisone (Sigma). All culture media were Total RNA was extracted using the RNEasy kit (Qiagen). Reverse supplemented with 10% fetal bovine serum (FBS) or human transcription reactions were performed on 1 mg of RNA with the serum (Millipore) where specified and 1% antibiotics (penicil- QuantiTect Reverse Transcription kit (Qiagen). The primers lin/streptomycin; Thermo Fischer Scientific). Cells were cultured sequences used for this study are listed in Supplementary Table S1. PCR amplifications were performed with the MESA GREEN at 37 C in a 5% CO2 atmosphere. qPCR MasterMix (Eurogentec) on an ABI Prism 7500 thermal Antibodies and reagents cycler. 2 fi The following antibodies were used: anti-BRD2, BRD3 and The gene screening was conducted with the RT Pro ler PCR BRD4 (Bethyl Laboratories), anti-Jagged1 and anti-a-tubulin EMT Array (Qiagen). (Santa Cruz Biotechnology), anti-phosphoY705-STAT3, total STAT3, and anti-Notch1 Val1744 (Cell Signaling Technology). Immunocytochemistry staining and confocal imaging HRP-conjugated secondary antibodies were purchased from Cells were fixed in absolute methanol for 5 minutes at À20 C Bio-Rad. Fluorochrome-conjugated secondary antibodies were and then permeabilized with 0.2% Triton X-100 in PBS buffer for obtained from The Jackson Laboratory. (R)- and (S)-JQ1 were 10 minutes. After saturation in blocking buffer (0.02% Triton purchased from Cayman Chemicals. Recombinant human IL6 X-100, 2% BSA in PBS) for 30 minutes, cells were incubated with and DAPT were purchased from Sigma. primary antibodies and then fluorochrome-conjugated secondary antibodies, both diluted in blocking buffer for 1 hours. Finally, Plasmids, siRNAs, and transfection coverslips were mounted with ProLong Gold with DAPI (Thermo fi Plasmids coding for His-tagged BET proteins, His-Jagged1, or Fischer Scienti c). Image acquisition was conducted using a Leica control vector pReceiver-M01 were purchased from GeneCo- SP5 confocal microscope. For z-stack acquisition, a step of 0.3 mm poeia. ON-TARGETplus BET protein siRNAs were obtained from was set. For calculation of NICD1 nuclear score, cells stained for Dharmacon and Jagged1 siRNA (sc-3702) from Santa Cruz Bio- NICD1 were entirely imaged in 3D and then z-stacks were technology. Cells were transfected with plasmids and siRNAs by projected into a single image by sum projection using ImageJ Lipofectamine 2000 reagent (Thermo Fischer Scientific). software (NIH). A mask based on DAPI staining was created to isolate the nucleus of each cell. Based on that mask, nuclear Migration and invasion assays background-corrected NICD1 intensity was measured. The cyto- For scratch assays, 100,000 cells were plated in 6-well plates. plasm area was determined by subtracting the nuclear mask from When confluence was reached, cell monolayers were scraped the entire cell area and cytoplasmic
Load more

Recommended publications
  • Functional Roles of Bromodomain Proteins in Cancer
    cancers Review Functional Roles of Bromodomain Proteins in Cancer Samuel P. Boyson 1,2, Cong Gao 3, Kathleen Quinn 2,3, Joseph Boyd 3, Hana Paculova 3 , Seth Frietze 3,4,* and Karen C. Glass 1,2,4,* 1 Department of Pharmaceutical Sciences, Albany College of Pharmacy and Health Sciences, Colchester, VT 05446, USA; [email protected] 2 Department of Pharmacology, Larner College of Medicine, University of Vermont, Burlington, VT 05405, USA; [email protected] 3 Department of Biomedical and Health Sciences, University of Vermont, Burlington, VT 05405, USA; [email protected] (C.G.); [email protected] (J.B.); [email protected] (H.P.) 4 University of Vermont Cancer Center, Burlington, VT 05405, USA * Correspondence: [email protected] (S.F.); [email protected] (K.C.G.) Simple Summary: This review provides an in depth analysis of the role of bromodomain-containing proteins in cancer development. As readers of acetylated lysine on nucleosomal histones, bromod- omain proteins are poised to activate gene expression, and often promote cancer progression. We examined changes in gene expression patterns that are observed in bromodomain-containing proteins and associated with specific cancer types. We also mapped the protein–protein interaction network for the human bromodomain-containing proteins, discuss the cellular roles of these epigenetic regu- lators as part of nine different functional groups, and identify bromodomain-specific mechanisms in cancer development. Lastly, we summarize emerging strategies to target bromodomain proteins in cancer therapy, including those that may be essential for overcoming resistance. Overall, this review provides a timely discussion of the different mechanisms of bromodomain-containing pro- Citation: Boyson, S.P.; Gao, C.; teins in cancer, and an updated assessment of their utility as a therapeutic target for a variety of Quinn, K.; Boyd, J.; Paculova, H.; cancer subtypes.
    [Show full text]
  • Function of Bromodomain and Extra-Terminal Motif Proteins (Bets) in Gata1-Mediated Transcription
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2015 Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription Aaron James Stonestrom University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Molecular Biology Commons, and the Pharmacology Commons Recommended Citation Stonestrom, Aaron James, "Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription" (2015). Publicly Accessible Penn Dissertations. 1148. https://repository.upenn.edu/edissertations/1148 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1148 For more information, please contact [email protected]. Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription Abstract Bromodomain and Extra-Terminal motif proteins (BETs) associate with acetylated histones and transcription factors. While pharmacologic inhibition of this ubiquitous protein family is an emerging therapeutic approach for neoplastic and inflammatory disease, the mechanisms through which BETs act remain largely uncharacterized. Here we explore the role of BETs in the physiologically relevant context of erythropoiesis driven by the transcription factor GATA1. First, we characterize functions of the BET family as a whole using a pharmacologic approach. We find that BETs are broadly required for GATA1-mediated transcriptional activation, but that repression is largely BET-independent. BETs support activation by facilitating both GATA1 occupancy and transcription downstream of its binding. Second, we test the specific olesr of BETs BRD2, BRD3, and BRD4 in GATA1-activated transcription. BRD2 and BRD4 are required for efficient anscriptionaltr activation by GATA1. Despite co-localizing with the great majority of GATA1 binding sites, we find that BRD3 is not equirr ed for GATA1-mediated transcriptional activation.
    [Show full text]
  • Bromodomain Protein BRDT Directs ΔNp63 Function and Super
    Cell Death & Differentiation (2021) 28:2207–2220 https://doi.org/10.1038/s41418-021-00751-w ARTICLE Bromodomain protein BRDT directs ΔNp63 function and super-enhancer activity in a subset of esophageal squamous cell carcinomas 1 1 2 1 1 2 Xin Wang ● Ana P. Kutschat ● Moyuru Yamada ● Evangelos Prokakis ● Patricia Böttcher ● Koji Tanaka ● 2 3 1,3 Yuichiro Doki ● Feda H. Hamdan ● Steven A. Johnsen Received: 25 August 2020 / Revised: 3 February 2021 / Accepted: 4 February 2021 / Published online: 3 March 2021 © The Author(s) 2021. This article is published with open access Abstract Esophageal squamous cell carcinoma (ESCC) is the predominant subtype of esophageal cancer with a particularly high prevalence in certain geographical regions and a poor prognosis with a 5-year survival rate of 15–25%. Despite numerous studies characterizing the genetic and transcriptomic landscape of ESCC, there are currently no effective targeted therapies. In this study, we used an unbiased screening approach to uncover novel molecular precision oncology targets for ESCC and identified the bromodomain and extraterminal (BET) family member bromodomain testis-specific protein (BRDT) to be 1234567890();,: 1234567890();,: uniquely expressed in a subgroup of ESCC. Experimental studies revealed that BRDT expression promotes migration but is dispensable for cell proliferation. Further mechanistic insight was gained through transcriptome analyses, which revealed that BRDT controls the expression of a subset of ΔNp63 target genes. Epigenome and genome-wide occupancy studies, combined with genome-wide chromatin interaction studies, revealed that BRDT colocalizes and interacts with ΔNp63 to drive a unique transcriptional program and modulate cell phenotype. Our data demonstrate that these genomic regions are enriched for super-enhancers that loop to critical ΔNp63 target genes related to the squamous phenotype such as KRT14, FAT2, and PTHLH.
    [Show full text]
  • Structure of the Human Clamp Loader Reveals an Autoinhibited Conformation of a Substrate-Bound AAA+ Switch
    Structure of the human clamp loader reveals an autoinhibited conformation of a substrate-bound AAA+ switch Christl Gaubitza,1, Xingchen Liua,b,1, Joseph Magrinoa,b, Nicholas P. Stonea, Jacob Landecka,b, Mark Hedglinc, and Brian A. Kelcha,2 aDepartment of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester MA 01605; bGraduate School of Biomedical Sciences, University of Massachusetts Medical School, Worcester MA 01605; and cDepartment of Chemistry, The Pennsylvania State University, University Park, PA 16802 Edited by Michael E. O’Donnell, HHMI and Rockefeller University, New York, NY, and approved July 27, 2020 (received for review April 20, 2020) DNA replication requires the sliding clamp, a ring-shaped protein areflexia syndrome (15), Hutchinson–Gilford progeria syn- complex that encircles DNA, where it acts as an essential cofactor drome (16), and in the replication of some viruses (17–19). It for DNA polymerases and other proteins. The sliding clamp needs is unknown whether loading by RFC contributes to PARD to be opened and installed onto DNA by a clamp loader ATPase of disease. the AAA+ family. The human clamp loader replication factor C Clamp loaders are members of the AAA+ family of ATPases (RFC) and sliding clamp proliferating cell nuclear antigen (PCNA) (ATPases associated with various cellular activities), a large are both essential and play critical roles in several diseases. De- protein family that uses the chemical energy of adenosine 5′- spite decades of study, no structure of human RFC has been re- triphosphate (ATP) to generate mechanical force (20). Most solved. Here, we report the structure of human RFC bound to AAA+ proteins form hexameric motors that use an undulating PCNA by cryogenic electron microscopy to an overall resolution ∼ spiral staircase mechanism to processively translocate a substrate of 3.4 Å.
    [Show full text]
  • BET Family Members Bdf1/2 Modulate Global Transcription Initiation and Elongation in Saccharomyces Cerevisiae Rafal Donczew*, Steven Hahn*
    RESEARCH ARTICLE BET family members Bdf1/2 modulate global transcription initiation and elongation in Saccharomyces cerevisiae Rafal Donczew*, Steven Hahn* Fred Hutchinson Cancer Research Center, Division of Basic Sciences, Seattle, United States Abstract Human bromodomain and extra-terminal domain (BET) family members are promising targets for therapy of cancer and immunoinflammatory diseases, but their mechanisms of action and functional redundancies are poorly understood. Bdf1/2, yeast homologues of the human BET factors, were previously proposed to target transcription factor TFIID to acetylated histone H4, analogous to bromodomains that are present within the largest subunit of metazoan TFIID. We investigated the genome-wide roles of Bdf1/2 and found that their important contributions to transcription extend beyond TFIID function as transcription of many genes is more sensitive to Bdf1/2 than to TFIID depletion. Bdf1/2 co-occupy the majority of yeast promoters and affect preinitiation complex formation through recruitment of TFIID, Mediator, and basal transcription factors to chromatin. Surprisingly, we discovered that hypersensitivity of genes to Bdf1/2 depletion results from combined defects in transcription initiation and early elongation, a striking functional similarity to human BET proteins, most notably Brd4. Our results establish Bdf1/2 as critical for yeast transcription and provide important mechanistic insights into the function of BET proteins in all eukaryotes. *For correspondence: [email protected] (RD); Introduction [email protected] (SH) Bromodomains (BDs) are reader modules that allow protein targeting to chromatin via interactions with acetylated histone tails. BD-containing factors are usually involved in gene transcription, and Competing interests: The their deregulation has been implicated in a spectrum of cancers and immunoinflammatory and neu- authors declare that no rological conditions (Fujisawa and Filippakopoulos, 2017; Wang et al., 2021).
    [Show full text]
  • Involvement of Brd4 in Different Steps of the Papillomavirus Life Cycle
    Virus Research 231 (2017) 76–82 Contents lists available at ScienceDirect Virus Research j ournal homepage: www.elsevier.com/locate/virusres Review Involvement of Brd4 in different steps of the papillomavirus life cycle a,∗ a b,c Thomas Iftner , Juliane Haedicke-Jarboui , Shwu-Yuan Wu , b,c,d,∗∗ Cheng-Ming Chiang a Division of Experimental Virology, Institute for Medical Virology, University Hospital Tübingen, Elfriede-Aulhorn-Str. 6, 72076 Tübingen, Germany b Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA c Department of Biochemistry, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA d Department of Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA a r t i a b s c t l e i n f o r a c t Article history: Bromodomain-containing protein 4 (Brd4) is a cellular chromatin-binding factor and transcriptional reg- Received 13 September 2016 ulator that recruits sequence-specific transcription factors and chromatin modulators to control target Received in revised form 2 December 2016 gene transcription. Papillomaviruses (PVs) have evolved to hijack Brd4’s activity in order to create a facili- Accepted 2 December 2016 tating environment for the viral life cycle. Brd4, in association with the major viral regulatory protein E2, is Available online 10 December 2016 involved in multiple steps of the PV life cycle including replication initiation, viral gene transcription, and viral genome segregation and maintenance. Phosphorylation of Brd4, regulated by casein kinase II (CK2) Keywords: and protein phosphatase 2A (PP2A), is critical for viral gene transcription as well as E1- and E2-dependent Brd4 origin replication.
    [Show full text]
  • BRD2-2 (His) (Bromodomain Containing Protein 2 (RING3), Bromodomain 2)
    BRD2-2 (His) (Bromodomain containing protein 2 (RING3), bromodomain 2) CATALOG NO.: RD-11-145 LOT NO.: DESCRIPTION: Human recombinant BRD2, bromodomain-2 (residues 344-454; Genbank Accession # NM_005104; MW = 15.7 kDa) expressed in E. coli with an N-terminal His-tag. BRD2, like other human members of the BET family of chromatin-binding proteins (BRD3, BRD4, BRDT), comprises two bromodomains (see reviews 1,2 ), protein modules that bind ε-N-acetyllysine residues 3,4 . When overexpressed in 293 cells, BRD2, along with BRD3, binds the hyperacetylated chromatin of transcribed genes, regions enriched in acetylated histone H4 lysine-5 (H4K5Ac), H4K12Ac, H3K14Ac, but deficient in H4K16Ac and H3K9me 5. A single H4K5AcK12Ac peptide can bind two copies of BRD2-2 (BRD2, bromodomain 2), each interacting with one of the two acetylated lysines 6. In an in vitro RNA polymerase II transcription system, binding of either BRD2 or BRD3 to a chromatin template assembled with hyperacetylated histones enabled transcription through the nucleosomes 5. Further, BRD2 displayed histone chaperone activity, catalyzing the transfer of histone octamers from hyperacetylated oligonucleosomes to a labeled 190 bp 5s rDNA fragment 5. Like BRD4, BRD2 is a ubiquitously expressed 7 transcriptional regulator 8 and atypical protein kinase 9, with functions in cell cycle progression 8 and embryogenesis 10,11 . BRD2 binds preferentially to hyperacetylated histone H4 in H2A.Z-containing nucleosomes and this interaction is required for activation of androgen receptor (AR)-regulated genes in prostate cancer cells 12 . In addition to prostate cancer, leukemia is a potential indication for specific BRD2 inhibition9,13 . BRD2 suppresses HIV transcription in latently infected cells and may therefore represent a target in therapeutic strategies involving viral reactivation 14 .
    [Show full text]
  • Quantification of Nuclear Protein Dynamics Reveals Chromatin Remodeling During Acute Protein Degradation
    bioRxiv preprint doi: https://doi.org/10.1101/345686; this version posted June 14, 2018. 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. Quantification of nuclear protein dynamics reveals chromatin remodeling during acute protein degradation Alexander J. Federation1, Vivek Nandakumar1, Hao Wang1, Brian Searle2, Lindsay Pino2, Gennifer Merrihew2, Sonia Ting2, Nicholas Howard1, Tanya Kutyavin1, Michael J. MacCoss2, John A. Stamatoyannopoulos1,2 1. Altius Institute for Biomedical Sciences; Seattle, WA 98121 2. University of Washington, Department of Genome Sciences; Seattle, WA 98195 Correspondence: John A. Stamatoyannopoulos [email protected] Michael J. MacCoss [email protected] bioRxiv preprint doi: https://doi.org/10.1101/345686; this version posted June 14, 2018. 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. Abstract Sequencing-based technologies cannot measure post-transcriptional dynamics of the nuclear proteome, but unbiased mass-spectrometry measurements of nuclear proteins remain difficult. In this work, we have combined facile nuclear sub-fractionation approaches with data-independent acquisition mass spectrometry to improve detection and quantification of nuclear proteins in human cells and tissues. Nuclei are isolated and subjected to a series of extraction conditions that enrich for nucleoplasm, euchromatin, heterochromatin and nuclear-membrane associated proteins. Using this approach, we can measure peptides from over 70% of the expressed nuclear proteome. As we are physically separating chromatin compartments prior to analysis, proteins can be assigned into functional chromatin environments that illuminate systems-wide nuclear protein dynamics.
    [Show full text]
  • Brd4-Bound Enhancers Drive Cell-Intrinsic Sex Differences in Glioblastoma
    Brd4-bound enhancers drive cell-intrinsic sex differences in glioblastoma Najla Kfourya,b,1, Zongtai Qic,d,1, Briana C. Pragere,f, Michael N. Wilkinsonc,d, Lauren Broestla,g, Kristopher C. Berretth, Arnav Moudgilc,d,g, Sumithra Sankararamanc,d, Xuhua Chenc,d, Jason Gertzh, Jeremy N. Riche,i, Robi D. Mitrac,d,2,3, and Joshua B. Rubina,j,2,3 aDepartment of Pediatrics, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110; bDepartment of Neurological Surgery, University of California San Diego, La Jolla, CA 92037; cDepartment of Genetics, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110; dCenter for Genome Sciences and Systems Biology, Washington University in St. Louis, St. Louis, MO 63110; eDivision of Regenerative Medicine, Department of Medicine, University of California San Diego, La Jolla, CA 92037; fCleveland Clinic Lerner College of Medicine, Cleveland, OH 44195; gMedical Scientist Training Program, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110; hDepartment of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT 84112; iDepartment of Neurosciences, University of California San Diego, La Jolla, CA 92037; and jDepartment of Neuroscience, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110 Edited by Rene Bernards, The Netherlands Cancer Institute, Amsterdam, The Netherlands, and approved December 25, 2020 (received for review August 21, 2020) Sex can be an important determinant of cancer phenotype, and cancer risk and progression, with the ultimate goal of incorporating exploring sex-biased tumor biology holds promise for identifying sex-informed approaches to treatment to improve survival of all novel therapeutic targets and new approaches to cancer treatment.
    [Show full text]
  • Renoprotective Effect of Combined Inhibition of Angiotensin-Converting Enzyme and Histone Deacetylase
    BASIC RESEARCH www.jasn.org Renoprotective Effect of Combined Inhibition of Angiotensin-Converting Enzyme and Histone Deacetylase † ‡ Yifei Zhong,* Edward Y. Chen, § Ruijie Liu,*¶ Peter Y. Chuang,* Sandeep K. Mallipattu,* ‡ ‡ † | ‡ Christopher M. Tan, § Neil R. Clark, § Yueyi Deng, Paul E. Klotman, Avi Ma’ayan, § and ‡ John Cijiang He* ¶ *Department of Medicine, Mount Sinai School of Medicine, New York, New York; †Department of Nephrology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China; ‡Department of Pharmacology and Systems Therapeutics and §Systems Biology Center New York, Mount Sinai School of Medicine, New York, New York; |Baylor College of Medicine, Houston, Texas; and ¶Renal Section, James J. Peters Veterans Affairs Medical Center, New York, New York ABSTRACT The Connectivity Map database contains microarray signatures of gene expression derived from approximately 6000 experiments that examined the effects of approximately 1300 single drugs on several human cancer cell lines. We used these data to prioritize pairs of drugs expected to reverse the changes in gene expression observed in the kidneys of a mouse model of HIV-associated nephropathy (Tg26 mice). We predicted that the combination of an angiotensin-converting enzyme (ACE) inhibitor and a histone deacetylase inhibitor would maximally reverse the disease-associated expression of genes in the kidneys of these mice. Testing the combination of these inhibitors in Tg26 mice revealed an additive renoprotective effect, as suggested by reduction of proteinuria, improvement of renal function, and attenuation of kidney injury. Furthermore, we observed the predicted treatment-associated changes in the expression of selected genes and pathway components. In summary, these data suggest that the combination of an ACE inhibitor and a histone deacetylase inhibitor could have therapeutic potential for various kidney diseases.
    [Show full text]
  • GSK973: a BD2 Selective Inhibitor of BRD2, BDR3, BRD4, BRDT
    GSK973: A BD2 selective inhibitor of BRD2, BDR3, BRD4, BRDT Version 1.0 (19th April 2021) Web link for more details: https://www.sgc-ffm.uni-frankfurt.de/#!specificprobeoverview/GSK973 Overview Proteins of the bromodomain and extra-terminal (BET) domain family – BRD2, BRD3, BRD4 and BRDT - are epigenetic readers that bind acetylated histones through their bromodomains to regulate gene transcription. BET family of bromodomains (BRDs) are well-known drug targets for many human diseases. The active pockets of the two tandem bromodomains BD1/BD2 are highly conserved (sequence similarity is about 95%), thus it is of great medical importance and still a significant challenge to develop BD1/BD2 selective inhibitors. Summary Chemical Probe Name GSK973 Negative control compound GSK943 Target(s) (synonyms) BRD2/ Bromodomain-containing protein 2/KIAA9001/RING3; BRD3/ Bromodomain-containing protein 3/KIAA0043/RING3L; BRD4/ Bromodomain- containing protein 4/HUNK1; BRDT/ Bromodomain testis-specific protein/CT9 Recommended cell assay Use at concentrations up to 10 µM. Test at various concentrations with a 9 concentration point curve starting from 10 µM down in 1/3 serial dilutions Suitability for in vivo use and Tested in rat and dog, shows excellent pharmacokinetics in dog with low recommended dose blood clearance, good oral bioavailability, and a moderate half-life. Publications PMID: 32832027 (compound 36) Orthogonal chemical probes GSK046, GSK620 In vitro assay(s) used to characterise TR-FRET, BROMOscan, SPR Cellular assay(s) for target- Cellular
    [Show full text]
  • Oxidized Phospholipids Regulate Amino Acid Metabolism Through MTHFD2 to Facilitate Nucleotide Release in Endothelial Cells
    ARTICLE DOI: 10.1038/s41467-018-04602-0 OPEN Oxidized phospholipids regulate amino acid metabolism through MTHFD2 to facilitate nucleotide release in endothelial cells Juliane Hitzel1,2, Eunjee Lee3,4, Yi Zhang 3,5,Sofia Iris Bibli2,6, Xiaogang Li7, Sven Zukunft 2,6, Beatrice Pflüger1,2, Jiong Hu2,6, Christoph Schürmann1,2, Andrea Estefania Vasconez1,2, James A. Oo1,2, Adelheid Kratzer8,9, Sandeep Kumar 10, Flávia Rezende1,2, Ivana Josipovic1,2, Dominique Thomas11, Hector Giral8,9, Yannick Schreiber12, Gerd Geisslinger11,12, Christian Fork1,2, Xia Yang13, Fragiska Sigala14, Casey E. Romanoski15, Jens Kroll7, Hanjoong Jo 10, Ulf Landmesser8,9,16, Aldons J. Lusis17, 1234567890():,; Dmitry Namgaladze18, Ingrid Fleming2,6, Matthias S. Leisegang1,2, Jun Zhu 3,4 & Ralf P. Brandes1,2 Oxidized phospholipids (oxPAPC) induce endothelial dysfunction and atherosclerosis. Here we show that oxPAPC induce a gene network regulating serine-glycine metabolism with the mitochondrial methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFD2) as a cau- sal regulator using integrative network modeling and Bayesian network analysis in human aortic endothelial cells. The cluster is activated in human plaque material and by atherogenic lipo- proteins isolated from plasma of patients with coronary artery disease (CAD). Single nucleotide polymorphisms (SNPs) within the MTHFD2-controlled cluster associate with CAD. The MTHFD2-controlled cluster redirects metabolism to glycine synthesis to replenish purine nucleotides. Since endothelial cells secrete purines in response to oxPAPC, the MTHFD2- controlled response maintains endothelial ATP. Accordingly, MTHFD2-dependent glycine synthesis is a prerequisite for angiogenesis. Thus, we propose that endothelial cells undergo MTHFD2-mediated reprogramming toward serine-glycine and mitochondrial one-carbon metabolism to compensate for the loss of ATP in response to oxPAPC during atherosclerosis.
    [Show full text]