DOCSLIB.ORG

Sterol Biosynthesis Pathway Is Part of the Interferon Host Defence Response

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Sterol Biosynthesis Pathway Is Part of the Interferon Host Defence Response Sterol biosynthesis pathway is part of the interferon host defence response Mathieu Blanc A thesis submitted in fulfilment of requirements for the degree of Doctor of Philosophy Division of Pathway Medicine, School of Biomedical Sciences The University of Edinburgh April 2010 Declaration I hereby declare that this thesis is of my own composition, and that it contains no material previously submitted for the award of any other degree. The work reported in this thesis has been executed by myself, except where due acknowledgement is made in the text. Mathieu Blanc ii Abstract Recently, cholesterol metabolism has been shown to modulate the infection of several viruses and there is growing evidence that inflammatory response to infection also modulates lipid metabolism. However little is known about the role of inflammatory processes in modulating lipid metabolism and their consequences for the viral infection. This study investigates host-lipid viral interaction pathways using mouse cytomegalovirus, a large double-stranded DNA genome, which represents one of the few models for a natural infection of its natural host. In this study, transcriptomic and lipidomic profiling of macrophages shows that there is a specific coordinated regulation of the sterol pathways upon viral infection or treatment with IFN or (but not TNFα, IL1 or IL6) resulting in the decrease of free cellular cholesterol. Furthermore, we show that pharmacological and RNAi inhibition of the sterol pathway augments protection against infection in vitro and in vivo and we identified that the prenylation branch of the sterol metabolic network was involved in the protective response. Finally, we show that genetic knock out of IFN results in a partial reduction while genetic knock out of Ifnar1 completely abolishes the reduction of the sterol biosynthetic activity upon infection. Overall these results support a role for part of the sterol metabolic network in protective immunity and show that type 1 IFN signalling is both necessary and sufficient for reducing the sterol metabolic network upon infection; thereby linking the sterol pathway with IFN defence responses. iii Acknowledgements A considerable number of people helped me during this project. First of all I would like to thank my supervisor Prof Peter Ghazal for giving me the opportunity of starting this project in his laboratory and for his guidance all along the difficult ways of my PhD. Thanks to Prof Rudolph Riemersma who has taken a considerable amount of his time to teach and advise me during the PhD. I am also grateful to Dr John McLauchlan for advises and Dr Ana Angulo for helping me during my time in Barcelona and for her helpful comments. I would also like to thank all the past and present members of the DPM for their help and especially, Garwin (who made me see science in a different way), Paul for our helpful discussions, Kai and Donal for their help with the writing process but also for the coffee time. Also a special thanks to Sara alias Dr Rodriguez Martin who has been here since I started and helped me to pass the difficult times inside and outside of the lab. I also would like to thank my friend Sanjay (alias Dr Shrew), who helped me with the corrections of the manuscript but also for all the good and bad times that we went through together. I am also very grateful to the British heart foundation and the Wellcome trust for funding my PhD. Of course, I would like to thanks my family: My parents who gave an unconditional support during my studies, my grand- parents who are a model for me and to whom I dedicate this thesis and finally my two brothers Damien and Hugo. And last but not least, I would like to thank Laura for her kindness, her patient and all the other qualities she needed to support me during this PhD, certainly I would not have finished without her. iv List of Abbreviations % Percentage ~ Approximately °C Degrees Celsius µ Micro µg Micrograms µl Microlitre A Absorbance Ad Adenovirus AIDs Acquired immunodeficiency syndrome ANOVA Analysis of variance ATCC American Type Culture Collection BAC Bacterial artificial chromosome BALB/c Laboratory mouse strain BMDMs Bone marrow derived macrophages bp Base pairs BSA Bovine serum albumin C57Bl/6 laboratory mouse strain CBP CREB‐binding protein cm2 Centimeters square CO2 Carbon dioxide CPE Cytopathogenic effect CS Calf serum DC Denditric cell dH2O Distilled water DMEM Dulbecco’s modified essential medium DMSO Dimethyl sulfoxide DNA deoxyribonucleic acid EBV Epstein‐Barr virus FACS fluorescent‐activated cell sorter FAM Carboxyfluorescein FCS Fetal calf serum FITC Fluorescein isothiocyanate gravitational force or grams, depending on g context h.p.i hrs post infection hCMV Human cytomegalovirus HCV Hepatitis C virus hour hr HRP horseradish peroxidase HSV1 Herpes simplex virus 1 i.p Intraperitoneal route IE Immediate early v IE3 Immediate early 3 protein IFN Interferon IFNAR Type I interferon receptor IFNGR I Type II interferon receptor Ig Immunoglobulin IL Interleukin IPA Ingenuity Pathway Analysis kb kilo bases kg kilogram LPS lipopolysaccharide LT Lymphotoxin M Molar MAPK Mitogen‐activated protein kinases mCMV murine cytomegalovirus mg milligram MHC Major histocompatibility complex MIEP Major Immediate Early Promoter min Minute ml Milliliter mM millimolar MOI Multiplicity of infection mRNA Messenger ribonucleic acid NIH 3T3 murine fibroblast cell line NK Natural Killer nm nanometer ORF Open reading frame PBMC Peripheral blood mononuclear cell PBS Phosphate buffered saline PCR Polymerase chain reaction pfu Plaque forming units RNA Ribonucleic acid RT Reverse transcriptase rt Room temperature Q‐PCR Quantitative polymerase chain reaction SD Standard deviation SFV1 semliki forest virus 1 Th T helper TNF Tumor necrosis factor tyk2 Tyrosine kinase 2 U Units UV Ultraviolet VV Vaccinia virus w/vol Weight per volume vi Table of contents 1 CHAPTER 1: INTRODUCTION..................................................................... 1 1.1 Characteristics of Cytomegalovirus ................................................................. 1 1.1.1 Overview of the Herpes virus family........................................................... 1 1.1.2 Characteristics of the Herpes virus life cycle............................................... 4 1.1.3 Latency......................................................................................................... 7 1.1.4 Immune evasion........................................................................................... 7 1.1.5 Biology of Cytomegalovirus........................................................................ 8 1.1.5.1 Pathogenesis of cytomegalovirus infection.......................................... 9 1.1.5.2 Diagnosis and therapy for hCMV infection....................................... 10 1.1.5.3 mCMV has a model for studying hCMV........................................... 11 1.1.5.4 mCMV characteristics........................................................................ 11 1.2 Innate immune response to CMV infection................................................... 13 1.2.1 Cells of the innate immune response to CMV infection............................ 13 1.2.2 CMV interactions with innate immune cells.............................................. 19 1.2.3 Initiation of the innate immune response................................................... 20 1.2.4 Type I Interferon response and CMV ........................................................ 22 1.3 Adaptive immune response and CMV infection ........................................... 26 1.3.1 Type II interferon response to mCMV infection........................................ 28 1.4 Cholesterol and host pathogen interaction .................................................... 29 1.4.1 Cholesterol metabolism.............................................................................. 30 1.4.1.1 Structure of cholesterol ...................................................................... 30 1.4.1.2 Cholesterol synthesis and prenylation pathway ................................. 31 1.4.1.3 Regulation of the cholesterol synthesis.............................................. 33 1.4.1.4 Cholesterol homeostasis..................................................................... 35 1.4.2 Involvement of cholesterol in viral lifecycle ............................................. 38 1.4.2.1 Cholesterol is essential for viral entry, assembly and budding of viruses ........................................................................................................... 38 1.4.2.2 Role of cholesterol synthesis in viral replication ............................... 39 1.4.2.3 Viral infection manipulates intra cellular cholesterol metabolism .... 40 1.4.2.4 Viral infection induced dyslipidemia in patient................................. 41 1.4.2.5 Herpes viruses and cholesterol metabolism ....................................... 41 1.5 Innate immunity and cholesterol metabolism ............................................... 44 1.5.1 Cytokines and chemokines modulate cholesterol metabolism................... 44 1.5.2 Cross talk between innate immunity and cholesterol metabolism: Involvement of TLRs and nuclear hormones receptor........................................... 45 vii 1.5.2.1 Innate immune response modulate cholesterol metabolism............... 45 1.5.2.2 Cholesterol modulates innate immune response................................ 46 1.5.2.3 Limitation of current studies .............................................................
Load more

Recommended publications
  • Hypoxia and Oxygen-Sensing Signaling in Gene Regulation and Cancer Progression
    International Journal of Molecular Sciences Review Hypoxia and Oxygen-Sensing Signaling in Gene Regulation and Cancer Progression Guang Yang, Rachel Shi and Qing Zhang * Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; [email protected] (G.Y.); [email protected] (R.S.) * Correspondence: [email protected]; Tel.: +1-214-645-4671 Received: 6 October 2020; Accepted: 29 October 2020; Published: 31 October 2020 Abstract: Oxygen homeostasis regulation is the most fundamental cellular process for adjusting physiological oxygen variations, and its irregularity leads to various human diseases, including cancer. Hypoxia is closely associated with cancer development, and hypoxia/oxygen-sensing signaling plays critical roles in the modulation of cancer progression. The key molecules of the hypoxia/oxygen-sensing signaling include the transcriptional regulator hypoxia-inducible factor (HIF) which widely controls oxygen responsive genes, the central members of the 2-oxoglutarate (2-OG)-dependent dioxygenases, such as prolyl hydroxylase (PHD or EglN), and an E3 ubiquitin ligase component for HIF degeneration called von Hippel–Lindau (encoding protein pVHL). In this review, we summarize the current knowledge about the canonical hypoxia signaling, HIF transcription factors, and pVHL. In addition, the role of 2-OG-dependent enzymes, such as DNA/RNA-modifying enzymes, JmjC domain-containing enzymes, and prolyl hydroxylases, in gene regulation of cancer progression, is specifically reviewed. We also discuss the therapeutic advancement of targeting hypoxia and oxygen sensing pathways in cancer. Keywords: hypoxia; PHDs; TETs; JmjCs; HIFs 1. Introduction Molecular oxygen serves as a co-factor in many biochemical processes and is fundamental for aerobic organisms to maintain intracellular ATP levels [1,2].
    [Show full text]
  • Phenotype Heterogeneity of Congenital Adrenal Hyperplasia Due
    Kor et al. BMC Medical Genetics (2018) 19:115 https://doi.org/10.1186/s12881-018-0629-2 CASE REPORT Open Access Phenotype heterogeneity of congenital adrenal hyperplasia due to genetic mosaicism and concomitant nephrogenic diabetes insipidus in a sibling Yılmaz Kor1†, Minjing Zou2†, Roua A. Al-Rijjal2, Dorota Monies2, Brian F. Meyer2 and Yufei Shi2* Abstract Background: Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21OHD) is an autosomal recessive disorder caused by mutations in the CYP21A2. Congenital nephrogenic diabetes insipidus (NDI) is a rare X- linked recessive or autosomal recessive disorder caused by mutations in either AVPR2 or AQP2. Genotype-phenotype discordance caused by genetic mosaicism in CAH patients has not been reported, nor the concomitant CAH and NDI. Case presentation: We investigated a patient with concomitant CAH and NDI from a consanguineous family. She (S- 1) presented with clitoromegaly at 3 month of age, and polydipsia and polyuria at 13 month of age. Her parents and two elder sisters (S-2 and S-3) were clinically normal, but elevated levels of serum 17-hydroxyprogesterone (17-OHP) were observed in the mother and S-2. The coding region of CYP21A2 and AQP2 were analyzed by PCR-sequencing analysis to identify genetic defects. Two homozygous CYP21A2 mutations (p.R357W and p.P454S) were identified in the proband and her mother and S-2. The apparent genotype-phenotype discordance was due to presence of small amount of wild-type CYP21A2 alleles in S-1, S-2, and their mother’s genome, thus protecting them from development of classic form of 21OHD (C21OHD).
    [Show full text]
  • Interplay Between Epigenetics and Metabolism in Oncogenesis: Mechanisms and Therapeutic Approaches
    OPEN Oncogene (2017) 36, 3359–3374 www.nature.com/onc REVIEW Interplay between epigenetics and metabolism in oncogenesis: mechanisms and therapeutic approaches CC Wong1, Y Qian2,3 and J Yu1 Epigenetic and metabolic alterations in cancer cells are highly intertwined. Oncogene-driven metabolic rewiring modifies the epigenetic landscape via modulating the activities of DNA and histone modification enzymes at the metabolite level. Conversely, epigenetic mechanisms regulate the expression of metabolic genes, thereby altering the metabolome. Epigenetic-metabolomic interplay has a critical role in tumourigenesis by coordinately sustaining cell proliferation, metastasis and pluripotency. Understanding the link between epigenetics and metabolism could unravel novel molecular targets, whose intervention may lead to improvements in cancer treatment. In this review, we summarized the recent discoveries linking epigenetics and metabolism and their underlying roles in tumorigenesis; and highlighted the promising molecular targets, with an update on the development of small molecule or biologic inhibitors against these abnormalities in cancer. Oncogene (2017) 36, 3359–3374; doi:10.1038/onc.2016.485; published online 16 January 2017 INTRODUCTION metabolic genes have also been identified as driver genes It has been appreciated since the early days of cancer research mutated in some cancers, such as isocitrate dehydrogenase 1 16 17 that the metabolic profiles of tumor cells differ significantly from and 2 (IDH1/2) in gliomas and acute myeloid leukemia (AML), 18 normal cells. Cancer cells have high metabolic demands and they succinate dehydrogenase (SDH) in paragangliomas and fuma- utilize nutrients with an altered metabolic program to support rate hydratase (FH) in hereditary leiomyomatosis and renal cell 19 their high proliferative rates and adapt to the hostile tumor cancer (HLRCC).
    [Show full text]
  • Mutant IDH, (R)-2-Hydroxyglutarate, and Cancer
    Downloaded from genesdev.cshlp.org on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW What a difference a hydroxyl makes: mutant IDH, (R)-2-hydroxyglutarate, and cancer Julie-Aurore Losman1 and William G. Kaelin Jr.1,2,3 1Department of Medical Oncology, Dana-Farber Cancer Institute, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02215, USA; 2Howard Hughes Medical Institute, Chevy Chase, Maryland 20815, USA Mutations in metabolic enzymes, including isocitrate whether altered cellular metabolism is a cause of cancer dehydrogenase 1 (IDH1) and IDH2, in cancer strongly or merely an adaptive response of cancer cells in the face implicate altered metabolism in tumorigenesis. IDH1 of accelerated cell proliferation is still a topic of some and IDH2 catalyze the interconversion of isocitrate and debate. 2-oxoglutarate (2OG). 2OG is a TCA cycle intermediate The recent identification of cancer-associated muta- and an essential cofactor for many enzymes, including tions in three metabolic enzymes suggests that altered JmjC domain-containing histone demethylases, TET cellular metabolism can indeed be a cause of some 5-methylcytosine hydroxylases, and EglN prolyl-4-hydrox- cancers (Pollard et al. 2003; King et al. 2006; Raimundo ylases. Cancer-associated IDH mutations alter the enzymes et al. 2011). Two of these enzymes, fumarate hydratase such that they reduce 2OG to the structurally similar (FH) and succinate dehydrogenase (SDH), are bone fide metabolite (R)-2-hydroxyglutarate [(R)-2HG]. Here we tumor suppressors, and loss-of-function mutations in FH review what is known about the molecular mechanisms and SDH have been identified in various cancers, in- of transformation by mutant IDH and discuss their im- cluding renal cell carcinomas and paragangliomas.
    [Show full text]
  • Characterization of the Ergosterol Biosynthesis Pathway in Ceratocystidaceae
    Journal of Fungi Article Characterization of the Ergosterol Biosynthesis Pathway in Ceratocystidaceae Mohammad Sayari 1,2,*, Magrieta A. van der Nest 1,3, Emma T. Steenkamp 1, Saleh Rahimlou 4 , Almuth Hammerbacher 1 and Brenda D. Wingfield 1 1 Department of Biochemistry, Genetics and Microbiology, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 0002, South Africa; [email protected] (M.A.v.d.N.); [email protected] (E.T.S.); [email protected] (A.H.); brenda.wingfi[email protected] (B.D.W.) 2 Department of Plant Science, University of Manitoba, 222 Agriculture Building, Winnipeg, MB R3T 2N2, Canada 3 Biotechnology Platform, Agricultural Research Council (ARC), Onderstepoort Campus, Pretoria 0110, South Africa 4 Department of Mycology and Microbiology, University of Tartu, 14A Ravila, 50411 Tartu, Estonia; [email protected] * Correspondence: [email protected]; Fax: +1-204-474-7528 Abstract: Terpenes represent the biggest group of natural compounds on earth. This large class of organic hydrocarbons is distributed among all cellular organisms, including fungi. The different classes of terpenes produced by fungi are mono, sesqui, di- and triterpenes, although triterpene ergosterol is the main sterol identified in cell membranes of these organisms. The availability of genomic data from members in the Ceratocystidaceae enabled the detection and characterization of the genes encoding the enzymes in the mevalonate and ergosterol biosynthetic pathways. Using Citation: Sayari, M.; van der Nest, a bioinformatics approach, fungal orthologs of sterol biosynthesis genes in nine different species M.A.; Steenkamp, E.T.; Rahimlou, S.; of the Ceratocystidaceae were identified.
    [Show full text]
  • Deletion of the Gene Encoding the Reductase Component of 3
    Yeh et al. Microbial Cell Factories 2014, 13:130 http://www.microbialcellfactories.com/content/13/1/130 RESEARCH Open Access Deletion of the gene encoding the reductase component of 3-ketosteroid 9α-hydroxylase in Rhodococcus equi USA-18 disrupts sterol catabolism, leading to the accumulation of 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid and 1,4-androstadiene-3,17-dione Chin-Hsing Yeh1, Yung-Shun Kuo1, Che-Ming Chang1, Wen-Hsiung Liu2, Meei-Ling Sheu3 and Menghsiao Meng1* Abstract The gene encoding the putative reductase component (KshB) of 3-ketosteroid 9α-hydroxylase was cloned from Rhodococcus equi USA-18, a cholesterol oxidase-producing strain formerly named Arthrobacter simplex USA-18, by PCR according to consensus amino acid motifs of several bacterial KshB subunits. Deletion of the gene in R. equi USA-18 by a PCR-targeted gene disruption method resulted in a mutant strain that could accumulate up to 0.58 mg/ml 1,4-androstadiene-3,17-dione (ADD) in the culture medium when 0.2% cholesterol was used as the carbon source, indicating the involvement of the deleted enzyme in 9α-hydroxylation of steroids. In addition, this mutant also accumulated 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid (Δ1,4-BNC). Because both ADD and Δ1,4-BNC are important intermediates for the synthesis of steroid drugs, this mutant derived from R. equi USA-18 may deserve further investigation for its application potential. Background generally believed to be carried out by cholesterol oxidase, Steroid drugs, including androgens, anabolic steroids, es- which catalyzes the oxidation of the 3β-hydroxyl moiety of trogens and corticosteroids, have been widely used for a sterols with the simultaneous isomerization of Δ5 to Δ4, variety of health purposes.
    [Show full text]
  • Protein Identities in Evs Isolated from U87-MG GBM Cells As Determined by NG LC-MS/MS
    Protein identities in EVs isolated from U87-MG GBM cells as determined by NG LC-MS/MS. No. Accession Description Σ Coverage Σ# Proteins Σ# Unique Peptides Σ# Peptides Σ# PSMs # AAs MW [kDa] calc. pI 1 A8MS94 Putative golgin subfamily A member 2-like protein 5 OS=Homo sapiens PE=5 SV=2 - [GG2L5_HUMAN] 100 1 1 7 88 110 12,03704523 5,681152344 2 P60660 Myosin light polypeptide 6 OS=Homo sapiens GN=MYL6 PE=1 SV=2 - [MYL6_HUMAN] 100 3 5 17 173 151 16,91913397 4,652832031 3 Q6ZYL4 General transcription factor IIH subunit 5 OS=Homo sapiens GN=GTF2H5 PE=1 SV=1 - [TF2H5_HUMAN] 98,59 1 1 4 13 71 8,048185945 4,652832031 4 P60709 Actin, cytoplasmic 1 OS=Homo sapiens GN=ACTB PE=1 SV=1 - [ACTB_HUMAN] 97,6 5 5 35 917 375 41,70973209 5,478027344 5 P13489 Ribonuclease inhibitor OS=Homo sapiens GN=RNH1 PE=1 SV=2 - [RINI_HUMAN] 96,75 1 12 37 173 461 49,94108966 4,817871094 6 P09382 Galectin-1 OS=Homo sapiens GN=LGALS1 PE=1 SV=2 - [LEG1_HUMAN] 96,3 1 7 14 283 135 14,70620005 5,503417969 7 P60174 Triosephosphate isomerase OS=Homo sapiens GN=TPI1 PE=1 SV=3 - [TPIS_HUMAN] 95,1 3 16 25 375 286 30,77169764 5,922363281 8 P04406 Glyceraldehyde-3-phosphate dehydrogenase OS=Homo sapiens GN=GAPDH PE=1 SV=3 - [G3P_HUMAN] 94,63 2 13 31 509 335 36,03039959 8,455566406 9 Q15185 Prostaglandin E synthase 3 OS=Homo sapiens GN=PTGES3 PE=1 SV=1 - [TEBP_HUMAN] 93,13 1 5 12 74 160 18,68541938 4,538574219 10 P09417 Dihydropteridine reductase OS=Homo sapiens GN=QDPR PE=1 SV=2 - [DHPR_HUMAN] 93,03 1 1 17 69 244 25,77302971 7,371582031 11 P01911 HLA class II histocompatibility antigen,
    [Show full text]
  • Farnesyl Phosphatase, a Corpora Allata
    Florida International University FIU Digital Commons Department of Biological Sciences College of Arts, Sciences & Education 8-5-2013 Farnesyl Phosphatase, a Corpora allata Enzyme Involved in Juvenile Hormone Biosynthesis in Aedes aegypti Pratik Nyati Department of Biological Sciences, Florida International University, [email protected] Marcela Nouzova Department of Biological Sciences, Florida International University, [email protected] Crisalejandra Rivera-Perez Department of Biological Sciences, Florida International University,, [email protected] Mark E. Clifton Department of Biological Sciences, Florida International University Jamie G. Mayoral Department of Biological Sciences, Florida International University See next page for additional authors Follow this and additional works at: https://digitalcommons.fiu.edu/cas_bio Part of the Biology Commons Recommended Citation Nyati P, Nouzova M, Rivera-Perez C, Clifton ME, Mayoral JG, et al. (2013) Farnesyl Phosphatase, a Corpora allata Enzyme Involved in Juvenile Hormone Biosynthesis in Aedes aegypti. PLoS ONE 8(8): e71967. doi:10.1371/journal.pone.0071967 This work is brought to you for free and open access by the College of Arts, Sciences & Education at FIU Digital Commons. It has been accepted for inclusion in Department of Biological Sciences by an authorized administrator of FIU Digital Commons. For more information, please contact [email protected]. Authors Pratik Nyati, Marcela Nouzova, Crisalejandra Rivera-Perez, Mark E. Clifton, Jamie G. Mayoral, and Fernando G. Noriega This article is available at FIU Digital Commons: https://digitalcommons.fiu.edu/cas_bio/131 Farnesyl Phosphatase, a Corpora allata Enzyme Involved in Juvenile Hormone Biosynthesis in Aedes aegypti Pratik Nyati, Marcela Nouzova, Crisalejandra Rivera-Perez, Mark E. Clifton, Jaime G. Mayoral, Fernando G.
    [Show full text]
  • The Role of Bile Acids in Overcoming Resistance to Chemotherapy By
    The Role of Bile Acids in Overcoming Resistance to Chemotherapy by Simon Chewchuk A thesis submitted as a partial fulfillment of the Requirements for the degree of Doctor of Philosophy (Ph.D.) in Biomolecular Sciences The Faculty of Graduate Studies Laurentian University Sudbury, Ontario, Canada © Simon Chewchuk, 2017 ii Abstract: In the context of cancer therapy, resistance to chemotherapy agents is a serious threat to patient welfare. In these circumstances, patients can either present with cancers that are naturally resistant to conventional therapy, referred to as innate resistance, or with cancers that become resistant following treatment, referred to as acquired resistance. In this thesis, we address the phenomenon of acquired drug resistance, involving cell lines selected for resistance to the anthracycline, doxorubicin. In the first study, we examined the role of the aldo-keto reductases AKR1C3 and AKR1B10 in doxorubicin resistance, enzymes that can hydroxylate doxorubicin to a less toxic form (doxorubicinol). Additionally, these enzymes can function to promote estrogen biosynthesis from estrone, which can have significant effects on cell growth and survival. We demonstrated in the first study that AKR1C3 and AKR1B10 are expressed at higher levels in doxorubicin resistant MCF-7 cells than their isogenic control counterparts. This change in expression correlated very well with increased estrogen synthesis. siRNA-mediated reduction in AKR1C3 and/or AKR1B10 transcript expression had no major effect on doxorubicin resistance, suggesting that these enzymes are not sufficient to mediate the doxorubicin resistance phenotype and that other mechanisms of doxorubicin resistance exist in these cells. We did, however, note that a pharmacological inhibitor of AKR enzymes (a bile acid termed β-cholanic acid) was effective in reversing doxorubicin resistance in doxorubicin-selected cell lines.
    [Show full text]
  • Biological and Translational Insights of Mutant IDH1/2 in Glioma
    Neurosurg Focus 34 (2):E2, 2013 ©AANS, 2013 From genomics to the clinic: biological and translational insights of mutant IDH1/2 in glioma GAVIN P. DUNN, M.D., PH.D.,1 OVIDIU C. ANDRONESI, M.D., PH.D.,2 AND DANIEL P. CAHILL, M.D., PH.D.1 Departments of 1Neurosurgery and 2Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts The characterization of the genomic alterations across all human cancers is changing the way that malignant disease is defined and treated. This paradigm is extending to glioma, where the discovery of recurrent mutations in the isocitrate dehydrogenase 1 (IDH1) gene has shed new light on the molecular landscape in glioma and other IDH- mutant cancers. The IDH1 mutations are present in the vast majority of low-grade gliomas and secondary glioblas- tomas. Rapidly emerging work on the consequences of mutant IDH1 protein expression suggests that its neomorphic enzymatic activity catalyzing the production of the oncometabolite 2-hydroxyglutarate influences a range of cellular programs that affect the epigenome, transcriptional programs, hypoxia-inducible factor biology, and development. In the brief time since its discovery, knowledge of the IDH mutation status has had significant translational implica- tions, and diagnostic tools are being used to monitor its expression and function. The concept of IDH1-mutant versus IDH1-wild type will become a critical early distinction in diagnostic and treatment algorithms. (http://thejns.org/doi/abs/10.3171/2012.12.FOCUS12355) KEY WORDS • isocitrate dehydrogenase 1 • isocitrate dehydrogenase 2 • glioblastoma • low-grade glioma • magnetic resonance spectroscopy HE comprehensive taxonomic encyclopedia of the significant because there are specific inhibitors that target genetic alterations underlying all cancers is rapidly their expressed proteins and lead to dramatic clinical re- becoming widely available.61,85,91 To date, this mo- sponse in patients with these cancers.
    [Show full text]
  • The Role of Regulated Necrosis in Endocrine Diseases
    PERSPECTIVES system results in the typical morphological features such as rapid shrinking of the cell, The role of regulated necrosis nuclear condensation, DNA fragmentation, exposure of phosphatidylserine and a in endocrine diseases process known as membrane blebbing11,12. Phosphatidylserine exposure functions Wulf Tonnus , Alexia Belavgeni , Felix Beuschlein , Graeme Eisenhofer, as an ‘eat me’ signal to macrophages13–15. Martin Fassnacht , Matthias Kroiss , Nils P. Krone, Martin Reincke , Importantly, the plasma membrane remains intact in apoptotically dying cells, Stefan R. Bornstein and Andreas Linkermann a mechanism that prevents the release of Abstract | The death of endocrine cells is involved in type 1 diabetes mellitus, intracellular content to the interstitial and/or autoimmunity, adrenopause and hypogonadotropism. Insights from research on extracellular space. Therefore, apoptosis is immunologically silent. The detection basic cell death have revealed that most pathophysiologically important cell death of apoptosis has been misinterpreted for is necrotic in nature, whereas regular metabolism is maintained by apoptosis decades by the TdT-mediated dUTP-biotin programmes. Necrosis is defined as cell death by plasma membrane rupture, which nick end- labelling (TUNEL) method (BOx 1). allows the release of damage- associated molecular patterns that trigger an Mechanistically, extrinsic apoptosis immune response referred to as necroinflammation. Regulated necrosis comes in is mediated by death receptors such as different forms, such as necroptosis, pyroptosis and ferroptosis. In this Perspective, tumour necrosis factor receptor 1 (TNFR1) or the FAS receptor (also known as with a focus on the endocrine environment, we introduce these cell death CD95)16. To kill a cell through a TNFR1 pathways and discuss the specific consequences of regulated necrosis.
    [Show full text]
  • Evidence for Cross Species Extrapolation of Mammalian-Based High-Throughput Screening Assay Results † † ‡ † Carlie A
    Article Cite This: Environ. Sci. Technol. 2018, 52, 13960−13971 pubs.acs.org/est Evidence for Cross Species Extrapolation of Mammalian-Based High-Throughput Screening Assay Results † † ‡ † Carlie A. LaLone,*, Daniel L. Villeneuve, Jon A. Doering, Brett R. Blackwell, § § ¶ † || Thomas R. Transue, Cody W. Simmons, Joe Swintek, Sigmund J. Degitz, Antony J. Williams, † and Gerald T. Ankley † Office of Research and Development, National Health and Environmental Effects Research Laboratory, Mid-Continent Ecology Division, US Environmental Protection Agency, 6201 Congdon Blvd., Duluth, Minnesota 55804, United States ‡ National Research Council, 6201 Congdon Blvd., Duluth, Minnesota 55804, United States § CSRA Inc., 109 T.W. Alexander Drive, Research Triangle Park, North Carolina 27711, United States ¶ Badger Technical Services, 6201 Congdon Blvd., Duluth, Minnesota 55804, United States || Office of Research and Development, National Center for Computational Toxicology, US Environmental Protection Agency, 109 T.W. Alexander Drive, Research Triangle Park, North Carolina 27711, United States *S Supporting Information ABSTRACT: High-throughput screening (HTS) and computational technologies have emerged as important tools for chemical hazard identification. The US Environmental Protection Agency (EPA) launched the Toxicity ForeCaster (ToxCast) Program, which has screened thousands of chemicals in hundreds of mammalian-based HTS assays for biological activity. The data are being used to prioritize toxicity testing on those chemi- cals likely to lead to adverse effects. To use HTS assays in predicting hazard to both humans and wildlife, it is necessary to understand how broadly these data may be extrapolated across species. The US EPA Sequence Alignment to Predict Across Species Susceptibility (SeqAPASS; https://seqapass.epa.gov/seqapass/) tool was used to assess conservation of the 484 protein targets represented in the suite of ToxCast assays and other HTS assays.
    [Show full text]