DOCSLIB.ORG

Fatty Acid Fate in Determining Oxidation and Inflammation in Adipose Tissue Emilio Patrick Mottillo Wayne State University

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Fatty Acid Fate in Determining Oxidation and Inflammation in Adipose Tissue Emilio Patrick Mottillo Wayne State University Wayne State University Wayne State University Dissertations 1-1-2013 Fatty Acid Fate In Determining Oxidation And Inflammation In Adipose Tissue Emilio Patrick Mottillo Wayne State University, Follow this and additional works at: http://digitalcommons.wayne.edu/oa_dissertations Part of the Endocrinology Commons, and the Medicine and Health Sciences Commons Recommended Citation Mottillo, Emilio Patrick, "Fatty Acid Fate In Determining Oxidation And Inflammation In Adipose Tissue" (2013). Wayne State University Dissertations. Paper 678. This Open Access Dissertation is brought to you for free and open access by DigitalCommons@WayneState. It has been accepted for inclusion in Wayne State University Dissertations by an authorized administrator of DigitalCommons@WayneState. FATTY ACID FATE IN DETERMINING OXIDATION AND INFLAMMATION IN ADIPOSE TISSUE by EMILIO PATRICK MOTTILLO DISSERTATION Submitted to the Graduate School of Wayne State University, Detroit, Michigan in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY 2013 MAJOR: PATHOLOGY Approved By: Advisor Date DEDICATION “Nel mezzo del cammin di nostra vita mi ritrovai per una selva oscura che' la diritta via era smarrita.” – Dante Alighieri La Divina Commedia – Inferno “...ma gia volgena il mio disio e'l velle si come rota ch'igualmente e mossa, l'Amor che muove il sole e l'altre stelle” – Dante Alighieri La Divina Commedia – Paradiso To my wife, Sabrina, I thank you for your love and support down this long and arduous road. Your love truly does move the sun and the stars. “e quindi uscimmo a riveder le stelle" – Dante Alighieri La Divina Commedia – Inferno To my children, Liliana and Cristiano, I thank you for bringing a smile to my face each and every day. You truly are the stars in my life. ii ACKNOWLEDGEMENTS First and foremost, I would like to thank my advisor, Dr. James Granneman for sharing his wealth of knowledge. Dr. Granneman’s mentorship has sculpted my growth as an independent researcher. He has taught me that one needs to a well rounded multi-disciplinary scientist, and I only hope that I can be as knowledgeable as him. I would like to thank my committee members for their support throughout this work. I thank Drs. Leff and Mackenzie for their insightful scientific conversations. I believe that my success is a reflection of the training that I received in the CIMER group. I thank Drs. Fridman and Sheng for pushing me to think beyond the details and consider the big picture. In addition, to past and current lab members, I thank them for their help and support along the way. To my colleague, Dr. Yun-Hee Lee, I thank you for your expertise and for advancing work in our laboratory. I am grateful to Dr. Meng-Jer Lee for providing mass-spec data on sphingolipids, and for his collaborative effort. I thank Dr. Diglio, my initial graduate advisor, for his help in the transition to becoming a graduate student. To Dr. Vickie Kimler, I thank you for your editing expertise. iii PREFACE This thesis is based upon the following published papers: 1. Mottillo E.P., X.J. Shen, J.G. Granneman. β3-adrenergic receptor induction of adipocyte inflammation requires lipolytic activation of p38 and JNK. BBA - Molecular and Cell Biology of Lipids. Sep;1801(9):1048-55 2. Mottillo E.P. and J.G. Granneman (2011). Intracellular fatty acids suppress β-adrenergic induction of PKA-targeted gene expression in white adipocytes. Am J Physiol Endocrinol Metab., 301(1), E122-31. 3. Mottillo E.P., Block A.E., Leff T. and J.G. Granneman (2012). Lipolytic products activate peroxisome proliferator-activated receptor (PPAR) α and δ in brown adipocytes to match fatty acid oxidation with supply. J. Biol. Chem. 287: 25038-48. 4. Lee Y.H*, Mottillo E.P.* and J.G. Granneman (2013). Adipose Tissue Plasticity from WAT to BAT and in between. BBA Molecular Basis of Disease. Invited review, in submission. *Equal contribution iv TABLE OF CONTENTS Dedication...................................................................................................................................... ii Acknowledgements....................................................................................................................... iii Preface.......................................................................................................................................... iv List of Figures.............................................................................................................................. viii Chapter 1. Introduction................................................................................................................ 1 1.1 Adipose tissue and obesity.......................................................................................... 1 1.2 Function of adipose tissue........................................................................................... 2 1.3 White and brown adipose tissue................................................................................. 3 1.4 Lipolysis and fatty acid fate in adipocytes.................................................................... 4 1.5 Obesity and inflammation............................................................................................ 6 1.6 Therapeutic remodelling of adipose tissue by targeting β3-Adrenergic Receptors.... 8 1.7 Peroxisome proliferator-activated receptors (PPARs)............................................... 10 1.8 Lipolysis and PPAR signaling...................................................................................... 12 1.9 Hypothesis and aim of the project............................................................................. 13 Chapter 2. Materials and Methods............................................................................................ 15 2.1 Animal studies............................................................................................................ 15 2.2 Cell culture and in vitro assays................................................................................... 16 2.3 Liquid chromatography–mass spectrometry (LC-MS)/MS quantification of ceramides and sphingolipids............................................................................................ 17 2.4 cAMP measurements................................................................................................. 18 2.5 siRNA knockdowns..................................................................................................... 19 2.6 Lentivirus Transduction.............................................................................................. 19 v 2.7 Reporter assays.......................................................................................................... 20 2.8 Generation of reporter constructs............................................................................. 21 2.9 Live cell imaging......................................................................................................... 21 2.10 Quantification of cellular FFAs................................................................................. 22 2.11 Quantification of FFAs and glycerol by biochemical methods................................. 23 2.12 RNA extraction and gene expression analysis.......................................................... 23 2.13 Protein isolation and western blot analysis............................................................. 24 2.14 Quantification of mitochondrial DNA content......................................................... 25 2.15 Measurement of mitochondrial respiration............................................................ 25 2.16 Statistical analysis.................................................................................................... 26 Chapter 3. Intracellular fatty acids promote the expression of inflammatory cytokines in white adipocytes.................................................................................................................... 27 3.1 Expression of inflammatory cytokines by β-AR activation is mediated by intracellular fatty acids......................................................................................................................... 27 3.2 The induction of PAI-1 is partly mediated by ceramides/sphingolipids..................... 29 3.3 Chapter 3 Discussion.................................................................................................. 31 Chapter 4. Intracellular fatty acids limit the induction of oxidative genes in WAT................. 33 4.1 Inhibition of HSL potentiates β3-AR gene expression in WAT................................... 33 4.2 Limiting lipolysis potentiates β-AR induction of PKA-targeted genes in 3T3-L1 adipocytes........................................................................................................................ 36 4.3 Intracellular fatty acids reduce β-AR-coupled gene expression in 3T3-L1 adipocytes........................................................................................................................ 39 4.4 Fatty acids blunt β-AR induction of cAMP by inhibiting adenylyl cyclase.................. 41 vi 4.5 Limiting lipolysis increases mitochondrial activity and promotes CL-induced fat loss................................................................................................................................... 44 4.6 Chapter 4 Discussion.................................................................................................. 46 Chapter 5. Lipolytic
Load more

Recommended publications
  • Remodeling of Lipid Droplets During Lipolysis and Growth in Adipocytes
    Remodeling of Lipid Droplets during Lipolysis and Growth in Adipocytes Margret Paar*1, Christian Jungst§1,2, Noemi A. Steinert, Christoph Magnes ~, Frank Sinner~, Dagmar Kolbll, Achim Lass*, Robert Zimmermann*, Andreas Zumbusch§, Sepp D. Kohlwein*, and Heimo Wolinski*3 From the *Institute of Molecular Biosciences, Lipidomics Research Center LRC Graz, University of Graz, 8070 Graz, Austria, the §Department of Chemistry, University of Konstanz, 78457 Konstanz, Germany, ~HEAL TH, Institute for Biomedicine and Health Sciences, Joanneum Research, 8036 Graz, Austria, and the IIlnstitute of Cell Biology, Histology and Embryology, and ZMF, Center for Medical Research, Medical University of Graz, 8070 Graz, Austria Background: Micro-lipid droplets (mLDs) appear in adipocytes upon lipolytic stimulation. LDs may grow by spontaneous, homotypic fusion. Results: Scavenging of fatty acids prevents mLD formation. LDs grow by a slow transfer of lipids between LDs. Conclusion: mLDs form due to fatty acid overflow. I.D growth is a controlled process. Significance: Novel mechanistic insights into LD remodeling are provided. Synthesis, storage, and turnover oftriacylglycerols (TAGs) in mation of large LOs requires a yet uncharacterized protein adipocytes are critical cellular processes to maintain lipid and machinery mediating LO interaction and lipid transfer. energy homeostasis in mammals. TAGs are stored in metaboli­ cally highly dynamic lipid droplets (LOs), which are believed to undergo fragmentation and fusion under lipolytic and lipogenic Most eukaryotic organisms deal with a typically fluctuating conditions, respectively. Time lapse fluorescence microscopy food supply by storing or mobilizing lipids as an energy source. showed that stimulation of lipolysis in 3T3-Ll adipocytes causes Malfunction of the synthesis or degradation of fat stores is progressive shrinkage and almost complete degradation of all linked to prevalent diseases, such as obesity, type II diabetes, or cellular LDs but without any detectable fragmentation into various forms of lipodystrophy (1).
    [Show full text]
  • Modulation of Triglyceride and Cholesterol Ester Synthesis Impairs
    THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 289, NO. 31, pp. 21276–21288, August 1, 2014 Author’s Choice © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A. Modulation of Triglyceride and Cholesterol Ester Synthesis Impairs Assembly of Infectious Hepatitis C Virus*□S Received for publication, May 20, 2014, and in revised form, June 8, 2014 Published, JBC Papers in Press, June 10, 2014, DOI 10.1074/jbc.M114.582999 Jolanda M. P. Liefhebber‡1, Charlotte V. Hague‡1, Qifeng Zhang§, Michael J. O. Wakelam§, and John McLauchlan‡2 From the ‡Medical Research Council-University of Glasgow Centre for Virus Research, 8 Church Street, Glasgow G11 5JR, Scotland, United Kingdom and §The Babraham Institute, Babraham Research Campus, Cambridge CB22 3AT, United Kingdom Background: Lipid droplets have been implicated in HCV virion assembly. Results: Inhibitors of the synthesis of triacylglycerol and cholesterol ester, the main components of lipid droplets, impair virion assembly. Conclusion: Production of infectious HCV is linked integrally with the biosynthesis of the major components of lipid droplets. Significance: Inhibitors of enzymes that generate acylglycerols and cholesterol esters have antiviral activity. In hepatitis C virus infection, replication of the viral genome including cirrhosis, hepatocellular carcinoma, and liver failure and virion assembly are linked to cellular metabolic processes. (1). Metabolic diseases including insulin resistance and steato- In particular, lipid droplets, which store principally triacylglyc- sis are particular features of chronic infection (2, 3), and there is Downloaded from erides (TAGs) and cholesterol esters (CEs), have been impli- now conclusive evidence that factors and pathways involved in cated in production of infectious virus.
    [Show full text]
  • Long-Chain Acyl-Coa Synthetase Is Associated with the Growth of Malassezia Spp
    Journal of Fungi Article Long-Chain Acyl-CoA Synthetase is Associated with the Growth of Malassezia spp. Tenagy, Kengo Tejima, Xinyue Chen , Shun Iwatani and Susumu Kajiwara * School of Life Science and Technology, Tokyo Institute of Technology, J3-7, 4259 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan; [email protected] (T.); [email protected] (K.T.); [email protected] (X.C.); [email protected] (S.I.) * Correspondence: [email protected]; Tel.: +81-45-924-5715; Fax: +81-45-924-5773 Received: 14 July 2019; Accepted: 9 September 2019; Published: 21 September 2019 Abstract: The lipophilic fungal pathogen Malassezia spp. must acquire long-chain fatty acids (LCFAs) from outside the cell. To clarify the mechanism of LCFA acquisition, we investigated fatty acid uptake by this fungus and identified the long-chain acyl-CoA synthetase (ACS) gene FAA1 in three Malassezia spp.: M. globosa, M. pachydermatis, and M. sympodialis. These FAA1 genes could compensate for the double mutation of FAA1 and FAA4 in Saccharomyces cerevisiae, suggesting that Malassezia Faa1 protein recognizes exogenous LCFAs. MgFaa1p and MpFaa1p utilized a medium-chain fatty acid, lauric acid (C12:0). Interestingly, the ACS inhibitor, triacsin C, affected the activity of the Malassezia Faa1 proteins but not that of S. cerevisiae. Triacsin C also reduced the growth of M. globosa, M. pachydermatis, and M. sympodialis. These results suggest that triacsin C and its derivatives are potential compounds for the development of new anti-Malassezia drugs. Keywords: Malassezia; acyl-CoA synthetase; fatty acid uptake 1. Introduction Malassezia spp.
    [Show full text]
  • Glucagon-Like Peptide 1 Stimulates Lipolysis in Clonal Pancreatic Я-Cells
    Glucagon-Like Peptide 1 Stimulates Lipolysis in Clonal Pancreatic ␤-Cells (HIT) Gordon C. Yaney, Vildan N. Civelek, Ann-Marie Richard, Joseph S. Dillon, Jude T. Deeney, James A. Hamilton, Helen M. Korchak, Keith Tornheim, Barbara E. Corkey, and Aubrey E. Boyd, III Glucagon-like peptide 1 (GLP-1) is the most potent incretin rather than a secretagogue (5). Activation of PKA physiological incretin for insulin secretion from the leads to phosphorylation of multiple ␤-cell proteins, many of pancreatic ␤-cell, but its mechanism of action has not which have been hypothesized to play a role in insulin secre- been established. It interacts with specific cell-surface tion (6–8). The nature of the endogenous substrates for PKA receptors, generates cAMP, and thereby activates pro- that may potentiate insulin secretion is unknown. Because the ␤ tein kinase A (PKA). Many changes in pancreatic -cell islet contains large stores of triglycerides (9), particularly in function have been attributed to PKA activation, but the diabetes (10), another possible role of the normal rise in contribution of each one to the secretory response is unknown. We show here for the first time that GLP-1 cAMP could be to stimulate lipolysis (via lipase activation), rapidly released free fatty acids (FFAs) from cellular thereby providing the cell with free fatty acids (FFAs). Recent research on hormone-sensitive lipase (HSL) in ␤-cells stores, thereby lowering intracellular pH (pHi) and stimulating FFA oxidation in clonal ␤-cells (HIT). Sim- yielded results consistent with that notion (11). The released ilar changes were observed with forskolin, suggesting FFAs may directly effect secretion, or they may do so indi- that stimulation of lipolysis was a function of PKA rectly via generation of other lipids, including the putative activation in ␤-cells.
    [Show full text]
  • Apolipoprotein O Is Mitochondrial and Promotes Lipotoxicity in Heart
    Apolipoprotein O is mitochondrial and promotes lipotoxicity in heart Annie Turkieh, … , Philippe Rouet, Fatima Smih J Clin Invest. 2014;124(5):2277-2286. https://doi.org/10.1172/JCI74668. Research Article Cardiology Diabetic cardiomyopathy is a secondary complication of diabetes with an unclear etiology. Based on a functional genomic evaluation of obesity-associated cardiac gene expression, we previously identified and cloned the gene encoding apolipoprotein O (APOO), which is overexpressed in hearts from diabetic patients. Here, we generated APOO-Tg mice, transgenic mouse lines that expresses physiological levels of human APOO in heart tissue. APOO-Tg mice fed a high-fat diet exhibited depressed ventricular function with reduced fractional shortening and ejection fraction, and myocardial sections from APOO-Tg mice revealed mitochondrial degenerative changes. In vivo fluorescent labeling and subcellular fractionation revealed that APOO localizes with mitochondria. Furthermore, APOO enhanced mitochondrial uncoupling and respiration, both of which were reduced by deletion of the N-terminus and by targeted knockdown of APOO. Consequently, fatty acid metabolism and ROS production were enhanced, leading to increased AMPK phosphorylation and Ppara and Pgc1a expression. Finally, we demonstrated that the APOO-induced cascade of events generates a mitochondrial metabolic sink whereby accumulation of lipotoxic byproducts leads to lipoapoptosis, loss of cardiac cells, and cardiomyopathy, mimicking the diabetic heart–associated metabolic phenotypes.
    [Show full text]
  • A Role for the Malonyl-Coa/Long-Chain Acyl-Coa
    A Role for the Malonyl-CoA/Long-Chain Acyl-CoA Pathway of Lipid Signaling in the Regulation of Insulin Secretion in Response to Both Fuel and Nonfuel Stimuli Raphae¨l Roduit,1 Christopher Nolan,1 Cristina Alarcon,2 Patrick Moore,2 Annie Barbeau,1 Viviane Delghingaro-Augusto,1 Ewa Przybykowski,1 Johane Morin,1 Fre´de´ric Masse´,1 Bernard Massie,3 Neil Ruderman,4 Christopher Rhodes,2,5 Vincent Poitout,2,6 and Marc Prentki1 The malonyl-CoA/long-chain acyl-CoA (LC-CoA) model between the metabolic coupling factor malonyl-CoA, the of glucose-induced insulin secretion (GIIS) predicts partitioning of fatty acids, and the stimulation of insulin that malonyl-CoA derived from glucose metabolism in- secretion to both fuel and nonfuel stimuli. Diabetes 53: hibits fatty acid oxidation, thereby increasing the avail- 1007–1019, 2004 ability of LC-CoA for lipid signaling to cellular processes involved in exocytosis. For directly testing the model, INSr3 cell clones overexpressing malonyl- CoA decarboxylase in the cytosol (MCDc) in a tetracy- lthough glucose is the most potent nutrient cline regulatable manner were generated, and INS(832/ secretagogue for the islet ␤-cell, its effective- 13) and rat islets were infected with MCDc-expressing ness as such can be markedly modulated by adenoviruses. MCD activity was increased more than other fuel stimuli, hormones, and neurotrans- fivefold, and the malonyl-CoA content was markedly A mitters (1–3). Of the nutrient modulators, fatty acids are diminished. This was associated with enhanced fat oxi- particularly important as, at one extreme, islet fatty acid dation at high glucose, a suppression of the glucose- induced increase in cellular free fatty acid (FFA) deprivation essentially abolishes glucose-induced insulin content, and reduced partitioning at elevated glucose of secretion (GIIS) (4–7), whereas at the other extreme, exogenous palmitate into lipid esterification products.
    [Show full text]
  • WO 2011/103516 A2 25 August 2011 (25.08.2011) PCT
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau I (10) International Publication Number (43) International Publication Date WO 2011/103516 A2 25 August 2011 (25.08.2011) PCT (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every A61K 31/536 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, (21) Number: International Application CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, PCT/US201 1/025558 DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, 18 February 20 11 (18.02.201 1) KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (25) Filing Language: English NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, (26) Publication Language: English SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (30) Priority Data: 61/305,862 18 February 2010 (18.02.2010) US (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (71) Applicant (for all designated States except US): THE GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, TRUSTEES OF PRINCETON UNIVERSITY ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, [US/US]; P.O.
    [Show full text]
  • Triacsin © 1
    Triacsin © 1. Discovery, producing organism and structures1-4) Microbial inhibitors of fatty acid metabolism were Fatty acid Chapter 1 screened by an assay system using acyl-CoA synthe- tase I (ACS-I)-deficient (L-7) and fatty acid synthase (FAS)-deficient(A-1) mutants of Candida lipolytica. ACS-I ACS-II Triacsins were isolated from the culture broth of the actinomycete strain SK-1894 and recognized as acyl- Lipid Acyl–CoA Acyl–CoA Chapter 2 CoA synthetase inhibitors since they showed inhib- itory activity against growth of mutant A-1, but not β-Oxidation against mutant L-72,3). Triacsins C and D were iden- FAS tified as WS-1228 A and B, respectively, originally Peroxisomers isolated as vasodilators4). No correlation was observed Chapter 3 Malonyl–CoA Acetyl–CoA between the two activities. The first total synthesis of ACC triacsin C (WS-1228A) was reported by Tanaka et al.5) Glucose Fatty acid metabolism of C. lipolytica Appendices NNNOH H3C Triacsin A NNNOH H C 3 Publications Triacsin B NNNOH H3C Triacsin C (WS-1228A) NNNOH H3C Triacsin D (WS-1228B) Streptomyces sp. SK-1894 2. Physical data (Triacsin C) 1) Yellow powder. C11H17N3O ; mol wt 207. Sol. in acetone, EtOAc, EtOH. Insol. in H2O. 3. Biological activity6) 1) Inhibition of long chain acyl-CoA synthetase IC50 (µM) Enzyme source Reference Triacsin A Triacsin B Triacsin C Triacsin D Pseudomonas fraji 26 > 150 17 >150 1 Pseudomonas aeruginosa 17 > 200 3.6 >200 5 Rat liver 18 > 200 8.7 >200 5 Raji cells 12 > 100 6.3 >100 6 HSDMICI cells Long chain ACS NT NT 0.48 NT 7 Arachidonoyl-CoA synthetase NT NT 8.5 NT 7 Candida lipolytica ACS –I 5.5 > 50 4.0 >50 － 323 － 2) N-Hydroxytriazene moiety is essential for acyl-CoA synthetase inhibition.
    [Show full text]
  • A Link Between Lipid Metabolism and Epithelial-Mesenchymal Transition Provides a Target for Colon Cancer Therapy
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 6, No. 36 A link between lipid metabolism and epithelial-mesenchymal transition provides a target for colon cancer therapy Ruth Sánchez-Martínez1, Silvia Cruz-Gil1,*, Marta Gómez de Cedrón1,*, Mónica Álvarez-Fernández2, Teodoro Vargas1, Susana Molina1, Belén García1, Jesús Herranz3, Juan Moreno-Rubio4,5, Guillermo Reglero1, Mirna Pérez-Moreno6, Jaime Feliu4, Marcos Malumbres2, Ana Ramírez de Molina1 1 Molecular Oncology and Nutritional Genomics of Cancer Group, IMDEA Food Institute, CEI UAM + CSIC, Madrid, Spain 2 Cell Division and Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain 3 Biostatistics Unit, IMDEA Food Institute, CEI UAM+CSIC, Madrid, Spain 4 Medical Oncology, La Paz University Hospital (IdiPAZ-UAM), Madrid, Spain 5 Precision Oncology Laboratory (POL), Infanta Sofía University Hospital, Madrid, Spain 6 Epithelial Cell Biology Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain * These authors have contributed equally to this work Correspondence to: Ana Ramírez de Molina, e-mail: [email protected] Keywords: colorectal cancer, lipid metabolism, acyl-CoA synthetases, stearoyl-CoA desaturase, epithelial-mesenchymal transition Received: June 08, 2015 Accepted: September 24, 2015 Published: October 05, 2015 ABSTRACT The alterations in carbohydrate metabolism that fuel tumor growth have been extensively studied. However, other metabolic pathways involved in malignant progression, demand further understanding. Here we describe a metabolic acyl-CoA synthetase/ stearoyl- CoA desaturase ACSL/SCD network causing an epithelial-mesenchymal transition (EMT) program that promotes migration and invasion of colon cancer cells. The mesenchymal phenotype produced upon overexpression of these enzymes is reverted through reactivation of AMPK signaling. Furthermore, this network expression correlates with poorer clinical outcome of stage-II colon cancer patients.
    [Show full text]
  • Download Product Insert (PDF)
    PRODUCT INFORMATION Triacsin C Item No. 10007448 CAS Registry No.: 76896-80-5 Formal Name: 2E,4E,7E-undecatrienal, nitrosohydrazone Synonyms: FR 900190, WS 1228A MF: C11H17N3O N N FW: 207.3 N O Purity: ≥95% H UV/Vis.: λmax: 301 nm Supplied as: A crystalline solid Storage: -20°C Stability: ≥2 years Item Origin: Bacterium/Streptomyces aureofaciens Information represents the product specifications. Batch specific analytical results are provided on each certificate of analysis. Laboratory Procedures Triacsin C is supplied as a crystalline solid. A stock solution may be made by dissolving the triacsin C in the solvent of choice. Triacsin C is soluble in organic solvents such as methanol and DMSO, which should be purged with an inert gas. The solubility of triacsin C in these solvents is approximately 5 mg/ml. Description Triacsin C, originally isolated from Streptomyces sp., is an inhibitor of long fatty acid acyl-CoA synthetase 1 (IC50 = 6.3 μM in Raji cells). It has been shown to interfere with lipid metabolism by inhibiting the de novo synthesis of triglycerides, diglycerides, and cholesterol esters.2 It is also known to act as a hypotensive vasodilator, modulating endothelial nitric oxide synthase repalmitoylation by limiting palmitoyl CoA availability.3 Triacsin C can inhibit pancreatic β-cell function by suppressing the mobilization of intracellular calcium.4 References 1. Tomoda, H., Igarashi, K., Cyong, J.C., et al. Evidence for an essential role of long chain acyl-CoA synthetase in animal cell proliferation. Inhibition of long chain acyl-CoA synthetase by triacsins caused inhibition of Raji cell proliferation.
    [Show full text]
  • Fatty Acid-Induced Я Cell Apoptosis
    Proc. Natl. Acad. Sci. USA Vol. 95, pp. 2498–2502, March 1998 Medical Science Fatty acid-induced b cell apoptosis: A link between obesity and diabetes MICHIO SHIMABUKURO*†,YAN-TING ZHOU*†,MOSHE LEVI†, AND ROGER H. UNGER*†‡ *Gifford Laboratories for Diabetes Research, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75235; and †Veterans Administration Medical Center, Dallas, TX 75216 Contributed by Roger H. Unger, January 12, 1998 ABSTRACT Like obese humans, Zucker diabetic fatty vivo in the prediabetic phase of the disease causes the com- (ZDF) rats exhibit early b cell compensation for insulin pensatory b cell hyperplasia and hyperinsulinemia; a further resistance (4-fold b cell hyperplasia) followed by decompen- increase in islet fat to ;50 times normal reverses the foregoing sation (>50% loss of b cells). In prediabetic and diabetic ZDF compensatory changes and causes b cell dysfunction, a reduc- islets, apoptosis measured by DNA laddering is increased 3- tion in the number of b cells, and diabetes (7, 8). In other and >7-fold, respectively, compared with lean ZDF controls. words, the surfeit of fat in islets is associated with a dose- Ceramide, a fatty acid-containing messenger in cytokine- related biphasic effect, initially enhancing insulin output by induced apoptosis, was significantly increased (P < 0.01) in stimulating hyperplasia (7, 8) but subsequently reversing these prediabetic and diabetic islets. Free fatty acids (FFAs) in compensatory changes when the fat content rises to extremely plasma are high (>1 mM) in prediabetic and diabetic ZDF high levels (7, 8). rats; therefore, we cultured prediabetic islets in 1 mM FFA.
    [Show full text]
  • Mechanistic Studies of the Long Chain Acyl-Coa Synthetase Faa1p from Saccharomyces Cerevisiae Hong Li Center for Metabolic Disease, Ordway Research Institute
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Biochemistry -- Faculty Publications Biochemistry, Department of 2007 Mechanistic Studies of the Long Chain Acyl-CoA Synthetase Faa1p from Saccharomyces cerevisiae Hong Li Center for Metabolic Disease, Ordway Research Institute Elaina M. Melton Center for Metabolic Disease, Ordway Research Institute Steven Quackenbush Center for Metabolic Disease, Ordway Research Institute Concetta C. DiRusso Center for Metabolic Disease, Ordway Research Institute, [email protected] Paul N. Black Center for Metabolic Disease, Ordway Research Institute, [email protected] Follow this and additional works at: http://digitalcommons.unl.edu/biochemfacpub Part of the Biochemistry Commons, Biotechnology Commons, and the Other Biochemistry, Biophysics, and Structural Biology Commons Li, Hong; Melton, Elaina M.; Quackenbush, Steven; DiRusso, Concetta C.; and Black, Paul N., "Mechanistic Studies of the Long Chain Acyl-CoA Synthetase Faa1p from Saccharomyces cerevisiae" (2007). Biochemistry -- Faculty Publications. 201. http://digitalcommons.unl.edu/biochemfacpub/201 This Article is brought to you for free and open access by the Biochemistry, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Biochemistry -- Faculty Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. NIH Public Access Author Manuscript Biochim Biophys Acta. Author manuscript; available in PMC 2008 September 1. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Biochim Biophys Acta. 2007 September ; 1771(9): 1246±1253. Mechanistic Studies of the Long Chain Acyl-CoA Synthetase Faa1p from Saccharomyces cerevisiae Hong Li1,2, Elaina M. Melton1,2, Steven Quackenbush1,2, Concetta C. DiRusso1,2, and Paul N.
    [Show full text]