FUP New insights into creatine transporter deficiency transporter creatine into insights New ANGELO MOLINARO ANGELO 2019 Biomedica UNIVERSITY FIRENZE PRESS PRE M IO TESI DOTTOR A TO FIRENZE UNIVERSITY PRESS UNIVERSITÀ DEGLI STUDI DI FIRENZE – and metabolic targets for treatment and metabolictargetsfor Identification ofneuropathological transporter deficiency insightsintocreatine New ANGELO MOLINARO PREMIO TESI DI DOTTORATO ISSN 2612-8039 (PRINT) | ISSN 2612-8020 (ONLINE) – 81 – PREMIO TESI DI DOTTORATO Commissione giudicatrice, anno 2019 Vincenzo Varano, Presidente della Commissione Tito Arecchi, Area Scientifica Aldo Bompani, Area delle Scienze Sociali Mario Caciagli, Area delle Scienze Sociali Franco Cambi, Area Umanistica Giancarlo Garfagnini, Area Umanistica Roberto Genesio, Area Tecnologica Flavio Moroni, Area Biomedica Adolfo Pazzagli, Area Biomedica Giuliano Pinto, Area Umanistica Vincenzo Schettino, Area Scientifica Maria Chiara Torricelli, Area Tecnologica Luca Uzielli, Area Tecnologica Graziella Vescovini, Area Umanistica 2 Angelo Molinaro New insights into creatine transporter deficiency Identification of neuropathological and metabolic targets for treatment Firenze University Press 2020 New insights into creatine transporter deficiency : identification of neuropathological and metabolic targets for treatment / Angelo Molinaro. – Firenze : Firenze University Press, 2020. (Premio Tesi di Dottorato ; 81) https://www.fupress.com/isbn/9788855180825 ISSN 2612-8039 (print) ISSN 2612-8020 (online) ISBN 978-88-5518-081-8 (print) ISBN 978-88-5518-082-5 (PDF) ISBN 978-88-5518-083-2 (XML) DOI 10.36253/978-88-5518-082-5 Graphic design: Alberto Pizarro Fernández, Lettera Meccanica SRLs *** FUP Best Practice in Scholarly Publishing (DOI 10.36253/fup_best_practice) All publications are submitted to an external refereeing process under the responsibility of the FUP Editorial Board and the Scientific Boards of the series. The works published are evaluated and approved by the Editorial Board of the publishing house, and must be compliant with the Peer review policy, the Open Access, Copyright and Licensing policy and the Publication Ethics and Complaint policy. Firenze University Press Editorial Board M. Garzaniti (Editor-in-Chief), M.E. Alberti, M. Boddi, A. Bucelli, R. Casalbuoni, F. Ciampi, A. Dolfi, R. Ferrise, P. Guarnieri, R. Lanfredini, P. Lo Nostro, G. Mari, A. 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Metadata license: all the metadata are released under the Public Domain Dedication license (CC0 1.0 Universal: https://creativecommons.org/publicdomain/zero/1.0/legalcode). © 2020 Author(s) Published by Firenze University Press Firenze University Press Università degli Studi di Firenze via Cittadella, 7, 50144 Firenze, Italy www.fupress.com This book is printed on acid-free paper Printed in Italy 4 Ai miei genitori Tae of contents Historica perspectie 9 hsioog of creatine and its transporter 11 Creatine transporter eicienc 15 ose oes of CCDS1 19 Aim o the study 23 aterias an ethos 25 Reslts 35 Discssion 47 Concsions 55 Abbreiations 57 igres 59 Taes 91 References 99 Ringraiaenti 111 6 7 Tae of contents Historica perspectie 9 hsioog of creatine and its transporter 11 Creatine transporter eicienc 15 ose oes of CCDS1 19 Aim o the study 23 aterias an ethos 25 Reslts 35 Discssion 47 Concsions 55 Abbreiations 57 igres 59 Taes 91 References 99 Ringraiaenti 111 Angelo Molinaro, New insights into creatine transporter deficiency. Identification of neuropathological and metabolic targets for treatment, © 2020 Author(s), content CC BY 4.0 International, metadata CC0 1.0 Universal, published by Firenze University Press (www.fupress. com), ISSN 2612-8020 (online), ISBN 978-88-5518-082-5 (PDF), DOI7 10.36253/978-88-5518-082-5 istorica perspectie In 1832 the French philosopher and scientist Michel Eugene Chevreul success- fully extracted creatine from meat (the muscles of mammalians) and named it “crea- tine” from the Greek word κρέας (kreas), meaning “meat”. After Chevreul’s discov- ery, Justus von Liebig, a German chemist, in 187 demonstrated that creatine concen- tration as higher in ild animals compared to domestic ones and he concluded that creatine concentration in muscular tissue as dependent upon the level of activity. In 1912, researchers at Harvard University shoed that ingestion of creatine could rise muscular creatine content and later studies demonstrated that creatine supplemen- tation induced nitrogen retention suggesting increasing protein content in muscle. This effect as reversible with the ithdraal of creatine (Haffernan, 2015). In 1923 the content of creatine in human body as measured in about 100 grams, 95 of hich stored in muscle tissue. In the light of all these findings, Alfred Cha- nutin, in 1926, for the first time studied the effect of creatine in humans. He adminis- tered 10 g of creatine a day for a wee and found increased creatine content in mus- cles, proposing that creatine have anabolic effects (Chanutin, 1926). In 1990s the use of creatine as anabolic agent to increase athletes’ performance began and currently creatine is one of the most popular supplements in sport (Balsom, Sderlund and Eblom, 199). In 199, in a patient ith extrapyramidal movement disorder and lo creatinine concentration in serum and urine, by using proton magnetic resonance spectroscopy, a generalized depletion of creatine in the brain as described for the first time (Stcler et al., 199). hen the Authors tried to treat the patient ith arginine (a creatine precursor), they found that the metabolite guanidine acetate increased in the brain, not paralleled by an increase in creatine, indicating that the defect as in the second of the two enzymes responsible for creatine synthesis (see belo). In 2001 another patient presenting ith developmental delay and hypotonia was found to have lo brain creatine content: but in this case serum and urine creatine levels were increased (Salomons et al., 2001). Genetic analysis revealed a nonsense mutation in gene coding for creatine transporter and lo uptae of creatine in fibro- blasts coming from the patient. This as the first description of the clinical syndrome due to genetic defects of the creatine transporter. 8 9 istorica perspectie In 1832 the French philosopher and scientist Michel Eugene Chevreul success- fully extracted creatine from meat (the muscles of mammalians) and named it “crea- tine” from the Greek word κρέας (kreas), meaning “meat”. After Chevreul’s discov- ery, Justus von Liebig, a German chemist, in 187 demonstrated that creatine concen- tration as higher in ild animals compared to domestic ones and he concluded that creatine concentration in muscular tissue as dependent upon the level of activity. In 1912, researchers at Harvard University shoed that ingestion of creatine could rise muscular creatine content and later studies demonstrated that creatine supplemen- tation induced nitrogen retention suggesting increasing protein content in muscle. This effect as reversible with the ithdraal of creatine (Haffernan, 2015). In 1923 the content of creatine in human body as measured in about 100 grams, 95 of hich stored in muscle tissue. In the light of all these findings, Alfred Cha- nutin, in 1926, for the first time studied the effect of creatine in humans. He adminis- tered 10 g of creatine a day for a wee and found increased creatine content in mus- cles, proposing that creatine have anabolic effects (Chanutin, 1926). In 1990s the use of creatine as anabolic agent to increase athletes’ performance began and currently creatine is one of the most popular supplements in sport (Balsom, Sderlund and Eblom, 199). In 199, in a patient ith extrapyramidal movement disorder and lo creatinine concentration in serum and urine, by using proton magnetic resonance spectroscopy, a generalized depletion of creatine in the brain as described for the first time (Stcler et al., 199). hen the Authors tried to treat the patient ith arginine (a creatine precursor), they found that the metabolite guanidine acetate increased in the brain, not paralleled by an increase in creatine, indicating that the defect as in the second of the two enzymes responsible for creatine synthesis (see belo). In 2001 another patient presenting ith developmental delay and hypotonia was found to have lo brain creatine content: but in this case serum and urine creatine levels were increased (Salomons et al., 2001). Genetic analysis revealed a nonsense mutation in gene coding for creatine transporter and lo uptae of creatine in fibro- blasts coming from the patient. This as the first description of the clinical syndrome due to genetic defects of the creatine transporter. Angelo Molinaro, New insights into creatine transporter deficiency. Identification of neuropathological and metabolic targets for treatment, © 2020 Author(s), content CC BY 4.0 International, metadata CC0 1.0 Universal, published by Firenze University Press (www.fupress. com), ISSN 2612-8020 (online),