Mitochondrial profile and amyloidogenic molecules in sporadic inclusion body miositis Marc Catalán García ADVERTIMENT. La consulta d’aquesta tesi queda condicionada a l’acceptació de les següents condicions d'ús: La difusió d’aquesta tesi per mitjà del servei TDX (www.tdx.cat) i a través del Dipòsit Digital de la UB (diposit.ub.edu) ha estat autoritzada pels titulars dels drets de propietat intel·lectual únicament per a usos privats emmarcats en activitats d’investigació i docència. No s’autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d’un lloc aliè al servei TDX ni al Dipòsit Digital de la UB. No s’autoritza la presentació del seu contingut en una finestra o marc aliè a TDX o al Dipòsit Digital de la UB (framing). Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora. ADVERTENCIA. 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Mitochondrial profile and amyloidogenic molecules in sporadic inclusion body myositis Thesis presented by Marc Catalán García For the Degree of Philosophy Doctor by the University of Barcelona Thesis directed by Josep Maria Grau and Glòria Garrabou School of Medicine and Health Science, University of Barcelona Barcelona, 2017 Als meus pares “Tu mente responderá más preguntas si aprendes a relajarte y a esperar la respuesta” William S. Burroughs “El cerebro es un universo en constante expansión y movimiento. Vamos en una cárcel racional que navega dentro de un loco.” Alejandro Jodorowsky DIRECTOR’S REPORT The thesis presented by the PhD-student Marc Catalán is a compendium of two publications. Marc Catalán has participated actively in the experimental process, data analysis and interpretation of the results and also in the drafting of both manuscripts. The Fundació Cellex financially supported the research not only in laboratory expenses but with the incorporation of Marc to the Muscle Research and Mitochondrial function Laboratory. The two publications included are the core of this thesis and should not be used for other thesis. Publication 1. Mitochondrial DNA disturbances and deregulated expression of OXPHOS and mitochondrial fusion proteins in sporadic inclusion body myositis. Marc Catalán-García, Glòria Garrabou, Constanza Morén, Mariona Guitart-Mampel, Adriana Hernando, Àngels Díaz-Ramos, Ingrid González-Casacuberta, Diana-Luz Juárez, Maria Bañó, Jennifer Enrich-Bengoa, Sonia Emperador, JC. Milisenda, P. Moreno, Ester Tobías, Antonio Zorzano, Julio Montoya, Francesc Cardellach, Josep Maria Grau. Clinical Science. IF: 5.016 (Q1). 2014 IF 5.60 D1; 2015 IF 4.99 Q1 Marc Catalán has contributed in the patient inclusion, sample processing, molecular experiments, data analysis and statistics, interpretation of results, manuscript draft and revision. Publication 2. BACE-1, PS-1 and sAPPβ levels are increased in plasma from sporadic inclusion body myositis patients: surrogate biomarkers among inflammatory myopathies. Marc Catalán-García, Glòria Garrabou, Constanza Morén, Mariona Guitart-Mampel, Ingrid González-Casacuberta, Adriana Hernando, Jose Gallego-Escuredo, Dèlia Yubero, Francesc Villarroya, Raquel Montero, Albert Selva O’Callahan, Francesc Cardellach, Josep Maria Grau. Molecular Medicine. IF: 3.530 (Q1). 2014 IF 4.51 Q1; 2015 IF 3.53 Q2 Marc Catalán has contributed in the study design, patient inclusion, sample processing, molecular experiments, data analysis and statistics, interpretation of results, manuscript draft and revision. Glòria Garrabou Josep M. Grau Postdoctoral Researcher Internal Medicine MD Muscle Research and Mitochondrial Function Laboratory CONTENTS 1. List of abbreviations 13 2. Introduction 15 2.1 Chapter 1: The disease: sporadic inclusion body myositis (sIBM) 17 2.1.1.-Introduction 17 2.1.2.- Epidemiology 17 2.1.3.- Clinical Features 18 2.1.4.- Diagnosis 19 2.1.5.- Pathogenesis 20 2.1.6.- Therapy 22 2.2 Chapter 2: The Mitochondrion 24 2.2.1.- Definition, History and the Endosymbiotic Theory 24 2.2.2.- Structure 24 2.2.3.- Mitochondrial Genome 28 2.2.4.- Oxidative Stress and Apoptosis 28 2.3 Chapter 3: Amyloidogenesis and Sibm 30 2.3.1.- β-Amyloid Accumulation and Alzheimer’s Disease 30 2.3.2.- Relationship between sIBM and Alzheimer’s Disease 30 3. Hypothesis and Objectives 33 4. Articles and Studies 37 Publication 1 : Mitochondrial DNA disturbances and deregulated expression of oxidative phosphorylation and mitochondrial fusion proteins in sporadic inclusion body myositis 39 Publication 2 : BACE-1, PS-1 and sAPPβ levels are increased in plasma from sporadic inclusion body myositis patients: surrogate biomarkers among inflammatory myopathies 53 5. Discussion 63 6. Conclusions 71 7. References 75 8. Aknowledgements 85 9. Annexes 95 - 11 - 1. LIST OF ABBREVIATIONS Aβ: amyloid-β OMM: outer mitochondrial membrane AD: alzheimer’s disease OPA1: optic atrophy 1 AIF: apoptotic inducing factor OXPHOS: oxidative phosphorilation APAF1: apoptotic protease activator factor 1 PARL: presenilins-associated rhomboid like APP: amyloid precursor protein protein ARF: ADP ribosilation factor PBMC: peripheral blood mononuclear cells ATF: activating transcription factor PM: polymyositis ATP: adenosine triphosphate PS1: presenilin 1 BACE1: beta-secretase 1 QMT: quantitative muscle testing CAD: caspase activated DNAse protein ROS: reactive oxygen species CK: creatin kinase RRF: ragged red fiber CN1A: cytosolic 5’ nucleotidase 1A rRNA: ribosomal sibonucleic acid CoQ: Coenzyme Q sAPPβ: amyloid precursor protein (β) COX: cytochrome C oxidase SDH: succinate dehydrogenase CS: citrate synthase SMAC: second mitochondria-derived CytC: cytochrome C activator DM: dermatomyositis sIBM: sporadic inclusion body myositis Drp1: dynamin-related protein 1 SOD1: superoxide dismutase EMG: electromyography TGF-β: transforming growth factor β ENMC: European neuromuscular centre TNF-α: tumour necrosis factor alpha ER: endoplasmic reticulum tRNA: transference ribonucleic acid FAD: flavin adenine dinucleotide Ubb+1: Ubiquitin-B +1 HAE: hydroxyalkenal UPR: unfolded protein response hIBM: hereditary inclusion body myiositis VDAC1: voltage-dependant anion channel 1 IBMFRS: inclusion body myositis functional rating scale IL-1: interleukin 1 IMM: inner mitochondrial membrane IVIG: immunoglobulin therapy kDa: kilodalton MHC: major histocompatibility complex MiD49: mitochondrial dynamics protein 49 MiD51: mitochondrial dynamics protein 51 MDA: malondialdehyde MDV: mitochondria derived vesicles MFF: mitochondrial fusion factor Mfn1: mitofusin 1 Mfn2: mitofusin 2 MHC: major histocompatibility complex MRC: mitochondrial respiratory chain mRNA: messenger ribonucleic acid mtDNA: mitochondrial deoxyribonucleic acid NADH: nicotidamide adenine dinucleotide NAM: necrotizing autoimmune myositis - 13 - 2. INTRODUCTION Chapter 1 2.1. The disease: Sporadic Inclusion Body Myositis (sIBM) 2.2.1.- INTRODUCTION Inflammatory myopathies constitute a group of potentially treatable myopathies in children and adults. Regarding their distinct clinicopathological features, they can be classified in four main subtypes: dermatomyositis (DM), polymyositis (PM), necrotizing autoimmune myositis (NAM) and sporadic inclusion body myositis (sIBM). The common features are proximal muscle wasting and increasing difficulty with tasks requiring these muscles such as lifting objects or climbing steps among others. In addition, neck extensors and pharyngeal muscles could be affected in all forms of inflammatory myopathies, causing dropped head or dysphagia, respectively. The identification of the current subtype is fundamental, because each one has a different prognosis and response to therapies. Typical differential features of the DM consist in skin manifestations such as periorbital heliotrope rash with edema or erythrematous rash. Regarding PM, is still a diagnosis