Novel Experimental Strategies to Treat Multiple Sclerosis
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NOVEL EXPERIMENTAL STRATEGIES TO TREAT MULTIPLE SCLEROSIS Presented by Alba Sánchez Fernández ACADEMIC DISSERTATION To obtain the degree of PhD in Neuroscience by the Universitat Autònoma de Barcelona (2019) NOVEL EXPERIMENTAL STRATEGIES TO TREAT MULTIPLE SCLEROSIS Presented by Alba Sánchez Fernández ACADEMIC DISSERTATION To obtain the degree of PhD in Neuroscience by the Universitat Autònoma de Barcelona (2019) Directed by: Tutorized by: Rubèn López Vales Xavier Navarro Acebes INDEX Summary.............................................................................................................................. 4 Introduction ....................................................................................................................... 5 1. Multiple sclerosis ............................................................................................... 5 2. The inflammatory response in Multiple Sclerosis ................................ 12 3. Animal models of Multiple Sclerosis .......................................................... 16 4. Current therapies for Multiple Sclerosis ................................................... 19 5. Interleukin-1 Family ........................................................................................ 22 6. Resolving the inflammation ........................................................................... 30 Objectives ............................................................................................................................ 44 Materials and methods ................................................................................................... 46 - Human samples ......................................................................................................... 46 - Mice ................................................................................................................................ 47 - Experimental Autoimmune Encephalomyelitis induction ........................ 48 - Experimental Autoimmune Encephalomyelitis evaluation ...................... 48 - Drug administration ................................................................................................ 49 - Real-time quantitative PCR ................................................................................... 50 - Histological analysis ................................................................................................ 51 - Cytokine protein expression assessment ........................................................ 52 - Fluorescence activated cell sorting ................................................................... 52 - Statistical analysis .................................................................................................... 55 Results .................................................................................................................................. - Chapter 1. OLT1177 ................................................................................................. 57 - Chapter 2. IL-37 ......................................................................................................... 81 - Chapter 3. Maresin-1 ............................................................................................... 109 General discussion ........................................................................................................... 131 Conclusions ......................................................................................................................... 142 References ........................................................................................................................... 144 Abbreviations .................................................................................................................... 162 SUMMARY Despite multiple sclerosis was described by Dr. Jean-Martin Charcot in the 19th century, it is currently a challenging neurological disorder. The etiology of this pathology is not fully understood but the degeneration that occurs in myelin sheaths and neurons in individual suffering from multiple sclerosis is linked to aberrant presence of immune cells in the lesioned central nervous system. For this reason, in the present thesis we propose to modulate the inflammatory response to ameliorate neurological decline and demyelination in experimental autoimmune encephalomyelitis, a murine model of multiple sclerosis. The data shown here reveals that the administration of anti-inflammatory approaches, such as OLT1177 and IL-37, or specialized pro-resolving mediators, such as Resolvin-D1 or Maresin-1, has therapeutic effects in mice with experimental autoimmune encephalomyelitis, even if these treatments are initiated at the onset of the disease. OLT1177, IL-37 and Maresin-1 exerts beneficial effects by reducing some common aspects of inflammation, such as the production pro-inflammatory cytokines and the accumulation of immune cells. IL-37 and Maresin-1 also modulate the inflammatory response in the periphery and alter the polarization of lymphocyte and macrophage towards a more anti-inflammatory or regulatory phenotype. Collectively, the results presented here provide novel data that could pave the way to develop more effective strategies to treat multiple sclerosis. INTRODUCTION 1. MULTIPLE SCLEROSIS More than 100 years has passed since Charcot, Carswell, Cruveilhier and others described the clinical and pathological characteristics of multiple sclerosis (MS). This disease has originally been defined as a chronic inflammatory disease of the central nervous system (CNS), leading to focal plaques of primary demyelination and axonal damage in the white matter of the brain and spinal cord. It manifests clinically as neurological deficit (Charcot, 1880). 40 years later it was categorized as an autoimmune disease (Wu and Alvarez, 2011; Baecher-Allan et al., 2018). However, the aetiology and the pathogenesis of this disease are not yet conclusively known (Reviewed at Dyment et al., 2004; Trapp and Nave, 2008; Wu and Alvarez, 2011; Tselis, 2012; Didonna and Oksenberg, 2017; Baecher-Allan et al., 2018). This enigmatic, relapsing and often eventually progressive disorder of the white matter of the CNS continues to challenge researchers trying to understand the pathogenesis of the disease and prevent its progression. MS is a chronic, slowly progressive, inflammatory, demyelinating and degenerative disease of the CNS characterized by an autoimmune inflammation(Malpass, 2012; Grigoriadis and van Pesch, 2015; Bjelobaba et al., 2017). The symptoms vary widely, and affected individuals can experience one or more effects of nervous system damage This pathology affects about 2.5 million people worldwide, ranking as one of the most prevalent neurodegenerative diseases (Wekerle, 2008). MS affects 50000 people in