Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2009. / Härtill 5 uppsatser.

Thesis (M.D.)--Boston University

Graduation date: 1994

Abstract Undefined severe neurological and multi-organ diseases are rare as single diseases, but as a group of diseases, they are responsible for significant morbidity, impaired quality of life and mortality, emphasizing the importance of neuroscience research and its translation into novel diagnostic and treatment strategies. Molecular karyotyping and whole-exome sequencing were used to identify three novel disease-causing genes, GLE1, NHLRC2 and MYH7B, in Northem Finnish families having children with undefined progressive neuromuscular diseases. Functional studies on GLE1, NHLRC2, and MYH7B were conducted in order to understand better the impact of these mutations. The studies revealed that the cellular localization of GLE1 was impaired due to a mutation in the coding gene. The NHLRC2 is involved in many biological processes and its dysfunction has a role in the development of a novel FINCA disease and in fibrosis. Furthermore, mutations in MYH7B in the myosin family have now been connected to encephalomyopathies. Mutations in GLE1, NHLRC2 and MYH7B are involved in encephalomyopathies and neurodegeneration, stressing the important role of these genes in normal psychomotor development Analyses of these previously uncharacterized disease-causing gene mutations provided new insights into the etiologies behind these diseases, representing a relevant starting point for resolving the pathomechanisms underpinning these disorders. The newly-discovered human disease-causing genes and the novel phenotypes of childhood onset neuromuscular diseases provide the possibility for offering the relevant families preclinical diagnostics and may be beneficial in the identification of similar clinical phenotypes all around the world. / Tiivistelmä Yksittäiset, määrittelemättömät, vaikeat neurologiset monielinsairaudet ovat harvinaisia. Sen sijaan neurologisten ja monielinsairauksien alle ryhmittyvät taudit ovat merkittävä syy useisiin sairauksiin, jotka heikentävät elämänlaatua ja aiheuttavat kuolleisuutta. Tästä johtuen neurotieteiden tutkimus ja saatujen tulosten soveltaminen diagnostiikassa ja hoitomuotojen kehittämisessä on hyvin tärkeää. Molekyylikaryotyypitys- ja eksomisekvensointi-menetelmiä hyödynnettiin etsittäessä taudin syytä eteneville neuromuskulaarisairauksille pohjoissuomalaisissa perheissä. Tutkimuksessa tehtiin lisäksi funktionaalisia kokeita GLE1-, NHLRC2- ja MYH7B-proteiineilla, jotta ymmärrettäisiin paremmin löydettyjen mutaatioiden vaikutus potilaiden sairauksiin. Havaittiin, että GLE1-mutaatio vaikutti proteiinin solunsisäiseen paikantumiseen. NHLRC2-proteiini puolestaan on mukana useissa solun biologisissa prosesseissa ja sen toiminnanhäiriö vaikuttaa FINCA-taudin ja fibroosin kehittymiseen. MYH7B-myosiinigeenimutaatio puolestaan yhdistettiin ensimmäistä kertaa enkefalomyopatiaan. Havaittujen tautigeenien; GLE1, NHLRC2 ja MYH7B, vaikutus enkefalomyopatioissa ja neurodegeneraatiossa kertoo, että kyseisillä geeneillä on hyvin todennäköisesti tärkeä rooli ihmisen kehityksessä. Kyseisten, aiemmin tuntemattomien sairautta-aiheuttavien geenimutaatioiden analysointi lisäsi tietoa sairauksien etiologiasta ja loi pohjan tautimekanismien ratkaisemiselle tulevaisuudessa. Työssä esitettyjä uusia sairautta-aiheuttavia geenejä ja uusia karakterisoituja lapsuusiän neuromuskulaarisairauksien ilmiasuja voidaan hyödyntää perheille tarjotun sikiödiagnostiikan lisäksi myös muiden potilaiden samankaltaisen taudinkuvan diagnosoinnissa maailmanlaajuisesti.

Cache Valley virus (CVV) induced malformations have been previously reproduced in ovine fetuses; however, no studies have established the CVV infection sequence of the cells targeted by the virus or the development of the antiviral response of the early, infected fetus that results in viral clearance before development of immunocompetency. To address these questions, ovine fetuses at 35 dg were inoculated in utero with CVV and euthanized at 7, 10, 14, 21 and 28 dpi. On postmortem examination arthrogryposis and oligohydramnios were observed in some infected fetuses. Morphologic studies showed necrosis in the central nervous system (CNS) and skeletal muscle of earlier infected fetuses and hydrocephalus, micromyelia and muscular loss in later infected fetuses. Using immunohistochemistry and in situ hybridization, intense CVV viral antigenic signal was detected in the brain, spinal cord, skeletal muscles and fetal membranes of infected fetuses. Viral signal decreased in targeted and infected tissues with the progression of the infection. To determine specific cell types targeted by CVV in the CNS, indirect immunofluorescence was applied to sections of the CNS using a double labeling technique with antibodies against CVV together with antibodies against neurons, astrocytes and microglia. CVV viral antigen was shown within the cytoplasm of neurons in the brain and spinal cord. No viral signal was observed in microglial cells; however, infected animals had marked microgliosis. The antiviral immune response in immature fetuses infected with CVV was evaluated. Gene expression associated with an innate, immune response was quantified by real-time, quantitative PCR. Upregulated genes in infected fetuses included ISG15, Mx1, Mx2, IL-1, IL-6, TNF-?, TLR-7 and TLR-8. The amount of Mx protein, an interferon stimulated GTPase capable of restricting growth of bunyaviruses, was elevated in the allantoic and amniotic fluid in infected fetuses. ISG15 protein expression was significantly increased in target tissues of infected animals. B lymphocytes and immunoglobulin-positive cells were detected in lymphoid tissues and in the meninges of infected animals. This demonstrated that the infected ovine fetus is able to stimulate an innate and adaptive immune response before immunocompetency that presumably contributes to viral clearance in infected animals.

Magister Artium (Psychology) - MA(Psych) / Arthrogryposis multiplex congenita or A.M.C. is a disorder, which is characterized by multiple contractures of the joints. The disorder presents with varied grades of severity. Despite the physical deformities, intelligence is intact. The aim of this research was to explore the social coping experiences of an adolescent living with Arthrogryposis. This research was a qualitative study. The research design was a case study. Phenomenology was the theoretical framework used. The data collection was done by using one sample, an adolescent living with Arthrogryposis who shared her social coping experiences. Thematic analysis was used to analyse the data. This research was a personal journey for the researcher because he too lives with Arthrogryposis. Objectivity was maintained. After securing the participant’s consent,I emphasized anonymity, confidentiality and the voluntary nature of the study to her.The significance of this research was to develop an understanding of and support for people living with Arthrogryposis and for the disabled in general. The major issues, which arose from this research was the role of transport, social relationships,communication with the non-disabled, the role of the community, making friends, the role of the parent in socialization and others’ perceptions of Arthrogryposis. There were limitations to the study, which included interviews, which were partially inaudible, the absence of the participants’ father, the shyness of the participant, the social inexperience of the participant, lack of other participants and complications with transportation.

Les arthrogryposes multiples congénitales (AMC), limitations articulaires multiples survenant au cours du développement et présentes à la naissance, sont un ensemble hétérogène de maladies dont le dénominateur commun est une diminution des mouvements fœtaux. Dans la majorité des cas, l'AMC est liée à un mécanisme impliquant le système neuromusculaire. Les causes sont dans un grand nombre de cas d'origine génétique. La connaissance de cette cause permet de proposer un conseil génétique avec une évaluation précise du risque de récurrence et éventuellement un diagnostic anténatal. La connaissance du mécanisme sous-jacent participe à l'évaluation du pronostic et permet d'élargir les connaissances sur la mise en place du système neuromusculaire. Mes travaux de thèse ont porté sur ces deux aspects. Dans un premier temps, j'ai étudié l'expression du récepteur de la ryanodine RyR1 dans le muscle squelettique fœtal humain. Les mutations de RYR1 sont responsables de myopathies congénitales. Dans les formes sévères précoces, une AMC peut être occasionnellement associée. L'expression de RyR1 est détectée dès 14 semaines d'aménorrhée dans le muscle fœtal humain. Ce résultat confirme l'expression précoce de RyR1 chez l'homme et permet d'expliquer la possibilité de limitations articulaires. Dans un second temps, j'ai étudié une famille consanguine avec trois enfants atteints d'arthrogrypose distale à la recherche de la cause génétique sous-jacente. L'analyse pangénomique m'a permis de lier pour la première fois le gène ECEL1, codant une endopeptidase, à une pathologie humaine. La recherche de mutations d'ECEL1 dans une cohorte de 20 patients a permis d'identifier cinq autres familles. Toutes les mutations sont transmises sur un mode autosomique récessif et conduisent à une perte de fonction de la protéine. L'ensemble des patients présentent un phénotype clinique et IRM semblable et distinct des autres tableaux d'arthrogrypose distale. Au total, ces travaux confirment l'expression précoce de RyR1 chez l'homme et identifient le gène ECEL1 comme une cause récurrente d'un type particulier d'arthrogrypose distale autosomique récessive. / Arthrogryposis multiplex congenita (AMC) is a heterogeneous group of disorders characterised by multiple joint contractures at birth due to diminished foetal movements. In most cases, the underlying mechanism involves the neuromuscular system. Genetic causes are frequent. Identifying the genetic cause is paramount for precise recurrent risk assessment, genetic counselling and prenatal diagnosis. Elucidating the underlying mechanism allows for prognostic evaluation and expands our knowledge on the development of the human neuromuscular system. My thesis focused on these two aspects. First I studied the expression of the ryanodine receptor 1 in human foetal skeletal muscle. RYR1 mutations cause congenital myopathies. AMC can occasionally be associated with severe forms of RYR1 related congenital myopathies. RyR1 is expressed at 14 weeks of gestational age in human skeletal muscle. Thus it confirms the early expression of RyR1 in human and can account for the occurrence of joint contractures. Second, in order to identify an underlying genetic cause, I studied a consanguineous family with three affected children showing a distal arthrogryposis phenotype. The genome wide linkage study allowed me to link for the first time the endopeptidase coding gene ECEL1 to a human disease. Five other families were shown to carry ECEL1 mutations after screening a cohort of 20 families with distal arthrogryposis. All mutations were recessive and predicted to lead to a loss of function of the protein. All patients showed a recognisable clinical and MRI phenotype that differed to that of currently known distal arthrogryposes. Altogether, these results confirm the early expression of RyR1 in human and identify ECEL1 as a recurrent cause of a distinct type of distal arthrogryposis.