Myoclonus-Dystonia (MD) is an inherited, rare, autosomal dominant movement disorder characterized by quick, involuntary muscle jerking or twitching (myoclonus) and involuntary muscle contractions that cause twisting and pulling movements, resulting in abnormal postures (dystonia). The first MD locus was mapped to 7q21-q31 and called DYT11; this locus corresponds to the SGCE gene. Our group previously identified a second MD locus (DYT15) which maps to a 3.18 Mb region on 18p11. Two patients were chosen to undergo next-generation sequencing, which identified 2,292 shared novel variants within the critical region. Analysis of these variants revealed a 3 bp duplication in a transcript referred to as CD108131, which is believed to be a long non-coding RNA. Characterization of this transcript determined that it is 863 bp in size, it is ubiquitously expressed, with high expression in the cerebellum, and it accounts for ~3% of MD cases.

Objectives: The objectives of this thesis were to: (i) understand the experiences of parents/caregivers of children with inherited metabolic diseases (IMDs), including perceptions of the health care system; and (ii) identify important patient/family-centred outcomes for measurement in future studies. Methods: A qualitative study used semi-structured interviews to gain in-depth insight into caregivers’ experiences. In an adapted meta-synthesis study, the qualitative findings were integrated with the results of related research to identify priority outcomes. Results: Twenty-one caregivers were interviewed. Participants described adjusting to the management of their child’s illness through specific coping strategies but reported stress related to social development. While generally satisfied with disease-specific care, participants described negative experiences with non IMD-specific health services. Health-related quality of life, parental coping, and specific experiences with health care emerged as high-priority outcomes. Conclusions: This project contributes to the limited published literature on caregiver experiences with pediatric IMD and informs future patient-centred research.

Background: Generating evidence to inform care for pediatric medium-chain acyl-CoA dehydrogenase (MCAD) deficiency requires sustainable and integrated measurement of priority outcomes. Methods: From an existing Canadian cohort study, we evaluated the quality of metabolic clinic chart-abstracted data for measuring core outcomes for pediatric MCAD deficiency. We then modelled variation in emergency department (ED) use, in association with disease severity, child age, and distance to care. Results: Children with MCAD deficiency visit the metabolic clinic at least annually on average but we identified data quality challenges related to inconsistent definitions of core outcomes and missing information in patient charts. Rates of ED use were highest among children aged 6 to 12 months, with more severe disease, and living closest to care. Conclusion: While measuring core outcomes through the metabolic clinic for children with MCAD deficiency is feasible, harmonized data collection is needed to evaluate care and further understand ED use.

京都大学 / 0048 / 新制・課程博士 / 博士(社会健康医学) / 甲第20057号 / 社医博第75号 / 新制||社医||9(附属図書館) / 京都大学大学院医学研究科社会健康医学系専攻 / (主査)教授 山田 亮, 教授 小泉 昭夫, 教授 松田 文彦 / 学位規則第4条第1項該当 / Doctor of Public Health / Kyoto University / DFAM

Introduction: The development of molecular genetic methods has in many fields necessitated their inclusion in routine clinical practice, including ophthalmology. The main aim of this thesis was detailed clinical characterization of Czech patients with suspected inherited corneal disorders, followed by genetic testing to determine or specify their clinical diagnosis and subsequently to use the knowledge gained in clinical and genetic counselling and to apply preventive measures in order to avoid loss of vision. Material and Methods: Individuals included in this research were either followed up or newly referred to the Cornea clinic of the Department of Ophthalmology, First Faculty of Medicine, Charles University and General University Hospital in Prague. Detailed clinical examination included corneal tomography, specular microscopy, spectral domain optical coherence tomography, biometry and genealogical analysis. DNA was extracted from peripheral blood leucocytes or buccal cells. Disease-causing variants were searched for using Sanger or massively parallel sequencing, variant pathogenicity was assessed in silico using various algorithms and by segregation analyses within the families. In some cases assessment of the functional impact on the pre-mRNA splicing process was performed. In patients with...

Introduction: Inherited retinal diseases (IRDs) are one of the most common causes of incurable blindness in children and young adults. In the Czech Republic, prior to the start of our work, these disorders had not been the subject of a systematic research. The aim of the study was to identify, clinically characterize and molecular genetically analyse Czech patients with monogenic IRDs and based on the knowledge gained subsequently implement preventive and therapeutic measures to clinical practice. Material and methods: We have performed a comprehensive clinical examination, genealogical analysis and molecular genetic investigation in patients with IRDs and their family members. Detailed ocular examination included spectral domain optical coherence tomography, high-resolution fundus photography and autofluorescence imaging. DNA was isolated from venous blood samples or buccal cells. Causal variants were searched for using Sanger and massively parallel sequencing, and their pathogenicity was evaluated in the context of previously published data, bioinformatical analysis and segregation in available family members. Results: In total, 103 individuals from 76 Czech families diagnosed with IRDs were characterized and their data published. Specifically, we have described clinical and molecular genetic...

Inherited Retinal Degenerations (IRDs) are a heterogenous group of diseases which lead to vision impairment and can be found both in humans and in dogs. About 1 in 1,380 humans is estimated to suffer from an autosomal recessive IRD, which would be 5.5 million people worldwide, and many more are estimated to be unaffected carriers. This makes autosomal recessive IRDs likely the most common group of Mendelian diseases in humans. Today, about 300 genetic mutations have been connected to cause retinal diseases in humans. Whilst in dogs only 32 genes have been identified, numerous eye conditions have been described where the genetic cause has not yet been identified. This suggests that there are much more genetic causes to discover in the dog genome. Additionally, the dog serves well as a model organism to investigate IRDs as it is sharing morphological and genetic similarities with humans. For these reasons, proper software, a canine reference genome of high quality, and smart implementation of bioinformatic tools and methods are a big advantage to increase chances of finding new causative genetic variants and subsequently enable faster detection of possible preventions of the disease or at least alleviating its symptoms via early diagnosis. In this project, a pre-existing pipeline consisting of Bash scripts was stepwise improved with the goal to increase its efficiency. After controlling whether previous data could still be reproduced with the old pipeline in a first step, the software was exchanged to more updated versions in a second step. A main change was the replacement of the mapping tool Burrows-Wheeler Aligner (BWA) from bwa mem to bwa-mem2 mem, and the update of deprecated Genome Analysis Toolkit (GATK) 3.7 to version 4.3 or 4.4. Thirdly, the scripts were adapted from using the older canine reference genome CanFam3.1 to CanFam4. In a fourth step, for automatization and fastening the running time, the pipeline steps were implemented into the workflow management system Nextflow. Additionally, this step was partly aiming to make the pipeline in concordance with the FAIR-principles. All steps were tested on the same test data set, a Labrador retriever family trio, in which one genetic cause for a canine form of the IRD Stargardt disease in a previous study had been detected, namely an insertion in the ABCA4 gene. Lastly, the workflow was also tested on a second data set of a novel IRD of unknown genetic origin on two sibling pairs of Chinese Crested Dogs (CCR). The adjustment of the pipeline shows similar results regarding the change of mapping tool. Introducing the new reference genome revealed a drop of average coverage by one read average for when using CanFam4, while other results were similar. Using the new reference genome increased the number of unknown variants compared to findings with CanFam3.1. However, the known causative variant for the canine form of Stargardt disease, an insertion in ABCA4 gene, could be found in all cases. The run with Nextflow produced identical results to when the respective steps were run with Bash scripts, but it reduced the running time. Running the workflow on the new data set (CCR) and subsequent annotation and filtering indicate new candidates which could be further investigated as a potential cause for this currently unknown cause for an IRD.

No description available.

Objectives: The aim of this study was to gain a thorough understanding of parents’ perceptions of and experiences with the care networks surrounding young children (<=12 years) with inherited metabolic diseases (IMDs). Methods: In this mixed methods study, parent participants created a ‘care map’ depicting their child’s network of care providers. We analyzed care maps using social network analysis. A subset of parents participated in a semi-structured interview. We analyzed interviews thematically and integrated quantitative and qualitative results narratively. Results: Sixty parents contributed care maps and 10 participated in interviews. Parent-drawn care networks were large with few connections between providers. Parents felt responsible for creating and maintaining care networks and for coordinating care. They valued providers who trusted them as part of their child’s health care team. Conclusions: Our findings highlight the complexity of care for children with IMDs and can inform the design of interventions to improve care.

Retinitis pigmentosa (RP) is the most common form of inherited retinal degeneration, heterogenous in its clinical presentation and genetic cause. Understanding the short-term disease mechanisms is pivotal for the development of new therapies. Upcoming clinical trials will take genotype-agnostic approaches; therefore, a comprehensive analysis of progression that encompasses many genetic factors will be needed. In this 10-year retrospective cohort study, rates of progression were measured, structurally and functionally, in 85 RP patients seen at the Ottawa Eye Institute. Parameters examined were the ellipsoid zone (EZ) length on an optical coherence tomography (OCT) image, Humphrey and Goldmann visual fields (VF), and full-field and multifocal electroretinograms (ERGs). RP is revealed to have a 1st order exponential decay pattern of loss, with mean rates of decline of 7.65 %/year for ellipsoid zone (EZ) length, 6.35%/year, 4.39%/year, and 1.57%/year for the Humphrey VF 30-2, 24-2, and 10-2 mean deviation (MD) respectively, and 5.22%/year, 7.77%/year, 6.77%/year, 6.80%/year, and 12.45%/year for Goldmann V4e, III4e, I4e, I3e, and I2e isopter lengths, respectively. In cases where different diagnostic tests were conducted within 3 months of each other, the data was analysed to determine if there was a positive correlation between the diagnostic tests. Ellipsoid zone length and Humphrey 24-2 mean deviation exhibited the strongest association with a coefficient of 0.99. The study reveals structural and functional changes in advanced retinitis pigmentosa and presents a protocol for assessing short-term progression.