No description available.

No description available.

PhD, Faculty of Health Sciences, University of the Witwatersrand / HIV/AIDS continues to be one of the greatest health challenges which South Africa faces. The epidemic in children is closely linked to that in women, the prevalence of which continues to grow according to antenatal statistics from the South African Department of Health (DOH). HIV is known to invade the central nervous system at the time of infection, and causes widespead damage. In children, this leads to a well-researched condition known as HIV encephalopathy, which affects all areas of neurodevelopment. The effects of timely initiation of antiretroviral therapy on alleviating the impact of encephalopathy have been well described. Neurodevelopmental delay is a stage four disease indicator according to the World Health Organisation (WHO), and therefore is a criterion for initiation of Highly Active Antiretroviral Therapy (HAART). HAART is often only administered according to the virologic and immunologic status of a child, as standardised neurodevelopmental assessment tools are not widely available in South African clinics. When HAART initation is dependent on immunologic status, it is often too late to prevent encephalopahy. To date, the only means of prevention of this condition is early initation of HAART, which has not been widely available in South Africa. Stringent guidelines for the commencement of this therapy according to the WHO, and the South African Department of Health (DOH) have had to be followed, leading to late initiation of HAART, and widespread central nervous system encephalopathy. Studies which have been carried out in South African clinics have demonstrated the high prevalence of this condition. Once there is evidence of encephalopathy, children should be referred for assessments in all facets of development, and where necessary, for rehabilitation. A standardised developmental screening tool which is suitable for use in a developing country is therefore necessary in order to screen for neurodevelopmental delays to allow for further assessment and referral to rehabilitation services, as well as providing an additional assessment criterion for initiation of HAART. Paediatric HIV clinics in developing countries are understaffed, and children may be seen by junior staff or screened by nurses due to the high numbers of clinic attendees. This often results in neurodevelopment being inadequately assessed and children are therefore not referred for intervention services. A standardised screening tool, which The Development of a Screening Tool to Evaluate Infants who are HIV Positive could be administered by clinic staff in order to ensure correct and timely referral of children for further assessment and intervention is therefore necessary. This is of importance both locally and internationally where a screening tool, which has been developed specifically for this purpose, does not exist. The aim of this study was therefore to evaluate the agreement between the Bayley-III Screening Test and the Bayley Scales of Infant Development (3rd version) in a population of HIV positive infants in order to evaluate its appropriateness for use in South Africa. The Bayley Scales of Infant Development have long been considered the ‘gold standard’ in infant developmental assessment, which is why this tool was chosen to evaluate the Bayley-III Screening Test against. The developmental scores in each facet (cognitive, motor or language) were evaluated to determine which should be included in an assessment tool for this population. Further objectives for the study were to adapt the screening tool to the needs of the population, or to develop a new screening tool should the Bayley-III Screening Test not prove suitable for use in this population. In order to meet the aims and objectives, a cross-sectional study was conducted where 112 HIV positive infants between the ages of six and eighteen months were assessed using the Bayley-III Screening Test and the Bayley Scales of Infant Development (3rd version) (BSID III). The infants were stratified into four age groups namely 6-8 months, 9- 12 months, 13-16 months, and 17-18 months. Children were recruited from Harriet Shezi Children’s Clinic at Chris Hani Baragwanath Hospital in Soweto. The agreement between the Bayley-III Screening Test and the Bayley Scales of Infant Development (3rd version) was analysed using Kappa, for the overall group, and for each age group. Overall agreement between the tools was as follows: K=0.58 for the Cognitive facet, K=0.82 for the Expressive Communication facet, K=0.76 for the Receptive Communication facet, K=0.44 for the Fine Motor facet and K=0.57 for the Gross Motor facet. These values indicate that the Bayley-III Screening Test is therefore not acceptable for clinical use, as excellent agreement (k≥0.75) in all facets would be necessary for this purpose. A new screening tool therefore had to be developed. The infant’s developmental scores from the BSID III were analysed to determine which facets of development were most severely affected, and therefore which facets should be included in a new screening tool. Gross motor function was demonstrated to be the area which was most severely affected, followed by cognitive function. A gross motor screening tool would therefore be suitable for use in this population, as no equipment would be necessary. Gross motor development is the most universally similar aspect of development, which is not completely dependent on cultural or socioeconomic factors which often have an influence on language and cognitive development. Item selection from the BSID III was undertaken to determine which items should be included in a brief screening tool. In each of the four age groups, item selection occurred as follows: Two items which discriminated the At-Risk, from Emerging and Competent groups (less than 20% in the At-Risk group, and 100% in the other groups) were selected. Two items, which discriminated between children in the ‘Emerging’ and ‘Competent’ categories on the BSID III were selected (0-5% of children who were At-Risk obtained credit, 30-50% of the Emerging group obtained credit, and 100% of the Competent group obtained credit). Lastly, two items were selected which discriminated the Competent group from the other two groups (100% or as high as possible in the Competent group, and 0% in the other groups). The new gross motor screening tool was assembled using the selected items, scoring was allocated, and it was tested against the scores obtained on the Gross Motor facet of the BSID III for the initial 112 infants. Agreement between the tools was analysed using Kappa, and refinements were made according to the discrepancies. This was done three times, until the Kappa value revealed excellent agreement between the tools (k = 0.87). A panel of experts was then invited to examine the new gross motor screening tool, and to comment on it, and further adjustments were made accordingly. Preliminary concurrent validity testing of the new gross motor screening tool was then carried out against the Gross Motor facet of the BSID III on 60 children, who were recruited from the Harriet Shezi Children’s Clinic at Chris Hani Baragwanath Hospital in Soweto. Statistical analysis revealed that the agreement between the BSID III and the new screening tool was excellent (k=0.85). The diagnostic properties of the new gross motor screening tool were as follows: sensitivity 97.4%, specificity 85.7%, positive predictive value 92.7%, and negative predictive value 94.7%. These values indicate that the statistical properties of the tool are excellent, and the tool will not be predisposed to underreferrals or over-referrals. Preliminary reliability testing was carried out on 15 children for test-retest/intrarater reliability and 15 children for interrater reliability. Interrater, test-retest and intrarater reliability were excellent (r=1, r=0.98, r=0.98 respectively). Further testing of reliablity and validity should be undertaken in order to establish these properties, and standardisation should also be carried out on healthy children. Given the need for an assessment tool of this nature in South Africa and other developing countries, and the statistical properties thus far, the tool may be used clinically for the purposes for which it was developed.

Intellectual disability (ID) is categorised by a significant reduction in cognitive function and adaptive abilities that begin in childhood. ID is part of a heterogeneous group of neurodevelopmental conditions associated with impairment in developmental domains and a cause of particularly adverse socioeconomic impact worldwide. There have been many recent advances in identifying causative genetic mutations in previously unexplained ID cases. With these advances comes an increasing demand for understanding mechanisms underpinning these pathogenic pathways. In this PhD thesis, I have studied rare monogenic novel neurodevelopmental disorders associated with ID. The objective of the thesis was to model a subset of mutations associated with novel neurodevelopmental disorders in mice to demonstrate a causal link between mutation and phenotype and to further understand the mechanisms by which these mutations result in human neurodevelopmental disorders. In order to achieve this, I adopted a multi-phase approach. Firstly, I designed a phenotyping platform, by combining behavioural and cognitive tests with morphometric brain analysis and genome-wide transcriptional analysis. I then used this approach to study KPTN-related syndrome, a novel developmental disorder that to date has not been characterised in mice, successfully recapitulating the main phenotypes described in the patients. Moreover, I gained further insight into the underlying pathogenic mechanisms associated with the disorder, opening the possibility of a therapy that could treat some aspects of cognitive and morphological impairments identified in the patients with KPTN-related syndrome. Lastly, I determined whether such an approach could be scaled-up to study multiple novel neurodevelopmental disorders, each with a mutation associated with a haploinsufficient novel neurodevelopmental disorder. I identified specific phenotypes for each of the four mouse lines under investigation, providing a platform for comparison between several developmental disorders. These refinements contributed to a larger five-year project starting at the Sanger Institute, aimed at characterising a wider diversity of human neurodevelopmental disorders.

No description available.

Children with Down syndrome (DS) display language deficits in expressive and receptive skills beyond what is predicted by nonlingusitic cognitive skills. Clinically, a ubiquitous presumption is that vocabulary taught in one modality will generalize incidentally to the other, untreated modality. Five children with DS (four male, one female, ages 3;6-5;1) were each taught three orthogonal sets of receptive and expressive vocabulary within a multiple probe single subject design. During each probe condition, vocabulary knowledge for trained and untrained modalities was probed. Cross modal generalization probes for all five children indicated moderate transfer from expressive modality to receptive and relatively low receptive generalization to the expressive modality. These results support expressive vocabulary interventions for children with DS provided that clinicians systematically test for generalization to receptive knowledge. Conversely, receptive vocabulary training is much less likely to generalize across modality

The recent history of autoimmune neurology is marked by the discovery of many central nervous system (CNS) antibody-mediated diseases. These disorders are caused by antibodies that target important proteins expressed in the neuronal surface, which are believed to be directly pathogenic. These antibodies are immunoglobulin G (IgG) isotype and, as such, have the potential to cross the placenta during gestation. Foetal exposure to CNS-targeting antibodies could alter developing neuronal circuits, leading to disease. However, the consequences of exposure to these antibodies during neurodevelopment has hardly been considered. To study the relationship between maternal antibodies towards neuronal surface proteins and neurodevelopmental disorders in the foetus a dual approach was undertaken. First, pregnancy serum samples from mothers of children later diagnosed with a neurodevelopmental disorder and from mothers of children with typical development were screened for the presence of neuronal surface antibodies. Next, the effects of pathogenic neuronal surface antibodies in the offspring were assessed in a maternal-to-foetal transfer mouse model. Antibodies to neuronal surface proteins in the gestational serum, particularly CASPR2 antibodies, were found to associate with an increased risk of mental retardation and disorders of psychological development in the progeny. The animal model showed that mice exposed in utero to CASPR2 antibodies have long term behavioural sequelae and histological findings suggestive of abnormalities in brain development. These findings support a model in which maternal antibodies towards foetal neuronal proteins cause long-term behavioural deficits and permanent abnormalities at the cellular and synaptic level in a subset of children with neurodevelopmental disorders.

No description available.

Autism spectrum disorder (ASD) encompasses a broad range of neurodevelopmental disorders, with two core symptoms: deficits in social communication, and restrictive interests and repetitive behaviors. Genetics is thought to play a large role in ASD and currently there are hundreds of associated genes. We first studied the thousand and one amino acid kinase gene (TAOK2), which plays an important role in neurodevelopment. We found that loss of TAOK2 causes deficits in neuron development and activity, leading to morphological changes in various mouse brain regions and ASD-related behaviors. We studied the impact of de novo mutations identified in TAOK2, which caused aberrant neuron dendritic arborization and formation of synapses. To elucidate how TAOK2 regulates neuron development we used a proximity-labeling proteomics technique (BioID) to identify its protein-protein interaction (PPI) network. We applied this same methodology to a total of 41 ASD-risk genes and observed multiple convergent biological processes, including the less-studied mitochondrial and metabolic pathways. ASD-risk genes, including TAOK2, associated with mitochondrial proteins were found to have altered cellular respiration. The shared ASD-risk gene PPI network enriched for other ASD-risk genes and was used to group genes based on their shared PPI networks. These gene groups showed correlation between the clinical behavior scores of individuals that had mutations within the distinct gene groups. Lastly, we identified changes in the PPI networks of multiple ASD-risk genes through BioID, which we validated with various functional assays. In summary, we developed a proximity-labeling proteomics method that identified multiple convergent biological pathways associated with ASD. Studying the function of TAOK2 revealed multiple disease-relevant pathologies associated with the disorder, however proximity labeling has the potential to categorize multiple ASD-risk genes and elucidate their shared signaling pathways, which together, can advance the development of robust treatments for ASD. / Thesis / Doctor of Philosophy (PhD) / Autism spectrum disorder (ASD) is a group of brain disorders that affect more than 1% of children. Genetic variants are thought to cause ASD pathology, however there are currently hundreds of genes that have not been studied. We studied how disruption of one of those genes, TAOK2, alters brain development in mice and identified TAOK2 variants in multiple children with ASD. We then used BioID to find the shared disease-related mechanisms between multiple ASD-risk genes, and found that mitochondrial function and activity were connected to many of these genes. We showed that BioID can be used to study the effect of mutations in multiple ASD-risk genes simultaneously. Last, we could group children with ASD with similar behavior test scores based on the shared mechanisms of ASD-risk genes. Together our findings could be used to advance the development of robust treatments or new diagnostic tools for ASD.

The basic helix-loop-helix PAS domain transcription factor gene NPAS3 is a risk factor for psychiatric disorders. A knockout mouse model also exhibits behavioural and adult neurogenesis deficits consistent with human illness. To define the location and mechanism of NPAS3 aetiopathology immunofluorescent and transcriptomic approaches were used. Npas3 was co-localised with Dcx, but not other neurogenesis markers, in the hippocampal subgranular zone - the site of adult neurogenesis. This implied that NPAS3 might be involved in maturing, rather than proliferating, neuronal precursor cells. Microarray analysis revealed that the transcriptional activities of NPAS3 and its truncated form (C-terminal deletion) in the HEK293 cell line are sensitive to circadian rhythm context. The most highly up-regulated NPAS3 target gene, VGF, encodes secretory peptides with established roles in neurogenesis, depression and schizophrenia. VGF was one of many NPAS3 target genes also shown to be regulated by the SOX family of transcription factors, suggesting an overlap in neurodevelopmental pathways. The transcriptional repression of multiple glycolytic genes indicated that NPAS3 has a second role in metabolic regulation. This finding was also confirmed by collaboration with a metabolomics research group at the University of Strathclyde. SOX11, a transcription factor known to play a role in neuronal and glial cell differentiation, was shown to be down-regulated by NPAS3. The set of genes targeted by SOX11 and their ontologies were deduced by a microarray analysis in a SOX11 overexpressing HEK293 cell line. Regulated genes include a previously established SOX11 target, known markers of neurogenesis as well as genes implicated in neuropsychiatric disorders. Multiple histone and zinc finger genes are regulated by SOX11, many of which were located in two clusters on chromosomes 6 and 19. The chromosome 6 cluster lies within a region of the genome showing the strongest genetic association with schizophrenia. SOX11 may alter localised expression competence and its targets induce a complex programme of chromatin remodelling and downstream gene expression changes to achieve the mature neuronal phenotype. This thesis details how transcription factors are involved in biological processes linked to psychiatric illness. The dual neurodevelopmental and metabolic aspects of NPAS3 activity described here increase our understanding of aspects of neurogenesis relevant to mental illness and may explain the innate and medication-induced susceptibility to diabetes reported in psychiatric patients.