Project title: Gene therapy and omics discovery platforms for developmental and epileptic encephalopathies
Developmental and epileptic encephalopathies (DEEs) are rare disorders that impact neural development and function, resulting in debilitating symptoms such as seizures, delayed development, impaired motor activity, cognitive deficits, and difficulty communicating. Although many DEEs have a clear genetic basis, these conditions are complex and remain poorly understood. As a result, current treatment options for DEEs are severely limited, primarily targeting symptom management without correcting the underlying biology. My research aims to provide deep insights into the pathophysiology of DEEs and leverage this understanding towards improved diagnostic and therapeutic outcomes, with a particular focus on Rett syndrome (caused by mutations in the methyl-CpG binding protein 2 (MECP2) gene) and syntaxin-binding protein 1 (STXBP1/MUNC18-1)-related conditions. Using high-throughput “omics” technologies, our group is gaining a wholistic understanding of DEE biology and identifying novel disease drivers, biomarkers, and potential therapeutic targets. We are also developing new gene therapy approaches for highly targeted, safe, and effective restoration of healthy gene function to prevent or reverse the symptoms of DEEs. Through these efforts, our research is addressing the critical needed for therapeutic options that substantially improve outcomes and quality of life for patients, their carers, and wider-community.
Project: Novel Biomarkers of Mitochondrial Dysfunction: Primary Mitochondrial Disease and Disorders Causing Secondary Mitochondrial Dysfunction
Primary and secondary mitochondrial disorders are both complex groups of diseases with many aetiologies, and diagnosis is challenging due to clinical and genetic heterogeneity. For many of these disorders, treatment is limited and often purely symptomatic. A significant impediment in determining the efficacy and viability of treatment is the lack of a clinical biomarker that is both diagnostic and can predict disease severity. Recently, the mitokines fibroblast growth factor 21 (FGF21) and growth differentiation factor 15 (GDF15) have been established as biomarkers of cellular stress and mitochondrial dysfunction in patients with primary mitochondrial disease. Our project aims to investigate the suitability of these mitokines as both diagnostic and monitoring biomarkers for a disorder causing secondary mitochondrial dysfunction, Rett syndrome (RTT). Firstly, we will determine if FGF21 and GDF15 can predict disease course in a Mecp2T158A RTT mouse model. Secondly, we will investigate if these mitokines are differentially expressed in the blood of human females with RTT (relative to a control population) and determine if they are prognostic biomarkers. Finally, we will utilise an integrative, multi-omics approach (proteomics, metabolomics, and transcriptomics) to better understand the pathophysiology of disease and identify novel biomarkers and drug targets. This ultimately will assist in identifying improved therapeutic options for individuals with RTT.
Project: Exon replacement gene editing therapy for Rett syndrome
Rett syndrome is a rare neurodevelopmental disorder which is highly complicated and regulated by the expression pattern of the Methyl-CpG-binding Protein 2 (MECP2) gene. My projects looks at editing the MECP2 gene at the native locus to correct pathogenic variants in the gene and restore functional activity of MECP2. I am testing this approach in vitro in patient cell lines, including fibroblasts and induced pluripotent stem cell (iPSC) derived cortical brain organoids, as well as in vivo in the T158A Rett syndrome mouse model.
Project: Functional Studies for SNAREopathies
The purpose of my project is to characterise a patient mutation occuring in the SNAP25 gene. Ultimately, the aim of the project is to develope a targetted gene therapy to treat disease caused by this mutation. The SNAP25 gene enodes one of the key proteins which complex to form the molecular machinery necessary for neurotransmitter release from brain cells. I will investigate the effects of the patient SNAP25 mutation in a 3D brain organoid model. Combining molecular, electrophysiological, and imaging techinques, I aim to devleope an understanding of how SNAP25 dysfunction contributes to pathology, and to validate a corrective treatment.
Project: Integrative omics: A novel approach to unravelling the complex panoramic landscape of Rett syndrome.
Rett syndrome is a rare neurodevelopmental disorder where treatment is purely symptomatic with no curative treatment. The paucity of any disease-modifying therapeutics entering the clinic is primarily due to the lack of useful clinical biomarkers and a complete understanding of the complex underlying disease pathophysiology and function of MeCP2. Recently, the application of high-throughput approaches known as “omics” has developed rapidly and revolutionised the diagnosis and the understanding of the pathophysiology of many neurological disorders. Additionally, cortical brain organoids are an emerging human 3D model that mimics various developmental features at the cellular and molecular levels and provides a powerful model for studying neurodevelopmental disorders. By harnessing the power of omics platforms along with advanced stem cell technology, my project aims to dissect the molecular complexity of Rett syndrome to identify genes, proteins, metabolites, and cellular pathways from the blood that are differentially dysregulated among patients in the cohort (relative to controls) in an unbiased approach. This integrated approach will identify the dysregulated pathways in Rett syndrome individuals, providing an impartial approach to therapeutic targets that will lead to preclinical validation, the initiation of transformative Phase I clinical trials and the translation of novel therapeutics from clinical trials into clinical care.
Project: Accelerating treatment for epilepsy in children with KIF1A-associated neurological disorders
Kif1a-associated neurological disorder (KAND) is a rare and severe neurodegenerative condition. It is caused by mutations in Kif1a - a neuronal molecular protein. One hallmark of this disorder is early life epilepsy which can exasperate or cause detriments to brian development. The goal of this study is to better understand KIF1a and identify FDA approved drugs that can decrease seizure activity by testing patient iPSC derivied brain organoids. By doing so, we hope to alleviate a major debilitating clinical feature of KAND.
Project: MECP2 Gene Addition Therapy for Rett syndrome
Rett syndrome is a rare neurodevelopmental disorder, affecting 1 in 10 000 children. It is characterised by mutations within the X-linked Methyl-CpG-binding Protein 2 (MECP2) gene that encodes for the MeCP2 protein. This project aims to develop a gene replacement therapy to restore physiological levels of functional MeCP2 expression. To test the gene therapy, MeCP2 expression in vitro cell cultures and behavioural tests of in vivo Rett mouse models will be used to determine and fine-tune treatment efficacy.
Project: Preclinical assessment of gene therapy for RARS2-related early onset epileptic encephalopathy
Pontocerebellar Hypoplasia Type 6 (PCH6) is a type of mitochondrial disease with early-onset encephalopathy caused by recessive mutations in mitochondrial arginyl-tRNAsynthetase gene (RARS2). Characteristic clinical features comprise of neonatal lactic acidosis, severe encephalopathy, intractable seizures, feeding problems and profound developmental delays. Most patients show typical neuroradiologic abnormalities including cerebellar hypoplasia and progressive pontocerebellar atrophy. There is currently no cure and therapeutic interventions are aimed at symptomatic relief. Significant advances have been made in our understanding of the pathophysiology of mitochondrial disease, however, due to large genetic heterogeneity and numerous missense variations of uncertain significance, the pathogenesis of PCH6 remains unclear. In this project, we aim to elucidate the mechanisms by which variants in RARS2 lead to the development of PCH6 and assess the efficacy of restoring RARS2 gene via gene therapy in preclinical models. Towards these aims, we will first characterise PCH6 disease phenotype in RARS2 variant-iPSC derived brain organoids. We will then design, and generate clinical cassettes containing RARS2 to transduce brain organoids. Following transduction, organoids will be evaluated for the safety and efficacy of gene therapy treatment.
Project: Understanding the role of inflammation and epigenetics in neurodevelopmental and neuropsychiatric disorders
Neurodevelopmental and neuropsychiatric disorders have severe social, educational, emotional, and economic impacts on affected children and their families. Current management plans range from behavioural therapy to antidepressant and neuroleptic medications which give rise to many adverse effects. There is increasing evidence that environmental factors, such as maternal immune activation, also play a key role in the pathophysiology of these disorders. Although these associations between MIA and risk of neurodevelopmental disorders are well-established, there is limited research in humans regarding the underlying mechanisms of how maternal inflammation might result in adverse offspring outcomes. My project aims to investigate the role of inflammation and epigenetic modulation in neurodevelopmental and neuropsychiatric disorders. The focus will be on how maternal inflammation influences epigenetic modulation and subsequently, immune function, in children with neurodevelopmental disorders.