Genetic analysis and investigation of molecular pathogenesis for mitochondrial diseases

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Whole exome sequencing of mitochondrial disease patients was performed in our laboratory. In fiscal year 2018, I investigated candidate mutations in 4 genes/gene cluster for their pathogenicity in 9 patients. Bioinformatic, molecular genetic and biochemical analyses including RNA sequencing, Sanger sequencing, DNA cloning, qPCR, SDS-PAGE and BN-PAGE western blotting, respiration rate analysis and OXPHOS enzyme assays were performed.We detected large chromosomal deletions and rearrangements of the gene region in several patients with confirmed or suspected mitochondrial disease. My research highlighted the technical challenge in elucidating the exact gDNA mutations in the gene cluster region. Molecular diagnosis was confirmed in 2 of the 5 patients investigated. This is an on-going research in collaboration with Prof. David Thorburn from Murdoch Children’s Research Institute in Australia.One of the novel disease genes was investigated in collaboration with Dr Diana Stojanovski from the University of Melbourne in Australia. As a result, the variant was excluded from further analysis in two patients due to the lack of supportive evidence for pathogenicity.Two novel DNA variants in a nuclear gene encoding an OXPHOS complex III subunit were identified in a patient with Leigh Syndrome. Genomic DNA, RNA and protein analyses were performed to identify evidence for pathogenicity caused by those 2 compound heterozygous variants in the patient. This study led to an on-going collaboration with Prof. David Thorburn. A joint-publication is in preparation.