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  • http://hdl.handle.net/10402/era.24749
  • Using 1H-NMR based metabolomics to investigate the pathological consequences of mitochondrial disease and human rabies infection
  • Reinke, Stacey N
  • English
  • metabolomics
    NMR
    mitochondria
    mitochondrial disease
    C. elegans
    S. cerevisiae
    rabies
    biological variability
  • Dec 15, 2011 3:29 PM
  • Thesis
  • English
  • Adobe PDF
  • 9940156 bytes
  • Mitochondrial diseases encompass a wide range of clinical phenotypes. The etiology of these disorders is extremely complex; mitochondria are central to energy metabolism and dysfunction can have a profound effect on global metabolism. The original objective of this thesis was to use 1H-NMR based metabolomics to investigate the metabolic mechanisms of mitochondrial disease. Metabolomics is the systematic study of metabolites and is a powerful approach to understanding disease. In the clinical realm, metabolomics is useful for identifying biomarkers of disease and for understanding disease mechanisms. The metabolome is extremely sensitive to a number of factors, such as genetic background, age, diet, gender and stress; it is therefore difficult to meaningfully interpret results from human metabolomic data. For this reason, we chose to employ two common laboratory models of mitochondrial disease: the nematode Caenorhabditis elegans and the yeast Saccharomyces cerevisiae. These model systems provide the opportunity to study mitochondrial disease in a well-defined genetic background under controlled environmental conditions. Our findings revealed that even in a highly-controlled experiment, an appreciable amount of biological variability still exists. In the laboratory, C. elegans can be fed two different strains of E. coli: OP50 for regular maintenance and HT115 for RNAi-mediated gene suppression. We discovered that the nematode metabotype, mtDNA copy number, brood size and lifespan are significantly altered by diet. We also investigated the metabolic consequences of Complex II dysfunction in yeast, as reflected in the extracellular metabolome or exometabolome. Metabolomic studies are frequently used to identify a small number of key biomarkers that discriminate individuals of different phenotypes. Our exometabolome data revealed that the entire metabolome contributes to discrimination between phenotypic classes. Amino acid metabolism, which is closely linked to energy metabolism, was profoundly affected by mitochondrial dysfunction in both model systems. I also investigated the pathogenic, molecular and metabolic consequences of a human rabies infection following an aggressive and controversial neuro-therapeutic treatment protocol, revealing a possible adverse and fatal consequence of this procedure.
  • Lamarre SG, Molloy AM, Reinke SN, Sykes BD, Brosnan ME, and Brosnan JT. (2011) Formate can differentiate between hyperhomocysteinemia due to impaired remethylation and impaired transsulfuration. Am J Physiol Endocrinol Metab. 2011 Sep 20
    Szeto SSW, Reinke SN, and BD Lemire. (2011) 1H-NMR based metabolic profiling reveals inherent biological variation in the yeast and nematode model systems. J. Biomol. NMR. 49: 245-54.
    Szeto SSW, Reinke SN, Sykes BD, and BD Lemire. (2010) Mutations in the Saccharomyces cerevisiae succinate dehydrogenase result in distinct metabolic phenotypes revealed through 1H-NMR based metabolic footprinting. J. Proteome Res. 9: 6729-39.
    Reinke SN, Hu X, Sykes BD and BD Lemire. (2010) Caenorhabditis elegans diet significantly affects metabolic profile, mitochondrial DNA levels, lifespan, and brood size. Mol. Genet. Metab. 100: 274-82.
    Friis RM, Wu BP, Reinke SN, Hockman DJ, Sykes BD, and MC Schultz. (2009) A glycolytic burst drives glucose induction of global histone acetylation by picNuA4 and SAGA. Nucleic Acids Res. 37(12): 3969-80.
    Szeto SSW, Reinke SN, Sykes BD and BD Lemire. (2007) Ubiquinone-binding site mutations in the Saccharomyces cerevisiae succinate dehydrogenase generate superoxide and lead to the accumulation of succinate. J. Biol. Chem. 282: 27518-27526.
  • Doctoral
  • Doctor of Philosophy
  • Department of Biochemistry
  • Spring 2012
  • Lemire, Bernard (Biochemistry)
    Sykes, Brian (Biochemistry)
  • Lemire, Bernard (Biochemistry)
    Sykes, Brian (Biochemistry)
    Baracos, Vickie (Oncology)
    Broadhurst, David (Medicine)
    Simpson, Myrna (Chemistry, University of Toronto)