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Trypanosoma brucei: Protein Expression Microarrays and Circulating miRNA during Infection

Lueong, Smiths Sengkwawoh

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The last decades have been galvanized by efforts to reduce the ever widening gap that exist between functional genomics and proteomics. Assigning molecular functions to, and breaking through the complex networks in which each and every individual protein of the cellular proteome is involved is the breathtaking task that needs to be overcome in order to understand the molecular and physiological basis underlying health and disease. System-wide analysis is an approach that can permit a better understanding of the proteome, and there is need for robust and reliable platforms for such analysis to be developed. To contribute to current efforts, I have developed and optimized methods for the production of functional protein microarrays in a miniaturized form from cDNA products and genomic DNA as template source. I have further shown that such arrays are very useful and reliable in various applications such as protein-protein interactions, protein-RNA interactions, Kinase substrate identification and antibody selection. I have validated some data obtained from these arrays in vivo in a model organism, Trypanosoma brucei. This confirms that the platform can well contribute to the already existing proteomic tools in generating reliable biological data. Moreover, using cDNA products allow for the analysis of disease-related and rare transcripts as well as other spliced variants. Current diagnostic tools for human African trypanosomiases are very invasive and in some cases are not sensitive enough, and this is compounded by a highly heterogeneous seropositive patient population that is difficult to classify. To address this, I have equally analyzed the miRNA and mRNA expression pattern in the peripheral blood of patients in search for new markers. Thirteen differentially expressed miRNAs were identified, three of which (miRNA-199a-3p, miRNA-27b and miRNA-126*) were highly selective (>95%). These miRNAs have also been reported to be differentially regulated in other diseases and miRNA-199a-3p for example is used as a diagnostic biomarker. They are therefore not suitable as specific biomarkers in sleeping sickness. I have however shown that there is deregulation of miRNA expression following T. brucei infection, and most of the differentially regulated miRNAs are related to immune responses to infectious agents and other inflammatory responses influencing disease outcome.

Item Type: Dissertation
Supervisor: Clayton, Prof. Dr. Christine E.
Date of thesis defense: 16 October 2014
Date Deposited: 30 Oct 2014 10:27
Date: 2014
Faculties / Institutes: The Faculty of Bio Sciences > Dean's Office of the Faculty of Bio Sciences
Controlled Keywords: Trypanosoma brucei, Protein expression microarrays, Circulating miRNA
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