Date of Award
Doctor of Philosophy in Clinical-Bioanalytical Chemistry
Gangliosides are a large subclass of glycosphingolipids that structurally characterized by the addition of mono- or poly-sialyated carbohydrate moieties onto the ceramide scaffold. They are presented at significant abundance in the central nervous system of vertebrates. Their interactions with trans-membrane receptors, especially receptor tyrosine kinase, are believed to play critical roles in modulating signal transduction events during cell proliferation, differentiation, migration, and adhesion. Furthermore, the disruption and dysfunction on their metabolism have been found to be associated with various neurodegenerative disorders, such as Alzheimer disease, Parkinson’s disease, and GM3 synthase deficiency. GM3 synthase deficiency (GSD) is a newly identified neurological disorder that has been prevalently found in the Amish population in the United States. Although the pathological mechanism remains to be understood, the condition is severe. It is characterized by infantile onset of severe irritability, developmental stagnation, profound intellectual disability and intractable seizures. In order to advance our understanding on the etiology of these neurodegenerative disorders, especially GSD, analytical assays with sufficient specificity, sensitivity, and throughput are urgently demanded to practically assist the implementation of relevant clinical studies. In this thesis, three projects have been conducted separately to benefit the clinical studies from individual aspects. In project 1, owing to the increased complexity of molecular assays in screening and diagnosing patients affected by GSD associated with increased numbers of identified nonsense/missense mutations, we have developed and validated a novel UPLC/MS/MS method with promising sensitivity, specificity, and throughput for four major ganglioside species, GM2, GM3, GD2, and GD3, in human plasma to facilitate the diagnosis and screening of GSD. In project 2, in order to obtain more informative results from GSD patients undergoing clinical trial in relation to the development of related therapeutic intervention, we have developed and validated a new UPLC/MS/MS method in human plasma for quantification of monosialogangliosides in combination with DMTMM&PAEA chemical derivatization for analysis with superior sensitivity and specificity. In project 3, we have implemented a clinical study aiming at developing a therapeutic intervention strategy based on oral administration of commercialized ganglioside-enrich formula to the affected patients.
Huang, Qianyang, "Development and Validation of UPLC/MS/MS Methods for Quantification of Gangliosides in the Clinical Study of Ganglioside GM3 Synthase Deficiency" (2016). ETD Archive. 927.