Metabolic profiling for biomarker discovery in biochemical genetics
File(s)
Author(s)
Robinette, Steven Lawrence
Type
Thesis
Abstract
Functional characterization of the phenotypic consequences of genetic variants
increasingly constitutes the rate-limiting step in the study of biochemical genetics.
This thesis presents the application of metabolic profiling technologies, including
Nuclear Magnetic Resonance (NMR) spectroscopy and Mass Spectrometry (MS), to
identify metabolic perturbations resulting from inborn errors of metabolism,
congenital diseases affecting the kidney, and animal models of both genetic mutations
and renal pathology. In the case of newborn screening for enzyme deficiencies, two
mass spectrometric assays, traditional tandem mass spectrometry (MS/MS) with
multiple reaction monitoring and direct injection nanospray high resolution mass
spectrometry (ns-HR-MS), were applied to profile dried blood spot (DBS) samples of
over 6,000 newborns and identify the metabolic perturbations resulting from 24
congenital disorders of metabolism.
To study genetic mutations with more complex phenotypic consequences
affecting the kidney, urinary metabolic profiles were evaluated for four congenital
kidney diseases. The natural history of cystinosis, showing changes in the urinary
profile of cystinosis patients over time and with age, glomerular filtration rate, drug
therapy, and transplantation, and a comparison of urinary perturbations seen in these
human Mendelian disorders to the perturbations induced by region specific
nephrotoxins characterize the urinary metabolic associations with these renal
phenotypes. Finally, new computational approaches for analyzing metabolic profiling
data are presented and evaluated, including for improving biomarker identification
with two-dimensional NMR and increasing metabolomic coverage with high resolution
mass spectrometry.Integrating metabolic and genetic diagnostics should
enhance understanding of the relationship between genes and health and mechanisms
by which genetic variations manifest as inherited disease.
increasingly constitutes the rate-limiting step in the study of biochemical genetics.
This thesis presents the application of metabolic profiling technologies, including
Nuclear Magnetic Resonance (NMR) spectroscopy and Mass Spectrometry (MS), to
identify metabolic perturbations resulting from inborn errors of metabolism,
congenital diseases affecting the kidney, and animal models of both genetic mutations
and renal pathology. In the case of newborn screening for enzyme deficiencies, two
mass spectrometric assays, traditional tandem mass spectrometry (MS/MS) with
multiple reaction monitoring and direct injection nanospray high resolution mass
spectrometry (ns-HR-MS), were applied to profile dried blood spot (DBS) samples of
over 6,000 newborns and identify the metabolic perturbations resulting from 24
congenital disorders of metabolism.
To study genetic mutations with more complex phenotypic consequences
affecting the kidney, urinary metabolic profiles were evaluated for four congenital
kidney diseases. The natural history of cystinosis, showing changes in the urinary
profile of cystinosis patients over time and with age, glomerular filtration rate, drug
therapy, and transplantation, and a comparison of urinary perturbations seen in these
human Mendelian disorders to the perturbations induced by region specific
nephrotoxins characterize the urinary metabolic associations with these renal
phenotypes. Finally, new computational approaches for analyzing metabolic profiling
data are presented and evaluated, including for improving biomarker identification
with two-dimensional NMR and increasing metabolomic coverage with high resolution
mass spectrometry.Integrating metabolic and genetic diagnostics should
enhance understanding of the relationship between genes and health and mechanisms
by which genetic variations manifest as inherited disease.
Version
Open Access
Date Issued
2013-06
Date Awarded
2013-10
Copyright Statement
Attribution NoDerivatives 4.0 International Licence (CC BY-ND)
Advisor
Nicholson, Jeremy
Gahl, William
Sponsor
Great Britain. Marshall Aid Commemoration Commission
National Science Foundation (U.S.)
National Institutes of Health
Publisher Department
Surgery and Cancer
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)