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Pilot & Feasibility Program Application Abstract

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Proteomic analysis of mouse plasma lipoproteins

Sean Davidson (Reading, OH)

Pilot & Feasibility Program

Diabetes has numerous pathological effects on lipoprotein metabolism. There is a general increase in atherogenic triglyceride-rich lipoproteins and remnants with a corresponding decrease in the cardioprotective high density lipoprotein (HDL) fraction. Recent proteomics studies have shown that each lipoprotein class can have various combinations of up to 80 different protein factors associated. These proteins, called apoproteins, modulate the metabolism, function and potential pathology of lipoproteins. Unfortunately, little is understood about how the “lipoproteome” is distributed among lipoproteins in either basal or diabetic states. It is highly likely that these proteins move between lipoproteins in order to facilitate key metabolic reactions at the proper time and place to satisfy the organism’s need for lipid delivery. We believe that the onset of diabetes will produce differences in these lipoproteomic patterns that will be useful for understanding the pathology of diabetic dyslipidaemia. In pursuit of our goal of developing high throughput technologies for characterizing and comparing lipoproteomic profiles, we have developed gel filtration-based lipoprotein separation techniques in conjunction with electrospray ionization mass spectrometry (ESI-MS) to derive a detailed lipoproteomic profile of human plasma. The aims of this proposal are: 1) to translate these technologies from the human to the mouse and, 2) to enhance the quantitative power of the technique using isobaric affinity tags (iTRAQ). We anticipate that this work will underpin a new MMPC test that will allow investigators to compare plasma apoprotein profiles between wild-type and genetically/pharmacologically induced diabetic mouse models. These can be correlated with other phenotypic data to help determine the underlying metabolic defects in diabetes that lead to CVD and vascular dysfunction.

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