Abstract 198: Robust Quantification Of Hdl Proteome For Mechanistic And Clinical Studies

• Published in: Arteriosclerosis, thrombosis, and vascular biology Vol. 42; no. Suppl_1; p. A198
• Main Authors: Silva, Amanda R, Souza Junior, Douglas R, Ronsein, Graziella E
• Format: Journal Article
• Language: English
• Published: Lippincott Williams & Wilkins 01.05.2022
• ISSN: 1079-5642; 1524-4636
• DOI: 10.1161/atvb.42.suppl_1.198

IntroductionEmerging studies have proposed novel roles of HDL in cardiovascular and noncardiovascular diseases. The understanding of how HDL participates in a pathophysiological process involves animal models, as well as clinical studies. A key issue involving HDL studies is the accurate quantification of its proteome. HDL proteome is complex, and its composition may be modulated by the subject’s health state. The main goal of this work is to establish a pipeline for accurate quantification of HDL proteome in clinical and animal studies. HypothesisThe knowledge of the composition of HDL proteome in clinical and animal studies is critical to understand its biological functions. MethodsWe first performed a deep proteomic analysis to identify proteins belonging to HDL of mice and humans. We then established a targeted, data independent acquisition (DIA) strategy that works for mouse and humans. Next, by using different software platforms, we developed a quantification methodology specific for each species. ResultsAlthough mice and humans share many of their HDL proteins, the protein sequence is unique for each species, and thus, HDL proteome quantification methods must be species specific. Using recent advances in mass spectrometry, we employed a single acquisition method that worked for both, mice and human samples. Differentiation occurred after acquisition, using spectral libraries specifically built for each species. Using different software platforms, we compared four strategies of quantification for HDL proteome, showing that CVs depend on the strategy employed. Thus, using quality control samples (n=8), we showed that for the same protein, CVs may vary up to 50 %. The quantification strategy was thus optimized in order to provide the best quantification with the least variance, reaching CVs lower than 15 % for 90 % of the proteins. ConclusionsUsing the same mass spectrometry acquisition method, it is possible to quantify HDL proteome of mice and humans in a consistent and accurate way. These advances may integrate animal models and clinical studies of HDL proteome.