In Vitro Biomarker Discovery for Atherosclerosis by Proteomics

Abstract

The purpose of this study was to identify in vitro and then prioritize a tractable set of protein biomarker candidates of atherosclerosis that may eventually be developed to measure the extent, progression, regression, and stability of atherosclerotic lesions. A study was conducted using an in vitro "foam cell" model based on the stimulation of differentiated THP1 cells with oxidized low-density lipoprotein (oxidized LDL) as compared with low-density lipoprotein (LDL). Analysis of the proteins contained in the cell supernatant using proteome scanning technology identified 59 proteins as being increased, 57 with no statistically measurable difference, and 17 decreasing in abundance following treatment with oxidized LDL, as compared with LDL. From the up-regulated list, proteins were prioritized based on their analytical confidence as well as their relevance to atherosclerosis pathways. Within the group of increased abundance, seven families of proteins were of particular interest: fatty acid-binding proteins, chitinase-like enzymes, cyclophilins, cathepsins, proteoglycans, urokinase-type plasminogen activator receptor, and a macrophage scavenger receptor.

Integrative Urinary Peptidomics in Renal Transplantation Identifies Biomarkers for Acute Rejection

“…We used the approach of ion mapping, 45,46 in which biomarker candidate MS peaks are selected on the basis of discriminant analysis and then targeted for MS/MS sequencing analysis to obtain protein identification. We have developed an informatics platform, "MASS-Conductor," 7 which contains an integrated suite of algorithms, statistical methods, and computer applications to allow for signal processing and statistical analysis in LC-MS-based urine peptide profiling.…”

Vascular proteomics

“…In order to look for potential biomarkers of atherosclerosis, an in vitro "foam cell" model based on the stimulation of differentiated THP1 cells with oxidized LDL (oxLDL) as compared with LDL was performed [44]. Analysis of the proteins contained in the cell supernatant, using proteome scanning technology, identified 59 proteins as being increased, 57 with no statistically measurable difference, and 17 decreasing in abundance, following treatment with oxLDL as compared with LDL.…”

Overview of Current Proteomic Approaches for Discovery of Vascular Biomarkers of Atherosclerosis

“…In this respect, cell culture systems could represent a useful tool to partially overcome drawbacks of tissue analyses, allowing researchers to study single aspects of the atherosclerotic process in very controlled conditions. In the last years, studies on proteome (the intracellular proteins) and secretome (the proteins released into the cell culture medium) by 2DE and MS of ECs (Bruneel et al, 2003;Chen et al, 2007;Tunica et al, 2009), VSMCs (McGregor et al, 2001Dupont et al, 2005;Lee et al, 2006), and monocytes/macrophages (Dupont et al, 2004;Fach et al, 2004;Slomianny et al, 2006;Zhang et al, 2007;Zhao et al, 2009) have been performed. Moreover, in the attempt to help in elucidating the mechanisms of atherogenesis, several proteomic studies have been carried out on vascular cells cultured in different experimental conditions (table 3).…”

“…The most applied proteomic methodologies were 1DE-2DE coupled to MS analyses. Very few studies applied gel free proteomic approaches such as LC MS/MS (Fach et al, 2004;X.L. Wang et al, 2007;Zhao et al, 2009;Tunica et al, 2009;Zimman et al, 2010) and microarrays (Sukhanov et al, 2005).…”