Plasma proteomic alterations in non-human primates and humans after chronic alcohol self-administration

Abstract

Objective diagnostics of excessive alcohol use are valuable tools in the identification and monitoring of subjects with alcohol use disorders. A number of potential biomarkers of alcohol intake have been proposed, but none have reached widespread clinical usage, often due to limited diagnostic sensitivity and specificity. In order to identify novel potential biomarkers, we performed proteomic biomarker target discovery in plasma samples from non-human primates that chronically self-administer high levels of ethanol. 2-dimensional in-gel electrophoresis (2D-DIGE) was used to quantify plasma proteins from within subject samples collected before exposure to ethanol and after three months of excessive ethanol self-administration. Highly abundant plasma proteins were depleted from plasma samples to increase proteomic coverage. Altered plasma levels of SAA4, RBP, ITIH4, clusterin, and fibronectin, identified by 2D-DIGE analysis, were confirmed in unmanipulated, whole plasma from these animals by immunoblotting. Examination of these target plasma proteins in human subjects with excessive alcohol consumption (and control subjects) revealed increased levels of SAA4 and clusterin and decreased levels of fibronectin compared to controls. These proteins not only serve as targets for further development as biomarker candidates or components of biomarker panels, but also add to the growing understanding of dysregulated immune function and lipoprotein metabolism with chronic, excessive alcohol consumption.

Monkey Alcohol Tissue Research Resource: Banking Tissues for Alcohol Research

“…Now these tissues and associated bioinformatics are widely available to the larger alcohol research community through the interactive MATRR website. Since its inception, the MATRR has contributed resources that have been cited in 70 peer-reviewed publications, including reports of the functional (Ariwodola et al, 2003; Budygin et al, 2003; Floyd et al, 2004; Alexander et al, 2006; Carden et al, 2006; Anderson et al, 2007; Cuzon-Carlson et al, 2012; Welsh et al, 2011), genomic (Hemby et al, 2006; Acosta et al, 2009; Burnett et al, 2012), proteomic (Freeman et al, 2006, 2010, 2011), biochemical (Ivester et al, 2003; 2007; Lebold et al, 2010) and cardiac (Cheng et al, 2010) consequences of long-term, excessive ethanol consumption. The pathophysiological effects of chronic EtOH on the hypothalamic-pituitary-adrenal axis have also been well-characterized (Morrow et al, 2006; Porcu et al, 2006a, b, 2010; Ferguson et al, 2012; Helms et al, 2012a, b, c, 2013).…”

Proteomic Profiling of Liver and Plasma in Chronic Ethanol Feeding Model of Hepatic Alcohol Dehydrogenase-Deficient Deer Mice

“…Serine protease inhibitor (inter-alpha-trypsin inhibitor heavy chain H4 produced in the liver) negatively regulates endopeptidase activity and its precursor protein (serine protease inhibitor A3A precursor). However, ethanol fed group showing no significant differential expression of serine protease inhibitor diminishes its utility as a potential biomarker (Freeman et al, 2011). Amongst many other plasma proteins, down regulation of clusterin or sulfated glycoprotein-2 isoform 2 or apolipoprotein E isoform CRA_e in ethanol fed group is also supported by data from nonhuman primate model (Freeman et al, 2011), although precise function of clusterin is not established yet.…”

Protein biomarkers of alcohol abuse

“…Altered levels of serum amyloid A4, retinol-binding protein, inter-α inhibitor H4, Apo J (clusterin) and fibronectin were detected and confirmed by immunoblotting. Investigation of these target proteins in human subjects with excessive alcohol intake revealed increased levels of serum amyloid A4 and Apo J and decreased levels of fibronectin compared with controls [99]. …”