Identification and Validation of Alcohol Biomarker Signatures by Proteomics

描述（由申请人提供）：三分之一的慢性酒精滥用者表现出心功能障碍，导致扩张性心肌病，最终导致充血性心力衰竭。功能上，酒精性心肌病（ACM）的特点是可逆和不可逆的改变。酒精中毒还会引起骨骼肌萎缩，称为酒精性肌病，这与ACM密切相关。骨骼肌无力的进展与左心室射血分数的下降并行。酒精对肌肉结构和功能的慢性影响已被证明会引起几种蛋白质和途径的变化。本拨款申请的目的是确定人类ACM的可靠诊断和预后蛋白生物标志物。该应用的具体目的是：1)鉴定长期饮酒大鼠心脏和血清中早发性acm相关蛋白生物标志物。因为在人类中，三角肌可以至少部分地作为心脏组织的替代物，ii)研究具有临床ACM和肌病的慢性酗酒者的肌肉活检和血清中的蛋白质表达，并将其与无症状的酗酒者和非酒精对照进行比较。

DESCRIPTION (provided by applicant): One third of chronic alcohol abusers display cardiac dysfunction, leading to dilated cardiomyopathy and eventually to congestive heart failure. Functionally, alcoholic cardiomyopathy (ACM) is characterized by reversible and irreversible alterations. Alcoholism also causes atrophy of skeletal muscle, termed alcoholic myopathy, which is closely correlated with ACM. Skeletal muscle weakness progresses in parallel with depression of the left ventricle ejection fraction. A chronic effect of alcohol on the structure and function of the muscles has been shown to induce changes in several proteins and pathways. The goal of this grant application is to identify reliable diagnostic and prognostic protein biomarkers of ACM in humans. The specific aims of this application are i) to identify early onset ACM-related protein biomarkers in heart and serum of rats fed alcohol chronically. Since in humans, the deltoid muscle may serve at least in part, as a surrogate for cardiac tissue, ii) to study protein expression in muscle biopsies and serum of chronic alcoholics with clinical ACM and myopathy and compare it to asymptomatic alcoholics and nonalcoholic controls. A unique heart tissue bank, from irreversibly brain damaged alcoholic patients whose hearts could not be transplanted and were frozen after termination of life support, will be investigated, iii) To discover ACM protein biomarkers by comparing protein profiles of the left ventricle of chronic alcoholics with and without ACM, and nonalcoholic controls. Additionally, corresponding muscle and serum samples from the same subjects will be evaluated for biomarkers. iv) To perform biostatistical and bioinformatics data mining for differential analysis of proteomic expression from rat and human samples, correlate global protein changes with clinical data to obtain disease-specific protein patterns for validation, integrate data in a relational database for future comparative studies with other cardiomyopathies, and map clusters of protein changes in affected pathways of the protein interactome in order to develop a mechanistic model of the pathogenesis of ACM and myopathy. v) To validate the specificity, accuracy and usefulness of emergent proteomic panels of biomarkers in new serum and muscle samples that will lead to clinical context validation and application of ACM biomarkers. The comprehensive search and discovery of protein biomarkers of ACM will encompass the proteomes of sera, and of subcellular components of tissues. A combination of 2D electrophoresis with multidimensional liquid chromatography and tandem mass spectroscopy will be employed to characterize protein changes, as well as, post-translational protein modifications, including those directly related to alcohol metabolites. Immunoassays will be also developed for validation purposes. There are 18 million alcohol abusers in the US. Many of these persons suffer from alcoholic cardiomyopathy, which is the single leading cause of dilated cardiomyopathy in which an etiology can be determined. Early diagnosis of this condition by specific biomarkers will offer the opportunity for treatment and prevention of heart enlargement, congestive heart failure and death.