Alcohol-specific modifications of glycated Hb: Novel biomarkers of alcohol abuse

描述(由申请者提供)：每年有数百万美国人受到长期酗酒的影响。最近的一项研究表明，酒精占全球疾病负担的4%，相当于烟草使用和高血压对世界范围的影响(发表在Room等人的评论中，兰瑟，2005年)。此外，酗酒还与沉重的社会和经济负担有关(每年约1850亿美元)。基于敏感性和特异性，酒精滥用最可靠的筛查技术是自我报告方法。显然，自我报告滥用者饮酒的主要缺点是漏报。有趣的是，目前还没有关于酗酒的生物标志物。我们的长期目标是开发可靠的方法来检测急性和慢性酒精滥用。 我们的中心假设是，在(A)酗酒和(B)慢性饮酒之后，会产生两个稳定的乙醛修饰的糖化血红蛋白(HB)加合物，它们分别代表(A)急性和(B)慢性酒精滥用的特异和丰富的标志。我们的初步数据显示：(A)乙醛修饰糖化的Hb形成稳定的加合物；(B)在健康人中，糖化的HbAo约占总Hb的20%(而糖化的HbA1c占3-5%)；以及(C)两种类型的Hb-乙醛加合物的形成取决于酒精代谢物暴露的时间和浓度。具体目标：我们建议(1)将乙醛加合物结合到蛋白质载体(分别代表酗酒和慢性饮酒后形成的加合物)上，以开发特异性抗体；(2)建立特异和灵敏的免疫分析方法，分别在急性/酗酒或慢性饮酒的大鼠实验模型上检测红细胞和血清/血浆中的这些生物标志物。 与公共健康相关：酒精滥用，包括慢性酒精滥用(或酒精中毒)和酗酒(定义为在过去一个月内一次饮酒等于或等于5英镑)是美国可预防死亡的第三大原因。在过去的几年里，酗酒现象显著增加，在接受采访的100名青少年中，有高达39人受到影响。虽然酒精治疗项目在帮助酗酒者方面取得了成功，但许多酗酒者没有得到治疗，因为我们缺乏识别酗酒者的好方法。目前，自我报告方法(问卷调查)被用来检测酒精滥用，因为我们没有更好的程序来可靠地确定使用血液、唾液或尿液来确定酒精滥用。我们早期的研究表明，在酗酒和长期饮酒后，乙醛(酒精在血液中的分解产物)与修饰的血红蛋白(红细胞中的主要蛋白质)结合，形成两种不同的标记。我们将开发特定的方法来识别急性或慢性酒精中毒大鼠血液/红细胞中的酗酒和慢性酒精特异性标记物。未来的研究将被设计用来测量在酗酒和长期饮酒后人类的这些酒精特异性标志物。

DESCRIPTION (provided by applicant): Chronic and binge drinking affect millions of Americans each year. A recent study revealed that alcohol contributes to 4% of the global burden of disease which is equivalent to the world-wide impact of tobacco use and hypertension (Reviewed in Room et al, Lancer 2005). In addition, alcohol abuse is associated with heavy social and economic burdens (approximately $185B annually). Based on sensitivity and specificity, the most reliable screening techniques for alcohol abuse are self-reporting methods. Clearly, the major shortcoming of self-reporting alcohol use by abusers is under-reporting. Interestingly, no biomarkers of binge drinking are available. Our long-term goal is to develop reliable methods to detect acute and chronic alcohol abuse. Our central hypothesis is that two stable acetaldehyde-modified glycosylated hemoglobin (Hb) adducts are produced following (A) binge and (B) chronic drinking and these represent specific and abundant markers of (A) acute and (B) chronic alcohol abuse, respectively. Our preliminary data show that (a) acetaldehyde modifies glycosylated Hb to form stable adducts; (b) glycosylated HbAo represents approximately 20% of the total Hb in healthy persons (while glycosylated HbA1c represents 3-5%); and (c) two types of Hb-acetaldehyde adducts form depending on the length and concentration of alcohol metabolite exposure. Specific aims: We propose to (1) conjugate acetaldehyde adducts onto protein carriers (that represent adducts formed following binge and chronic drinking, respectively) for the development of specific antibodies and (2) develop specific and sensitive immunoassay methods for detecting these biomarkers in the red blood cells and serum/plasma using rat experimental models of acute/binge drinking or chronic alcohol administration, respectively. Relevance to public health: Alcohol abuse, including chronic alcohol abuse (or alcoholism) and binge drinking (defined as having equal to or >5 drinks on one occasion within the past month) is the third leading cause of preventable death in the US. Binge drinking has significantly increased in the past few years, affecting up to 39 out of 100 teens and young adults interviewed. Although alcohol treatment programs are successful for helping alcohol abusers, many alcohol abusers are not treated because we lack good methods for identifying alcohol abusers. Currently, self-reporting methods (questionnaires) are used to detect alcohol abuse because we do not have better procedures that reliably determine alcohol abuse using blood, saliva, or urine. Our early studies show that acetaldehyde (alcohol's breakdown product in the blood) binds to modified hemoglobin (the major protein found in red blood cells) following binge and chronic drinking to form two different 'markers.' We will develop specific methods for identifying binge and chronic alcohol-specific markers in the blood/red blood cells of rats administered acute or chronic alcohol. Future studies will be designed to measure these alcohol-specific-markers in humans following binge and chronic alcohol intake.