Modifications of glycated Hb:Novel biomarkers of alcohol

描述（由申请人提供）：慢性和酗酒每年影响数百万美国人。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的全球影响（在Room等人，Lancer 2005中综述）。此外，酒精滥用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，最可靠的酒精滥用筛查技术是自我报告方法。显然，滥用者自我报告酒精使用情况的主要缺点是报告不足。有趣的是，没有酗酒的生物标志物可用。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，两种稳定的乙酰丙酮酸修饰的糖化血红蛋白（Hb）加合物在（A）狂饮和（B）慢性饮酒后产生，它们分别代表（A）急性和（B）慢性酒精滥用的特异性和丰富的标志物。我们的初步数据显示：（a）乙醛修饰糖基化Hb形成稳定的加合物;（B）糖基化HbAo约占健康人总Hb的20%（而糖基化HbA 1c占3-5%）;（c）根据酒精代谢物暴露的时间和浓度，形成两种类型的Hb-乙醛加合物。具体目标：我们建议（1）将乙醛加合物偶联到蛋白质载体（分别代表狂饮和慢性饮酒后形成的加合物）上，用于开发特异性抗体;（2）开发特异性和灵敏度高的免疫测定方法，分别使用急性/狂饮或慢性酒精给药的大鼠实验模型检测红细胞和血清/血浆中的这些生物标志物。 与公共卫生的相关性：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有一次等于或>5杯）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名青少年和年轻人中，有39人受到影响。虽然酒精治疗计划在帮助酒精滥用者方面取得了成功，但许多酒精滥用者没有得到治疗，因为我们缺乏识别酒精滥用者的好方法。目前，自我报告的方法（问卷）被用来检测酒精滥用，因为我们没有更好的程序，可靠地确定酒精滥用使用血液，唾液或尿液。我们早期的研究表明，乙醛（酒精在血液中的分解产物）在狂饮和慢性饮酒后与修饰的血红蛋白（红细胞中发现的主要蛋白质）结合，形成两种不同的标记物。“我们将开发特定的方法来识别急性或慢性酒精大鼠血液/红细胞中的狂欢和慢性酒精特异性标志物。未来的研究将被设计来测量这些酒精特异性标志物在人类酗酒和慢性酒精摄入后。

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.