Pilot Project I: Cross-Sectional Analysis of Areca Alkaloids in Buccal Cells and Hair from Areca Nut Chewers as Candidate Biomarkers for Short- and Long-Term Areca Nut Exposure

项目总结 咀嚼槟榔是一种令人上瘾的致癌行为，数以亿计的 世界各地的人们。这些咀嚼者中的大多数是服务不足的少数民族，他们居住在 西太平洋地区，包括关岛。因此，迫切需要停止试验来解决这个问题 严重的健康差距问题。然而，迫切需要客观的生物标记物来验证短期和 目前尚不能获得此类试验的长期AN暴露。应用创新的质量 光谱技术，我们首次提出建立可靠的生物标志物，以客观地 衡量短期和长期的风险敞口。这些生物标志物可以用来评估咀嚼状态 并客观地确定停药试验中的依从性和有效性。正如我们和其他人所发现的， 槟榔碱槟榔碱、槟榔碱、古瓦林和古瓦卡因是AN所特有的。因此，这些 如果在人体样本中能检测到生物碱，生物碱可以作为接触的可靠生物标志物。 嚼了一口之后。口腔细胞(BCS)和头皮毛发可以无创地收集并提供广泛的 根据长度，曝光几天(BCS)和几个月(头发)的日历。在这 研究中，我们将使用一种创新的方法，将BCS和头皮中测量的槟榔生物碱水平关联起来 咀嚼者的毛发到自我报告的短期(前一周)和长期(前一个月或六个月) 几个月前)使用经过验证的调查问卷进行暴露。作为一个新的概念，我们将表达一个 咀嚼者的暴露是他们的“平均负荷量”(“AANL”)。使用来自调查问卷的数据，我们 将计算咀嚼者的AANL，方法是将每次咀嚼的量乘以平均值 咀嚼时间和平均每天咀嚼频率，并考虑AN的时间 留在嘴里。作为额外的新鲜感，我们将利用前一周的咀嚼者AANL， 前一个月和前六个月生成的算法回溯地近似于 从测量的生物碱水平中暴露出来。这些算法将分别为两个主要的 关岛的咀嚼者类型：单独咀嚼的人(1类咀嚼者)和作为咀嚼的人 槟榔馅料，以槟榔叶为主要原料，裹以熟石灰和 烟草(‘2级咀嚼者’)。停药试验是减少AN诱导的最有效的方法 癌症。然而，这些试验迫切需要可靠的生物标志物来确定短期和长期试验 合规性。我们提出的工作旨在建立这样的生物标记物。这将有助于取得全面的成功 在全球范围内进行戒断试验，从而降低全球艾滋病的患病率 咀嚼及其相关的健康差距，特别是口腔癌。

PROJECT SUMMARY Areca nut (AN) chewing is an addictive cancer-causing behavior practiced by hundreds of millions of people worldwide. Most of these AN chewers are underserved minorities who reside in countries of the Western Pacific Region including in Guam. Thus, AN cessation trials are urgently needed to address this severe health disparity problem. However, critically required objective biomarkers to verify short- and long-term AN exposure for such trials are currently not available. Applying an innovative mass spectrometry technique, we propose to establish for the first time reliable biomarkers to objectively measure short- and long-term AN exposure. These biomarkers can be used to assess chewing status and objectively determine compliance and efficacy in AN cessation trials. As found by us and others, the areca alkaloids arecoline, arecaidine, guvacoline, and guvacine are specific to the AN. Thus, these alkaloids can serve as reliable biomarkers for AN exposure if they can be detected in human specimens after AN chewing. Buccal cells (BCs) and scalp hair can be non-invasively collected and provide a broad calendar of AN exposure of several days (BCs) and several months (hair) depending on length. In this study, we will use an innovative approach of correlating areca alkaloid levels measured in BCs and scalp hair of AN chewers to self-reported short-term (previous week) and long-term (previous month or six months prior) AN exposure using validated AN questionnaires. As a novel concept, we will express a chewer’s AN exposure as their "average AN load" (“AANL”). Using data from the AN questionnaires, we will calculate the chewer’s AANL by multiplying the amount of AN chewed per session by the average chewing time and the average daily chewing frequency and taking into consideration the time the AN is retained in the mouth. As an additional novelty, we will utilize the chewer’s AANL for the previous week, previous month, and six months prior to generate algorithms that retrospectively approximate AN exposure from measured alkaloid levels. These algorithms will be generated separately for the two main types of chewers in Guam: those who chew AN alone (‘class 1 chewers’) and those who chew AN as a betel quid consisting of AN as the primary ingredient wrapped in a betel leaf with slaked lime and tobacco (‘class 2 chewers’). AN cessation trials are the most effective approach to reduce AN-induced cancers. However, these trials urgently require reliable biomarkers to determine short- and long-term trial compliance. Our proposed work aims to establish such biomarkers. This will assist in the overall success of AN cessation trials worldwide and consequently in the reduction of the global prevalence of AN chewing and its associated health disparities, particularly oral cancer.