Minimally Invasive Molecular Approaches for the Detection of Barrett’s Esophagus and Esophageal Adenocarcinoma

项目摘要/摘要 食管腺癌(EAC)是一种预后较差的致命性癌症(5年生存率和20%)。 在症状出现后诊断，但早期诊断的存活率很高。肠化生， 或Barrett‘s食道(BE)，是唯一已知的EAC前体，并通过发展为EAC 发育不良。EAC可以通过异型增生的内窥镜治疗来预防。因此，内窥镜筛查BE和 推荐使用内窥镜监测以发现异型增生和EAC。然而，60%的流行BE 仍未确诊，90%的EAC病例是在BE监测计划之外诊断的。少校 筛查的障碍是内窥镜检查的侵袭性和高昂的费用。此外，建议进行筛查。 仅限于慢性胃食道反流(GERD)患者，尽管50%的BE/EAC患者没有报告 热病症状。内窥镜监测漏掉33%流行的EAC和异型增生，原因是斑块 异型增生/EAC的分布，以及活检样本不足。因此，内窥镜检查的整体效果 监控也严重受损。 我们使用简化代表性亚硫酸氢盐测序(RRBS)来鉴定一组甲基化的DNA标记 (MDMS)BE和异型增生/EAC，然后进行验证。MDM小组具有很高的鉴别力(AUCS&GT；0.9) 对于BE和普遍的异型增生/EAC。对通过海绵获得的食道细胞学样本进行化验 在两个病例对照研究中，在串(SOS)设备上高精度地检测到BE(AUCS 0.97-1.0 在转诊人群中。FDA批准的SOS设备(Capnostics，Doylestown，PA)是一种25 mm的聚氨酯 压缩在可溶胶囊外壳中的海绵，胶囊外壳被压缩后在胃中膨胀成球状。 吞下去了。当通过连接的绳子从嘴里拉出时，对整个食道进行采样 实现了粘膜。护士管理的SOS测试是安全的，耐受性好，参与率高 (65%)。因此，我们的中心假设是通过食道细胞学检测的新的判别MDM 通过SOS设备获得的标本将能够在筛查中准确地检测BE和异型增生/EAC 患有和不患有慢性GERD的人群。我们将通过三个具体目标来检验这一假设。 在具体目标1中，我们将在筛查中测量SOS BE测试的阳性和阴性预测价值 梅奥卫生系统初级保健诊所的合格人口，并将这些值在 无论有没有GERD。在具体目标2中，我们将确定临床和人口因素，特别是GERD， 影响精度的是一个预定的SOS BE测试预测算法。在具体目标3中，我们将衡量 使用SOS装置检测BE异型增生/EAC的MDM的准确性。 利用一种创新的微创(非内窥镜)和分子方法，这项提议将 积极影响BE检测和监测，实现有效治疗，改善EAC结果。

PROJECT SUMMARY/ABSTRACT Esophageal adenocarcinoma (EAC) is a lethal cancer with poor outcomes (5 year survival <20%), when diagnosed after the onset of symptoms, but survival is excellent when diagnosed early. Intestinal metaplasia, or Barrett’s esophagus (BE), is the only known precursor of EAC, and progresses to EAC via development of dysplasia. EAC can be prevented by endoscopic therapy of dysplasia. Hence endoscopic screening for BE and endoscopic surveillance to detect dysplasia and EAC are recommended. However, > 60% of prevalent BE remains undiagnosed and 90% of all EAC cases are diagnosed outside a BE surveillance program. The major barrier to BE screening is the invasiveness and high cost of endoscopy. Further, screening is recommended only in those with chronic gastroesophageal reflux (GERD), despite 50% of BE/EAC patients not reporting GERD symptoms. Endoscopic surveillance misses 33% of prevalent EAC & dysplasia, due to the patchy distribution of dysplasia/EAC, and inadequate biopsy sampling. Hence the overall effectiveness of endoscopic surveillance is also severely compromised. We used reduced representation bisulfite sequencing (RRBS) to identify a panel of methylated DNA markers (MDMs) of BE and dysplasia/EAC followed by validation. MDM panels were highly discriminant (AUCs > 0.9) for BE and prevalent dysplasia/EAC. When assayed on esophageal cytology specimens obtained via a sponge on string (SOS) device, BE was detected with high accuracy (AUCs 0.97-1.0) in two case control studies done in referral populations. The FDA approved SOS device (Capnostics, Doylestown, PA) is a 25 mm polyurethane sponge compressed in a dissolvable capsule shell, which expands into a sphere in the stomach after being swallowed. When pulled out through the mouth via an attached string, sampling of the entire esophageal mucosa is achieved. The nurse-administered SOS test is safe and well tolerated with high participation rates (65%). Hence our central hypothesis is that novel discriminant MDMs assayed on esophageal cytology specimens obtained via the SOS device will enable accurate BE and dysplasia/EAC detection, in a screening population with and without chronic GERD. We will test this hypothesis by three specific aims. In specific Aim 1, we will measure the positive and negative predictive value of the SOS BE test in a screening eligible population from primary care clinics in the Mayo Health System and compare these values in those with and without GERD. In specific Aim 2 we will identify clinical and demographic factors, particularly GERD, influencing the accuracy a predetermined SOS BE test prediction algorithm. In Specific Aim 3, we will measure the accuracy of MDMs for the detection of dysplasia/EAC in BE, using the SOS device. Utilizing an innovative, minimally invasive (non-endoscopic) and molecular approach, this proposal will favorably impact BE detection and surveillance, enabling effective treatment, and improved EAC outcomes.