Biomarker Detection Using NAPPA Tumor Antigen Arrays

描述（由申请人提供）：该项目的总体目标是鉴定血清中的自身抗体生物标志物，这些标志物可以很容易地用于癌症的早期检测。抗体是由肿瘤特异性的蛋白质表达、突变、降解或定位改变诱导的，具有高特异性。我们开发了一种新的可编程蛋白微阵列技术（NAPPA），该技术使用打印的cDNA编码肿瘤抗原，并在体外翻译。在过去的四年里，我们获得了EDRN生物标志物开发实验室的资助，开发NAPPA作为临床研究工具，并将其用于检测乳腺癌的自身抗体生物标志物，并筛选了700多种血清，鉴定出32种新的乳腺癌和23种卵巢生物标志物，这些生物标志物已进行了盲法验证研究。在这里，我们建议增加NAPPA阵列的特征密度，以适应我们的cDNA收集规模的增加，现在接近10,000个独特的基因（目标1），并开发一种生产糖蛋白组阵列的方法（目标2）。这两种方法都将通过增加筛选抗原的数量和质量来增强自身抗体生物标志物的发现。目标3将利用目标1和目标2的结果，扩大已确定的顶级乳腺癌生物标志物，重点关注更难检测的特定乳腺癌亚型，包括Her2+乳腺癌、三阴性（ER-PR-Her2-）乳腺癌和发生在高乳腺密度环境中的癌症。探讨抗体检测与肿瘤抗原表达的关系。目的4将使用多中心临床研究中收集的血清评估II期研究中顶级乳腺癌生物标志物组合的性能特征。目标5将侧重于这些生物标志物在临床诊断前检测疾病的效用。在这些实验的最后，我们的目标是根据EDRN生物标志物发现指南，确定EDRN CEVCs在进一步的III期独立验证研究中用于乳腺癌早期检测的自身抗体血清特征。

DESCRIPTION (provided by applicant): The overall goal of this project is to identify autoantibody biomarkers in sera that can be readily used for the early detection of cancer. Antibodies are induced by tumor-specific alterations in protein expression, mutation, degradation, or localization, with high specificity. We have developed a novel programmable protein microarray technology (NAPPA), which uses printed cDNA encoding tumor antigens that are translated in vitro. We have been funded for the past four years as an EDRN Biomarker Development Laboratory to develop NAPPA as a clinical research tool and to employ it for the detection of autoantibody biomarkers for breast cancer, and have screened over 700 sera and identified 32 novel breast and 23 ovarian biomarkers that have undergone blinded validation studies. Here, we propose to increase the feature density of NAPPA arrays in order to accommodate the increasing size of our cDNA collection, which now approaches 10,000 unique genes (Aim 1) and to develop a method for producing glycoproteome arrays (Aim 2). Both of these methods will enhance autoantibody biomarker discovery by increasing the number and quality of the antigens screened. Aim 3 will use the results of Aims 1 and 2 to expand the top breast cancer biomarkers that have been identified to focus on particular subtypes of breast cancer that are more difficult to detect including Her2+ breast cancers, triple-negative (ER-PR-Her2-) breast cancers, and cancers that occur in the setting of high breast density. Association of antibody detection and tumor antigen expression will be explored. Aim 4 will assess the performance characteristics of the combined set of top breast cancer biomarkers in Phase II studies using sera collected in multicenter clinical studies. Aim 5 will focus on the utility of these biomarkers to detect disease prior to clinical diagnosis. At the end of these experiments, our goal is to identify autoantibody serum signatures that can be used by EDRN CEVCs in further Phase III independent validation studies for the early detection of breast cancer according to the EDRN biomarker discovery guideline.