Urine Biomarker Discovery for Early Detection of Liver Cancer

描述（由申请人提供）：用于早期检测肝癌的尿液生物标志物发现。尿液中含有大分子，可用于早期检测非泌尿道部位发生的疾病和癌症。鉴于在生物标志物监测中使用尿液具有许多优势，该项目的目标是开发用于发现尿液中大分子（DNA 和聚糖）的生物标志物的系统，最终可能改善此类疾病的预后。将使用处于不同健康和疾病阶段直至诊断肝癌的个体的尿液样本对 DNA 修饰和聚糖种类（宿主和适当情况下的病毒）进行比较分析。因此，该项目将为 HCC 以及最终使用尿液作为生物液体的其他癌症提供新方法和新生物标志物。这些目标将通过以下目标来实现。目标 1：检测以前与尿液中 HCC 发生相关的基因或表观遗传修饰 DNA 标记，作为早期检测 HCC 的潜在尿液生物标记。将开发检测与 HCC 相关的遗传和表观遗传修饰的方法来分析尿液 DNA。将检测不同阶段肝病患者和作为对照的健康受试者尿液中的每种 DNA 标记物，以评估每种标记物作为 HCC 尿液衍生生物标记物组的候选者。目标 2：确定从人尿液中分离的游离聚糖是否可用于生物标志物发现和 HCC 的早期检测。该目标将确定尿液中存在的非复杂聚糖（大概是糖蛋白的残余物）是否可以作为癌症生物标志物的来源。从个体尿液中分离出的聚糖种类，作为疾病阶段的函数，将通过 HPLC 进行比较分析。与肝病诊断相关的聚糖结构将被表征，并将作为可能的生物标志物进行进一步探索。从目标 1 和 2 中获得的有前景的尿液衍生生物标志物将用于提出另一阶段的研究，其中可以开展更大规模的预验证研究，以确定每种生物标志物的敏感性和特异性以及单个生物标志物或生物标志物组合对 HCC 早期检测的预测价值。这些知识最终可以用来构建一组用于早期检测 HCC 的多种尿液来源的生物标志物。该项目完成后，我们将开发出使用尿液作为生物样本液的生物标志物发现技术平台，然后可应用于其他癌症。 公共健康相关性：该提案旨在使用尿液作为生物样本液体来发现用于早期检测肝癌的生物标志物。使用尿液既是适当的又是有利的，因为尿液可以含有来自非尿路部位的生物标志物，并且尿液的使用将是非侵入性的。该项目完成后，我们将开发用于发现尿液中生物标志物的技术平台，然后将其应用于其他癌症。

DESCRIPTION (provided by applicant): Urine Biomarker Discovery for the Early Detection of Liver Cancer. Urine contains macro-molecules that can be used for the early detection of diseases and cancers that occur at non-urinary tract sites. Given the many advantages of using urine in biomarker monitoring, the goal of this project is to develop systems for the biomarker discovery of macromolecules (DNA and glycan) in urine that may ultimately improve the prognosis for such a disease. Comparative analysis of DNA modifications and glycan species (host and, where appropriate, viral) will be performed using urine samples from individuals at different stages of health and disease up to the diagnosis of liver cancer. This project will thus deliver new methods and new biomarkers for HCC and ultimately other cancers using urine as the biological fluid. These goals will be achieved by the following Aims. Aim 1: Detection of genetically or epigenetically modified DNA markers that have previously been associated with the development of HCC in urine as potential urine biomarkers for the early detection of HCC. The assays for detecting genetic and epigenetic modifications associated with HCC will be developed for analyzing urine DNA. The detection of each DNA marker in the urine of patients with different stages of liver disease and healthy subjects as the control will be performed to evaluate each marker as a candidate for a HCC, urine-derived biomarker panel. Aim 2: Determine if free glycan, isolated from human urine, can be used for biomarker discovery and the early detection of HCC. This Aim will determine if uncomplex glycans (presumably remnants of glycoproteins) present in urine can be a source for cancer biomarkers. The species of glycan, isolated from the urine of individuals, as a function of disease stage will be comparatively analyzed by HPLC. Glycan structures that appear to correlate with a diagnosis of liver disease will be characterized, and will be further explored as possible biomarkers. The promising urine-derived biomarkers obtained from Aim 1 and 2 will be used to propose another phase of study in which a larger scale, pre-validation study can be instituted to determine the sensitivity and specificity of each biomarker and the predicative value of a single biomarker or a combination of biomarkers for the early detection of HCC. This knowledge can ultimately be used so that a panel of multiple urine-derived biomarkers for the early detection of HCC can be constructed. Upon accomplishment of this project, we will have developed the technology platform for biomarker discovery that uses urine as the biological sample fluid, which can then be applied to other cancers. PUBLIC HEALTH RELEVANCE: This proposal aims to discover biomarkers for the early detection of liver cancer using urine as the biological sample fluid. The use of urine would be both appropriate and advantageous because urine can contain biomarkers from non- urinary tract sites, and the use of urine would be non-invasive. Upon accomplishment of this project, we will have developed the technology platform for biomarker discovery in urine, which can then be applied to other cancers.