Urine Biomarker Discovery for Early Detection of Liver Cancer

描述(申请人提供)：尿液生物标志物的发现用于肝癌的早期诊断。尿液中含有大分子物质，可用于早期发现发生在非尿路部位的疾病和癌症。鉴于尿液用于生物标志物监测的诸多优点，该项目的目标是开发尿液中大分子(DNA和多糖)的生物标志物发现系统，最终可能改善此类疾病的预后。对DNA修饰和糖链种类(宿主和适当情况下，病毒)的比较分析将使用健康和疾病直到诊断为肝癌的不同阶段的个人尿样进行。因此，该项目将提供新的方法和新的生物标记物，用于肝癌以及最终使用尿液作为生物液体的其他癌症。这些目标将通过以下目标实现。目的1：检测尿液中与肝细胞癌发生相关的遗传或表观遗传修饰的DNA标志物，作为早期发现肝细胞癌的潜在尿液生物标志物。检测与肝癌相关的遗传和表观遗传修饰的分析方法将被开发用于分析尿液DNA。对不同阶段的肝病患者和健康受试者尿液中的每个DNA标志物进行检测，以评估每个标志物作为肝细胞癌尿源性生物标志物小组的候选者。目的2：确定从人尿液中分离的游离多糖是否可用于发现生物标记物和肝癌的早期发现。这一目标将确定尿液中存在的简单多糖(可能是糖蛋白的残留物)是否可以成为癌症生物标记物的来源。从个体尿液中分离出的不同种类的葡聚糖将通过高效液相色谱进行比较分析。似乎与肝病诊断相关的糖链结构将被表征，并将作为可能的生物标记物进行进一步探索。从目标1和目标2获得的有前景的尿源性生物标志物将被用于提出另一阶段的研究，在该阶段中，可以开展更大规模的预验证研究，以确定每个生物标志物的敏感性和特异性，以及单一生物标志物或生物标志物组合对肝细胞癌早期检测的预测价值。这一知识最终可以用于构建一组用于肝细胞癌早期检测的尿液衍生生物标记物。该项目完成后，我们将开发出以尿液为生物样本液体的生物标记物发现技术平台，并可应用于其他癌症。 公共卫生相关性：这项建议旨在利用尿液作为生物样本液体，发现用于肝癌早期检测的生物标记物。使用尿液将是适当和有利的，因为尿液可以包含来自非尿路部位的生物标志物，并且使用尿液将是非侵入性的。该项目完成后，我们将开发尿液中生物标记物发现的技术平台，然后可以应用于其他癌症。

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.