Urine Protein Biomarker Exploration in Ovarian Cancer

描述(申请人提供)：大多数卵巢癌患者在晚期出现疾病，存活率很低。通过更有效的筛查方法来改善这一点的努力显然是可取的。传统上，卵巢癌的筛查侧重于成像或使用血清标志物。然而，高昂的成像成本以及20年来未能发现比CA125更好的生物标志物的研究表明，一种新的方法将是值得的。因此，除了使用血清样本来发现生物标记物外，我们还将探索使用尿液作为替代体液来发现潜在的多肽标记物的选择，这最终可能转化为可靠的家庭筛查测试。这个试点项目将测试尿肽可能是卵巢癌有用的尿标记物的假设，尿肽通常尺寸小，热力学稳定，相互作用较少。在这个项目中，我们建议：1.)应用创新的蛋白质组学技术，如多维液相色谱(MD-LC)、质谱学、二维差示凝胶电泳法(2D-DGE)的荧光标记(2D-DGE)，从分类的卵巢癌亚型和年龄匹配的健康女性中生成尿肽图谱。2.)使用生物信息学工具来选择、鉴定和表征新的尿多肽，包括与卵巢癌特异相关的修饰。3.)建立可靠的尿液免疫分析和归一化方法，并将尿标志物组合应用于一项大型研究，包括术前尿液标本和正常对照。如果该项目成功，将从根本上改变卵巢癌早期检测的方法，并显著提高检测到的病例的存活率。

DESCRIPTION (provided by applicant): A majority of ovarian cancer patients presenting with diseases at late stages have poor survival rates. Efforts to improve this with more effective screening methods are clearly desirable. Traditionally, screening for ovarian cancer has focused on imaging or the use of serum markers. However, the high cost of imaging and the failure of 20 years of research to discover any better biomarkers than CA125, suggests a new approach would be worthwhile. Therefore, besides using serum samples for biomarker discovery, we will explore the option of using urine as an alternative body fluid to discover the potential peptide markers, which may ultimately be transformed into a reliable home screening test. This pilot project will test the hypothesis that urinary peptides, which normally are small size, thermodynamically stable and less interactive, could be useful urinary markers for ovarian cancer. In this project, we propose to: 1.) Apply innovative proteomic technologies such as multidimensional liquid chromatography (MD-LC), mass spectrometry, fluorescent labeling with two-dimensional differential gel electrophoresis (2D-DIGE) to generate urinary peptide profiles from the classified ovarian cancer subtypes and the age-matched healthy women. 2.) Using bio-informatic tools to select, identify and characterize the novel urinary peptides including modifications, which are specifically associated with ovarian cancers. 3.) Develop the reliable urine immunoassays and normalization methods, and apply the panel of urinary markers for validation in a large study including pre-operative urine specimens and normal controls. This project, if successful, could fundamentally shift the approach to the early detection of ovarian cancer and significantly improve the survival rate of cases detected.