Discovery and validation of novel serological biomarkers of colon cancer

结肠癌新型血清学生物标志物的发现和验证

• 批准号: 7212781
• 负责人: DAVID W. SPEICHER
• 金额: $ 40.54万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-29 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7212781
• 关键词: biomarker; clinical research; colon neoplasms; human; proteins; serum

DESCRIPTION (provided by applicant): The overall goal of this proposal is to identify novel serological biomarkers of human colon cancer and evaluate their capacities to predict disease occurrence or clinical outcome. To achieve this goal, we will use a powerful new 4-D protein profiling method recently developed in our lab that can identify many low abundance serum proteins. This method utilizes three tandem orthogonal protein separations consisting of: major protein depletion, solution IEF and 1-D SDS gels. Each gel lane is cut into uniform slices, and digested with trypsin. Each tryptic digest, which still contains many proteins, is then analyzed by LC-MS/MS using a nanocapillary reverse phase column coupled to a high performance linear ion trap mass spectrometer. Candidate human biomarkers in SCID mice bearing human tumors will be identified by distinguishing human and mouse proteins based on species sequence differences. Validation of candidate biomarkers will be conducted in two stages. Initially, candidate human biomarkers will be tested using medium throughput quantitative mass spectrometer assays and Western blots to evaluate sera from a small group of colon cancer patients and matched controls. Higher throughput sandwich ELISA assays will then be developed for the most promising biomarkers, and these assays will be used to systematically test sera from a larger number of early and late stage cancer patients and matched controls. Levels of individual biomarkers as well as groups of biomarkers will be evaluated for their capacity to predict disease occurrence and clinical outcome. This ambitious but feasible five year project involves the following Specific Aims: 1) Identify candidate human colon cancer biomarkers in a SCID mouse xenograph model system using a novel multi-dimensional protein profiling method; 2) Validate candidate serum biomarkers in patient and control sera using medium throughput assays; 3) Develop high throughput quantitative immunoassays for the most promising biomarkers from initial patient screens; and 4) Compare patient and control serum using high throughput immunoassays. Relevance to public health. Discovery of novel protein biomarkers of colon cancer that can be measured using a simple blood test has great potential to decrease the suffering and loss of life associated with this disease. Recently developed, powerful mass spectrometry-based methods provide a unique opportunity to systematically discovery groups of colon cancer biomarkers. It is most likely that groups of biomarkers (biomarker signatures) will have greater power to monitor cancer than individual biomarkers.

描述（由申请人提供）：该提案的总体目标是鉴定人类结肠癌的新型血清学生物标志物并评估其预测疾病发生或临床结果的能力。为了实现这一目标，我们将使用我们实验室最近开发的强大的新型 4-D 蛋白质分析方法，该方法可以识别许多低丰度血清蛋白质。该方法利用三种串联正交蛋白质分离，包括：主要蛋白质去除、溶液 IEF 和一维 SDS 凝胶。将每个凝胶泳道切成均匀的切片，并用胰蛋白酶消化。然后，使用与高性能线性离子阱质谱仪联用的纳米毛细管反相柱，通过 LC-MS/MS 分析仍含有许多蛋白质的每个胰蛋白酶消化物。将根据物种序列差异区分人类和小鼠蛋白质，从而鉴定携带人类肿瘤的 SCID 小鼠中的候选人类生物标志物。候选生物标志物的验证将分两个阶段进行。最初，将使用中等通量定量质谱仪测定和蛋白质印迹来测试候选人类生物标志物，以评估一小群结肠癌患者和匹配对照的血清。然后，将为最有希望的生物标志物开发更高通量的夹心 ELISA 测定，并且这些测定将用于系统地测试来自大量早期和晚期癌症患者和匹配对照的血清。将评估个体生物标志物以及生物标志物组的水平预测疾病发生和临床结果的能力。这个雄心勃勃但可行的五年项目涉及以下具体目标：1）使用新型多维蛋白质分析方法在 SCID 小鼠异种模型系统中识别候选人类结肠癌生物标志物； 2) 使用中等通量测定验证患者和对照血清中的候选血清生物标志物； 3) 针对最初患者筛选中最有希望的生物标志物开发高通量定量免疫测定方法； 4) 使用高通量免疫测定法比较患者血清和对照血清。与公共卫生的相关性。发现可以使用简单的血液测试来测量的新型结肠癌蛋白质生物标志物，具有减少与这种疾病相关的痛苦和生命损失的巨大潜力。最近开发的强大的基于质谱的方法为系统地发现结肠癌生物标志物组提供了独特的机会。生物标记物组（生物标记物特征）很可能比单个生物标记物具有更大的癌症监测能力。