Differential Capture Proteomics for Cardiovascular Disease Biomarker Discovery

描述（由申请人提供）：在美国，心血管疾病（CVD）每年造成约140万人死亡，占所有死亡人数的近40%。2007年心脏病和中风的预计费用为4318亿美元。新的生物标志物的发现对于这种毁灭性疾病的筛查、诊断、预后、治疗反应预测和治疗监测方面急需的进展至关重要。该项目第一阶段的目标是在人类血浆中优化一种新的蛋白质组学技术，差分捕获蛋白质组学stm (DCP)，该技术可以识别两种生物样品之间蛋白质组成的差异。该技术还可以同时为每种鉴定的不同蛋白质创建亲和试剂。差分蛋白质组学公司已经获得了DCP的专利。后续II期研究的目标是将新优化的DCP应用于人类血浆，利用合适的患者血浆样本发现CVD的生物标志物，从而实现新的蛋白质分离和鉴定、新的亲和试剂创建、测定配方和临床验证。在我们的实验室优化DCP技术的研究已经成功地集中在一个模型系统上。然而，DCP在患者血浆样本中发现CVD生物标志物的能力尚未得到优化研究的证明。本文提出了这样一项研究，在该研究中，将测试掺杂CVD生物标志物（代表CVD模型患者样本）在临床样本水平浓度下的可检测性。拟议的研究将使用五种特定的CVD蛋白抗原，这些抗原有商业上可用的免疫测定方法。I期研究的成功将导致II期项目，对多个CVD患者[血浆]样本进行测试，以发现新的生物标志物、生成分析方法和验证。特异性目的1：确认三个汇集的随机肽噬菌体文库实际上包含针对所选的五种CVD抗原的结合噬菌体。特异性目标2：展示差异捕获蛋白质组学如何很好地检测两种人类血浆样品之间的差异，其中一种血浆样品掺杂了五种CVD抗原的混合物。特异性目的3：建立亲和捕获试剂的特异性，这些试剂是该过程的一部分。公共卫生相关性：由于差异捕获蛋白质组学将比目前的方法更具成本效益和时间效益，它有可能为生物标志物的综合发现、验证和验证提供一个突破性的系统。新的心血管疾病生物标志物的发现将对这种毁灭性疾病的筛查、诊断、预后、治疗反应预测和治疗监测方面急需的进展至关重要。

DESCRIPTION (provided by applicant): In the United States, cardiovascular disease (CVD) accounts for approximately 1.4 million deaths annually, representing nearly 40% of all deaths. The projected cost of heart disease and stroke for 2007 is $431.8 billion. The discovery of new biomarkers will be critical for much-needed advances in screening, diagnosis, prognosis, prediction of therapeutic response and therapeutic monitoring for this devastating disease. The goal for Phase I of this Project is to optimize in human plasma a new proteomics technology, Differential Capture ProteomicsTM (DCP), which can identify the differences in protein composition between two biological samples. This technology also simultaneously creates affinity reagents for each of the identified difference proteins. A patent for DCP has been issued to Differential Proteomics Inc. The goal for a subsequent Phase II study is to apply DCP, newly optimized for human plasma, to the discovery of biomarkers of CVD using appropriate patient [plasma] samples to enable novel protein isolation and identification, novel affinity reagent creation, assay formulation, and clinical validation. Studies in our laboratory to optimize the DCP technology have successfully focused on a model system. However, the capabilities of DCP for CVD biomarker discovery in patient plasma samples have yet to be demonstrated with an optimization study. Such a study is proposed here, in which doped CVD biomarkers (representing a model CVD patient sample) will be tested for detectability at clinical sample level concentrations. The proposed study will use five specific CVD protein antigens, for which there are commercially available immunoassays. The success of this Phase I study will lead to a Phase II program for testing multiple CVD patient [plasma] samples for novel biomarker discovery, assay generation, and validation. Specific Aim 1: Confirm that three pooled Random Peptide Phage Libraries actually contain binding phage species against each of the five CVD antigens chosen. Specific Aim 2: Show how well Differential Capture Proteomics detects the differences between two human plasma samples, one of which is doped with the mixture of five CVD antigens. Specific Aim 3: Establish the specificity of the affinity capture reagents that are generated as part of the process. PUBLIC HEALTH RELEVANCE: Since Differential Capture Proteomics will be significantly more cost- and time-effective than current approaches, it potentially offers a breakthrough system for the integrated discovery, verification, and validation of biomarkers. The discovery of new biomarkers of cardiovascular disease will be critical for much-needed advances in screening, diagnosis, prognosis, prediction of therapeutic response and therapeutic monitoring for this devastating disease.