Refinement and Discovery of Nuclear Matrix Protein Markers for Colorectal Cancer

描述（由申请人提供）：本申请的重点是继续开发一组独特的结肠癌相关核基质蛋白（NMPs），该蛋白是在我们之前的edrn资助的生物标志物开发实验室资助下发现和开发的。nmp为结直肠癌的早期检测提供了潜在的基于血清的生物标志物。这项工作已经取得了实质性进展，从测试非盲法单一机构便利样本一直到测试盲法，EDRN赞助的多机构参考集。标志物在不同的实验室环境、癌症亚型和患者群体特征中进行了测试。这些数据为临床人群的应用提供了令人信服的潜力。目前CCSA-2和CCSA-4两种标记物正准备进行i -ll期验证研究。我们建议在检测开发专家的帮助下，进一步完善这些检测方法，使其能够用于最终的临床测试。此外，我们将进一步研究与肝转移相关的NMP标志物。两个主要目标1。优化和完善CCSA-2和CCSA-4检测方法，并在一系列小规模实验中对其进行测试，以便在EDRN机制中对其进行最终评估。具体而言，我们的目标是(a)开发准确和精确测量CCSA-2和CCSA-4的稳健分析；(b)在不同的临床环境和条件下测试测定法，以了解和预测在大规模测试测定法时可能遇到的错误来源和实施中的潜在问题。第一个目标是提供两种检测方法，通过充分的初步测试进行验证，以最大限度地提高EDRN项目成功的潜力。2. 建立肝转移的NMP标志物。我们已经在肝转移组织标本中发现了既不存在于邻近未受累肝脏中也不存在于正常供者肝脏中的蛋白质。这些蛋白是晚期疾病标志物检测开发的主要候选蛋白。我们建议(a)分离、表征和开发用于血清检测肝转移蛋白的抗体，(b)在适当的临床标本中评估这些抗体，以便最终交付EDRN验证。

DESCRIPTION (provided by applicant): The focus of this proposal is to continue development of a unique panel of colon cancer associated nuclear matrix proteins (NMPs), discovered and developed during our previous EDRN-sponsored, Biomarker Development Laboratory funding. NMPs offer the potential serum-based biomarkers for the early detection of colorectal cancer. This work has progressed substantially from testing of un-blinded single institution convenience samples all the way to testing of blinded, EDRN sponsored multi-institution reference sets. Markers were tested in different laboratory settings, cancer subtypes, and patient population characteristics. These data provide compelling potential for application in clinical populations. Currently two markers, CCSA-2 and CCSA-4 are poised for Phase ll-lll validation studies. We propose, with the help of established experts in assay development, to further refine those assays to position them for definitive clinical testing. In addition, we will advance NMP markers related to liver metastasis. The two major objectives 1. To OPTIMIZE AND REFINE the CCSA-2 and CCSA-4 assays and test them in a series of small scale experiments to position them for definitive evaluation within the EDRN mechanism. In specific, we aim to (a) develop robust assays for the accurate and precise measurement of CCSA-2 and CCSA-4; (b) test the assays under a variety of clinical circumstances and conditions to understand, and anticipate sources of error and potential problems in implementation that might be encountered when testing the assays on a larger scale. The goal of this first objective is to deliver two assays to validation with ample preliminary testing to maximize the potential for success in the EDRN program. 2. To develop NMP markers for liver metastasis. We have identified proteins in liver metastasis tissue specimens that are neither present in adjacent uninvolved liver nor in liver from normal donors. These proteins are prime candidates for assay development for markers of advanced disease. We propose (a) to isolate, characterize and develop antibodies for serum detection of liver metastasis proteins, (b) to evaluate these antibodies in appropriate clinical specimens for eventual delivery for EDRN validation.