Protein-Chip Diagnostic Assay Detection for Cancer

描述（由申请人提供）：肿瘤产生循环因子，有可能被用作肿瘤存在的诊断标记。这些标记物可能是肿瘤类型特异性、血管生成因子或参与肿瘤免疫反应的细胞因子。仅基于一个标记的诊断会产生太多的假阳性。需要的是通过蛋白质组学分析许多标记物（多达100个标记物）的能力，然后在标记物的存在与肿瘤的存在之间建立准确的相关性，从而消除假阳性。我们建议使用增强免疫分析（IA/MPD）和蛋白芯片（P-chips/MPD）的超敏多光子检测（MPD）来检测浓度范围为0.01 pg/ml至500 pg/ml的肿瘤标志物。这项技术将产生一种更敏感、更具体的诊断方法，可能使肿瘤在发展的早期阶段就被发现。该提案的I期目标是开发血管生成因子，成纤维细胞生长因子-1 （FGF-1）的超敏感IA/MPD检测技术，这些因子已被发现在许多肿瘤类型中表达。然后筛选临床相关的血清样本，检测FGF-1、FGF-2、血管内皮生长因子（VEGF）、肝细胞生长因子（HGF）、il -1、IL-6、IL-10、IL-11、IL-12和tnf - α，以及前列腺血清抗原等肿瘤标志物。目前存在针对这些因素的超灵敏IA/MPD检测方法。临床相关的血清样本将包括癌症治疗前和治疗后的患者以及健康对照者。治疗后将对癌症患者进行一段时间的监测，看看标志物水平随时间的变化是否可以预测肿瘤复发。结果将被检查，以确定是否一个特定模式的标志物水平与肿瘤的存在相关。第二阶段的目标是开发MPD增强型p芯片。P-chips/MPD将适用于大量标记物，包括PSA、CEA、CA-125、甲胎蛋白、细胞因子和与肿瘤存在相关的血管生成因子。还将研究肿瘤中发现的其他因子，但目前相对较低灵敏度的诊断方法无法检测到这些因子。扩大临床相关样本检测。一旦同时测量这些生物标志物的重要性得到确认，我们将开发专用的超敏感p芯片，用于定量多达100个目标标记蛋白，检测范围为0.01至500 pg/ml。因此，低成本的癌症诊断和治疗监测检测将成为可能，并且可能提出早期肿瘤形成的潜在机制，因为一些标记物可能仅在早期阶段以较低水平表达。

DESCRIPTION (provided by applicant): Tumors produce circulating factors that have the potential to be used as diagnostic markers for the presence of the tumor. These markers may be tumor type specific, angiogenic factors, or cytokines involved in the immune response to the tumor. The diagnostic based on just one marker yield too many false positives. What is needed is the ability to assay many markers (up to 100 markers) by proteomics and then develop an accurate correlation between the presence of the markers and the presence of the tumor, thus eliminating false positives. We propose use of the supersensitive MultiPhoton Detection (MPD) enhanced to immunoassays (IA/MPD) and protein-chips (P-chips/MPD), to detect tumor markers in concentration ranges from 0.01 pg/ml level to 500 pg/ml. The technology will result in a more sensitive and specific diagnostic method, which may enable the detection of the tumors in an earlier stage of development. The objective of Phase I of this proposal is to develop the supersensitive IA/MPD assay techniques for the angiogenesis factors, Fibroblast Growth Factor-1 (FGF-1) that have been found to be expressed in many tumor types. Then clinically relevant serum samples will be screened for FGF-1, along with FGF-2, Vascular Endothelial Growth Factor (VEGF), Hepatocyte Growth Factor (HGF), IL-lbeta, IL-6, IL-10, IL-11, IL-12, and TNF-alpha, and tumor markers like Prostate Serum Antigen. Ultrasensitive IA/MPD assays currently exist for many of these factors. The clinically relevant serum samples will include pre- and post- cancer treatment patients, and healthy control subjects. Post treatment cancer patients will be monitored over time to see if marker level variations over time can predict tumor recurrence. Results will be examined to determine if a particular pattern of marker levels correlates with the presence of a tumor. The goal of Phase II is the development of the MPD enhanced P-chip. The P-chips/MPD will be adapted to a large set of markers including PSA, CEA, CA-125, and alpha-fetoprotein, cytokines and angiogenesis factors that correlate with the presence of a tumor. Additional factors, which are found expressed by tumors, but can not be detected by current relatively low sensitivity diagnostics will also be studied. Testing on clinically relevant samples will be expanded. Once the importance of concurrent measurement of several of these biomarkers are confirmed, we will develop dedicated supersensitive P-chips for quantifying up to 100 targeted marker proteins with detection ranges from 0.01 to 500 pg/ml. Thus, low cost cancer diagnostics and therapy monitoring assay will become available and a potential mechanism(s) may be suggested for earlier stage tumor formation since some of the markers may only be expressed at lower levels at the early stages.