EARLY DETECTION OF CANCER BY AFFINITY MASS SPECTROMETRY

前列腺癌和乳腺癌是美国男性和女性最常见的癌症。 目前的诊断测试不够敏感或特异，无法在肿瘤细胞数量较少且治愈或控制机会最大的早期阶段检测癌症。 此外，没有单一的生物标志物可能改善这些癌症的早期检测。 我们需要的是更好的工具来确定肿瘤的特征，这是一个决定早期检测，治疗选择和治疗效果并预测预后的特征。 本研究的目的是确定发出癌症存在信号的独特蛋白质指纹，长期目标是开发临床检测方法，以改善前列腺癌和乳腺癌的早期检测。 新的蛋白质组学技术，指定的表面增强激光电离/解吸（SELDI）蛋白质芯片质谱，将用于实现这一目标。第一个目标是致力于发现显微切割癌细胞中的癌前和癌性特征蛋白。将对来自所有发育阶段（即正常、肿瘤前、增生、原发性和转移性病变）的纯细胞群进行SELDI分析，以鉴定独特的癌症蛋白指纹。 第二个目标将确定这些独特的癌症相关蛋白中的哪些在体液中分泌/脱落。 通过比较单个样本的蛋白质谱与癌症蛋白质谱的复合参考图谱，将在AIM 3中确定SELDI蛋白质谱作为诊断/预后检测的潜在临床用途。与癌前病变和癌症病变相关的蛋白质将被纯化和测序，并在目标4中制备重组蛋白（用于新蛋白质）。独特蛋白质的抗体将用于目标5，以开发SELDI免疫测定法，用于定量体液中的癌症相关蛋白质。 由于相同的平台用于发现、鉴定和诊断测定开发，因此SELDI技术相对于其他现有的蛋白质组学技术具有明显的优势。 预计将开发SELDI蛋白质谱分析或SELDI免疫测定，这将改善前列腺癌和乳腺癌的早期检测。

Prostate and breast cancer are the most common cancers in men and women in the United States. Current diagnostic tests are not sensitive or specific enough to detect cancers in their early stages when the number of tumor cells is small and the chance for cure or control is greatest. Furthermore, no single biomarker is likely to improve early detection of these cancers. What are needed are better tools to determine the traits of the tumor, a profile that dictates early detection, treatment choice, and efficacy of treatment and predicts prognosis. The objective of this study is to identify the unique protein fingerprints that signal the presence of cancer with the long term goal of developing clinical assays to improve the early detection of prostate and breast cancers. The novel proteomic technology, designated Surface Enhanced Laser Ionization/Desorption (SELDI) ProteinChip mass spectrometry, will be used to meet this goal. The first Aim is devoted to discovery of the pre-cancerous and cancerous signature proteins in microdissected cancer cells. Pure population of cells from all developmental stages, i.e. normal, pre-neoplastic, hyperplatic, primary and metastatic lesions, will be SELDI analyzed to identify the unique cancer protein fingerprints. The second aim will identify which of these unique cancer-associated proteins are secreted/shed in body fluids. By comparing protein profiles of individual samples with composite reference maps of the cancer protein profiles, the potential clinical use of SELDI protein profiling as a diagnostic/prognostic test will be determined in AIM 3. Proteins that correlate with pre-cancer and cancer lesions will be purified and sequenced, and recombinant proteins made (for novel proteins) in Aim 4. Antibodies to the unique proteins will be used in Aim 5 to develop SELDI immunoassays for quantitation of the cancer-associated proteins in body fluids. Since the same platform is used for discovery, identification, and diagnostic assay development, the SELDI technology has a distinct advantage over other existing proteomic technologies. It is expected that either a SELDI protein profiling or SELDI immunoassay will be developed that will improve the early detection of prostate and breast cancer.