Cancer detection technology

描述（由申请人提供）：I期：越来越清楚的是，实体癌的转移潜力与原发肿瘤细胞迁移的性质密切相关。这些迁移细胞进入循环系统，因此可以很容易地恢复使用一个简单的和微创的程序。然而，这些细胞的分子分析在技术上仍然具有挑战性，因为迁移的转移性肿瘤细胞相对于大量的正常血细胞来说数量很少，并且因为在外周血中发现的许多肿瘤细胞在任何给定时刻都是死亡或死亡的。目前检测外周血肿瘤细胞的技术包括结合细胞大小和密度或阳性和阴性选择免疫富集步骤。然而，这些方案只恢复了潜在可用肿瘤细胞的一小部分，并且所得到的细胞群仅仅是丰富的，即，它们部分由非肿瘤上皮细胞以及死亡和垂死的肿瘤细胞组成。在I期，我们建议通过优化一种新的细胞分离技术，开发一种临床有用的方法来鉴定和分离循环胃肠道（GI）肿瘤细胞。我们的实验将使用细胞粘附基质（CAM）来构建一个模拟肿瘤血管内微环境的血液过滤单元，从而实现循环肿瘤细胞的功能亲和纯化。我们预计CAM富集将能够完成一个步骤，从晚期胃肠道癌症患者的外周血中富集100万倍的活的人类肿瘤细胞。流式细胞术、免疫细胞化学和使用细胞谱系标记的实时RT-PCR将用于验证cam回收的细胞。拟议的I期研究将测量CAM技术用于血液中癌细胞的II期病理研究和分子分析的可行性和效率。最终，转移性肿瘤细胞富集和特异性胃肠道肿瘤分子靶点的结合将创造一种技术，临床医生可以用它来建立分期，预测预后，并确定多种类型上皮肿瘤的治疗反应性。

DESCRIPTION (provided by applicant): Phase I: It has become increasingly clear that the metastatic potential of solid cancer best correlates with the nature of the cells emigrating from the primary tumors. These emigrating cells enter the circulatory system and thus can readily be recovered using a simple and minimally invasive procedure. However, molecular analysis of these cells remains technically challenging because the emigrating metastatic tumor cells are few in number relative to the enormous numbers of normal blood cells, and because many of the tumor cells found in peripheral blood are dead or dying at any given moment. Current technology to detect tumor cells in peripheral blood involves a combination of cell size and density or positive- and negative-selecting immuno-enrichment steps. However, these protocols recover only a small fraction of the potentially available tumor cells, and the resulting cell populations are merely enriched, i.e., they are composed partially of non-tumor epithelial cells as well as dead and dying tumor cells. In Phase I, we propose to develop a clinically useful assay for the identification and isolation of circulating gastro-intestinal (GI) tumor cells by optimizing a novel cell separation technology. Our assay will use a cell adhesion matrix (CAM) to build a blood filtration unit that mimics the tumor intravascular microenvironment and thus will enable function-affinity purification of the circulating tumor cells. We anticipate the CAM enrichment will be able to accomplish a single step, one million-fold enrichment of viable human tumor cells from the peripheral blood of patients with late stage GI cancers. Flow cytometry, immunocytochemistry and real-time RT-PCR using cell lineage markers will be used to validate the CAM-recovered cells. The proposed Phase I study will measure the feasibility and efficiency of the CAM technology for use in the Phase II pathological study and molecular analysis of cancer cells in the blood. Ultimately, the combination of metastatic tumor cell enrichment and molecular targets acquired for specific GI cancers will create a technology that clinicians can use to establish staging, predict prognosis, and determine therapeutic responsiveness for multiple types of epithelial tumors.