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从原发性肿瘤迁移的细胞的性质最相关。这些迁移的细胞进入循环系统，因此可以使用简单且微创的程序容易地回收。然而，这些细胞的分子分析在技术上仍然具有挑战性，因为相对于大量的正常血细胞，迁移的转移性肿瘤细胞的数量很少，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的或正在死亡的。目前检测外周血中肿瘤细胞的技术涉及细胞大小和密度或阳性和阴性选择免疫富集步骤的组合。然而，这些方案仅回收一小部分潜在可用的肿瘤细胞，并且所得细胞群仅被富集，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。 在第一阶段，我们建议开发一种临床上有用的检测方法，通过优化一种新的细胞分离技术来识别和分离循环胃肠道（GI）肿瘤细胞。我们的试验将使用细胞粘附基质（CAM）来构建血液过滤单元，该单元模拟肿瘤血管内微环境，从而能够对循环肿瘤细胞进行功能亲和纯化。我们预计CAM富集将能够完成一个步骤，从晚期GI癌症患者的外周血中富集100万倍的活人类肿瘤细胞。将使用流式细胞术、免疫细胞化学和使用细胞谱系标志物的实时RT-PCR来验证CAM回收的细胞。拟议的第一阶段研究将衡量CAM技术用于第二阶段病理学研究和血液中癌细胞分子分析的可行性和效率。最终，转移性肿瘤细胞富集和针对特定GI癌症获得的分子靶点的组合将创造一种技术，临床医生可以使用该技术来建立分期，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.