Gene Expression of Viable Ovarian Cancer Cells

描述(由申请人提供)： 基因表达的分子图谱是一种强大的工具，可用于对人类肿瘤进行分期，并根据预测的预后和治疗反应对其进行分类。这项技术已被最有效地应用于白血病和淋巴瘤，对于这些疾病来说，获得高度浓缩的肿瘤细胞池相对容易。类似的研究实体肿瘤的方法不太成功，因为实体肿瘤组织样本包含许多非致瘤类型的细胞，因为在其中发现的许多肿瘤细胞在任何给定的时刻都是死亡或死亡的，而且因为低丰度的转移细胞正在向外迁移。目前在手术时从腹水中浓缩转移性卵巢癌细胞的技术包括结合使用离心法和正负选择免疫亲和力步骤。这一繁琐的方案产生了不到50%纯度和活性的转移细胞池，这些转移细胞池被非肿瘤白细胞和上皮细胞以及死亡和垂死的肿瘤细胞污染。在这项STTRI期应用中，我们建议优化和验证一种新的、快速的细胞分离技术，用于从卵巢癌患者的腹水中分离出有活力的转移肿瘤细胞。该方法有望产生99%以上纯度的活肿瘤细胞。同时，我们还将提纯肿瘤相关的白细胞，以进行表达谱比较。这将衡量浓缩程序的效率，并对拟议的分析方法进行原则证明。在第二阶段，第一阶段优化的集成技术将转化为卵巢和子宫癌的广泛基因表达研究：我们将进行微阵列分析，以确定在转移性卵巢和子宫肿瘤细胞中从血液和腹水中上调的基因，并与非致癌卵巢和子宫上皮细胞以及原代(原位)卵巢和子宫肿瘤细胞进行比较。最终，特定的转移性肿瘤细胞浓缩和正确靶向的分子图谱的结合将创造出一种能够对多种类型的上皮性肿瘤进行分期和预测预后以及治疗反应的技术。

DESCRIPTION (provided by applicant): Molecular profiling of gene expression is a powerful tool that can be used to stage human tumors and categorize them with respect to projected prognosis and therapeutic responsiveness. This technology has been applied most effectively to leukemias and lymphomas for which it is relatively easy to obtain highly enriched pools of tumor cells. Similar approaches to study solid tumors have been less successful because solid tumor tissue samples contain many non-tumorigenic types of cells, because many of the tumor cells found in them are dead or dying at any given moment, and because of the low abundance emigrating metastatic cells. Current technology to enrich for metastatic ovarian cancer cells from ascites at the time of surgery involves using a combination of centrifugation and positive- and negative-selecting immuno-affinity steps. This cumbersome protocol yields pools of metastatic cells less than 50% pure and viable that are contaminated by non-tumor leukocyte and epithelial cells and dead and dying tumor cells. In this STTR Phase I application, we propose to optimize and validate a novel and rapid cell separation technology for enrichment of viable metastatic tumor cells from the ascites of patients with ovarian cancer. The approach is anticipated to yield a collection of viable tumor cells greater than 99% pure. In parallel, we will also purify tumor-associated leukocytes to enable expression profiling comparisons. This will provide a measure of the efficiency of the enrichment procedure and a proof-of-principle demonstration of the proposed profiling methods. In Phase II, the integrated technologies optimized in this first phase will be translated into broad gene expression studies on ovarian and uterine cancer: We will perform microarray analysis to identify genes up-regulated in metastatic ovarian and uterine tumor cells from blood and ascites in comparison to non-tumorigenic ovarian and uterine epithelial cells and primary (in situ) ovarian and uterine tumor cells. Ultimately, the combination of specific metastatic tumor cell enrichment and correctly targeted molecular profiling will create a technology capable of staging and predicting prognosis and therapeutic responsiveness for multiple types of epithelial tumors.