MOLECULAR CHARACTERIZATION OF METASTATIC BREAST CELLS

转移性乳腺细胞的分子特征

• 批准号: 2102273
• 负责人: JAMES F. LEARY
• 金额: $ 21.49万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-15 至 1997-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2102273
• 关键词: biomarker; bone marrow; bone marrow purging; breast neoplasms; female; flow cytometry; human subject; immunocytochemistry; in situ hybridization; metastasis; molecular oncology; neural information processing; polymerase chain reaction

Metastatic breast cancer in women is presently treated with a variety of modalities of therapy. Autologous bone marrow transplantation (ABMT) prior to chemotherapy, while potentially effective, suffers from the problem that it is difficult to identify and remove all metastatic cells from the bone marrow (BM). Metastatic cells in peripheral blood (PB) are also difficult to identify and remove. An important aspect of this problem is to identify and characterize the metastatic cells present before and after cleansing techniques. These metastatic cells are so rare that they are undetectable by most means. What properties of the rare metastatic cells significantly differ from those of the primary lesion? Can these differences be exploited to help remove them by other techniques directly from peripheral blood and/or bone marrow? If successful, ABMT when combined with chemotherapy would be an important and effective systemic therapy. Using new special high-speed (greater than 100,000 cells/sec) flow cytometric detection of ultra-rare (less than 10-6 rare cell frequencies) it should now be possible to detect and isolate rare metastatic breast cells in BM and in PB at frequencies below one metastatic cell per million normal cells. Special high-speed sorting techniques will be used with subsequent image analysis and PCR analysis for subsequent molecular characterization of single or small numbers of these rare metastatic cells. Attempts will be made to purge them from BM. Such immunocytochemical and molecular characterizations of these cells could provide improved understanding of the underlying causes of metastatic breast cancer and may suggest new therapies. Rare metastatic cells in PB and BM will be identified on the basis of multiple positive selection markers including (but not limited to) epithelial and breast cancer cell surface markers PHM-5, 2G3, 9C6, 741F8, and/or intracellular markers GCDFP-15 and cytokeratins AE1/AE3 and multiple negative selection markers. No single marker has thus far proved unique either for positive or negative selection, but there may well exist a combination of such positive and negative markers sufficient to perform either a single-cell multi-tube sort or an impure sort with subsequent further processing by image analysis with additional markers and cloning either for growth or for "cookie-cutter" re-sorting for PCR characterization with one or more molecular markers. Comparisons of molecular characterizations of metastatic cells with those in the primary lesion will be performed using PCR sequences for a variety of genes frequently amplified or over- expressed in metastatic breast cancer cells including c-erbB2, c-myc, int- 2 and mts1.

女性转移性乳腺癌目前采用多种治疗方法 治疗方式。自体骨髓移植（ABMT）之前 化疗虽然可能有效，但也存在问题 很难识别并去除所有转移细胞 骨髓（BM）。外周血（PB）中的转移细胞也 难以识别和去除。这个问题的一个重要方面是 识别和表征前后存在的转移细胞 清洁技术。这些转移细胞非常罕见，因此 大多数方法都无法检测到。罕见转移细胞有什么特性 与原发病灶有显着差异吗？这些可以吗 利用差异来帮助通过其他技术直接消除它们 来自外周血和/或骨髓？如果成功，ABMT 时 联合化疗将是一种重要且有效的系统治疗 治疗。 采用新型特殊高速（大于100,000个细胞/秒）流 超稀有细胞的细胞计数检测（稀有细胞频率少于 10-6） 现在应该可以检测和分离罕见的转移性乳房 BM 和 PB 中的细胞频率低于百万分之一转移细胞 正常细胞。将使用特殊的高速分拣技术 后续分子影像分析和PCR分析 单个或少量这些罕见转移的特征 细胞。我们将尝试将它们从 BM 中清除。这样的 这些细胞的免疫细胞化学和分子特征可以 更好地了解转移的根本原因 乳腺癌，可能会建议新的疗法。 PB中罕见的转移细胞 并在多重正选的基础上确定BM 标记物包括（但不限于）上皮细胞和乳腺癌细胞 表面标记 PHM-5、2G3、9C6、741F8 和/或细胞内标记 GCDFP-15 和细胞角蛋白 AE1/AE3 以及多个负选择标记。 迄今为止，还没有任何一个标记被证明是唯一的，无论是阳性还是阴性。 负选择，但很可能存在这种组合 阳性和阴性标记足以执行单细胞 多管排序或不纯排序并随后进一步处理 使用附加标记进行图像分析并进行克隆以进行生长或 用于“千篇一律”的 PCR 表征重排序，使用一种或多种 分子标记。分子特征比较 转移细胞与原发灶中的细胞将使用 多种基因的 PCR 序列经常被扩增或过度扩增 在转移性乳腺癌细胞中表达，包括 c-erbB2、c-myc、int- 2 和 mts1。