MOLECULAR CHARACTERIZATION OF METASTATIC BREAST CELLS

转移性乳腺细胞的分子特征

• 批准号: 3204922
• 负责人: JAMES F. LEARY
• 金额: $ 19.87万
• 依托单位: UNIVERSITY OF ROCHESTER
• 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 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3204922
• 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.

目前，女性转移性乳腺癌的治疗方法有多种 治疗方式。自体骨髓移植 化疗虽然可能有效，但也存在这个问题 很难识别并移除所有的转移细胞 骨髓(BM)。外周血(PB)中的转移细胞也 难以辨认和移除。这个问题的一个重要方面是 鉴定和鉴定前后存在的转移细胞 清洁技术。这些转移细胞是如此罕见，以至于它们 大多数情况下都是无法察觉的。罕见的转移细胞有什么特性 与原发灶有显著差异吗？这些能不能 通过其他技术直接利用差异来帮助消除这些差异 外周血和/或骨髓？如果成功，则ABMT何时 联合化疗将是一个重要而有效的全身 心理治疗。 使用新的特殊高速(大于100,000个信元/秒)流 超稀有细胞(低于10-6稀有细胞频率)的细胞学检测 现在应该可以检测和分离罕见的转移性乳房 骨髓和外周血中的细胞频率低于每百万个转移细胞中的一个 正常细胞。特殊的高速分拣技术将用于 后续图像分析和后续分子的聚合酶链式反应分析 这些罕见转移瘤中单个或少数的特征 细胞。将尝试将它们从BM中清除。是这样的 这些细胞的免疫细胞化学和分子特征可能 更好地了解转移的潜在原因 乳腺癌，并可能建议新的治疗方法。外周血中罕见的转移细胞 并在多重正面选择的基础上确定BM 标志物包括(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、细胞角蛋白AE1/AE3和多个阴性选择标记。 到目前为止，还没有一个单一的标记被证明是唯一的，无论是阳性还是 负选择，但很可能存在这样的组合 阳性和阴性标记物足以进行单个细胞 多管排序或不纯排序，后续进一步处理 使用其他标记和克隆进行图像分析以实现生长或克隆 用于对具有一个或多个 分子标记。两种分子表征的比较 转移细胞与原发灶中的细胞将使用 各种基因的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-1。 2和mts1.