Isolation of human tumor cells in dormant cancer

描述(申请人提供)：早期诊断的乳腺癌通常是治愈的，但很大一部分患者会经历转移性复发，尽管治疗方法有所进步，但这种复发是无法治愈的。在某些情况下，复发的时间间隔很长，这表明癌症处于休眠状态，但对转移的生理学和控制临床复发发生和时间的决定因素知之甚少。我们已经证明，在乳腺癌患者中存在循环肿瘤细胞(CTC)，这些患者在乳腺癌切除后7-22年，没有疾病证据(NED)，因此很可能处于肿瘤休眠状态。我们对其中40名患者的初步数据显示，使用非常严格的标准，包括细胞形态、免疫表型和肿瘤细胞，18例患者患有CTC。19名正常女性的血液中没有符合CTC标准的细胞。所有参与程序/口译的人员都失明了。该程序包括免疫磁性纯化，然后使用上述标准对载玻片上的CTCs进行免疫荧光分析。根据初步观察，四氯化碳的半衰期估计只有几个小时。由于13例CTCs患者是乳腺癌术后13-22年的NED患者，而且CTCs的半衰期如此之短，我们推测乳腺癌中的休眠状态可以代表细胞复制和细胞死亡之间的精确平衡。我们计划将我们的研究扩展到大量患者(150例)，包括乳腺癌NED术后8年或更长时间的患者，年龄匹配的正常女性的对照组(50例)，以及50例转移性乳腺癌患者。将通过计数、免疫表型和基因分型对CTCs进行深入研究，以确定来自休眠组的CTCs是否与转移性乳腺癌中的CTCs不同，以及是否正在复发。潜伏期肿瘤和转移性肿瘤组的初始肿瘤及其治疗也将分析差异。将进行探索性实验，以确定导致这种精确平衡的机制。提供休眠状态的具体证据，并能够表征CTCs的动力学、基因和表型以及它们的调控机制，对癌症控制和治疗具有巨大的意义。

DESCRIPTION (provided by applicant): Breast cancer diagnosed at an early stage is usually cured, but a significant percentage of patients will experience a metastatic recurrence that is not curable despite advances in therapy. The very long interval of time to recurrence seen in some cases suggests the phenomenon of cancer dormancy, but very little is known about the physiology of metastasis and the determinants that govern the occurrence and timing of clinical recurrence. We have demonstrated the presence of circulating tumor cells (CTCs) in patients with prior breast cancer who are 7-22 years post-mastectomy and who have no evidence of disease (NED), thus very likely to be in a tumor dormancy state. Our preliminary data from 40 of these patients indicate that 18 have CTCs, using very stringent criteria including cytomorphology, immunophenotyping and aneusomy. Nineteen normal women had no cells in the blood that met the criteria for CTCs. All personnel involved in the procedures /interpretation were blinded. The procedure involved immunomagnetic purification followed by immunofluorescence analysis for CTCs on slides using the above criteria. From preliminary observations, the half-life of CTCs was estimated to be only several hours. Since 13 of the patients with CTCs are 13- 22 years post mastectomy with NED and the half-life of the CTCs is so short, we postulate that dormancy in breast cancer can represent a precise balance between cell replication and cell death. We plan to extend our studies to a large number (150) of patients 8 or more years after removal of a breast carcinoma with NED, a control group (50) of age-matched normal women, and 50 patients with metastatic breast carcinoma. The CTCs will be studied in-depth by counting, immunophenotyping and genotyping to determine if the CTCs from the dormant group differ from CTCs in metastatic breast carcinomas and whether a relapse is developing. The initial tumors and their treatments in the dormant vs. the metastatic tumor group will also be analyzed for differences. Exploratory experiments to determine the mechanisms responsible for this precise balance will be done. Providing concrete evidence of a dormancy state and being able to characterize the kinetics, genotype and phenotype of CTCs and the mechanisms by which they are regulated have enormous implications in cancer control and treatment.