Prognostic Marker Analysis of Disseminated Cancer Cells

播散性癌细胞的预后标志物分析

• 批准号: 6968924
• 负责人: Rebecca L. Aft
• 金额: $ 13.16万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-21 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6968924
• 关键词: biomarker; bone marrow; breast neoplasms; cancer risk; cell population study; clinical research; female; human tissue; immunomagnetic separation; metastasis; neoplasm /cancer chemotherapy; neoplastic cell; patient oriented research; prognosis

DESCRIPTION (provided by applicant): Thirty percent of all women newly diagnosed with breast cancer and no clinical evidence of metastatic disease will have disseminated tumor cells (DTC) detectable in their bone marrow at the time of diagnosis. The presence of DTC is associated with an increased risk of metastatic disease development as well as cancer death. Chemotherapy can eliminate DTC in some breast cancer patients. However, those women with persistent DTC after chemotherapy are five times more likely to die from their breast cancer. This data suggests that chemotherapy-resistant DTC are likely to be precursor cells capable of forming metastatic foci. Biologically, DTC are a heterogeneous population of cells and clinical studies suggest that only a subset have the ability to form metastases. Identification of specific DTC that correlate with poor clinical outcome would result in a new prognostic marker and identify women at high risk for developing metastatic disease and breast cancer death. We propose that persistent disseminated tumor cells present after chemotherapy represent a unique subpopulation of all DTC, are predictors of a poor response to chemotherapy, and correlate with poor clinical outcome. We hypothesize that chemotherapy-resistant DTC can be identified by their expression of a unique constellation of tumor marker proteins which may be similar to those expressed by breast cancer stem cells. In this proposal, our specific aims are: 1) characterize tumor markers expressed by DTC which are present after chemotherapy, 2) compare the expression of these markers to that on DTC detected prior to chemotherapy, 3) correlate expression of the defined tumor markers on DTC with clinical outcome of breast cancer patients to identify those markers that are predictive of disease recurrence, 4) Utilize biomarkers identified in Specific Aims 1 and 2 to isolate purified DTC for further molecular analysis. Specimens to be analyzed are from the Siteman Cancer Center Tissue Procurement Center where processed and cryopreserved bone marrow cells and cytological slides have been prepared from women with locally advanced breast cancer before and after treatment with neoadjuvant chemotherapy. Tumor marker expression by DTC will be determined by double immunocytochemistry using anti-cytokeratin antibodies and a panel of antibodies directed against cell surface proteins known to be associated with metastatic potential, chemotherapy resistance, and breast cancer stem cell markers. Information on biomarker expression from Specific Aims 1 and 2 will be utilized for immunomagnetic isolation of DTC and further molecular analysis of these cells. We have successfully used both double immunostaining and immunomagnetic separation to detect breast cancer cells in model systems of bone marrow containing DTC. The results of the proposed experiments will identify a unique constellation of tumor makers expressed by DTC persistent after chemotherapy and associated with the metastatic potential of the cells. Identification of biologically important DTC will allow us to stratify women newly diagnosed with breast cancer into high and low risk groups for the development of metastatic disease. Furthermore, defining tumor markers expressed by DTC and performing molecular analysis of these cells will lead to new insights into the biology of the disseminated tumor cells and to the development of new targeted therapies.

描述（由申请人提供）：在所有新诊断为乳腺癌且无转移性疾病临床证据的女性中，有30%在诊断时骨髓中可检测到播散性肿瘤细胞（DTC）。DTC的存在与转移性疾病发展以及癌症死亡的风险增加有关。化疗可以消除某些乳腺癌患者的DTC。然而，那些化疗后持续DTC的女性死于乳腺癌的可能性是其他女性的五倍。这些数据表明，化疗耐药DTC可能是能够形成转移灶的前体细胞。在生物学上，DTC是一种异质性细胞群，临床研究表明，只有一个子集具有形成转移的能力。识别与不良临床结果相关的特定DTC将导致新的预后标志物，并识别发展转移性疾病和乳腺癌死亡的高风险女性。我们认为化疗后持续播散的肿瘤细胞是所有DTC的一个独特亚群，是化疗反应不良的预测因子，并与不良的临床结局相关。我们假设，化疗耐药DTC可以通过其表达的一个独特的星座的肿瘤标志物蛋白，这可能是类似的乳腺癌干细胞表达。在这项建议中，我们的具体目标是：1）表征化疗后存在的由DTC表达的肿瘤标志物，2）将这些标志物的表达与化疗前检测到的DTC上的表达进行比较，3）将DTC上定义的肿瘤标志物的表达与乳腺癌患者的临床结果相关联，以鉴定预测疾病复发的那些标志物，4）利用特定目的1和2中鉴定的生物标志物分离纯化的DTC用于进一步的分子分析。 待分析的样本来自Siteman癌症中心组织采购中心，在该中心，已从新辅助化疗治疗前后的局部晚期乳腺癌女性患者制备了经处理和冷冻保存的骨髓细胞和细胞学切片。将使用抗细胞角蛋白抗体和一组针对已知与转移潜能、化疗耐药性和乳腺癌干细胞标志物相关的细胞表面蛋白的抗体，通过双重免疫细胞化学测定DTC的肿瘤标志物表达。特异性目的1和2的生物标志物表达信息将用于DTC的免疫磁性分离和这些细胞的进一步分子分析。我们已经成功地使用双重免疫染色和免疫磁性分离检测乳腺癌细胞的模型系统的骨髓含有DTC。 所提出的实验的结果将确定一个独特的星座的肿瘤标记物表达的DTC持续化疗后，并与细胞的转移潜力。识别生物学上重要的DTC将使我们能够将新诊断为乳腺癌的妇女分为高风险组和低风险组，以发展转移性疾病。此外，定义DTC表达的肿瘤标志物并对这些细胞进行分子分析将导致对播散性肿瘤细胞生物学的新见解和新靶向治疗的发展。