BIO-POD: PALLET MICRO-ARRAY FOR RARE CELL ANALYSIS

BIO-POD：用于稀有细胞分析的托盘微阵列

• 批准号: 8362628
• 负责人: TATIANA B KRASIEVA
• 金额: $ 0.51万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362628
• 关键词: Antibodies; Antigens; Biotechnology; Breast Cancer Cell; Cell Line; Cell surface; Cells; Color; Detection; Diagnosis; Elements; Epithelial Cells; Excision; Feasibility Studies; Fine needle aspiration biopsy; Flow Cytometry; Funding; Grant; Individual; Inflammatory Infiltrate; Lasers; Mammary Neoplasms; Methodology; Methods; Microdissection; Molecular Profiling; Myoepithelial cell; Nanotechnology; National Center for Research Resources; Patients; Primary Neoplasm; Principal Investigator; Research; Research Infrastructure; Resources; Sampling; Source; Specimen; Stem cells; Technology; Testing; Time; Treatment Efficacy; Tumor Tissue; United States National Institutes of Health; base; cancer stem cell; cost; design; fluorescence imaging; malignant breast neoplasm; optical imaging; outcome forecast; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have developed a pallet array nanotechnology that permits isolation of individual adherent cells. Combining this nanotechnology with antibody-based detection methods, advanced optical imaging and fine needle aspirate (FNA) sampling of breast tumors, we are piloting a methodology that 1) overcomes limitations to existing technologies, such as laser microdissection, 2) could be available to patients at the time of diagnosis vs. after tumor resection, 3) permits enumeration of various cellular elements present within a tumor potentially yielding important information for prognosis or predictive of therapeutic efficacy, 4) is designed for high throughput automated analyses, and 4) has the potential to assess the molecular profile of individual cellular compartments, which could provide additional information for the design of tailored individualized therapy. The HYPOTHESIS for these studies is that the pallet array nanotechnology will permit the identification, enumeration, and isolation of the following individual cellular elements from primary breast tumors: breast cancer stem cells, endothelial progenitor cells, myoepithelial cells, and inflammatory infiltrate. This hypothesis will be tested by pursuing the following Specific Aims: 1. Establish the detection threshold for identifying rare adherent cells. We will employ mixtures of cells expressing unique combinations of cell surface molecules in varying proportions, confirmed by flow cytometry, applied to the pallet array. As detection of multiple tumor cellular subsets will require multicolor/multi-antigen detection, we will perform multi-color fluorescence imaging to establish the detection threshold for rare cells. Additionally, we will refine the design of the pallet array. 2. Apply this methodology to primary breast cancer cells using Fine Needle Aspirate (FNA) sampling. Pilot feasibility studies will be conducted on breast tumor specimens using FNAs performed immediately after resection, in order to test the ability to generalize the findings from cell lines to primary tumor tissues. These studies will establish isolation, identification, and detection methods for the enumeration of breast cancer stem cells, endothelial progenitor cells, myoepithelial cells, tumor epithelial cells, and inflammatory infiltrates.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 我们已经开发了一种托盘阵列纳米技术，允许分离单个贴壁细胞。 将这种纳米技术与基于抗体的检测方法、先进的光学成像和乳腺肿瘤的细针抽吸（FNA）取样相结合，我们正在试验一种方法，该方法1）克服了现有技术的局限性，如激光显微切割，2）可以在诊断时与肿瘤切除后提供给患者，3）允许对肿瘤内存在的各种细胞成分进行计数，可能产生预后或预测治疗功效的重要信息，4）设计用于高通量自动化分析，和4）具有评估个体细胞区室的分子谱的潜力，这可以为定制的个体化治疗的设计提供额外的信息。 这些研究的假设是托盘阵列纳米技术将允许从原发性乳腺肿瘤中识别、计数和分离以下单个细胞成分：乳腺癌干细胞、内皮祖细胞、肌上皮细胞和炎性浸润。 将通过追求以下具体目标来检验这一假设： 1.建立鉴别罕见贴壁细胞的检测阈值。 我们将采用表达不同比例的细胞表面分子的独特组合的细胞混合物，通过流式细胞术确认，应用于托盘阵列。 由于多个肿瘤细胞亚群的检测将需要荧光/多抗原检测，我们将进行多色荧光成像，以建立罕见细胞的检测阈值。 此外，我们将完善托盘阵列的设计。 2.使用细针抽吸（FNA）取样将此方法应用于原发性乳腺癌细胞。 将使用切除后立即进行的FNA对乳腺肿瘤标本进行初步可行性研究，以测试将结果从细胞系推广到原发性肿瘤组织的能力。 这些研究将建立乳腺癌干细胞、内皮祖细胞、肌上皮细胞、肿瘤上皮细胞和炎性浸润细胞的分离、鉴定和检测方法。