Application of a Novel Nanotechnology for Molecular Profiling of Tumor Cellular E

新型纳米技术在肿瘤细胞 E 分子谱分析中的应用

• 批准号: 7368621
• 负责人: EDWARD L. NELSON
• 金额: $ 18.3万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-20 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7368621
• 关键词: Address; Adjuvant Chemotherapy; Antibodies; Arts; Aspirate substance; Be++ element; Behavior; Beryllium; Biological; Biomedical Engineering; Biopsy; Cancer Patient; Cell Line; Cell surface; Cells; Cellular biology; Clinical; Clinical Oncology; Confocal Microscopy; Cytotoxic agent; Detection; Diagnosis; Diagnostic; Dissection; Elements; Epithelial; Epithelium; Evaluation; Excision; Exhibits; Fine needle aspiration biopsy; Flow Cytometry; Future; Gene Expression; Gene Expression Profile; Genetic; Heterogeneity; Histologic; Histology; Immunofluorescence Immunologic; Immunology; Individual; Inflammatory; Lasers; Malignant - descriptor; Malignant Neoplasms; Mammary Neoplasms; Methodology; Methods; Molecular; Molecular Analysis; Molecular Genetics; Molecular Profiling; Molecular Target; Myoepithelial; Myoepithelial cell; Myoepithelioma; Nanotechnology; Needles; Neoadjuvant Therapy; Neoplasm Metastasis; None or Not Applicable; Operative Surgical Procedures; Optics; Palliative Care; Patients; Phenotype; Polymers; Population; Primary Neoplasm; Recovery; Relative (related person); Resected; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Slide; Specimen; Stem cells; Surface; Technology; Testing; Therapeutic; Time; Toxic effect; Treatment Efficacy; Tumor Biology; Tumor Tissue; angiogenesis; base; cancer stem cell; clinical application; concept; design; desire; innovation; malignant breast neoplasm; neoplastic cell; novel; optical imaging; outcome forecast; progenitor; prospective; response; stem; therapy resistant; tumor

DESCRIPTION (provided by applicant): Our increasing appreciation of 1) tumor genetic and cellular heterogeneity, 2) the recent descriptions of cancer stem, endothelial progenitor, & myoepithelial cells, among others, and 3) disparate responses to treatment even for histologically similar tumors; raise fundamental questions as to the relative contributions of various tumor cellular subsets to the biologic behavior of a tumor. However, technologies permitting the prospective characterization of discrete tumor cellular elements and recovery of selected viable cells from a tumor have yet to be developed. We have developed a novel nanotechnology consisting of an array of microfabricated SU8 polymer elements that permits the isolation and recovery of individual adherent cells. This advanced nanotechnology combined with multicolor immunofluorescence and advanced confocal microscopy enables us to propose application of this technology to the simultaneous identification, recovery, and evaluation of selected molecular profiles from viable primary adherent cell populations representing the various cellular elements within individual tumors. The HYPOTHESIS for these studies is that the pallet array nanotechnology will permit identification, enumeration, and recovery of the following individual cellular tumor elements: cancer stem, endothelial progenitor, myoepithelial, epithelial, and inflammatory cells leading to the molecular characterization of these cellular subsets within individual tumors and will be tested by pursuing the following Specific Aims: AIM #1. Refine the pallet array for maximum cell capture, detection, and recovery of tumor cellular elements. AIM #2 Apply pallet array to Fine Needle Aspirate (FNA) samples of primary breast tumors to identify and isolate individual cells from discrete tumor cellular elements. AIM #3 Establish feasibility of molecular analysis of recovered individual rare cells, e.g. single cell RT-PCR. These studies represent a convergence of biomedical engineering, advanced laser optics, cell biology, immunology, and clinical oncology and will drive future studies to address fundamental biological and clinical questions.

描述（由申请人提供）：我们越来越认识到 1) 肿瘤遗传和细胞异质性，2) 最近对癌症干细胞、内皮祖细胞和肌上皮细胞等的描述，以及 3) 即使对于组织学相似的肿瘤，对治疗的不同反应；提出了关于各种肿瘤细胞亚群对肿瘤生物学行为的相对贡献的基本问题。然而，尚未开发出能够对离散肿瘤细胞成分进行前瞻性表征并从肿瘤中回收选定的活细胞的技术。我们开发了一种新型纳米技术，由一系列微加工的 SU8 聚合物元件组成，可以分离和恢复单个贴壁细胞。这种先进的纳米技术与多色免疫荧光和先进的共聚焦显微镜相结合，使我们能够建议应用该技术来同时识别、恢复和评估来自代表单个肿瘤内各种细胞元素的存活原代贴壁细胞群的选定分子谱。这些研究的假设是托盘阵列纳米技术将允许识别、计数和恢复以下单个细胞肿瘤元件：癌干细胞、内皮祖细胞、肌上皮细胞、上皮细胞和炎症细胞，从而对单个肿瘤内这些细胞亚群进行分子表征，并将通过追求以下特定目标进行测试：AIM#1。优化托盘阵列，以最大程度地捕获、检测和回收肿瘤细胞成分。目标#2 将托盘阵列应用于原发性乳腺肿瘤的细针抽吸 (FNA) 样本，以从离散的肿瘤细胞成分中识别和分离单个细胞。目标 #3 建立对回收的单个稀有细胞进行分子分析的可行性，例如单细胞RT-PCR。这些研究代表了生物医学工程、先进激光光学、细胞生物学、免疫学和临床肿瘤学的融合，并将推动未来的研究解决基本的生物学和临床问题。