Analysis of scant cancer cells in fine needle aspirates

细针抽吸物中少量癌细胞的分析

• 批准号: 9023623
• 负责人: RALPH WEISSLEDER, MD, PHD
• 金额: $ 43.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-02 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9023623
• 关键词: 1-Phosphatidylinositol 3-Kinase; Address; Antibodies; Biological Markers; Breast Cancer Patient; Cell Count; Cells; Cleaved cell; Clinical; Clinical Research; Clinical Trials; Clinical Trials Design; Complex; Core Biopsy; Cytometry; DNA; DNA analysis; Detection; Devices; Diagnostic; Drug Targeting; Emerging Technologies; Failure; Fine needle aspiration biopsy; Funding; Goals; Harvest; Heterogeneity; Human; Immunohistochemistry; Kinetics; Lesion; Magnetism; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Messenger RNA; Metastatic breast cancer; Methods; Microfluidic Microchips; Microfluidics; Morbidity - disease rate; Optics; Other Genetics; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphoproteins; Preparation; Procedures; Process; Protein Analysis; Proteins; Reaction; Recurrent disease; Reproducibility; Research Infrastructure; Residual Tumors; Resolution; Sampling; Signal Transduction; Source; Specialist; Specificity; Specimen; System; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Tumor Biology; Work; anticancer research; base; cancer cell; clinical practice; cost; cost effective; genetic analysis; innovation; insight; minimally invasive; mutational status; next generation; novel therapeutics; point of care; prospective; protein profiling; public health relevance; response; single cell analysis; single cell proteins; tool; treatment response; tumor; tumor heterogeneity

 DESCRIPTION (provided by applicant): Comprehensive analysis of key cancer proteins and pathway markers in clinical samples remains challenging yet is critical in assessing efficacy of molecularly targeted drugs in clinical trial and in understanding complex tumor biology. Currently, the number of markers being studies is often limited (<10) and requires time- consuming analyses of tissue sections harvested by large core biopsies which carry a not insignificant morbidity. We developed a technology that allows simultaneous analysis of hundreds of proteins in cancer cells harvested from fine needle aspirates (FNA). The method capitalizes on DNA-barcoded antibody sensing where barcodes are photo-cleaved and digitally detected without any amplification steps. In a recent proof-of- concept study (Sci Transl Med 2014;6: 219ra9) this method showed high reproducibility, achieved single cell sensitivity and was able to identify pathway responses to molecularly targeted drugs, even in single cells. Compared to existing technology (immunohistochemistry, cytometry and mass spectrometry) it: i) allows hundreds of markers to be detected simultaneously, ii) works well in single cells or small numbers of cells, iii) does not destroy valuable samples, iv) is fast and inexpensive and v) can be combined with mRNA and DNA analytical techniques. The goal of this R33 application is to further the technology by integrating it with i) on-chip microfluidics for cancer cell enrichmen and single/bulk cell harvesting and ii) combined protein, mRNA and DNA analysis. We will then expand and rigorously test this next generation device for point-of-care analyses of single cells in two clinical studies to broadly demonstrate its broad cancer utility: i) in a clinical diagnosti study to compare cancer cell protein profiles in breast cancer patients and ii) in a drug trial of PI3K inhibition to determine treatment response/failure over time. The proposed integrated profiling method has the potential to transform cancer research and clinical practice. It will allo inexpensive, more extensive and robust profiling of cellular markers in scant and valuable materials from clinical trials.

 描述（由申请人提供）：临床样本中关键癌症蛋白和途径标志物的综合分析仍然具有挑战性，但对于评估临床试验中分子靶向药物的疗效和理解复杂的肿瘤生物学至关重要。目前，正在研究的标记物的数量通常是有限的（<10），并且需要对通过大的核心活检收获的组织切片进行耗时的分析，所述组织切片具有不小的发病率。我们开发了一种技术，可以同时分析从细针抽吸物（FNA）中收获的癌细胞中的数百种蛋白质。该方法利用DNA条形码化抗体感测，其中条形码被光切割并在没有任何扩增步骤的情况下进行数字检测。在最近的概念验证研究（Sci Transl Med 2014; 6：219ra9）中，该方法显示出高再现性，实现了单细胞灵敏度，并且能够鉴定对分子靶向药物的途径响应，即使在单细胞中。与现有技术（免疫组织化学、细胞计量术和质谱法）相比，它：i）允许同时检测数百种标志物，ii）在单个细胞或少量细胞中工作良好，iii）不破坏有价值的样品，iv）快速且便宜，以及v）可以与mRNA和DNA分析技术组合。该R33应用的目标是通过将其与i）用于癌细胞富集和单个/大量细胞收获的芯片上微流体和ii）组合的蛋白质、mRNA和DNA分析相集成来进一步发展该技术。然后，我们将在两项临床研究中扩展并严格测试这种下一代设备，用于单细胞的即时分析，以广泛证明其广泛的癌症效用：i）在临床诊断研究中比较乳腺癌患者的癌细胞蛋白质谱，ii）在PI3K抑制的药物试验中确定随时间的治疗反应/失败。提出的综合分析方法有可能改变癌症研究和临床实践。它将允许廉价，更广泛和强大的分析细胞标记物的稀缺和有价值的材料从临床试验。