Analysis of scant cancer cells in fine needle aspirates

细针抽吸物中少量癌细胞的分析

基本信息

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
项目类别：
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

项目摘要

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）：对临床样本中关键癌症蛋白和通路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功效和了解复杂的肿瘤生物学至关重要。通常有限（<10），并且需要对通过大核心活检采集的组织切片进行耗时的分析，这具有不显着的发病率。我们开发了一种技术，可以同时分析从细针抽吸物中采集的癌细胞中的数百种蛋白质。 (FNA)。该方法利用 DNA 条形码抗体传感，无需任何扩增步骤即可对条形码进行光切割和数字检测（Sci Transl Med 2014；6：219ra9）。与现有技术（免疫组织化学、细胞计数和质量）相比，即使在单细胞中，也能实现单细胞敏感性并能够识别分子靶向药物的通路反应。光谱法）它：i）允许同时检测数百个标记，ii）在单细胞或少量细胞中效果良好，iii）不会破坏有价值的样品，iv）快速且便宜，v）可以与 mRNA 结合该 R33 应用的目标是通过将其与 i) 用于癌细胞富集和单/大细胞收获的片上微流体以及 ii) 组合蛋白质、mRNA 和 DNA 分析相结合来进一步发展该技术。扩大和在两项临床研究中严格测试这种用于单细胞即时护理分析的下一代设备，以广泛证明其广泛的癌症用途：i) 在临床诊断研究中比较乳腺癌患者的癌细胞蛋白谱，ii) 在PI3K 抑制药物试验以确定随着时间的推移治疗反应/失败。所提出的综合分析方法有可能改变癌症研究和临床实践，它将允许对来自临床的稀缺和有价值的材料进行廉价、更广泛和稳健的细胞标记物分析。试验。