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（适用提供）：临床样本中关键癌症蛋白和途径标记的全面分析仍然挑战，但对于评估临床试验中分子靶向药物的有效性至关重要，并且在理解复杂的肿瘤生物学方面。当前，研究标记的数量通常受到限制（<10），需要对大型核心活检收获的组织切片的耗时分析，而大型核心活检的发病率并不微不足道。我们开发了一项技术，可以简单地分析从细针吸入（FNA）收获的癌细胞中数百种蛋白质。该方法在DNA-ardarcoded抗体感测上大写，其中条形码是照片切割并在没有任何扩增步骤的情况下进行了数字检测的。在最近的概念验证研究（SCI Transic Med 2014; 6：219RA9）中，该方法表现出很高的可重现性，达到了单细胞敏感性，并且即使在单个细胞中，也能够识别对分子靶向药物的途径响应。与现有技术（免疫组织化学，细胞术和质谱法）相比：i）i）简单地检测到数百个标记，ii）ii）在单个细胞或少量细胞中效果很好，iii）不破坏有价值的样品，iv），iv），iv）是快速且便宜的，v）可以与mRNA和DNA分析技术结合使用。该R33应用的目的是通过将技术与i）芯片微流体进行整合，以用于癌细胞富集和单/散装细胞收集以及II）组合蛋白，mRNA和DNA分析。然后，我们将在两项临床研究中扩展和严格测试下一代单个细胞的即将到来的单个细胞分析，以广泛证明其广泛的癌症实用性：i）在一项临床诊断研究中，以比较乳腺癌患者的癌细胞蛋白谱和ii）在PI3K抑制药物抑制药物中，以确定治疗反应反应/失败。提出的综合分析方法具有改变癌症研究和临床实践的潜力。从临床试验中，它将对细胞标志物进行廉价，更广泛和更强大的细胞标记分析。