Nanotechnologies for Comprehensive Single Cancer Cell Analysis

项目概述：针对表皮生长因子受体的癌症治疗 EGFR在肺癌中表现出巨大的潜力;然而，这些药物仅在一个子集中有效。 病人。此外，最初具有响应性的肿瘤随着时间的推移经常获得抗性。虽然它 可直接测量分子（DNA、RNA、蛋白质）和生物物理（质量、密度、电荷） 肿瘤的特点，在散装，最近的研究表明，广泛的细胞到细胞 变异性以及描述变异性在估计患者预后中的重要性^。我们假设循环细胞的分子和生物物理特征可以区分对治疗有反应的细胞和耐药细胞。 当分析从循环或从其他体液（例如，胸腔积液、腹水），通常只有少量细胞可用。为了评估这有限数量的细胞的细胞间异质性，我建议开发和应用定量的、全面的单细胞分析装置来评估DNA。 基因组（例如，单核苷酸多态性、融合、缺失）、RNA表达、蛋白质丰度（细胞表面、细胞内和分泌组丰度）和单细胞的生物物理特性，用于预测患者对EGFR靶向治疗的可能反应和监测患者对EGFR靶向治疗的耐药性的获得的双重目的（图N3.3.1）。我们为综合分析平台的开发、测试和应用提出了两个具体目标（表N3.3.1）。

Project Overview: Cancer therapeutics targeted against the Epidermal Growth Factor Receptor (EGFR) have demonstrated great potential in lung cancer; however, these agents are effective in only a subset of patients. Furthermore, tumors tliat are initially responsive frequently acquire resistance over time. Though it is straightforward to measure molecular (DNA, RNA, protein) and biophysical (mass, density, charge) characteristics of tumors in bulk, recent studies have shown wide cell-tocell variability and the importance of characterizing that variability in estimating patient outcome^^^. We hypothesize that molecular and biophysical characterizations of circulating cells can discriminate cells that are responsive to therapy from those that are resistant. When analyzing cells collected from the circulation, or from other bodily fluids (e.g., pleural effusions, ascites), typically only a small number of cells are available. To asses the cell-to-cell heterogeneity of this limited number of cells, ive propose to develop and to apply quantitative, comprehensive single-cell analysis devices for assessing the DNA genome (e.g., single nucleotide polymorphisms, fusions, deletions), RNA expression, protein abundance (cell surface, intracellular, and secretome abundance), and biophysical properties of single cells for the dual purposes of predicting a patient's likely response to EGFR-targeted therapies and for monitoring a patient's acquisition of resistance to EGFR-targeted therapies (Fig. N3.3.1). We propose two specific aims for the development, testing, and application of our comprehensive analysis platform (Table N3.3.1).