Developing a single cell growth monitor for classifying therapeutic response

开发用于对治疗反应进行分类的单细胞生长监测器

• 批准号: 8046340
• 负责人: PARAG Kumar MALLICK
• 金额: $ 14.19万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8046340
• 关键词: Antibodies; Biochemical; Biological Assay; Biological Markers; CCNE1 gene; Cancer Biology; Cancer Cell Growth; Cancer Center; Cancer cell line; Cell Count; Cell Line; Cell Surface Proteins; Cells; Clinical Oncology; Coupled; DNA copy number; Detection; Devices; Ensure; Environment; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Fluorescence; Gefitinib; Growth; Immunoassay; In Vitro; Intervention; Kinetics; Malignant Neoplasms; Mammalian Cell; Measurement; Measures; Methods; Molecular; Monitor; Nanotechnology; Optics; Pathway interactions; Phenotype; Phosphotransferases; Physiological; Proteins; Proteomics; Resolution; Shapes; Silicon; Structure; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Time; Validation; Work; Xenograft Model; base; cancer cell; cancer therapy; cell growth; density; design; in vivo; instrument; novel strategies; oncology; protein expression; response; single cell proteins; small molecule

DESCRIPTION (provided by applicant): Clinical oncology and cancer biology are challenged by the lack of assay platforms for measuring changes in cancer-cells' growth kinetics in response to therapeutic intervention. Cell growth kinetics can be measured in a number of ways, such as by DNA copy number, volume, mass, density, shape, or by expression of particular proteins. Here we define growth kinetics as changes in a cell's mass and density over time. We propose to develop an instrument for concurrently monitoring single cell growth kinetics and cell surface protein expression. We hypothesize that changes in growth kinetics and in cell surface protein expression can be used as a surrogate for response to pathway-directed therapeutic agents. As a validation of the instrument, we will monitor mass, density and cell-surface protein expression (determined by fluorescence) in single A431 cells in response to intervention with the pathway targeted therapy gefitinib, a small molecule inhibitor of the epidermal growth factor receptor (EGFR), and appropriate controls. Single cell mass and density will be measured by a previously validated device known as the suspended microchannel resonator (SMR). The SMR can measure the mass of a mammalian cell with a resolution near 0.01% (1 Hz bandwidth). In order to achieve a continuous measurement of mass and density, silicon posts will be used for capturing a single cell within the sensitive region of the microchannel resonator. To enable simultaneous detection of protein expression, by fluorescence of coupled antibodies, the SMR will be modified to have optical transparency within the region where the cell is captured. The proposed instrument will leverage our prior proteomics work with the Center for Cancer Nanotechnology Excellence focused on Therapeutic Response (CCNE-TR) where we used proteomic methods to discover cell-surface protein biomarkers indicative of therapeutic response. Single-cell measurement of the abundance of these proteins, concurrently with cell mass and density kinetics will provide a new approach for characterizing and monitoring cell response to therapy on a physiological and molecular level.

描述（由申请人提供）：缺乏测量癌症细胞生长动力学的变化的临床肿瘤学和癌症生物学对治疗干预的响应。细胞生长动力学可以通过多种方式进行测量，例如通过DNA拷贝数，体积，质量，密度，形状或通过特定蛋白质的表达。在这里，我们将生长动力学定义为细胞质量和密度随着时间的变化的变化。我们建议开发一种同时监测单细胞生长动力学和细胞表面蛋白表达的仪器。我们假设生长动力学和细胞表面蛋白表达的变化可以用作对途径定向治疗剂的反应的替代物。作为对仪器的验证，我们将在单个A431细胞中监测质量，密度和细胞表面蛋白表达（由荧光确定），以响应靶向靶向治疗的靶向治疗吉法替尼的干预，Gefitinib是表皮生长因子受体（EGFR）的小分子抑制剂（EGFR）的小分子抑制剂，以及适当的对照。单细胞质量和密度将通过称为悬浮的微通道谐振器（SMR）的先前验证的设备进行测量。 SMR可以测量分辨率接近0.01％（1 Hz带宽）的哺乳动物细胞的质量。为了实现质量和密度的连续测量，硅柱将用于捕获微通道谐振器敏感区域内的单个单元。为了通过耦合抗体的荧光同时检测蛋白质表达，将修改SMR以在捕获细胞的区域内具有光学透明度。该提出的仪器将利用我们先前的蛋白质组学与卓越癌症纳米技术中心的工作，以治疗反应（CCNE-TR）使用蛋白质组学方法来发现细胞表面蛋白质生物标志物，指示具有治疗反应的细胞表面蛋白。这些蛋白质丰度的单细胞测量，同时与细胞质量和密度动力学同时测量，将为您在生理和分子水平上表征和监测细胞对治疗的反应的新方法。

项目成果

Continuous and long-term volume measurements with a commercial Coulter counter.

• DOI: 10.1371/journal.pone.0029866
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Bryan AK;Engler A;Gulati A;Manalis SR
• 通讯作者: Manalis SR

Mass sensors with mechanical traps for weighing single cells in different fluids.

• DOI: 10.1039/c1lc20736a
• 发表时间: 2011-11
• 期刊: Lab on a chip
• 影响因子: 6.1
• 作者: Yaochung Weng;Francisco Feijó Delgado;S. Son;T. Burg;S. Wasserman;S. Manalis