SPECTRAL IMAGING FOR PHENOTYPE ANALYSIS OF CANCER CELLS

描述：(申请人描述) 这项提议寻求开发一种新的筛查系统来支持癌症 基于基因测量的诊断、肿瘤分期、预后 在RNA水平上的表达。肿瘤进展的潜在假设 因为基因变化的积累，以及基因变化 通过一种或几种改变的、可测量的表达来表现自己 基因。目前的技术将RNA表达水平的测量限制在 每个实验只有很少的基因。敏感的肿瘤细胞检测和预后 将需要同时对RNA表达水平进行分析的方法 五个或五个以上疑似癌基因/抑癌基因。此外，如果一个 癌基因过度表达，则其GEM剂量也应确定。 对几个标记基因的定量分析对敏感性是至关重要的 肿瘤细胞检测和准确的细胞分类以及 开发新的抗肿瘤策略。这条信息应该会有帮助 鉴别良性肿瘤和恶性肿瘤，并定义预后标记物。 我们将发展多色荧光原位杂交(FISH) 与Spectral Ima(SIM)一起执行这些测量。现有 SIM仪器可以记录从nm到1100 nm的荧光光谱 分辨率约为10 nm。这允许对Edna进行独特的标记和检测 使用商用荧光显色剂的探头。SIM技术可以 改良以研究癌基因表达与肿瘤的相关性 进步。空间共定位和光谱重叠将通过 数字记录图像的软件处理，称为“光谱分解” (太阳)。 使用现有甲状腺肿瘤和乳腺细胞的人工混合物 癌细胞株，我们将开发所需的软件模块进行测量 同一细胞中多个RNA物种的胞内水平并确定 检测的灵敏度、准确性和重复性。这项工作将是 扩展到甲状腺和乳腺肿瘤组织，增加了 杂交靶点和基因重排/扩增与 基因表达。这项技术，当应用于细针活检组织时 抽吸物，可以帮助诊断和预测可疑细胞在 快速、廉价、微创的方式。

DESCRIPTION: (Applicant's Description) This proposal seeks to develop a novel screening system to support cancer diagnosis, tumor staging, prognostication based on the measurement of gene expression at the RNA level. The underlying hypothesis hat tumors progress because of accumulation of genetic changes, and that the genetic changes manifest themselves in altered and measurable expression of one or several genes. Current techniques limit the measurement of RNA expression levels to a few genes per experiment. Sensitive tumor cell detection and prognostication will require methods that simultaneously profile the RNA expression levels of five or more suspected oncogenes/tumor suppressor genes. Furthermore, if an oncogene is overexpressed, then its gem dosage should be determined as well. The quantitative analysis of several marker genes is crucial for sensitive tumor cell detection and accurate cell classification as well as the development of novel anti-tumor strategies. This information should help discriminate benign vs. malignant neoplasms and define prognostic markers. We will develop multicolor fluorescence in situ hybridization (FISH) in conjunction with Spectral Ima (SIm) to perform these measurements. Existing SIm instrumentation can record fluorescence spectra from nm to 1100 nm with about 10 nm resolution. This allows the unique labeling and detection of eDNA probe with commercially available fluorochromes. SIm technology can be modified to investigate the correlation cancer gene expression with tumor progression. Spatial co-localization and spectral overlap will be add by software processing of digitally recorded images, termed "spectral un-mixing" (SUN). Using artificial mixtures of cells from existing thyroid tumor and breast cancer cell lines, we will develop the software modules needed to measure intracellular levels of multiple RNA species in the same cell and to determine the assay sensitivity, accuracy, and reproducibility. The work will be extended to thyroid and breast tumor tissues increasing the number of hybridization targets and correlating gene rearrangements/amplifications with gene expression. This technology, when applied biopsy tissue from fine needle aspirates, could help diagnose and predict the course of suspicious cells in a rapid, inexpensive, and minimally invasive manner.