SPECTRAL IMAGING FOR PHENOTYPE ANALYSIS OF CANCER CELLS

描述：（申请人的描述） 该提案旨在开发一种新的筛查系统，以支持癌症 诊断、肿瘤分期、基于基因测量的鉴别诊断 在RNA水平表达。肿瘤进展的基本假设 因为基因变化的积累， 表现在一个或几个改变和可测量的表达 基因.目前的技术限制了RNA表达水平的测量， 每个实验的基因数。灵敏的肿瘤细胞检测和鉴定 将需要同时分析RNA表达水平的方法， 5个或更多疑似癌基因/肿瘤抑制基因。此外，如果 如果癌基因过度表达，那么也应该确定其gem剂量。 几个标记基因的定量分析对于敏感的 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。这些信息应该会有所帮助 区分良性与恶性肿瘤并定义预后标志物。 我们将开展荧光原位杂交技术（FISH）， 结合光谱图像（SIm）来执行这些测量。现有 SIM仪器可以记录从nm到1100 nm的荧光光谱， 约10 nm分辨率。这使得eDNA的独特标记和检测成为可能 探针与市售荧光染料。SIM技术可以 研究癌基因表达与肿瘤的相关性 进展空间共定位和光谱重叠将通过以下方式添加： 数字记录图像的软件处理，称为“光谱解混” （SUN）. 利用现有甲状腺肿瘤和乳腺癌细胞的人工混合物 癌细胞系，我们将开发测量所需的软件模块， 同一细胞中多种RNA种类的细胞内水平，并确定 测定灵敏度、准确度和重现性。这项工作将 扩展到甲状腺和乳腺肿瘤组织， 杂交靶点和相关基因重排/扩增， 基因表达。这项技术，当应用活检组织从细针 抽吸物，可以帮助诊断和预测可疑细胞的过程， 快速、廉价和微创的方式。

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.