Transcriptomics from Single Circulating Tumor Cells

描述（由申请人提供）：本申请涉及广泛的研究领域：06，使能技术和挑战主题：06-HG-102* 从复杂组织中的单个活细胞获得基因组学、蛋白质组学和代谢组学数据的技术。 将开发分析单个癌细胞基因表达的方法，使研究达到前所未有的灵敏度。 我们提出了测量从癌症患者血液中分离的单个循环肿瘤细胞（CTC）的RNA转录的技术。 通过访问已确认的癌细胞，从正常细胞的大背景中分离出来，将有可能获得有关疾病过程及其通过转移在体内传播的未探索的知识。 几项关键技术的最新进展使这种方法变得可行：1）Scripps研究所的研究人员已经开发出荧光标记CTC的方法，使得它们可以从人类血液样品中鉴定出来，2）分离单细胞的技术已经与J.克雷格文特尔研究所的核酸扩增方法相结合，使得能够首次证明来自单细胞的DNA测序，和3）Life Technologies（Invitrogen/Applied Biosystems）已经将从单细胞合成cDNA和全基因组表达谱分析的方法与其下一代SOLiD测序平台相结合。 这三个机构将合作分离单个癌细胞，合成cDNA并通过SOLiD方法对其进行测序。 目标是在最基本的单细胞水平上观察癌症。 公共卫生相关性：我们建议开发一种方法来测量从人类血液中获得的单个癌细胞中的基因表达。 这些循环癌细胞（CTCs）非常罕见，但可能是癌症通过转移扩散到全身的原因。 利用荧光标记CTC的新策略，可以分离它们。其他新技术能够确定单个细胞的mRNA表达水平，从而了解基因表达的变化。 对CTC的生理学以及它们如何导致转移的新理解将是可能的。

DESCRIPTION (provided by applicant): This application addresses the broad Research Area: 06, Enabling Technologies And Challenge Topic: 06-HG-102* Technologies for obtaining genomic, proteomic, and metabolomic data from individual viable cells in complex tissues. Methods will be developed to analyze gene expression from single cancer cells enabling studies at an unprecedented level of sensitivity. We propose techniques to measure RNA transcription from individual circulating tumor cells (CTCs) isolated from the blood of cancer patients. By accessing confirmed cancer cells, separated from the large background of normal cells, it will be possible to gain unexplored knowledge about the disease process and its spread through the body by metastasis. Recent advances in several key technologies have made this approach feasible: 1) Researchers at the Scripps Research Institute have developed the means to fluorescently label CTCs so that they can be identified from human blood samples, 2) Technologies for isolating single cells have been combined with nucleic acid amplification methods at the J. Craig Venter institute enabling the first demonstration of DNA sequencing from single cells, and 3) Life Technologies (Invitrogen/Applied Biosystems) have combined methods for cDNA synthesis from single cells and whole genome expression profiling with their next generation SOLiD sequencing platform. These three institutions will collaborate to isolate single cancer cells, synthesize cDNA and sequence it by the SOLiD method. The goal is to observe cancer at the most basic level of the single cell. PUBLIC HEALTH RELEVANCE: We propose development of a method to measure gene expression in individual cancer cells obtained from human blood. These circulating cancer cells (CTCs) are very rare but can be responsible for the spread of cancer throughout the body by metastasis. With new strategies to fluorescently label CTCs it is possible to isolate them. Other new technology enables determination of mRNA expression levels from a single cell giving an understanding of the alterations to gene expression. A new understanding of the physiology of CTCs and how they lead to metastasis will be possible.