Whole Genome Amplification and Sequencing of Single Cancer Cells

描述（由申请人提供）：该提案汇集了两个在单细胞操作，扩增策略，测序和翻译方面具有专业知识的研究团队。 癌症研究，以推进技术，以解决癌症生物学中的重要问题。它响应RFA-CA-12-003，并提出基于我们在单细胞全基因组扩增方面的最新进展，创建和验证用于人类样本稳健单细胞测序的平台。近期目标是将联合收割机不同的技术方法结合起来，然后用它们来确定个体癌细胞在群体中的差异。我们的重点将是对外周血中的单个癌细胞进行测序，以及通过细针穿刺对原发性和转移性癌组织进行测序。目标1将开发和验证综合方法，使免疫细胞化学分析可以告知决定哪些细胞测序。目标2将侧重于将单细胞扩增和测序技术应用于目标1中实现表型鉴定的原代人类样本。总的来说，我们的目标是适应和进一步开发一套最近开发的和非常有前途的单细胞分析技术，以了解原代和脱落癌细胞的基因组变异。该项目的成功将产生一个综合平台，将有助于克服普遍存在的问题， 癌症研究的障碍。该研究预计将对基础研究、临床实践以及新兴抗癌药物的开发产生广泛影响。

DESCRIPTION (provided by applicant): This proposal brings together two research teams with expertise in single cell manipulation, amplification strategies, sequencing, and translational cancer research to advance technologies to tackle important questions in cancer biology. It responds to RFA-CA-12-003, and proposes to create and validate a platform for robust single cell sequencing of human samples based on our most recent advance in single cell whole genome amplification. The immediate goal is to combine different technical approaches and then use them to determine how individual cancer cells differ in populations. Our focus will be on sequencing single cancer cells in peripheral blood, as well as from primary and metastatic cancer tissues by fine needle aspiration. Aim 1 will develop and validate integrated approaches so that immunocytochemical analyses can inform decisions on which cells to sequence. Aim 2 will focus on applying the single cell amplification and sequencing technology to primary human samples with phenotypic identification achieved in Aim 1. Overall, our goal is to adapt and further develop a suite of recently developed and highly promising single cell analytical technologies to gain an understanding of genomic variations in primary and shed cancer cells. Success of the project will result in an integrated platform that will serve to overcome prevailing impediments in cancer research. The research is expected to have a broad impact on basic research, clinical practice, and the development of emerging anti- cancer drugs.