Microfluidic PCR System for Single Cell Transcriptional Analysis

用于单细胞转录分析的微流控 PCR 系统

• 批准号: 8446702
• 负责人: Evan T Keller
• 金额: $ 24.17万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446702
• 关键词: Address; Advisory Committees; Area; Award; Bioinformatics; Biological Assay; Biology of Aging; Cancer Center; Cells; Commit; Dental Schools; Disease; Ensure; Equipment; Funding; Goals; Health; Image; Individual; Location; Maintenance; Malignant Neoplasms; Michigan; Microfluidics; Neoplasm Metastasis; RNA; Reaction; Research; Research Personnel; Research Project Grants; Running; System; TNFRSF5 gene; Time; Training; Transcript; Tumor Cell Biology; United States National Institutes of Health; Universities; age related; base; bone; cancer stem cell; cost; digital; experience; improved; medical schools; member; nanolitre; stem cell biology; therapeutic angiogenesis

DESCRIPTION (provided by applicant): The objective of this proposal is to acquire a PCR system that is sensitive enough for single cell transcriptional analysis. The system, called the Biomark HD System from Fluidigm is capable of real- time, end-point and digital PCR. The key aspects of this system are the use of integrated microfluidic channels on one chip that allow for simultaneous PCR reactions in nanoliter volumes and picogram amounts of RNA. In concrete terms, transcripts (up to 96 per run) from individual cells (up to 96 per run) can be assayed in a single run. This unique capability will greatly expand the research questions that can be addressed at University of Michigan for multiple investigative directions. Specific examples include metastasis studies such as disseminated tumor cell biology or cancer stem cell biology. This application supports 14 major users who together have 20 R01s, 4 projects in one P01, 3 projects in two different P50 SPORES, 2 projects in U01 consortia and 1 each of a NIH Director's Award and a U54 award for a total of 31 NIH-funded projects. The group consists of members of multiple Departments and the Medical and Dental Schools. Currently, there is not capability to perform single cell transcriptional analysis at University of Michigan and this equipment will enable us to expand studies into this very important area. The equipment will be placed in a central location in the Cancer Center Building so that it is accessible to many investigators. The projects already planning to use this equipment will utilize 86.8% of the available equipment time indicating the high demand for this equipment. Research projects are directly related to NIH goals of improving health and include studies on cancer metastasis, cancer therapeutics, angiogenesis, bone biology and age- related disease. The equipment will be supported by an experienced technical staff, a bioinformatics expert and an internal advisory committee. A system to attract and train new users is in place so that other NIH-funded investigators will benefit from the equipment. Significant institutional support for both maintenance and technical costs has been committed for the initial three years to ensure long- term viability of the system (Total of $100,500 over 3 years). A user-based recharge system will be put into place to continue its long-term use. Additionally, there is institutional commitment of space for the equipment. In summary access to this cutting edge imaging system will greatly accelerate the pace of research at University of Michigan for a large number of NIH-funded investigators.

描述（由申请人提供）： 该建议的目的是获取足够敏感的PCR系统，足以用于单细胞转录分析。该系统称为Fluidigm的Biomark HD系统，能够实时，终点和数字PCR。该系统的关键方面是在一个芯片上使用集成的微流体通道，该通道允许在纳米素体积和rNA的皮克图中同时进行PCR反应。用具体的术语，可以在一次运行中分析单个单元格（每次运行最多96个）的成绩单（每次运行96个）。这种独特的能力将大大扩大密歇根大学可以解决多个调查方向的研究问题。具体的例子包括转移研究，例如传播肿瘤细胞生物学或癌症干细胞生物学。该应用程序支持14个主要用户，他们共有20个R01，一个P01中的4个项目，两个不同的P50孢子中的3个项目，U01财团中的2个项目和NIH董事奖的1个项目和1个U54奖项，共有31个NIH资助的项目。该小组由多个部门以及医学和牙科学校的成员组成。当前，在密歇根大学没有能力进行单细胞转录分析，该设备将使我们能够将研究扩展到这个非常重要的领域。该设备将放置在癌症中心大楼的中心位置，以便许多调查人员可以使用。已经计划使用此设备的项目将利用86.8％的可用设备时间，表明对该设备的需求很高。研究项目与改善健康的目标直接相关，包括有关癌症转移，癌症治疗，血管生成，骨骼生物学和与年龄相关的疾病的研究。该设备将得到经验丰富的技术人员，生物信息学专家和内部咨询委员会的支持。吸引和培训新用户的系统已经到位，因此其他NIH资助的调查人员将从设备中受益。在最初的三年中，已经投入了对维护和技术成本的大量机构支持，以确保系统的长期可行性（在3年内总计100,500美元）。将建立基于用户的充电系统，以继续其长期使用。此外，设备的机构承诺。总而言之，该尖端成像系统将大大加快密歇根大学的研究速度，以实现大量NIH资助的研究人员。

项目成果

Poor Prognosis Indicated by Venous Circulating Tumor Cell Clusters in Early-Stage Lung Cancers.

• DOI: 10.1158/0008-5472.can-16-2072
• 发表时间: 2017-09-15
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Murlidhar V;Reddy RM;Fouladdel S;Zhao L;Ishikawa MK;Grabauskiene S;Zhang Z;Lin J;Chang AC;Carrott P;Lynch WR;Orringer MB;Kumar-Sinha C;Palanisamy N;Beer DG;Wicha MS;Ramnath N;Azizi E;Nagrath S
• 通讯作者: Nagrath S