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）： 本提案的目的是获得一个对于单细胞转录分析足够敏感的 PCR 系统。该系统名为 Fluidigm 的 Biomark HD 系统，能够进行实时、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微流体通道，允许同时进行纳升体积和皮克量 RNA 的 PCR 反应。具体而言，可以在单次运行中分析单个细胞（每次运行最多 96 个）的转录本（每次运行最多 96 个）。这种独特的能力将极大地扩展密歇根大学可以解决的多个研究方向的研究问题。具体例子包括转移研究，例如播散性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，他们总共拥有 20 个 R01、一个 P01 中的 4 个项目、两个不同 P50 SPORES 中的 3 个项目、U01 联盟中的 2 个项目以及 NIH 主任奖和 U54 奖各 1 个，总共 31 个 NIH-资助的项目。该小组由多个部门以及医学院和牙科学校的成员组成。目前，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一非常重要的领域。该设备将放置在癌症中心大楼的中心位置，以便许多研究人员可以使用。已经计划使用该设备的项目将利用 86.8% 的可用设备时间，这表明对该设备的需求很高。研究项目与 NIH 改善健康的目标直接相关，包括癌症转移、癌症治疗、血管生成、骨生物学和年龄相关疾病的研究。该设备将由经验丰富的技术人员、生物信息学专家和内部咨询委员会提供支持。吸引和培训新用户的系统已经到位，以便其他 NIH 资助的研究人员能够从该设备中受益。最初三年承诺对维护和技术成本提供大量机构支持，以确保系统的长期可行性（三年总计 100,500 美元）。将建立基于用户的充值系统以继续其长期使用。此外，还有机构对设备空间的承诺。总之，使用这一尖端成像系统将大大加快密歇根大学大量 NIH 资助的研究人员的研究步伐。

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