A Microfluidic System for Image-Guided RNA-Seq of Single Cells in Glioblastoma

用于胶质母细胞瘤单细胞图像引导 RNA 测序的微流体系统

• 批准号: 9129717
• 负责人: Peter Alan Sims
• 金额: $ 17.28万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9129717
• 关键词: Address; Area; Award; Behavior; Beryllium; Bioinformatics; Biological; Biological Assay; Biomedical Research; Brain Neoplasms; Buffers; Cell Count; Cell Cycle Stage; Cell Line; Cells; Cellular Morphology; Clinical; Collaborations; Communication; Complementary DNA; Complex; Cytolysis; Deposition; Development; Development Plans; Disease; Engineering; Epigenetic Process; Exhibits; Gene Expression; Gene Expression Profile; Genetic Materials; Genetic Transcription; Genomic Instability; Glioblastoma; Glioma; Goals; Grant; Guide RNA; Heterogeneity; Human; Image; Immunofluorescence Immunologic; Individual; Infection; Label; Lasers; Malignant - descriptor; Mammalian Cell; Mentors; Mentorship; Microfluidics; Molecular Profiling; Noise; Normal Cell; Phenotype; Population; Process; RNA; RNA library; Reading; Reporter; Research; Research Personnel; Resource Sharing; Sampling; Surface; Suspension substance; Suspensions; System; Systems Analysis; Techniques; Technology; Testing; Therapeutic; Tissues; Training; Transcript; Translating; Universities; Writing; base; career; career development; cellular imaging; chemical genetics; deep sequencing; design; direct application; effective therapy; genome-wide; genome-wide analysis; image guided; improved; interest; laser tweezer; live cell imaging; neoplastic cell; optical imaging; optical traps; pluripotency; programs; research and development; research study; response; seal; self-renewal; single cell analysis; skills; targeted imaging; tissue culture; tool; transcriptome; transcriptome sequencing; transcriptomics; tumor; tumorigenic

DESCRIPTION (provided by applicant): The Candidate proposes a five-year career development and research program under the mentorship of Prof. Andrea Califano. The Candidate's career goal is to build an independent research program at Columbia University focusing on engineering new tools for system-wide interrogation of single cells with direct applications in biomedicine. The research component involves the development of a microfluidic array platform that allows highly parallel optical imaging and RNA capture from single cells. Importantly, the array elements are individually addressable so that RNA libraries from individual cells-of-interest can be extracted for RNA-Seq. This system will allow the Candidate and his collaborators to address key questions about tumor composition and progression in glioblastoma, a common and unusually heterogeneous type of brain tumor. Although the Candidate has a strong technology-oriented background, he requires significant training in order to carry out the biomedical aspects of the proposal. Hence, the career development component focuses on acquiring skills related to 1) tissue handling, dispersing single cell suspensions from tissue, and culturing tissue-derived cells; 2) labeling, FACS, and laser capture of tissue-derived cells; and 3) RNA stabilization/extraction and bioinformatics. In addition, the career development plan includes specific coursework in these areas as well as mentoring in grant-writing and scientific communication and hypothesis development in biomedical research. Columbia has well-established research programs in these areas and substantial shared resources which will be leveraged in various parts of the proposal. This award will facilitate the candidate's transitio as an independent biomedical researcher and the formation of valuable collaborations.

描述（由申请人提供）：候选人将在Andrea Califano教授的指导下进行为期五年的职业发展和研究计划。候选人的职业目标是在哥伦比亚大学建立一个独立的研究项目，专注于在生物医学中直接应用单细胞全系统审讯的工程新工具。研究部分涉及微流体阵列平台的开发，该平台允许高度平行的光学成像和单细胞RNA捕获。重要的是，阵列元素是可单独寻址的，因此可以从感兴趣的单个细胞中提取RNA库用于RNA- seq。该系统将允许候选人和他的合作者解决胶质母细胞瘤的肿瘤组成和进展的关键问题，胶质母细胞瘤是一种常见的和异常异质性的脑肿瘤。虽然候选人有很强的技术背景，但他需要大量的培训才能执行提案的生物医学方面。因此，职业发展部分侧重于获得以下技能：1)组织处理，从组织中分散单细胞悬浮液，培养组织来源的细胞；2)组织源性细胞的标记、FACS和激光捕获；3) RNA稳定/提取和生物信息学。此外，职业发展计划还包括这些领域的具体课程，以及在生物医学研究的拨款写作、科学交流和假设发展方面的指导。哥伦比亚大学在这些领域有完善的研究项目和大量的共享资源，这些资源将在提案的各个部分得到利用。该奖项将促进候选人作为独立生物医学研究人员的过渡，并形成有价值的合作。