IDBR: Collaborative Research: Real time secretion: single cell analyzer

IDBR：协作研究：实时分泌：单细胞分析仪

• 批准号: 1152030
• 负责人: James Glazier
• 金额: $ 58.02万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1152030&HistoricalAwards=false
• 关键词: IDBR; Collaborative; Research; Real; time

IDBR: Collaborative Research: Real time secretion: single cell analyzerIntellectual merit: The cells of every organ or organism release (secrete) a wide variety of molecules into their surroundings. Secretion changes over time as cells grow, change structure and function, interact with other cells, and when they become damaged or diseased. Molecules produced by cells play key roles in the growth of embryos, infection, immune response and healing, and in regulating internal body functions despite fluctuating environmental conditions, aging, injury and illness. A tool to measure the types and numbers of molecules secreted by individual cells and their changes in space and time would help researchers understand how cells and organisms function and malfunction, in biomedical, biological and bio-industrial contexts. The tool must be able to sort individual cells based on their secretion behaviors and recover them undamaged for further use, e.g. for tissue engineering or medical treatment. Currently, no technology is simultaneously: 1) Sensitive enough to quantitatively measure many important molecules secreted by single cells at the very low concentrations typical in living organisms, 2) Fast enough to measure changes in extracellular molecular concentrations over periods of minutes, 3) Able to sort and recover live cells for further use, 4) Able to use cells as they occur naturally, without needing genetic modification or an external supply of chemical labels, 5) Capable of quick extension to conduct many different measurements simultaneously on each cell or the same measurement on many cells (high-throughput), 6) Inexpensive enough for wide use in research laboratories, clinics, and industry. The proposed Real Time Secretion-Single Cell Analyzer (RTS-SCA) will be a significant breakthrough that will enable researchers to directly measure secretion dynamics, investigate the degree of variation in secretion behavior within populations of cells and the selection of cells with desired properties. RTS-SCA will find applications in biological and chemical engineering, developmental, immune and cell biology, computational biology, pharmacology and medicine. The technology to be developed is a critical first step towards developing micro- or nano-probes that could later be used as diagnostic tools in living tissues.Broader impact: Bioengineering, developmental and cell biology, biomedical research, and applied bioagriculture and bioindustry all require a currently unavailable capability to dynamically quantify secretion at the single cell or tissue level. This ability will advance basic understanding of cell functions and interactions, and aid tissue engineering, therapy development and drug discovery. The technology will allow improved versions of common research techniques, such as flow cytometry, polymerase chain reaction, gel electrophoresis, gene sequencing or microarray analysis.Immediate applications of the RTS-SCA include the analysis of the key signaling molecules in embryonic development, immunology, wound healing, tissue function regulation, and diseases like cancer, in which both cancer and host-tissue cells send complex chemical messages to each other. Bioengineering and pharmacology need to identify and select particular cells that produce molecules of interest at high levels, and to determine optimal stimulation and culture conditions for molecule production. The RTS-SCA could help in optimizing cell-secretion behaviors, essential for the development of tissue regeneration therapies. The RTS-SCA?s ability to quantify the complex kinetics of insulin release, which involves disrupted pulsatile secretion from pancreatic tissues, could accelerate research in treatments for type-2 diabetes, a disease which accounts for 10% of the U.S. health-care expenditure. Since periodic insulin secretion is an indicator of healthy pancreas function, RTS-SCA would help in selecting tissue fragments for transplant therapy for type-1 diabetes. Dynamic analysis of molecules is especially important in immune system responses and diseases, many of which are mediated by currently unmeasurable time-varying secreted molecules, examples include atherosclerosis, allergies, rheumatoid arthritis and multiple sclerosis. The RTS-SCA will help understand how immune system cells sense and respond to external challenges facilitating design of more effective treatments and vaccines.Dissemination Plan: The scientists from Indiana University-Bloomington (IUB) and University of California San Diego (UCSD) have established collaborations with two major research centers, The IUB Center for Genomics and Bioinformatics and The Moores UCSD Cancer Center Microscopy Shared Resource, covering research needs in the Midwest, West Coast, and across the USA. To encourage the RTS-SCA's wide use, each center will receive an operating RTS-SCA. These centers will train undergraduates, graduate students, postdoctoral researchers, faculty and external partners to use the RTS-SCA for their research, maintain the instrument, advertise the instrument to outside collaborators, and provide appropriate outreach support through their networks.

IDBR：合作研究：实时分泌：单细胞分析仪智力优势：每个器官或生物体的细胞释放（分泌）各种各样的分子到它们周围的环境中。随着细胞的生长、结构和功能的改变、与其他细胞的相互作用以及细胞受损或患病，分泌物会随着时间的推移而变化。细胞产生的分子在胚胎的生长、感染、免疫反应和愈合中发挥关键作用，并在不受环境条件波动、衰老、损伤和疾病影响的情况下调节体内功能。一种测量单个细胞分泌的分子类型和数量及其在空间和时间上的变化的工具将有助于研究人员了解细胞和生物体在生物医学、生物学和生物工业环境中的功能和故障。该工具必须能够根据单个细胞的分泌行为进行分类，并将其完好无损地恢复以供进一步使用，例如用于组织工程或医疗。目前，没有一种技术是同时：1)足够灵敏，可以定量测量生物体中单细胞在极低浓度下分泌的许多重要分子；2)足够快，可以在几分钟内测量细胞外分子浓度的变化；3)能够分类和回收活细胞以供进一步使用；4)能够使用自然产生的细胞，无需基因改造或外部化学标签的供应。5)能够快速扩展，在每个细胞上同时进行许多不同的测量或在许多细胞上进行相同的测量（高通量），6)足够便宜，可广泛用于研究实验室，诊所和工业。提出的实时分泌-单细胞分析仪（RTS-SCA）将是一个重大突破，它将使研究人员能够直接测量分泌动态，研究细胞群体内分泌行为的变化程度以及选择具有所需特性的细胞。RTS-SCA将在生物和化学工程、发育、免疫和细胞生物学、计算生物学、药理学和医学等领域得到应用。这项即将开发的技术是开发微型或纳米探针的关键的第一步，这些探针以后可以用作活组织的诊断工具。更广泛的影响：生物工程、发育和细胞生物学、生物医学研究以及应用生物农业和生物工业都需要目前尚不具备的在单细胞或组织水平上动态量化分泌的能力。这种能力将促进对细胞功能和相互作用的基本理解，并有助于组织工程、治疗开发和药物发现。该技术将改进常用的研究技术，如流式细胞术、聚合酶链反应、凝胶电泳、基因测序或微阵列分析。RTS-SCA的直接应用包括分析胚胎发育、免疫学、伤口愈合、组织功能调节和癌症等疾病中的关键信号分子，在这些疾病中，癌症和宿主组织细胞相互发送复杂的化学信息。生物工程和药理学需要鉴定和选择产生高水平分子的特定细胞，并确定分子产生的最佳刺激和培养条件。RTS-SCA可以帮助优化细胞分泌行为，这对组织再生疗法的发展至关重要。RTS-SCA吗?研究人员有能力量化胰岛素释放的复杂动力学，这涉及胰腺组织脉搏性分泌的中断，这可能会加速2型糖尿病治疗的研究，这种疾病占美国医疗保健支出的10%。由于周期性胰岛素分泌是健康胰腺功能的一个指标，RTS-SCA将有助于选择用于1型糖尿病移植治疗的组织片段。分子的动态分析在免疫系统反应和疾病中尤为重要，其中许多疾病是由目前无法测量的时变分泌分子介导的，例如动脉粥样硬化、过敏、类风湿关节炎和多发性硬化症。RTS-SCA将有助于了解免疫系统细胞如何感知和应对外部挑战，从而促进设计更有效的治疗方法和疫苗。传播计划：来自印第安纳大学布卢明顿分校（IUB）和加州大学圣地亚哥分校（UCSD）的科学家已经与两个主要研究中心建立了合作关系，IUB基因组学和生物信息学中心和Moores UCSD癌症中心显微镜共享资源，涵盖中西部，西海岸和整个美国的研究需求。为了鼓励RTS-SCA的广泛使用，每个中心都将获得一个运行中的RTS-SCA。这些中心将培训本科生、研究生、博士后研究人员、教师和外部合作伙伴使用RTS-SCA进行研究，维护仪器，向外部合作者宣传仪器，并通过其网络提供适当的外展支持。