Living Cell Arrays for Real Time Functional Genomics

描述（由申请人提供）：新的工具正在彻底改变生物医学研究，使有关基因，蛋白质及其在正常和患病状态下的结构和功能的数据采集呈指数增长。在这些进展中，大规模监测基因图谱的能力是值得注意的。然而，通常难以将在正常和异常状态下观察到的趋势和关系与由基因表达谱产生的表型相关联。对于涉及代谢紊乱的生理状态，基因表达谱不能完全解释所涉及的复杂分子机制。因此，为了全面了解代谢状态，必须了解控制代谢物变化的基因表达事件和细胞质事件。拟议的生物工程研究伙伴关系旨在开发一种新的功能基因组学方法，用于研究基因表达：使用完整细胞，以大规模并行，高通量格式使用水母发光蛋白型荧光蛋白（AFP）同时进行多个基因的时间表达谱分析。具体目标是：（2）设计微流体系统以动态控制输入刺激以及原代大鼠肝细胞和H35肝癌报告细胞系阵列的荧光响应;（3）表征细胞因子信号转导的动力学以及脂肪变性和不同代谢状态对这些细胞的急性期反应的影响。这一项目将由三个不同的研究小组进行，它们将进行广泛的互动。主要研究者Martin Yarmush博士（MGH）将监督行政和技术核心，并领导细胞生理学和成像小组，该小组将整合其他PI开发的技术，并进行大部分细胞生理学实验。Mehmet Toner博士是MGH微尺度工程设施的主任，他将领导微制造和微流体小组，专注于微制造设备的开发。Jeffrey Morgan博士（布朗大学，普罗维登斯，RI），分子生物学家和基因治疗专家，将领导分子生物学和细胞分析小组专注于报告细胞系的发展。

DESCRIPTION (provided by applicant): New tools are revolutionizing biomedical research, enabling an exponential growth in the acquisition of data regarding genes, proteins, and their structures and functions in normal and diseased states. Among these advances, the ability to monitor profiles of genes on a large scale is notable. However, it is often difficult to correlate the trends and relationships observed in normal and abnormal states to the phenotype resulting from the gene expression profile. For physiological states involving metabolic derangements, gene expression profiling does not fully explain the complex molecular mechanisms involved. Thus, in order to develop a comprehensive understanding of metabolic states, it is essential to understand both the gene expression events as well as the cytoplasmic events, which control changes in metabolites. The proposed Bioengineering Research Partnership seeks to develop a new functional genomics approach for studying gene expression: the use of intact cells for the simultaneous temporal expression profiling of multiple genes using aequorin-type fluorescent proteins (AFP) in a massively parallel, high throughput format. The specific aims are: (1) to generate and characterize a panel of reporter cell lines that monitors the major events in the inflammatory cytokine signaling cascades; (2) to design a microfluidic system to dynamically control the input stimulus as well as the fluorescent response of an array of primary rat hepatocytes and H35 hepatoma reporter cell lines; (3) to characterize the dynamics of cytokine signal transduction and the impact of steatosis and different metabolic states on the acute phase response of these cells. This project will be carried out by three distinct research groups, which will interact extensively. Dr. Martin Yarmush (MGH), the Principal Investigator, will oversee the Administrative and Technical Core, and lead the Cell Physiology and Imaging group, which will integrate the technologies developed by the other Pl's and carry out the bulk of the cell physiology experiments. Dr. Mehmet Toner, Director of the Microscale Engineering Facility at the MGH, will lead the Microfabrication and Microfluidics group focusing on the development of microfabricated devices. Dr. Jeffrey Morgan (Brown Univ., Providence, RI), a molecular biologist and expert in gene therapy, will lead the Molecular Biology and Cell Analysis group focusing on reporter cell line development.