Cellular logic of phenotype

表型的细胞逻辑

• 批准号: 8306904
• 负责人: JAMES H EBERWINE
• 金额: $ 77.96万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8306904
• 关键词: Cell Differentiation process; Cell Polarity; Cell Survival; Cells; Coupled; Data; Destinations; Diffuse; Gene Expression Profile; Genetic Transcription; Genomics; Goals; Laboratories; Lasers; Light; Logic; Memory; Phenotype; Population; Procedures; Proteins; RNA; Relative (related person); Technology; Therapeutic; cell type; insight; novel; theories

The Cellular Logic of Phenotype The goal of this application is to develop a strategy for predictably and reproducibly altering the phenotype of primary cells in culture. Differentiated cell types differ from each other in their RNA profiles (relative as well as absolute abundances of the RNAs they express). I hypothesize that, by the transferring entire RNA profiles from donor to recipient cells in a way that makes the recipient cells' survival dependent on donor RNA, the donor RNA will change the recipient into a destination phenotype that mimics the donor cell phenotype. This procedure is called Transcriptome Induced Phenotype Remodeling (TIPeR). Having the ability to transfer cell phenotypes between cells would provide important new insights into mechanisms controlling cell differentiation. The theory and technical strategies to accomplish this are being developed in my laboratory. Specifically, using laser light induced phototransfection (developed in my lab), we transiently produce pores in the host primary cell, through which RNA populations (in which RNA species and abundances are carefully controlled), can diffuse. Preliminary data shows that donor cell RNA populations carry "memory functions" in that, donor RNA can induce long-term changes in genomic transcription of the host cells thereby changing the functional phenotype of the host cells to that of the destination phenotype. This is due in part to the activity and abundances of the specific proteins made from the host cell RNA mixture. Through developing various high- throughput quantitative "Omics" level phenotyping technologies coupled with the TIPeR procedure it is anticipated that the "genomics logic" of phenotype will be discerned. An understanding of this logic will permit the creation of specific cell types at will. The ability to selectively and rationally create cellular phenotypes promises to provide important insights into the fundamental mechanisms underlying cellular polarity, functioning and phenotype stability and may yield novel "individualized medicinal therapeutics".

表型的细胞逻辑 该应用程序的目标是制定一种可预测和可预测的策略 可重复地改变培养物中原代细胞的表型。分化的细胞类型 它们的 RNA 谱彼此不同（RNA 的相对丰度和绝对丰度） 它们表达的RNA）。我假设，通过从 受体细胞的供体使受体细胞的存活依赖于 供体 RNA，供体 RNA 会将受体改变为目标表型 模仿供体细胞表型。这个过程称为转录组诱导 表型重塑（TIPeR）。具有在不同细胞之间转移细胞表型的能力 细胞将为控制细胞的机制提供重要的新见解 差异化。实现这一目标的理论和技术策略正在研究中 在我的实验室开发的。具体来说，使用激光诱导光转染 （在我的实验室开发），我们在宿主原代细胞中瞬时产生孔，通过 哪些 RNA 群体（其中 RNA 种类和丰度受到仔细控制）， 可以扩散。初步数据显示供体细胞RNA群体携带“记忆” 功能”，因为供体 RNA 可以诱导基因组转录的长期变化 从而将宿主细胞的功能表型改变为 目的地表型。这部分是由于特定的活性和丰度 由宿主细胞 RNA 混合物制成的蛋白质。通过开发各种高 通量定量“组学”水平表型技术与 TIPeR 相结合 预计表型的“基因组逻辑”将被辨别。一个 理解这一逻辑将允许随意创建特定的细胞类型。能力 有选择地、合理地创造细胞表型有望提供重要的 深入了解细胞极性、功能和功能的基本机制 表型稳定性并可能产生新的“个体化药物疗法”。