Cellular logic of phenotype

表型的细胞逻辑

• 批准号: 7693741
• 负责人: JAMES H EBERWINE
• 金额: $ 78.75万
• 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/7693741
• 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 本应用程序的目标是开发一种可预测的策略， 可重复地改变培养物中原代细胞的表型。分化的细胞类型 在它们的RNA谱中彼此不同（相对以及绝对丰度）， 它们表达的RNA）。我假设，通过转移整个RNA图谱， 使受体细胞的存活依赖于 供体RNA，供体RNA将受体改变为目的表型， 模仿供体细胞表型。这个过程被称为转录组诱导 表型重塑（TIPeR）。有能力将细胞表型在 细胞将提供重要的新的见解机制控制细胞 分化实现这一目标的理论和技术策略正在 在我的实验室里开发的。具体地，使用激光诱导的光转染， （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， 宿主细胞从而将宿主细胞的功能表型改变为宿主细胞的功能表型。 目的表型这部分是由于活动和丰富的具体 由宿主细胞RNA混合物制成的蛋白质。通过发展各种高... 通量定量“组学”水平表型分型技术与TIPeR相结合 程序，预期将辨别表型的“基因组学逻辑”。一个 对这种逻辑的理解将允许随意产生特定的细胞类型。的能力 选择性地和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 表型稳定性，并可能产生新的“个性化药物治疗”。