Analysis and engineering of cell signaling

细胞信号传导分析与工程

• 批准号: 10612418
• 负责人: Casim Sarkar
• 金额: $ 37.36万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612418
• 关键词: Agonist; Amino Acids; Antibody Binding Sites; Area; Biochemical; Biological; Biological Assay; Biology; Cell Surface Receptors; Cell Therapy; Cells; Clinical; Complex; Computer Models; Development; Disease; Engineering; Environment; Experimental Models; Goals; Health; Hot Spot; Human; Individual; Laboratories; Ligands; Maps; Methodology; Methods; Molecular; Outcome; Pharmaceutical Preparations; Process; Proteins; Public Health; Receptor Signaling; Research; Signal Transduction; Specificity; Vision; antagonist; conflict resolution; design; effective therapy; implicit bias; insight; lens; novel; programs; protein protein interaction; receptor; receptor binding; response; tool; transmission process

PROJECT SUMMARY This application builds upon the expertise in my laboratory to develop and apply foundational methods for analyzing and engineering cell signaling in human health. The overall vision of our research program is to create tools that enable more detailed mechanistic understanding of signaling processes that can further facilitate the design of more effective therapies. The broad goals for the next five years are to develop tools that will provide novel insights into the molecular mechanisms of receptor-ligand interactions and to analyze signaling dynamics that arise when conflicting receptor signals are simultaneously activated. In receptor biology, there is an implicit bias in what receptors are biochemically characterized: only those proteins that can be stably expressed in functional form are amenable to established biochemical assays that are used to identify the molecular basis for interaction with ligands. However, there are major classes of receptors that are biologically and clinically important but have proven highly recalcitrant in their heterologous expression. We propose to develop alternative methodologies to overcome conventional expression challenges, facilitate paratope mapping, and enable receptor re-engineering to alter ligand specificity. In ligand biology, there are established high-throughput methods for parsing the contributions of individual amino acids to receptor binding but not corresponding approaches for determining whether amino acids contribute to signal transmission. We propose to develop a methodologies that will enable mapping of a signaling ‘hot spot’ on a ligand and enable rapid engineering of novel agonists and antagonists. Finally, the initiation of signals via cell-surface receptors must be viewed through a holistic lens in order to understand the ultimate effect on cell response. This is most effectively illustrated by considering the simultaneous application of two ligands that, individually, drive mutually exclusive outcomes on a given cell, which is an underappreciated, but common, problem that cells in the body most resolve. Through a combination of quantitative experiments and computational modeling, this cellular ‘conflict resolution’ will be analyzed, providing new insights into the robust functioning of signal transduction networks. Collectively, advances in all of these areas will provide essential new tools for answering fundamental biological questions relevant to human health while also facilitating the design of novel molecular and cellular therapeutics for the treatment of a range of diseases.

项目摘要 该应用程序建立在我的实验室中的专业知识上，以开发和应用基础方法 人类健康中的分析和工程细胞信号传导。我们研究计划的总体愿景是创建 能够对信号过程进行更详细的机械理解的工具，以进一步促进 设计更有效的疗法。接下来五年的广泛目标是开发将提供的工具 对接收器配体相互作用的分子机制的新见解并分析信号传导动力学 当相互冲突的受体信号被简单地激活时，就会产生。在受体生物学中，有一个隐式 对哪种受体的生化表征偏见：只有那些可以稳定表达的蛋白质 功能形式适合已建立的生化测定，用于识别分子基础的基础 与配体的相互作用。但是，有一些主要的接收器在生物学上和临床上是 重要但在其异源表达中被证明是高度顽固的。我们建议开发替代方案 克服常规表达挑战的方法 受体重新设计以改变配体特异性。在配体生物学中，建立了高通量 解析单个氨基酸对受体结合的贡献但不相应的方法 确定氨基酸是否有助于信号传输的方法。我们建议开发一个 可以在配体上绘制信号“热点”并实现快速工程的方法 新颖的激动剂和对手。最后，必须通过细胞表面受体的信号主动性通过 整体镜头以了解对细胞反应的最终影响。这最有效地说明了 考虑到两种配体的简单应用，这些配体单独使用相互排斥的结果 一个给定的细胞，这是一个不足的但常见的问题，即体内细胞最能解决的问题。通过一个 定量实验和计算建模的结合，这种细胞“冲突解决”将是 进行了分析，提供了有关信号转导网络鲁棒功能的新见解。共同 所有这些领域的进步将为回答基本生物学问题提供重要的新工具 与人类健康相关，同时还支持新型分子和细胞疗法的设计 治疗一系列疾病。