CAREER: Signal processing in the Erk pathway: dynamics, decisions, and development

职业：Erk 通路中的信号处理：动态、决策和发展

• 批准号: 1750663
• 负责人: Jared Toettcher
• 金额: $ 130.83万
• 依托单位: Princeton University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1750663&HistoricalAwards=false
• 关键词: CAREER; Signal; processing; Erk; pathway

One of the most exciting and provocative ideas in modern biology is the notion that cells can be "reprogrammed" and engineered to perform a desired function. Yet the ability to manipulate cellular responses is still quite limited in two key ways. First, researchers have not deciphered the instructions used by cells to adopt a particular response. Second, even if researchers perfectly knew which instructions to provide, they lack the tools to deliver them to a particular cell of interest, at a precise time, without affecting its neighbors. This research will address both of these key challenges by studying a protein, Erk, that plays a fundamental role in organizing cell decisions in all multicellular organisms. This project will rely on a combination of cutting-edge tools: live-cell biosensors to measure natural Erk responses and engineered, light-sensitive proteins to alter Erk activity at precise times and locations in a tissue. This comprehensive understanding of how Erk activity patterns control responses at the cell, tissues, and whole organism scales and how one can alter those patterns to control cell fate. These studies will be integrated with an education plan to train the next generation of quantitative biologists and to foster early exposure to research for historically underrepresented students.The long-term goal of the researcher's lab is to address two fundamental problems in cellular systems biology. How are a small number of highly conserved signaling pathways repurposed in different cell types and organisms to encode information about the external world? Which features of the signals sent by these pathways are sufficient to specify cell-, tissue-, and organism-level phenotypes? Fulfilling these two aims will help to bridge the gap from a static genotype to complex multicellular phenotypes and define the computations that are carried out by the groups of proteins that work together to process cellular information. The research plan is focused on an extensively studied signaling pathway, the Erk pathway, which plays essential roles across eukaryotic organisms. Although the individual proteins comprising the pathway have been thoroughly studied by genetic and biochemical approaches, live-cell Erk biosensors have recently revealed exquisite spatiotemporal patterns of activity: oscillations in individual cells, propagating waves across tissues, and a fast succession of spatial patterns during embryogenesis. The work will dissect these ornate patterns using a combination of quantitative measurements and highly controlled optogenetic inputs that can be used to activate Erk signaling in any spatial or temporal pattern. In particular, the research seeks to: (1) dissect the mechanism that generates pulses of Erk activity in a laboratory model of the epidermis; (2) determine how Erk dynamics control gene expression and cell fate in epidermal cells; and (3) unravel how Erk signaling affects tissue morphogenesis in the developing fly embryo, where exquisite signaling patterns appear over time and in 3D space throughout the organism. The research is complemented by an education plan directed at broadening the early-stage research opportunities available to undergraduates, particularly historically underrepresented students, and at disseminating the tools of quantitative biology and optogenetics to the next generation of cell and developmental biology trainees.The Project is jointly funded by Molecular and Cellular Biosciences and Integrative Organismal Systems, with additional support provided by the Directorate's Rule of Life Venture Fund.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

现代生物学中最令人兴奋和最具煽动性的想法之一是细胞可以被“重新编程”并被设计为执行所需功能的概念。然而，操纵细胞反应的能力在两个关键方面仍然非常有限。首先，研究人员还没有破译细胞采用特定反应的指令。其次，即使研究人员完全知道提供哪些指令，他们也缺乏工具来在精确的时间将它们传递到特定的感兴趣的细胞，而不会影响它的邻居。这项研究将通过研究一种名为Erk的蛋白质来解决这两个关键挑战，Erk在所有多细胞生物中组织细胞决策方面发挥着重要作用。该项目将依赖于尖端工具的组合：活细胞生物传感器来测量天然Erk反应，以及工程化的光敏蛋白质来在组织中的精确时间和位置改变Erk活性。这是对Erk活动模式如何控制细胞，组织和整个生物体规模的反应以及如何改变这些模式以控制细胞命运的全面理解。这些研究将与教育计划相结合，以培养下一代定量生物学家，并促进历史上代表性不足的学生早期接触研究。研究人员实验室的长期目标是解决细胞系统生物学中的两个基本问题。在不同的细胞类型和生物体中，少数高度保守的信号通路是如何被重新利用来编码外部世界的信息的？由这些通路发送的信号的哪些特征足以指定细胞、组织和生物体水平的表型？ 实现这两个目标将有助于弥合从静态基因型到复杂多细胞表型的差距，并定义由蛋白质组共同处理细胞信息所进行的计算。该研究计划的重点是广泛研究的信号通路，Erk通路，它在真核生物中起着重要作用。虽然个别蛋白质组成的途径已被彻底研究的遗传和生物化学方法，活细胞Erk生物传感器最近揭示了精致的时空模式的活动：振荡在个别细胞，传播波在组织中，和一个快速连续的空间模式在胚胎发生。这项工作将使用定量测量和高度控制的光遗传学输入的组合来剖析这些华丽的模式，这些输入可用于在任何空间或时间模式中激活Erk信号传导。特别是，该研究旨在：（1）剖析在表皮的实验室模型中产生Erk活性脉冲的机制;（2）确定Erk动力学如何控制表皮细胞中的基因表达和细胞命运;（3）解开Erk信号传导如何影响发育中的苍蝇胚胎中的组织形态发生，其中精致的信号模式随着时间的推移和整个生物体的3D空间出现。该研究由一项教育计划补充，旨在扩大本科生，特别是历史上代表性不足的学生的早期研究机会，并向下一代细胞和发育生物学学员传播定量生物学和光遗传学工具。该项目由分子和细胞生物科学和整合有机系统联合资助，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Engineering combinatorial and dynamic decoders using synthetic immediate-early genes

使用合成的早期基因设计组合和动态解码器

• DOI: 10.1038/s42003-020-01171-1
• 发表时间: 2020
• 期刊: Communications Biology
• 影响因子: 5.9
• 作者: Ravindran, Pavithran T.;Wilson, Maxwell Z.;Jena, Siddhartha G.;Toettcher, Jared E.
• 通讯作者: Toettcher, Jared E.

Optogenetic control of the Bicoid morphogen reveals fast and slow modes of gap gene regulation.

• DOI: 10.1016/j.celrep.2022.110543
• 发表时间: 2022-03-22
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Singh, Anand P.;Wu, Ping;Ryabichko, Sergey;Raimundo, Joao;Swan, Michael;Wieschaus, Eric;Gregor, Thomas;Toettcher, Jared E.
• 通讯作者: Toettcher, Jared E.

Optimizing photoswitchable MEK

• DOI: 10.1073/pnas.1912320116
• 发表时间: 2019-12
• 期刊: Proceedings of the National Academy of Sciences
• 作者: Aleena L. Patel;E. Yeung;S. McGuire;Andrew Y Wu;Jared E. Toettcher;R. Burdine;S. Shvartsman

Putting in the Erk: Growth factor signaling and mesoderm morphogenesis

放入 Erk：生长因子信号传导和中胚层形态发生

• 发表时间: 2022
• 期刊: Current Topics in Developmental Biology
• 作者: McFann, Sarah E.;Shvartsman, Stanislav Y.;Toettcher, Jared E.
• 通讯作者: Toettcher, Jared E.

Signaling Dynamics Control Cell Fate in the Early Drosophila Embryo

• DOI: 10.1016/j.devcel.2019.01.009
• 发表时间: 2019-02-11
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Johnson, Heath E.;Toettcher, Jared E.
• 通讯作者: Toettcher, Jared E.