Optogenetic dissection of Erk signal interpretation in early embryogenesis

早期胚胎发生中 Erk 信号解释的光遗传学解析

基本信息

批准号：
10679294
负责人：
Emily Kolenbrander Ho
金额：
$ 6.95万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679294
关键词：
Address Affect Anterior Biosensor Cancer Etiology Cells Cellular biology Complex Decision Making Development Developmental Biology Diffusion Disease Dissection Dose Drosophila genus Drosophila melanogaster Ectoderm Embryo Embryonic Development Endoderm Event Extracellular Signal Regulated Kinases Gene Expression Gene Expression Profile Genes Genetic Genetic Transcription Genomic approach Genomics Goals Growth Head Human Development Laboratories Light Malignant Neoplasms Measures Mediating Microscopy Midgut Molecular Mutation Output Pathway interactions Pattern Physiologic pulse Play Positioning Attribute Proteins Reporter Research Role Signal Pathway Signal Transduction Spatial Distribution Specific qualifier value Structure Tail Time Tissue Model Tissues Training Travel Work career gastrulation genetic manipulation imaging approach in vivo in vivo Model innovation optogenetics programs response tool transcription factor transcriptome sequencing tumor progression wound healing

项目摘要

PROJECT SUMMARY/ABSTRACT Signaling pathways pattern cell fate choices in the developing embryo, but how do only a limited number of pathways control the complex events of development? Spatial signaling gradients as well as temporal signaling dynamics are known to be important for directing multiple fates with the same pathway, yet the mechanisms cells use to interpret these signaling events remains largely uncharacterized. One pathway which plays diverse roles throughout embryogenesis, wound healing, and cancer progression is the highly conserved extracellular signal-regulated kinase (Erk) signaling network. The early Drosophila embryo is a classic in vivo model of tissue patterning by Erk signaling: a spatial gradient of Erk signal at the poles specifies anterior and posterior structure and dictates endoderm versus ectoderm fate decisions. Although these patterns are well-defined, recent work in the Toettcher laboratory using an optogenetic tool to manipulate the Erk signal in vivo with exquisite spatial and temporal precision has revealed major open questions about the mechanisms by which Erk signals are interpreted. Here, I propose to define the mechanisms by which the Drosophila embryo interprets differences in Erk dose to create spatial patterns of gene expression and make decisions about cell fate. To address this question, I will use a combination of cutting-edge optogenetic tools, live-cell biosensors, genomic analysis, and classical genetic perturbations. In Aim 1, I will examine how spatial patterns of gene expression are robust to major corruption of the Erk gradient. In Aim 2, I will explore how only two transcription factors can control multiple distinct fates over a wide range of Erk signal strength. Altogether, using precision tools for manipulating signaling inputs and quantifying expression outputs, this proposal will reveal how Erk signals are measured and interpreted by downstream genes, addressing a fundamental question in developmental biology which will surely have relevance for other signaling pathways and cellular contexts for Erk signaling. Through this research and the accompanying training plan I will obtain exceptional training in optogenetics, advanced microscopy, and genomics, positioning me for an impactful career at the interface of cell and developmental biology.

项目概要/摘要信号通路决定了发育中胚胎的细胞命运选择，但只有有限数量的细胞如何做出选择？途径控制发育的复杂事件？空间信号梯度以及时间信号众所周知，动力学对于通过同一途径引导多种命运非常重要，但其机制用于解释这些信号事件的细胞在很大程度上仍然未知。一条道路发挥多种作用高度保守的细胞外蛋白在整个胚胎发生、伤口愈合和癌症进展中发挥着重要作用信号调节激酶（Erk）信号网络。早期果蝇胚胎是经典的体内模型通过 Erk 信号形成组织图案：两极 Erk 信号的空间梯度指定前部和后部结构并决定内胚层与外胚层的命运决定。尽管这些模式定义明确， Toettcher 实验室最近的工作使用光遗传学工具来操纵体内的 Erk 信号精确的空间和时间精度揭示了有关机制的重大悬而未决的问题 Erk 信号被解释。在这里，我建议定义果蝇胚胎的机制解释 Erk 剂量的差异以创建基因表达的空间模式并做出有关细胞的决策命运。为了解决这个问题，我将结合使用尖端光遗传学工具、活细胞生物传感器、基因组分析和经典的遗传扰动。在目标 1 中，我将研究基因的空间模式如何表达式对 Erk 梯度的严重损坏具有鲁棒性。在目标 2 中，我将探讨如何仅使用两个转录因素可以在广泛的 Erk 信号强度范围内控制多种不同的命运。总而言之，使用精度操纵信号输入和量化表达输出的工具，该提案将揭示 Erk 如何信号由下游基因测量和解释，解决了一个基本问题发育生物学肯定与其他信号通路和细胞环境相关埃尔克发出信号。通过这项研究和随附的培训计划，我将获得以下方面的特殊培训：光遗传学、先进显微镜学和基因组学，使我能够在以下领域从事有影响力的职业生涯细胞和发育生物学。