Chemical tools for developmental biology

发育生物学化学工具

基本信息

批准号：
10369652
负责人：
JAMES K CHEN
金额：
$ 77.56万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10369652
关键词：
Biological Biology Cell Proliferation Chemicals Complex Congenital Abnormality Detection Development Developmental Biology Directed Molecular Evolution Dynein ATPase Erinaceidae Floor Genetic Goals Image Imaging Device Ions Kinetics Laboratories Lanthanoid Series Elements Lead Life Light Medial Metals Methods Modeling Molecular Morphogenesis Natural regeneration Oligonucleotides Oncogenes Optics Pathway interactions Pattern Photochemistry Physiological Process Property Proteins RNA Research Resolution Signal Pathway Signal Transduction Somites Technology Tissues Whole Organism Zebrafish base cell behavior chemical genetics design genetic manipulation imaging modality in vivo in vivo imaging insight migration notochord novel optogenetics programs response small molecule inhibitor spatiotemporal tool transcription factor tumorigenesis

项目摘要

Tissue morphogenesis and regeneration requires dynamic, robust control of cell proliferation, differentiation, and migration. To coordinate these complex cell behaviors, developmental signaling pathways are actuated with spatiotemporal precision, and their dysregulation can lead to congenital birth defects or tumorigenesis later on in life. While developmental biologists have largely relied on genetic tools to deconstruct these processes, our laboratory has taken a different approach. Over the past five years, we have explored how chemical technologies and high-throughput biology can deepen our understanding of developmental signaling and tissue patterning. Over the past five years, we have invented caged morpholino oligonucleotides that can be activated by light or enzymatically triggered, and we have used these chemical tools to gain new insights into notochord, somite, and medial floor plate development. We established methods for the ultrasensitive imaging of lanthanide-based probes, allowing their unique photophysical properties to be fully exploited for autofluorescence-free in vivo imaging. We have also discovered novel regulators of the Hedgehog pathway, including ARHGAP36, a non-canonical GLI transcription factor activator and oncogene, and the first specific small-molecule inhibitors of cytoplasmic dyneins. We now seek to build upon these accomplishments and push the boundaries of in vivo chemical biology, focusing on the photochemistry of metal ions, synthetic compounds, and proteins. We envision that developmental biology would benefit from new optically controlled technologies that match the cellular resolution and rapid kinetics of patterning mechanisms, including both graded and switch-like responses. Imaging modalities that enable the detection of RNAs, proteins, and their activities at physiological concentrations would be equally transformative. Our research plans for the next five years include the synthesis of photoactivatable morpholinos with greater dynamic and spectral range, directed evolution of optogenetic regulators for key developmental signaling pathways, and design of lanthanide-based tools for imaging biological molecules in whole organisms. We will apply these technologies in zebrafish models, taking advantage of their optical transparency and amenability to chemical and genetic manipulations. Our long-term goal is to use these new experimental capabilities to perturb and observe in vivo biology in unprecedented ways, changing how we study and understand the molecular mechanisms that give rise to multicellular form.

组织形态发生和再生需要动态，强大控制细胞增殖，分化和迁移。为了协调这些复杂的细胞行为，发育信号通路以时空精度驱动，其失调可能导致先天性先天缺陷或肿瘤发生后来生活。发展生物学家主要依靠遗传工具来解构在这些过程中，我们的实验室采取了不同的方法。在过去的五年中，我们探索了化学技术和高通量生物学如何加深我们对发展的理解信号传导和组织模式。在过去的五年中，我们发明了笼中的莫菲利诺寡核苷酸可以通过光或酶触发来激活，我们已经使用这些化学工具来获得新的洞悉脊索，节和内侧板块的发展。我们建立了基于灯笼的探针的超敏感成像，使其独特的光物理特性完全在体内成像中利用无自荧光。我们还发现了刺猬的新型调节器途径，包括ARHGAP36，一种非典型的GLI转录因子激活剂和癌基因，以及第一个特异性小分子抑制剂的细胞质动力蛋白。现在，我们试图以这些成就为基础，并突破体内化学生物学的界限，专注于金属离子，合成化合物和蛋白质的光化学。我们设想发育生物学将受益于与细胞相匹配的新的光学控制技术模式机制的分辨率和快速动力学，包括分级和类似开关的响应。能够检测RNA，蛋白质及其在生理学上的活动的成像方式浓度将同样变化。我们未来五年的研究计划包括具有更大动态和光谱范围的光活化形态学的合成，定向演变用于关键发育信号途径的光遗传学调节器，以及基于灯笼的工具的设计成像整个生物体中的生物分子。我们将在斑马鱼模型中应用这些技术，采用它们的光学透明度以及对化学和遗传操作的合理性的优势。我们的长期目标是使用这些新的实验能力来扰动和观察史无前例的体内生物学改变我们研究和理解产生多细胞形式的分子机制的方式。