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Extracellular optogenetic control in the study of epithelial collective cell signaling

上皮集体细胞信号传导研究中的细胞外光遗传学控制

基本信息

批准号：
10213975
负责人：
Agnieszka Gil
金额：
$ 1.09万
依托单位：
PRINCETON 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：
2018-06-01 至 2020-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10213975
关键词：
Affect Animals Bacteriophage M13 Behavior Binding Biological Biological Assay Biological Models Biological Process Biosensor Cell Communication Cell Culture Techniques Cell Differentiation process Cell Proliferation Cell physiology Cells Chimeric Proteins Complex Development Dissociation EGF gene Engineering Ensure Environment Epidermal Growth Factor Epidermal Growth Factor Receptor Epidermis Epithelial Epithelium Equilibrium Event Extracellular Protein Future Generations Genetic Genome engineering Growth Growth Factor Growth Factor Receptors In Vitro Individual Laboratories Lead Libraries Life Ligand Binding Ligands Light Location Mammals Maps Measures Mitogen-Activated Protein Kinases Modeling Modification Monitor Movement Mus Organ Organism Phage Display Phase Phase Transition Phosphotransferases Photosensitivity Physiologic pulse Process Property Protein Tyrosine Kinase Proteins Reaction Receptor Protein-Tyrosine Kinases Receptor Signaling Regulation Reporting Role Scientist Signal Pathway Signal Transduction Site Skin Skin Cancer Structure Supervision System Technology Testing Therapeutic Tissues Travel Work base cell growth cell motility cell type experimental study extracellular gain of function high throughput technology in vivo inhibitor/antagonist interest keratinocyte nanobodies novel novel therapeutics optogenetics prevent protein aggregation receptor repaired tissue repair tool transmission process tumor tumorigenesis wound healing

项目摘要

Project Summary Cell signaling in multicellular tissues is extraordinarily complex, ensuring that cells coordinate their proliferation, survival and movement for proper tissue function. One model context for multicellular signaling is the skin, an important organ that remains highly proliferative throughout the life of the animal and must balance this constant proliferation and repair with tight regulation to prevent tumorigenesis. Recent work using a live-cell Erk biosensor has revealed complex tissue-level signaling dynamics in epidermal tissues, both in cell culture and in living mammals. Taken together with previous studies showing that Erk signaling is both necessary and sufficient for wound healing and skin cell proliferation suggests that cells share crucial information through the local transmission of complex, dynamic growth factor signals. The current proposal aims to shed new light on growth factor signaling by developing optogenetic tools to activate and inhibit growth factor receptors with high precision. Current optogenetic approaches modulate signaling dynamics by altering intracellular protein activity, which requires substantial genome engineering and thus has limited use in vivo or as a therapeutic approach. In contrast, we have devised two new photosensitive tools to modulate cell signaling from the extracellular environment. We will engineer light-controllable ligands and inhibitors against receptor tyrosine kinases, to enable spatial and temporal control over the signals transmitted between cells. We will then use these tools to dissect complex Erk dynamics in primary mouse keratinocytes, a model system for the mammalian epidermis. Achieving a better understanding of the complex signals transmitted between cells will enable future studies to dissect their roles in wound healing and skin cancer. Additionally, the development of new optogenetic systems for manipulating extracellular binding events could lead to highly generalizable tools with the potential to control many other cellular processes.

项目概要多细胞组织中的细胞信号传导极其复杂，确保细胞协调其增殖、存活和运动以获得适当的组织功能。多细胞信号传导的一种模型背景是皮肤是一个重要的器官，在动物的整个生命周期中都保持高度增殖性，并且必须保持平衡这种不断的增殖和修复受到严格的调节，以防止肿瘤发生。最近使用活细胞的工作 Erk 生物传感器揭示了细胞培养物中表皮组织复杂的组织水平信号动力学以及在活的哺乳动物中。与之前的研究一起表明，Erk 信号传导既是必要的，也是足以促进伤口愈合和皮肤细胞增殖表明细胞通过复杂的动态生长因子信号的本地传输。目前的提案旨在通过开发光遗传学工具来阐明生长因子信号传导高精度激活和抑制生长因子受体。当前的光遗传学方法可以调节通过改变细胞内蛋白质活性来改变信号动力学，这需要大量的基因组工程和因此在体内或作为治疗方法的用途有限。相比之下，我们设计了两种新的感光材料调节来自细胞外环境的细胞信号传导的工具。我们将设计光控配体和受体酪氨酸激酶抑制剂，以实现对信号的空间和时间控制细胞间传递。然后我们将使用这些工具来剖析原代小鼠中复杂的 Erk 动力学角质形成细胞，哺乳动物表皮的模型系统。更好地理解复杂性细胞之间传递的信号将使未来的研究能够剖析它们在伤口愈合和皮肤中的作用癌症。此外，开发用于操纵细胞外结合事件的新光遗传学系统可能会产生高度通用的工具，并有可能控制许多其他细胞过程。