Cellular Integration of Information in the Detection and Response to Epithelial Damage

上皮损伤检测和反应中的细胞信息整合

• 批准号: 9893174
• 负责人: M. Shane Hutson
• 金额: $ 21.5万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893174
• 关键词: Address; Adopted; Animals; Area; Behavior; Calcium; Calcium Signaling; Caliber; Cell Culture System; Cell Proliferation; Cell model; Cells; Cellular biology; Communication; Conflict (Psychology); Confusion; Data; Detection; Development; Diffuse; Distal; Drosophila genus; Epithelial; Epithelial Cells; Flare; Frequencies; Gap Junctions; Genetic; Genetic Transcription; Hour; Image; Individual; Knowledge; Lead; Literature; Location; MAPK8 gene; Malignant Neoplasms; Mechanics; Mediating; Membrane; Modeling; Molecular; Organism; Pathology; Pathway interactions; Pattern; Phylogeny; Process; Proliferating; Publishing; Pupa; Radial; Reporting; Role; Running; Side; Signal Transduction; Spatial Behavior; Stereotyping; System; Tissues; Work; Wound Healing; calcium indicator; cell behavior; cell injury; experience; extracellular; genetic manipulation; in vivo; mathematical model; migration; millisecond; programs; receptor; recruit; repaired; response; skills; tool; wound

Project Summary When an epithelial tissue is wounded, the first cellular response is a dramatic increase in cytosolic calcium levels, beginning immediately upon tissue damaged. This calcium increase is observed not just in cells at the wound margin but in large domain of cells surrounding the wound. With the advent of genetically encoded calcium indicators like GCaMP, this wound-induced calcium response has been observed in living organisms across the animal kingdom, yet conflicting mechanisms have been proposed to explain the induction of calcium, such that there is no sense of a conserved fundamental process. Our collaborative team of biophysicists and developmental geneticists recently published work identifying a major underlying obstacle: there are several contemporaneous mechanisms underlying the wound-induced calcium response, which we have been able to tease apart with our combination of highly quantitative approaches and our genetic manipulations. Without understanding the multiple mechanisms inducing calcium responses, it has been impossible to draw parallels across the literature and across wounding models, and it has impossible to fully block calcium responses to analyze the downstream consequences for wound healing. Our analysis tools identified stereotyped calcium responses, with different oscillatory patterns or signatures evident at different radial distances from the wound. We hypothesize that these patterned calcium responses inform the cell about its distance from the wound and determine its downstream cell behaviors. Like the calcium responses, wound-induced cell behaviors and transcriptional identities are patterned according to distance from the wound, with migratory cells and JNK signaling near the margin and proliferative cells and JAK-STAT signaling in a more distal ring. In the first Aim, we investigate the mechanisms of how calcium signaling is initiated in each of these patterns, working both with individual pathways and developing mathematical models for how these calcium patterns are integrated. In the second Aim, we perturb specific aspects of calcium patterns and ask how downstream cell behavior and identity are altered. We are able to achieve these Aims because of our unique collaborative skill-set, and because we have developed an unparalleled wounding model that allows genetic manipulation with high temporal control on one side of the wound only. Because it is internally controlled, comparing the two sides allows precise quantification and detection of even small changes, in both calcium signaling and wound-induced cell behaviors. At the completion of this project, we expect to have generated a high-precision model of how cells detect tissue damage at a distance, and how they interpret this information to select a spatially-appropriate repair program. This fundamental knowledge will be important to many areas of cell biology, to wound-healing studies, and to pathologies like cancer where wound-healing programs are inappropriately activated.

项目摘要 当上皮组织受伤时，第一个细胞反应是胞质的急剧增加 钙水平，立即开始损坏。观察到这种钙增加不仅在 伤口缘的细胞，但在伤口周围的大细胞中。随着遗传的出现 在生活中观察到了诸如GCAMP之类的编码钙指标，这种伤口诱导的钙反应已被观察到 已经提出了整个动物王国的生物体，但已提出相互矛盾的机制来解释 钙的诱导，使得没有一个保守的基本过程。我们的合作团队 生物物理学家和发展遗传学家最近发表的工作，确定了一个主要的潜在障碍： 伤口诱导的钙反应是有几种同时的机制 我们的高度定量方法和我们的遗传的结合都可以逗弄 操纵。不理解诱导钙反应的多种机制，它已经是 不可能在文学和伤害模型中划分相似之处，这是不可能完全的 阻断钙反应以分析伤口愈合的下游后果。 我们的分析工具确定了刻板印象的钙反应，具有不同的振荡模式或 在与伤口不同的径向距离处明显的签名。我们假设这些图案化的钙 响应将其与伤口的距离告知细胞，并确定其下游细胞行为。喜欢 钙反应，伤口引起的细胞行为和转录身份是根据 距伤口的距离，边缘和增殖细胞附近的迁移细胞和JNK信号传导以及 远端环中的jak-stat信号传导。在第一个目标中，我们研究了钙的机制 在每种模式中都启动了信号传导 这些钙模式如何集成的数学模型。在第二个目标中，我们特定于扰动 钙模式的各个方面，并询问下游细胞的行为和身份如何改变。我们能够 之所以实现这些目标，是因为我们具有独特的协作技能，并且因为我们开发了一个 无与伦比的伤害模型，允许在遗传操作的一侧具有高时间控制的基因操纵 仅受伤。因为它是内部控制的，所以比较双方可以精确量化，并且 在钙信号传导和伤口诱导的细胞行为中，甚至都检测小变化。 该项目完成后，我们希望已经生成了一个高精度模型 检测距离的组织损伤，以及他们如何解释此信息以选择适合空间的信息 维修计划。这种基本知识对细胞生物学的许多领域至关重要，对伤口愈合 研究，以及诸如癌症治疗计划之类的病理学，不适当地激活了伤口治疗计划。