Intercellular communication and cell regulation in airway epithelial ensembles in regeneration and disease

再生和疾病中气道上皮群的细胞间通讯和细胞调节

• 批准号: 9770564
• 负责人: JAYARAJ RAJAGOPAL
• 金额: $ 62.81万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9770564
• 关键词: Ablation; Antibodies; Architecture; Asthma; Basal Cell; Behavior; Biological Models; Bronchitis; Cell Differentiation process; Cells; Chronic Obstructive Airway Disease; Clinical; Complement; Cystic Fibrosis; Diphtheria Toxin; Disease; Epithelial; Epithelial Cells; Epithelium; Feedback; Generations; Genetic; Heterogeneity; Infection; Injury; Intervention; Ligands; Lung diseases; Mediating; Metaplasia; Mucous body substance; Natural regeneration; Normal tissue morphology; Pathway interactions; Patients; Physiological; Population; Process; Publishing; Reagent; Receptor Signaling; Recovery; Regenerative response; Regulation; Role; Secretory Cell; Signal Transduction; Source; Stem cells; Testing; Tissues; Work; airway epithelium; base; body system; cell type; feeding; injured; intercellular communication; notch protein; novel; prevent; respiratory; response; response to injury; stem cell population; tool

Project Summary: In published work, we used diphtheria toxin-induced genetic cellular ablation to understand how normal tissue architecture is restored after the loss of a single airway epithelial cell type. We discovered 4 new phenomenon not previously described (1) that a fully mature vertebrate cell can dedifferentiate into a stem cell when stem cells are ablated, (2) that the ablation of the most terminally differentiated cell type in the airway epithelium does not engender a regenerative response suggesting that there is no feedback injury signal emanating from the ciliated cell to guide stem cell-based regeneration, (3) that basal cells are not merely sources of new cells, but that they send feed-forward signals to secretory epithelial cells to actively orchestrate whole tissue behavior, and finally (4) that the basal cells are not a homogeneous population of stem cells. We further identified Notch signaling as the mechanistic basis for both the novel stem cell feed-forward signaling mechanism and as the basis of basal cell heterogeneity. In this application, we propose to continue our use of precise genetic cellular ablation studies to interrogate the regulatory circuitry of the airway epithelium, and to define how Notch signaling orchestrates the behavior of specific populations of airway epithelial cells. We now propose to directly extend our prior work examining the steady state airway epithelium, and deploy our model systems to study physiologically relevant- injury. We have three general hypotheses that we intend to verify or refute using our now well-developed tools for cellular ablation and cell type-specific Notch signaling modulation. First, we hypothesize that the basal stem cell is a central actor during the regenerative response, and that it actively makes use of Notch ligands to fundamentally regulate the process of mucous metaplasia in multiple cell types. Thus, rather than simply serving to supply new cells to replace injured ones, stem cells are hypothesized to orchestrate whole tissue behaviors. Secondly, we hypothesize that distinct components of both the Notch signal sending (Notch ligands) and receiving (Notch receptors) pathways are modulated differentially in response to differing degrees and types of injury. Thirdly we postulate that a feedback signal regulating regeneration must be present to complement the novel feed forward signaling mechanism that we have recently demonstrated. Furthermore, since this signal seems absent from ciliated cells, we hypothesize that the feedback signal must emanate from the secretory cells. This work has taken on added importance, as antibody reagents for Notch modulation are now being considered as clinical interventions.

项目摘要： 在已发表的工作中，我们使用白喉毒素诱导的遗传细胞消融来了解 在单一气道上皮细胞类型丧失后恢复正常的组织结构。我们 发现了4个以前没有描述过的新现象（1）完全成熟的脊椎动物细胞 当干细胞被消融时，可以去分化为干细胞，（2）大多数干细胞的消融 气道上皮中的终末分化细胞类型不会产生再生的 这表明纤毛细胞没有发出反馈损伤信号， 引导基于干细胞的再生，（3）基底细胞不仅是新细胞的来源， 它们向分泌上皮细胞发送前馈信号， 组织行为，最后（4）基底细胞不是干细胞的同质群体 细胞我们进一步确定了Notch信号传导作为新的干细胞 前馈信号传导机制，并作为基础细胞异质性。 在本申请中，我们建议继续使用精确的遗传细胞消融研究， 询问气道上皮的调节回路，并确定Notch信号传导如何 协调气道上皮细胞的特定群体的行为。我们现建议 直接扩展我们先前的工作，检查稳态气道上皮，并部署我们的 研究生理相关的模型系统-损伤。我们有三个基本假设， 我们打算使用我们现在开发良好的细胞消融和细胞消融工具来验证或反驳 特定类型的Notch信号调制。首先，我们假设基底干细胞是一种 在再生反应过程中的中央演员，它积极利用Notch配体， 从根本上调节多种细胞类型的粘液化生过程。因此， 干细胞不仅仅是提供新的细胞来取代受伤的细胞， 协调整个组织的行为。其次，我们假设， Notch信号发送（Notch配体）和接收（Notch受体）途径是 对不同程度和类型的损伤有不同的调节。第三，我们假设 必须存在调节再生的反馈信号以补充新的进料 我们最近展示的前向信号机制。此外，由于这 信号似乎缺席纤毛细胞，我们假设，反馈信号必须发出 从分泌细胞。这项工作具有额外的重要性，作为抗体试剂， Notch调制现在被认为是临床干预措施。