Tissue repair, extracellular vesicular biogenesis, and the control of immune responses

组织修复、细胞外囊泡生物发生和免疫反应的控制

• 批准号: 10387452
• 负责人: Brian P Eliceiri
• 金额: $ 21.96万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-20 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10387452
• 关键词: Address; Adoptive Transfer; Biogenesis; Biological; Cells; Environment; Flow Cytometry; Goals; Immune; Immune response; In Vitro; Inflammation; Intercellular Communication Process; Kinetics; Knockout Mice; Mediating; Nucleic Acids; Phase; Population; Proteins; Resolution; Role; Skin repair; Skin wound healing; Therapeutic; Vesicle; diabetic; extracellular; extracellular vesicles; genetic approach; in vivo; insight; mouse model; programs; skin wound; tissue repair; wound closure; wound healing

SUMMARY Resolution in skin wound healing is coordinated by intercellular communication processes that include extracellular vesicles (EVs) containing biologically active protein and nucleic acid. While EV biogenesis has been studied in vitro, systematic approaches to identify their in vivo relevance are limited. Having established that the expression of TBC/RabGAPs controls EV biogenesis and ‘re-programs’ EV content to uncouple the pro-reparative activities of EVs in inflammation, we propose to use a genetic approach to target the secretory machinery implicated in EV biogenesis (i.e. Rabs 5, 11, and 27A) to determine how these targets of TBC/RabGAPs regulate the payload of EVs. We will determine how EVs, and changes in EVs, impact the resolution of inflammation, and gain insight into the role of EVs in wound healing. In Aim 1, we will use vesicle flow cytometry to define EV subsets, followed by adoptive transfer of EVs to determine their capacity to activate specific immune cell populations in diabetic and immunodeficient knockout mouse models that mediate cutaneous wound closure. Aim 2 will study specific Rabs to determine how these Rabs impact EV biogenesis and content. In addition, using the EVs formed in the altered Rab environment, we will establish the mechanism of action and effect of these EVs on the kinetics of resolution of specific immune responses. Together, these studies will establish the relevance of EVs as potential therapeutics and address the over-arching goal of systematically creating therapeutic EVs that are specifically optimized for specific phases of skin repair.

概括 皮肤伤口愈合的解决是通过细胞间通讯过程来协调的 包括含有生物活性蛋白和核酸的细胞外囊泡（EV）。尽管 EV 生物发生已在体外进行了系统性研究，以确定其体内相关性 是有限的。确定 TBC/RabGAP 的表达控制 EV 生物发生并 “重新编程”EV内容以解开炎症中EV的促修复活动，我们 提议使用遗传方法来靶向与 EV 生物发生有关的分泌机制 （即 Rabs 5、11 和 27A）以确定 TBC/RabGAP 的这些目标如何调节有效负载 电动汽车。我们将确定 EV 以及 EV 的变化如何影响炎症的消退， 并深入了解电动汽车在伤口愈合中的作用。在目标 1 中，我们将使用囊泡流 通过细胞计数来定义 EV 子集，然后通过 EV 的过继转移来确定其容量 激活糖尿病和免疫缺陷基因敲除小鼠的特定免疫细胞群 介导皮肤伤口闭合的模型。目标 2 将研究特定的 Rabs 以确定如何 这些 Rabs 影响 EV 的生物发生和内容。此外，使用在改变后形成的电动汽车 Rab 环境中，我们将建立这些 EV 对 特异性免疫反应的解决动力学。这些研究将共同​​建立 EV 作为潜在疗法的相关性，并解决系统性的总体目标 创建专门针对皮肤修复特定阶段进行优化的治疗性 EV。