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

基本信息

批准号：
10413234
负责人：
Brian P Eliceiri
金额：
$ 37.96万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/10413234
关键词：
Address Adoptive Transfer Affect Animal Model Autologous Biogenesis Biological Biological Response Modifiers Cells Complement Environment Excision Family Flow Cytometry Gene Delivery Goals Human Immune Immune response Immunocompetent Immunodeficient Mouse Impaired wound healing Implant In Vitro Inflammation Intercellular Communication Process Kinetics Knockout Mice Liquid substance Location Mediating Mediator of activation protein Monomeric GTP-Binding Proteins Mus Mutant Strains Mice Nucleic Acids Phase Population Porifera Process Proteins Resolution Role Signal Transduction Site Skin Skin injury Skin repair Skin wound healing Source Surface Testing Therapeutic Thick Time Vesicle Xenograft procedure cell type diabetic extracellular extracellular vesicles genetic approach immune activation in vivo insight intercellular communication interdisciplinary approach knock-down member mouse model overexpression patient derived xenograft model programs recruit repaired skin wound subcutaneous tissue repair vesicular release wound wound bed wound closure wound healing

项目摘要

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.

概括皮肤伤口愈合中的分辨率通过细胞间沟通过程协调包括含有生物活性蛋白和核酸的细胞外蔬菜（EV）。尽管 EV生物发生已在体外研究，系统地识别其体内相关性有限。确定TBC/Rabgap的表达控制EV生物发生和 “重新编程”的EV含量以揭示电动汽车在炎症中的依rever徒活动，我们提出使用遗传方法来针对EV生物发生实施的秘密机械的提议（即RABS 5、11和27A）确定这些目标如何调节有效载荷电动汽车。我们将确定电动汽车和电动汽车的变化如何影响炎症的分辨率，并深入了解电动汽车在伤口愈合中的作用。在AIM 1中，我们将使用囊泡流定义EV子集的细胞仪，然后自适应转移EV以确定其容量激活糖尿病和免疫缺陷敲除小鼠中特定的免疫细胞群体介导皮肤伤口闭合的模型。 AIM 2将研究特定的RAB，以确定如何这些RAB会影响EV生物发生和含量。另外，使用更改中形成的电动汽车 Rab环境，我们将建立这些电动汽车对这些电动汽车的作用机理解决特定免疫反应的动力学。这些研究将共同确定电动汽车与潜在的治疗学的相关性，并解决系统的超级目标创建针对皮肤修复特定阶段专门优化的治疗性电动汽车。