Endothelial cell exosomes and fibroblast function

内皮细胞外泌体和成纤维细胞功能

• 批准号: 10092812
• 负责人: Anna Salapatas
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092812
• 关键词: Affect; Appearance; Architecture; Biological Assay; Biological Process; Blood Vessels; Blood capillaries; Caliber; Cell Communication; Cell Cycle; Cell Proliferation; Cells; Cicatrix; Collagen; Communication; Complex; Custom; Data; Deposition; Dermal; Development; Disease; Elastin; Endothelial Cells; Exposure to; Extracellular Matrix; Extracellular Structure; Fibroblasts; Fibrosis; Flow Cytometry; Gene Expression; Gene Targeting; Generations; Genomics; Goals; Growth; Human; Impairment; In Vitro; Infiltration; Inflammatory; Investigation; Laboratories; Lead; Liver; Lung; Measures; Mediating; Medical; Methods; MicroRNAs; Microscopy; Myofibroblast; Nutrient; Outcome; Oxygen; Pathway Analysis; Pathway interactions; Phase; Phenotype; Physiological; Polymerase Chain Reaction; Process; Protein Analysis; Research; Research Personnel; Research Training; Resources; Sirius Red F3B; Site; Skin; Small RNA; Smooth Muscle Actin Staining Method; Stains; Time; Tissues; Training; angiogenesis; autocrine; bioinformatics tool; career development; cell motility; cell type; craniofacial; design; differential expression; exosome; extracellular vesicles; healing; improved; in vivo; insight; intradermal injection; migration; mouse model; novel therapeutics; paracrine; psychologic; repaired; response; second harmonic; skeletal; tissue repair; tool; transcriptome sequencing; transcriptomics; two-photon; uptake; wound; wound bed; wound healing

PROJECT SUMMARY/ABSTRACT This application proposes a customized research training plan designed to promote the development of the applicant into an independent investigator. The plan includes advanced training in laboratory experimentation, along with tailored professional and career development opportunities. The training plan is supported by the outstanding availability of local and institutional resources at UIC. The proposed research will examine cellular communication mechanisms that control scar formation, a common result of the healing response. In most tissues, the end result of tissue repair is a fibrous scar containing altered amounts and structure of the extracellular matrix components. Scarring and fibrosis occurs in numerous tissues and can create serious functional problems such as limited mobility, restricted skeletal growth, and weakened tissue strength that may lead to wound dehiscence. In the craniofacial region, scarring can impair both functionality as well as appearance, leading to a variety of physiologic and psychologic problems. Robust angiogenesis, a prominent feature of wound repair, includes the creation and then pruning of vessels. Angiogenesis is associated with increased fibrosis in numerous tissues including skin, lung, and liver. The goal of the proposed studies is to examine how the endothelial cells that arise during wound angiogenesis communicate with surrounding fibroblasts and how they influence their function and subsequent collagen deposition. One way that endothelial cells might communicate with fibroblasts is via small extracellular vesicles, called exosomes. The research plan utilizes traditional wound healing assays, methods for exosome purification and application, and small RNA and RNA sequencing tools to study exosome-mediated communication between these two cell types and to identify the resultant phenotypic changes in fibroblast activity. The central hypothesis of this research is that exosomes secreted from endothelial cells influence the phenotype of fibroblasts during wound healing and fibrosis. Our long-term goal is to understand how communication between endothelial cells and fibroblasts affects the scarring phenotype. Aim 1 will examine the effects of endothelial cell exosomes on fibroblast activity and fibrosis in vitro and in vivo. In vitro wound healing assays will be employed, and fibroblasts exposed to endothelial cell exosomes will be assessed for changes in migration, cell cycle phase, proliferation, and gene expression. An in vivo mouse model will be used to assess fibrosis and collagen architecture after intradermal injection on endothelial cell exosomes. Aim 2 will utilize bioinformatics tools to identify and characterize the specific miRNA cargo of endothelial cell exosomes that is mostly likely to impact fibroblast function at sites of scarring and fibrosis. Together, the Aims will lead to a better understanding of the mechanisms by which endothelial cells might modulate fibroblast function, and may lead to the development of novel therapeutics to treat fibrotic diseases.

项目总结/摘要 本申请提出了一个定制的研究培训计划，旨在促进发展的 申请人成为独立调查员。该计划包括实验室实验的高级培训， 沿着量身定制的专业和职业发展机会。培训计划由 在UIC的地方和机构资源的出色可用性。这项拟议中的研究将检查细胞 控制疤痕形成的通信机制，愈合反应的常见结果。在大多数 组织，组织修复的最终结果是含有改变的量和结构的纤维瘢痕， 细胞外基质成分。瘢痕和纤维化发生在许多组织中， 功能性问题，如活动受限、骨骼生长受限和组织强度减弱， 导致伤口裂开。在颅面区域，疤痕可以损害功能以及 外观，导致各种生理和心理问题。强大的血管生成， 伤口修复的特征，包括血管的创建和修剪。血管生成与 包括皮肤、肺和肝在内的许多组织中的纤维化增加。拟议研究的目标是 研究在伤口血管生成过程中产生的内皮细胞如何与周围环境沟通， 成纤维细胞以及它们如何影响它们的功能和随后的胶原沉积。内皮细胞 细胞与成纤维细胞之间的联系是通过小的细胞外囊泡，称为外泌体。研究 计划利用传统的伤口愈合试验，外泌体纯化和应用的方法，以及小 RNA和RNA测序工具来研究这两种细胞类型之间的外泌体介导的通讯， 以鉴定成纤维细胞活性的表型变化。这项研究的中心假设是， 内皮细胞分泌的外来体影响伤口愈合过程中成纤维细胞的表型， 纤维化我们的长期目标是了解内皮细胞和成纤维细胞之间的通讯 影响瘢痕形成表型。目的1将检测内皮细胞外泌体对成纤维细胞活性的影响 和纤维化。将采用体外伤口愈合测定，并且将暴露于 将评估内皮细胞外泌体在迁移、细胞周期、增殖和基因表达方面的变化。 表情体内小鼠模型将用于评估皮内注射后的纤维化和胶原结构。 注射内皮细胞外泌体。目标2将利用生物信息学工具， 内皮细胞外泌体的特异性miRNA货物，其最可能影响成纤维细胞功能， 疤痕和纤维化。这些目标将有助于更好地理解 内皮细胞可能调节成纤维细胞功能，并可能导致开发新的治疗方法， 治疗纤维化疾病。