Endothelial cell exosomes and fibroblast function

项目摘要/摘要 这项申请提出了一个定制的研究培训计划，旨在促进 申请人成为一名独立的调查员。该计划包括实验室实验方面的高级培训， 以及量身定做的职业和职业发展机会。该培训计划由 UIC的当地和机构资源的突出可得性。这项拟议的研究将检查细胞 控制疤痕形成的通讯机制，这是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纤维瘢痕，包含改变的数量和结构的 细胞外基质成分。疤痕形成和纤维化发生在许多组织中，可以造成严重的 功能问题，如行动能力受限，骨骼生长受限，组织力量减弱，可能 导致伤口裂开。在头面部，疤痕既会损害功能，也会损害 外貌，导致各种生理和心理问题。强大的血管生成，一个突出的 伤口修复的特点，包括创建和修剪血管。血管生成与 包括皮肤、肺和肝脏在内的许多组织中的纤维化增加。拟议研究的目标是 检查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。一种方法是内皮细胞 细胞与成纤维细胞之间的交流可能是通过称为外体的细胞外小泡实现的。这项研究 PLAN使用传统的伤口愈合分析、外切体纯化和应用的方法，以及小的 RNA和RNA测序工具来研究外切体介导的这两种细胞类型之间的通讯 鉴定成纤维细胞活性的表型变化。这项研究的中心假设是 内皮细胞分泌的外切体在伤口愈合和修复过程中影响成纤维细胞的表型 纤维化症。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 会影响疤痕形成的表型。目的1将研究内皮细胞外切体对成纤维细胞活性的影响 以及体内和体外的纤维化。将采用体外伤口愈合试验，并将成纤维细胞暴露于 将评估内皮细胞外切体在迁移、细胞周期时相、增殖和基因方面的变化 表情。活体小鼠模型将用于评估皮内注射后的纤维化和胶原结构 内皮细胞外切体注射。目标2将利用生物信息学工具来识别和表征 内皮细胞外体的特异性miRNA载体，最有可能影响成纤维细胞功能的部位 结疤和纤维化。综合起来，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的功能，并可能导致新的治疗方法的开发 治疗纤维性疾病。

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.