Corneal Epithelial-Stromal Interactions During Regeneration and Fibrosis

再生和纤维化过程中角膜上皮-基质相互作用

• 批准号: 9893920
• 负责人: Joseph B. Ciolino
• 金额: $ 49.25万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893920
• 关键词: 3-Dimensional; Adherent Culture; Affect; Basement membrane; Cancer Cell Growth; Cell Communication; Cell Death; Cells; Communication Methods; Confocal Microscopy; Cornea; Corneal Diseases; Corneal Injury; Cultured Cells; DNA; Data; Descemet' s membrane; Development; Epithelial; Epithelial Cells; Epithelial-Stromal Communication; Epithelium; Excision; Fibroblasts; Fibrosis; Generations; Growth Factor; Homeostasis; In Vitro; Lead; Left; Lipids; Malignant Neoplasms; Messenger RNA; Methods; MicroRNAs; Modeling; Movement; Myofibroblast; Natural regeneration; Pathologic Processes; Pharmaceutical Preparations; Physiological Processes; Process; Production; Property; Proteins; Proteomics; Public Health; Research; Role; Site; Specificity; Stress; Stromal Cells; Subgroup; Techniques; Therapeutic; Therapeutic Agents; Tissues; Topical application; Transmission Electron Microscopy; Travel; Ulcer; Vesicle; base; cancer cell; corneal epithelial wound healing; corneal epithelium; cytokine; epithelial wound; exosome; extracellular vesicles; healing; in vivo; in vivo Model; interest; migration; prevent; response; response to injury; three dimensional cell culture; uptake; virtual; wound; wound healing

PROJECT SUMMARY Cell-cell communication is an important component of most functions of the cornea, including fibrosis, regeneration, and homeostasis. To a great extent, cell-cell communication has been thought to consist of the release of numerous soluble growth factors and cytokines to direct corneal wound repair; however an underlying question remains as to how these factors perfuse with any specificity from one cell to an adjacent cell. Recently, interest in another method of cell-communication has boomed. This method involves the secretion and uptake of small vesicles, termed Extracellular Vesicles (EVs). EVs can migrate long and short distances to be taken up by other cells. This crosstalk between cells is notable in that EVs carry “cargo” consisting of proteins, mRNA, miRNA, and DNA that can influence many of the functions in the recipient cell. Indeed, recent findings indicate that development of cancer involves crosstalk with EVs, where cancer cells produce EVs that act on the host's cells to produce a microenvironment that promotes growth of cancer cells. We propose to examine if corneal wound healing involves a similar mechanism, where epithelial cells release EVs that alter the properties of keratocytes. We have found that epithelial EVs can stimulate keratocytes to become myofibroblasts and that epithelial EVs from wounded cells have an even greater effect. This has lead us to the hypothesis that the epithelial EV cargo is altered as cells become migratory to heal a wound, subsequently causing the keratocyte to become myofibroblasts, which produce a microenvironment that promotes their persistence. Because of its clarity and accessibility, the cornea provides an outstanding model to examine movement of EVs through matrix in in vivo or ex vivo models. Techniques to be used include EV isolation, proteomics. miRNA isolation and characterization, transmission electron microscopy, confocal microscopy, and 3D cultures. Relevance to Public Health—EVs are involved in cell-cell communication in virtually all physiological and pathological processes. Because of their ability to travel long distances and interact with specific cells, EVs are widely being examined for usefulness to deliver therapeutics. We will determine if topical application of EVs to the cornea is useful as a therapeutic approach. Potentially, any corneal injury or disease could be treated with EVs.

项目概要 细胞间通讯是角膜大多数功能的重要组成部分，包括纤维化、 在很大程度上，细胞间的通讯被认为由以下组成： 释放大量可溶性生长因子和细胞因子来指导角膜伤口修复； 根本问题仍然是这些因素如何以任何特异性从一个细胞灌注到相邻细胞 最近，人们对另一种细胞通讯方法的兴趣日益浓厚。 分泌和摄取的小囊泡（称为细胞外囊泡（EV））可以长短迁移。 电池之间的串扰很明显，因为电动汽车携带“货物”。 由蛋白质、mRNA、miRNA 和 DNA 组成，可以影响受体细胞的许多功能。 事实上，最近的研究结果表明，癌症的发展涉及与 EV 的串扰，其中癌细胞 产生的 EV 作用于宿主细胞，产生促进癌细胞生长的微环境。 我们建议检查角膜愈合是否涉及类似的机制，即上皮细胞释放 改变角膜细胞特性的 EV 我们发现上皮 EV 可以刺激角膜细胞。 变成肌成纤维细胞，并且来自受伤细胞的上皮细胞外囊泡具有更大的影响。 我们假设上皮细胞外囊泡货物是细胞迁移以愈合伤口时， 随后导致角膜细胞变成肌成纤维细胞，从而产生一个微环境 由于其清晰度和可及性，角膜提供了一个出色的模型。 通过体内或离体模型中的矩阵检查 EV 的运动，所使用的技术包括 EV。 分离、蛋白质组学、miRNA 分离和表征、透射电子显微镜、共聚焦。 显微镜和 3D 培养与公共健康的相关性——EV 参与细胞间的通讯。 几乎所有的生理和病理过程都因为它们能够长距离传播。 由于 EV 与特定细胞相互作用，因此我们正在广泛检查 EV 是否有助于提供治疗。 确定将 EV 局部应用于角膜是否可以作为一种潜在的治疗方法。 角膜损伤或疾病可以用 EV 治疗。