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.

项目总结 细胞间的通讯是角膜大部分功能的重要组成部分，包括纤维化， 再生和动态平衡。在很大程度上，细胞间的通讯被认为是由 释放大量的可溶性生长因子和细胞因子来指导角膜伤口的修复；然而 潜在的问题仍然是这些因素如何以任何特异性从一个细胞向相邻的细胞灌流 手机。最近，人们对另一种细胞通讯方式的兴趣大增。此方法涉及 小泡的分泌和摄取，称为细胞外小泡(EVS)。电动汽车可以迁移多头和空头 其他单元格要占用的距离。电池之间的这种串扰是值得注意的，因为电动汽车携带的是“货物”。 由蛋白质、信使核糖核酸、微小核糖核酸和脱氧核糖核酸组成，可以影响受体细胞的许多功能。 事实上，最近的发现表明，癌症的发展与EVS有关，在EVS中，癌细胞 产生作用于宿主细胞的电动汽车，以产生促进癌细胞生长的微环境。 我们建议检查角膜伤口愈合是否涉及类似的机制，即上皮细胞释放。 EVS可以改变角质形成细胞的特性。我们发现上皮EVS可以刺激角质形成细胞 成为肌成纤维细胞，来自受伤细胞的上皮EV具有更大的影响。这件事具有先导性 我们接受了这样的假设，即当细胞迁移到伤口愈合时，上皮EV货物会发生变化， 随后导致角质形成细胞成为肌成纤维细胞，从而产生一个微环境， 提升他们的毅力。由于其清晰度和可及性，角膜提供了一个出色的模型 研究EVS在体内或体外模型中通过基质的运动情况。将使用的技术包括电动汽车 隔离，蛋白质组学。MiRNA的分离和鉴定、透射电子显微镜、共聚焦 显微镜和3D培养。与公共卫生的相关性-电动汽车参与细胞间的通讯 几乎所有的生理和病理过程。因为它们能够长途旅行，而且 在与特定细胞相互作用的过程中，电动汽车正在被广泛检测是否有用处来提供治疗药物。我们会 确定将EVS局部应用于角膜是否作为一种治疗方法有用。潜在的，任何 角膜损伤或疾病可以用EVS治疗。