Identify mechanisms of dedifferentiation during limbal stem cell niche reconstruction.

确定角膜缘干细胞生态位重建期间的去分化机制。

• 批准号: 9902499
• 负责人: Roy Wollman
• 金额: $ 15.93万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902499
• 关键词: Ablation; Address; Biology; Blindness; Cells; Charge; Clinical Trials; Communication; Complex; Cornea; Corneal Neovascularization; Custom; Development; Epithelial; Epithelial Cells; Epithelium; Excision; Future; Generations; Goals; Homeostasis; Image; Individual; Knowledge; Light; Microscope; Microscopy; Modeling; Mus; Operative Surgical Procedures; Organ Culture Techniques; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; Physiological Processes; Play; Process; Recovery; Role; Series; Side; Signal Pathway; Signal Transduction; Source; Stem cell transplant; Testing; Transgenic Mice; Transplantation; Work; alternative treatment; base; cell motility; conjunctiva; corneal epithelium; corneal regeneration; corneal repair; cost; experimental study; imaging modality; insight; limbal; migration; mouse model; movie; neovascularization; patient oriented; reconstruction; recruit; restoration; self-renewal; stem cell niche; stem cell population; stem cells; stemness; success; synergism; wound healing; Ablation; Address; Biology; Blindness; Cells; Charge; Clinical Trials; Communication; Complex; Cornea; Corneal Neovascularization; Custom; Development; Epithelial; Epithelial Cells; Epithelium; Excision; Future; Generations; Goals; Homeostasis; Image; Individual; Knowledge; Light; Microscope; Microscopy; Modeling; Mus; Operative Surgical Procedures; Organ Culture Techniques; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; Physiological Processes; Play; Process; Recovery; Role; Series; Side; Signal Pathway; Signal Transduction; Source; Stem cell transplant; Testing; Transgenic Mice; Transplantation; Work; alternative treatment; base; cell motility; conjunctiva; corneal epithelium; corneal regeneration; corneal repair; cost; experimental study; imaging modality; insight; limbal; migration; mouse model; movie; neovascularization; patient oriented; reconstruction; recruit; restoration; self-renewal; stem cell niche; stem cell population; stem cells; stemness; success; synergism; wound healing

Project Summary The corneal limbal stem cells (LSC) play a vital function in homeostasis and wound healing. Damage to the LSC and other pathologies associated LSC deficiency (LSCD) cause cornea neovascularization, corneal opacification, and vision loss. Current treatment paradigm is based on the transplantation of LSC into patients to restore the LSC niche. Substantial effort over the last few years aims at identifying sources of LSC for transplantation with varying degree of success. However, our recent work indicates that transplantation of LSC might not be the the only useful treatment paradigm for LSCD. We showed that committed epithelial cells that have lost the stem cell marker K15 are able to dedifferentiate and repopulate the stem cell niche. If this process can be controlled and manipulated it could provide alternative treatment that can avoid the need for complex stem cell transplantation. We have shown that the niche is important for the process of dedifferentiation demonstrating that dedifferentiation and repopulation of the niche require communication between cells that remain in the niche and corneal epithelial cells. Here we will use cutting edge long term multi-day imaging of corneal organ culture to investigate both sides of the communication between the niche and epithelial cells. In Aim #1 we will take a pharmacological approach and screen 99 different compounds and acquire long timelapse movies of the restoration of the LSC niche in the presence of different drugs. The high content analysis will provide key information on the different signaling pathways used by the niche to recruit and induce the dedifferentiation of epithelial cells. In Aim #2 we will focus on the receiver side of the communication and analyze the source of cells used to repopulate the niche. Using advanced light sheet microscopy approaches we will track individual cells and discover the identity of the cells that are capable of dedifferentiation. Single cell tracking will show whether the ability to dedifferentiate is ubiquitous or whether only a small subset of cells maintain the ability to dedifferentiate. The successful completion of these aims will provide key insights into the physiological process of LSC niche recovery and will pave the way to the development of new treatments to LSCD.

项目摘要 角膜缘干细胞（LSC）在体内平衡和伤口愈合中起着至关重要的功能。 损害LSC和其他与LSC缺乏症相关的病理损害导致角膜 新生血管形成，角膜无情和视力丧失。当前的治疗范例是基于 在将LSC移植到患者中以恢复LSC生态位。大力努力 最近几年的目的是确定LSC的来源以不同程度的移植 成功。但是，我们最近的工作表明，LSC的移植可能不是 仅适用于LSCD的有用治疗范例。我们表明，承诺的上皮细胞 丢失的干细胞标记K15能够去分化和重新填充干细胞生态位。如果 可以控制和操纵此过程，它可以提供可以提供的替代治疗方法 避免需要复杂的干细胞移植。我们已经证明利基很重要 对于去分化的过程，证明了对 利基需要在生境和角膜上皮细胞中的细胞之间进行通信。 在这里，我们将使用角膜器官培养的长期多日成像进行研究 利基和上皮细胞之间通信的两面。在AIM＃1中，我们将 药理学方法和屏幕99种不同的化合物，并获得长时间解脱 在存在不同药物的情况下，LSC利基恢复的电影。高素质 分析将提供有关利基用于不同信号通路的关键信息 募集并诱导上皮细胞的去分化。在AIM＃2中，我们将专注于接收器 通信的一面并分析用于重新占用利基的细胞来源。使用 高级光片显微镜接近我们将跟踪单个细胞并发现 能够去分化的细胞的身份。单细胞跟踪将显示是否 去分化的能力无处不在，或者仅一小部分细胞维持 去分化的能力。这些目标的成功完成将为您提供关键的见解 LSC利基恢复的生理过程，将为发展铺平道路 LSCD的新疗法。