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

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.

项目概要 角膜缘干细胞（LSC）在体内平衡和伤口愈合中发挥着重要作用。 LSC 损伤和其他与 LSC 缺乏 (LSCD) 相关的病理会导致角膜 新生血管形成、角膜混浊和视力丧失。目前的治疗范式基于 将LSC移植到患者体内以恢复LSC生态位。付出了巨大的努力 过去几年的目标是确定用于移植的 LSC 来源，并有不同程度的 成功。然而，我们最近的工作表明，LSC 移植可能不是解决问题的方法。 LSCD 唯一有用的治疗范例。我们发现定型上皮细胞具有 失去干细胞标记 K15 的人能够去分化并重新填充干细胞生态位。如果 这个过程可以被控制和操纵，它可以提供替代治疗，可以 避免复杂的干细胞移植的需要。我们已经证明利基市场很重要 去分化的过程表明，去分化和再增殖 生态位需要保留在生态位中的细胞和角膜上皮细胞之间的通信。 在这里，我们将使用角膜器官培养的尖端长期多日成像来研究 微环境和上皮细胞之间通讯的两侧。在目标#1中，我们将采取 药理学方法和筛选 99 种不同的化合物并获得长延时 在不同药物存在下恢复 LSC 生态位的电影。含量高 分析将提供有关利基所使用的不同信号通路的关键信息 募集并诱导上皮细胞去分化。在目标 #2 中，我们将重点关注接收者 通信的一侧并分析用于重新填充生态位的细胞的来源。使用 先进的光片显微镜方法我们将追踪单个细胞并发现 能够去分化的细胞的身份。单细胞追踪将显示是否 去分化的能力是普遍存在的，还是只有一小部分细胞维持这种能力 去分化的能力。这些目标的成功完成将为以下方面提供重要见解： LSC生态位恢复的生理过程，将为LSC的发展铺平道路 LSCD 的新疗法。

项目成果

Discrete limbal epithelial stem cell populations mediate corneal homeostasis and wound healing.

• DOI: 10.1016/j.stem.2021.04.003
• 发表时间: 2021-07-01
• 期刊: Cell stem cell
• 影响因子: 23.9
• 作者: Altshuler A;Amitai-Lange A;Tarazi N;Dey S;Strinkovsky L;Hadad-Porat S;Bhattacharya S;Nasser W;Imeri J;Ben-David G;Abboud-Jarrous G;Tiosano B;Berkowitz E;Karin N;Savir Y;Shalom-Feuerstein R
• 通讯作者: Shalom-Feuerstein R