Development of cell-free approaches to the treatment of limbal stem cell deficiency

开发治疗角膜缘干细胞缺陷的无细胞方法

• 批准号: 10737842
• 负责人: Yuzuru Sasamoto
• 金额: $ 24.9万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737842
• 关键词: ATP-Binding Cassette Transporters; Address; Admixture; Allogenic; Area; Autologous; Autologous Transplantation; BMP2 gene; Bilateral; Binding; Blindness; CAV1 gene; Cell Maintenance; Cell Separation; Cell Survival; Cells; Characteristics; Clinical; Clinical Trials; Contralateral; Cornea; Development; Disease model; Doctor of Philosophy; Experimental Models; Eye; FDA approved; FGF7 gene; FGFR2 gene; Fibroblast Growth Factor; Genes; Genetic Engineering; Goals; Homologous Transplantation; Human; In Vitro; Instruction; Maintenance; Mentorship; Modeling; Molecular; Mus; Natural regeneration; Ophthalmologist; Ophthalmology; Organ; Pathway interactions; Patients; Phase; Phenotype; Receptor Protein-Tyrosine Kinases; Regenerative Medicine; Regulation; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Stem Cell Development; Stem cell transplant; Testing; Therapeutic; Tissues; Topical application; Training; Undifferentiated; Vocational Guidance; cell regeneration; clinical training; clinical translation; conjunctiva; corneal epithelium; experience; fascinate; improved; in vivo; induced pluripotent stem cell; interest; limbal; member; morphogens; novel; pre-clinical; recruit; regeneration potential; restoration; single-cell RNA sequencing; stem cell biology; stem cell biomarkers; stem cell homeostasis; stem cell niche; stem cell population; stem cell self renewal; stem cells; success; therapeutic evaluation

PROJECT SUMMARY (See instructions): Limbal stem cells (LSCs) give rise to the entire corneal epithelium and are known to reside in the border area between the cornea and conjunctiva called limbus. Loss of LSCs or destruction of the LSC niche can result in Limbal Stem Cell Deficiency (LSCD) – a common cause of vision loss in the world. While transplantation of the autologous limbal tissues removed from the contralateral eye can cure patients with unilateral LSCD, bilateral LSCD patients have no autologous limbal tissues available. These patients often require transplantation of allogeneic donor limbal grafts; however, their success is highly variable. Moreover, the worldwide corneal donor shortage poses significant challenges for the availability of allogeneic LSCs for the treatment of bilateral LSCD patients. Thus, the overarching goal of this project is to develop cell-free LSCD therapies through the discovery of novel mechanisms of LSC maintenance and regeneration. Our lab has discovered an ATP-binding cassette (ABC) superfamily member B5 (ABCB5) as a novel LSC marker. ABCB5-positive LSCs isolated from human donors were capable of the long-term corneal restoration in pre-clinical LSCD models. Clinical trials are currently on the way to address the therapeutic potential of this stem cell population in human patients. Our most recent studies aimed to explore the cellular hierarchy within ABCB5-positive LSCs using single-cell RNA-sequencing revealed a novel LSC subpopulation that could be differentiated from the other LSC clusters by low expression levels of the cornea-specific genes. Here we hypothesized that this subpopulation possesses the most primitive stem cell characteristics with the highest regenerative potential. Further in-depth analyses revealed that these cells preferentially expressed the molecules involved in FGF, BMP, and AXL signaling cascades. We posit that these molecular pathways are essential for the maintenance of the undifferentiated LSC phenotype and can be employed for de-novo LSC induction and restoration of the LSC niche in the setting of bilateral LSCD. The two Aims of this proposal will: mechanistically dissect the role of FGF7, BMP2 and AXL in the LSC maintenance using murine and human genetically engineered experimental models (Aim 1) and will test the therapeutic potential of targeting these pathways for the treatment of LSCD in pre-clinical murine disease models (Aim 2). Successful completion of this study will further advance our understanding of LSC development, maintenance, and regulation with significant implications for clinical translation

项目摘要（请参阅说明）： 边缘干细胞（LSC）产生整个角膜上皮，众所周知居住在边界区域 在角膜和结膜之间称为边缘。 LSC的损失或LSC利基市场的破坏可能导致边缘 干细胞缺乏（LSCD） - 世界上视力丧失的常见原因。同时自体移植 从对侧眼中去除的边缘组织可以治愈单侧LSCD，双侧LSCD患者的患者 没有可用的自体膜层组织。这些患者经常需要移植同种异体供体的边缘 移植物；但是，他们的成功是高度可变的。此外，全球角膜供体短缺位置显着 同种异体LSC可用于治疗双侧LSCD患者的挑战。那是总体 该项目的目标是通过发现LSC的新机制来开发无细胞的LSCD疗法 维护和再生。我们的实验室发现了一个ATP结合盒（ABC）超家族成员B5 （ABCB5）作为一种新颖的LSC标记。从人类捐助者中分离出的ABCB5阳性LSC能够长期 临床前LSCD模型中的角膜修复。临床试验目前正在解决该疗法 该干细胞种群在人类患者中的潜力。我们最近的研究旨在探索细胞 使用单细胞RNA测序的ABCB5阳性LSC内的层次结构显示出一种新型LSC亚群， 可以通过角膜特异性基因的低表达水平与其他LSC簇区分。我们在这里 假设该亚群具有最原始的干细胞特征 再生潜力。进一步的深入分析表明，这些细胞优先表达了分子 参与FGF，BMP和AXL信号级联。我们肯定这些分子途径对于 维护未分化的LSC表型，可用于DE-NOVO LSC诱导和恢复 在双侧LSCD的环境中的LSC利基市场。该提议的两个目标将：机械地剖析角色 使用鼠和人类基因工程实验的LSC维护中的FGF7，BMP2和AXL 模型（AIM 1），并将测试针对这些途径以治疗LSCD的治疗潜力 临床前鼠病模型（AIM 2）。成功完成这项研究将进一步发展我们的 了解LSC的开发，维护和调节，对临床翻译产生重大影响