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: 10039310
• 负责人: Yuzuru Sasamoto
• 金额: $ 10.93万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 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 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10039310
• 关键词: ATP-Binding Cassette Transporters; Address; Admixture; Allogenic; Area; Autologous; Autologous Transplantation; BMP2 gene; Bilateral; Binding; Blindness; CAV1 gene; Cell Maintenance; 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; Institutes; Lead; Maintenance; Mentorship; Modeling; Molecular; Mus; Natural regeneration; Ophthalmologist; Ophthalmology; Organ; Pathway interactions; Patients; 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; base; cell regeneration; clinical translation; conjunctiva; corneal epithelium; experience; fascinate; improved; in vivo; interest; limbal; member; morphogens; novel; pre-clinical; recruit; regenerative; restoration; single-cell RNA sequencing; stem; stem cell biology; stem cell homeostasis; stem cell niche; stem cell population; stem cell therapy; stem cells; success; therapeutic evaluation

Title: Development of cell-free approaches to the treatment of limbal stem cell deficiency PROJECT SUMMARY/ABSTRACT 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. Building on the PI’s training in regenerative medicine, the PI will gain extensive experience in corneal stem cell biology under the mentorship of Dr. Frank, a world-class leader in the fields of LSC biology and ABC transporters, and a member of Harvard Stem Cell Institute. Furthermore, this research will provide an important career guidance on his path of becoming an independent investigator.

标题：开发无细胞干细胞缺乏治疗的方法 项目摘要/摘要 石灰干细胞（LSC）产生整个角膜上皮，已知存在于边界区域 在角膜和结膜之间称为边缘。 林瓦尔干细胞缺乏症（LSCD） - 世界上视力丧失的常见原因。 从对侧眼睛治疗单侧LSCD，双侧的自体limval组织从 LSCD患者没有可用的自体膜层组织。 同种异体供体的limval移植物； 供体短缺对同种异体LSC的可用性构成了重大挑战 LSCD患者。 发现LSC维护和再生的新型机制。 纸盒（ABC）超家族成员B5（ABCB5）作为NO新颖的LSC标记。 人类捐助者能够长期进行角膜恢复 目前，正在解决人类患者的干细胞种群的治疗潜力 最新的研究旨在探索使用lel-cell rna- 测序揭示了一种新型的LSC亚群，可以通过低点区分其他LSC簇 角膜特异性基因的表达水平。 最原始的干细胞特征具有最高的再生潜力。 人们对细胞优先表达涉及FGF的分子和AXL信号传导的启发。 我们认为分子途径对于维持未分化至关重要 LSC表型，可用于De-Novo LSC指示和恢复LSC的位置 双边LSCD的设置。 使用鼠和人类遗传引擎实验模型在LSC维护中的AXL（AIM） 1）并将测试靶向靶向LSCD的途径的治疗潜力 鼠类疾病模型（AIM 2）。 LSC开发，维护和调节对临床翻译具有重要意义。 在PI在再生医学方面的培训的基础上，PI将获得角膜干细胞的丰富经验 在LSC Biology和ABC领域的世界一流领导者Frank博士的指导下生物学 转运蛋白和哈佛干细胞研究所的成员。 关于他的独立调查员的道路的职业指导。