Development of small-molecule Wnt mimetics for corneal epithelial cell regeneration

摘要 角膜缘干细胞缺乏(LSCD)是导致严重视力丧失的主要原因，无论是原发还是继发性。 在许多常见的角膜疾病中出现失明。自体角膜缘干细胞移植 在组织培养中扩大，成功地恢复了视力，并使LSCD的治疗发生了革命性变化。一个更高的 干细胞/祖细胞群体在培养中的扩增效率对应于更大的可能性 移植物的长期存活率。我们研究的长期目标是开发能够控制LSC的小分子 自我更新和分化，并作为干细胞治疗LSCD的治疗试剂- 相关的障碍。最有效的扩张方法需要饲养单元提供适当的 支持LSCs生长的微环境。在饲养层细胞提供的外部信号中 LSCS是WNT信令。这个项目的中心假设是小分子可以被开发出来 以模拟Wnt蛋白并激活细胞中的Wnt信号。我们进一步建议，这些小的 分子将提高体外扩增有功能的人类LSC的效率。这样做的目的是 应用是使用基于结构的药物发现方法来开发有效的WNT模拟小分子 并测试其提高LSC体外扩增效率的能力。因为茎的维护 培养扩增过程中的细胞特性是眼表成功的关键 重建，这个项目中产生的小分子将作为发展的平台 治疗其他角膜上皮疾病的新型药物试剂。

Abstract Limbal stem cell deficiency (LSCD) is a major cause, either primary or secondary, of significant visual loss and blindness in many common corneal disorders. Transplantation of autologous limbal stem cells (LSCs) expanded in tissue culture has successfully restored vision and revolutionized the treatment of LSCD. A higher expansion efficiency of the stem/progenitor cell population in culture corresponds to a greater probability of long-term graft survival. The long-term goal of our study is to develop small molecules that can govern LSC self-renewal and differentiation and be used as therapeutic reagents for stem cell-based treatments of LSCD– related disorders. The most efficient expansion method requires feeder cells that provide a proper microenvironment to support the growth of LSCs. Among the external signaling that the feeder cells provide to the LSCs is the Wnt signaling. The central hypothesis of this project is that small molecules can be developed to mimic the Wnt proteins and activate Wnt signaling in the cells. We further propose that these small molecules will increase the efficiency of ex vivo expansion of functional human LSCs. The goal of this application is to use a structure-based drug discovery approach to develop potent Wnt mimics small molecule and to test their ability to increase the efficiency of ex vivo LSC expansion. Because the maintenance of stem cell characteristics in the process of culture expansion is essential for the success of ocular surface reconstruction, the small molecules generated in this project will serve as a platform for the development of novel pharmaceutical reagents for treating other corneal epithelial disorders.