Using single cell transcriptomic analysis to uncover genetic pathways for de novo generation of dental epithelial progenitors

项目总结 有效利用器官特异性躯体干细胞修复受损组织或生物工程器官将 革新疾病治疗，缓解因衰老和创伤造成的众多问题。然而，它 精确地提取体细胞并高效地扩增仍然是一个巨大的挑战 用于临床应用。技术障碍在很大程度上是由于我们对基因的不完全理解 在发育过程中控制祖细胞命运的调节，以及成年细胞的可塑性。 作为成年人，牙齿为进一步了解这些方面并应用于临床提供了一个很好的测试案例。 牙齿不能维持牙齿上皮干细胞，缺乏再生能力。老鼠的牙齿是一颗 强大的模型系统，研究器官发生和基于成体干细胞的再生，并服从于 包括体内遗传学研究和体外操作，以研究祖细胞功能。利用这一点 卓越的实验系统，并结合尖端的单细胞转录分析，这 提案将提供对遗传程序和转录变化的深入理解 牙上皮祖细胞的形成。这一知识将使我们能够识别一个基因网络和关键的 监管机构要求明确牙科命运并提供蓝图，通过区分来派生牙科祖细胞 基因路径上的多能干细胞。同时，该项目将测试IRX1/2的功能和使用情况 和YAP通过口腔上皮细胞分化诱导牙祖细胞的形成 成釉细胞分别去分化。我们将比较诱导的单细胞转录本和 胚胎和成年祖细胞，以确定IRX1/2和YAP是否可以激活牙齿遗传程序。 基于这些数据，我们还将能够解决干细胞生物学中的一个重要问题，即如何 胚胎和成体中不同的祖细胞类型在转录上彼此相似。的主要创新之处 该项目源于基因组技术和小鼠遗传模型的整合，以了解 牙祖细胞和干细胞形成的遗传调控，研究不足。这样的知识将形成 未来研究和赠款申请的基础，并使发展原则驱动的方法能够 牙齿生物工程。

PROJECT SUMMARY Effective utilization of organ-specific somatic stem cells to repair injured tissues or to bioengineer organs will revolutionize disease treatment and relieve numerous problems caused by aging and trauma. However, it remains significantly challenging to derive somatic stem cells with precision and expand them with high efficiency for clinical applications. The technical hurdles are in large part due to our incomplete understanding of the genetic regulation that controls fate specification of progenitors during development, as well as cell plasticity in adults. Teeth provide an excellent test case to further understand these aspects and apply clinically, as adult human teeth do not maintain dental epithelial stem cells and lack the capability to regenerate. The mouse tooth is a powerful model system to study both organogenesis and adult stem cell-based regeneration, and amenable for both in vivo genetic studies and ex vivo manipulations to investigate progenitor cell functions. Leveraging this remarkable experimental system and combining it with cutting-edge single cell transcriptomic analysis, this proposal will deliver an in depth understanding of the genetic program and transcriptional changes during the formation of dental epithelial progenitors. This knowledge will allow us to identify a genetic network and critical regulators required to specify the dental fate and provide a blueprint to derive dental progenitors by differentiating pluripotent stem cells along a genetic path. In parallel, this project will test the function and utilization of IRX1/2 and YAP in inducing the formation of dental progenitors through differentiation of oral epithelium and dedifferentiation of ameloblasts respectively. We will compare single cell transcriptomes between induced, embryonic, and adult progenitors to determine whether IRX1/2 and YAP can activate a dental genetic program. Based on these data, we will also be able to address an important question in stem cell biology, which is how different progenitor types in embryos and adults resemble each other transcriptionally. The main innovation of the project arises from the integration of genomic techniques and mouse genetic models to understand the genetic regulation of dental progenitor and stem cell formation, which is understudied. Such knowledge will form the basis of future research and grant applications, and enable developmental principle-driven approaches to tooth bioengineering.