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

使用单细胞转录组分析揭示牙上皮祖细胞从头生成的遗传途径

• 批准号: 10428476
• 负责人: Jimmy Kuang-Hsien Hu
• 金额: $ 15.6万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-14 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10428476
• 关键词: Address; Adhesives; Adult; Affect; Aging; Ameloblasts; Applications Grants; Biological Models; Biomedical Engineering; Cell Differentiation process; Cell physiology; Cells; Chronic; Clinical; Data; Dental; Derivation procedure; Development; Developmental Process; Disease; Ectoderm Cell; Embryo; Epigenetic Process; Epithelial; Epithelial Cells; Foundations; Future; Gene Expression Profile; Generations; Genes; Genetic; Genetic Identity; Genetic Models; Genetic Transcription; Genetic Variation; Genetic study; Genomics; Human; Incisor; Kidney; Knowledge; Learning; Life Style; Maintenance; Maps; Methods; Molecular; Mus; Natural regeneration; Oral; Organ; Organogenesis; Pathway interactions; Phenotype; Pluripotent Stem Cells; Population; Primordium; Process; Proliferating; Regenerative Medicine; Regulation; Research Project Grants; Role; Signal Pathway; Signal Transduction; Solid; Specific qualifier value; System; Techniques; Testing; Tissues; To specify; Tooth Loss; Tooth structure; Trauma; adult stem cell; base; capsule; cell type; clinical application; dental genetics; differential expression; embryonic stem cell; epithelial stem cell; genetic manipulation; genetic testing; imaging approach; in vivo; injury and repair; innovation; mouse genetics; oral cavity epithelium; progenitor; programs; single-cell RNA sequencing; stem cell biology; stem cell function; stem cells; three dimensional cell culture; tool; transcription factor; transcriptome; transcriptomics

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.

项目摘要 有效利用器官特异性体干细胞修复受伤的组织或生物工具器官 彻底改变疾病治疗，并缓解衰老和创伤引起的许多问题。但是，它 精确地得出躯体干细胞并以高效率扩展它们仍然很有挑战 用于临床应用。技术障碍在很大程度上是由于我们对遗传的不完全理解 调节在发育过程中控制祖细胞的命运规范以及成人的细胞可塑性。 牙齿提供了一个很好的测试案例，可以进一步了解这些方面并在临床上应用，因为成人人类 牙齿不能保持牙齿上皮干细胞，并且缺乏再生能力。鼠标牙齿是 强大的模型系统研究器官发生和基于干细胞的再生，并且适合 体内遗传学研究和实体操作，以研究祖细胞功能。利用这个 显着的实验系统，并将其与尖端的单细胞转录组分析相结合，这 提案将对遗传计划和转录变化深入了解 牙齿上皮祖细胞的形成。这些知识将使我们能够确定遗传网络和关键 指定牙齿命运并提供蓝图以通过区分来得出牙齿祖细胞所需的调节器 沿遗传路径的多能干细胞。同时，该项目将测试IRX1/2的功能和利用 通过分化口服上皮和 分别对成成成成成成叶。我们将比较诱导的单细胞转录组 胚胎和成年祖细胞确定IRX1/2和YAP是否可以激活牙科遗传程序。 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，这就是如何 胚胎和成年人中的不同祖细胞类型在转录上相互类似。主要创新 该项目源于基因组技术和小鼠遗传模型的整合，以了解 牙齿祖细胞和干细胞形成的遗传调节，这些调节已被研究。这样的知识将形成 未来研究和授予申请的基础，并实现发展原则驱动的方法 牙科工程。

项目成果

Understanding the development of oral epithelial organs through single cell transcriptomic analysis.

• DOI: 10.1242/dev.200539
• 发表时间: 2022-08-15
• 期刊: Development (Cambridge, England)

Nail mesenchyme: Tipping the hand on regeneration.

指甲间充质：促进再生。

• DOI: 10.1016/j.celrep.2022.111960
• 发表时间: 2023
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Hu,JimmyK