The role of Hippo and YAP/TAZ in dental renewal

Hippo 和 YAP/TAZ 在牙齿更新中的作用

• 批准号: 8984766
• 负责人: Ophir D Klein
• 金额: $ 39.63万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8984766
• 关键词: 3-Dimensional; Ablation; Actins; Address; Adult; Affect; Animals; Apoptosis; Atomic Force Microscopy; Binding; Biochemical; Biological; Biological Assay; Biological Models; Biomedical Engineering; Bromodeoxyuridine; Cell Death; Cell Differentiation process; Cell Line; Cell Proliferation; Cell Therapy; Cell division; Cell physiology; Cervical; Communities; Complex; Cues; Cytoskeleton; Data; Dental; Dental caries; Enhancers; Environment; Epithelial; Epithelial Cells; Epithelium; Equilibrium; Foundations; Gene Expression; Genetic; Goals; Human; In Situ Hybridization; In Vitro; Incisor; Knock-out; Lasers; Learning; Libraries; Life; Luciferases; Maintenance; Measures; Mechanics; Mediating; Microarray Analysis; Modeling; Molecular; Morphology; Mus; Natural regeneration; Nature; Nuclear; Oral cavity; Organ; Pathway interactions; Phenotype; Physical environment; Physiologic pulse; Play; Population; Process; Property; Proteins; Public Health; RNA Interference; Regenerative Medicine; Research Personnel; Rodent; Role; Signal Pathway; Signal Transduction; Sirolimus; Solid; Stem cells; Syndrome; TdT-Mediated dUTP Nick End Labeling Assay; Testing; Tissues; Tooth Diseases; Tooth Loss; Tooth structure; adult stem cell; base; craniofacial; follow-up; human LATS1 protein; in vivo; interest; loss of function; microCT; mineralization; mutant; novel; overexpression; public health relevance; reconstruction; regenerative; repaired; research study; rho GTP-Binding Proteins; small hairpin RNA; stem cell biology; stem cell niche; stem cell population; tool

 DESCRIPTION (provided by applicant): The utilization of adult stem cells for organ renewal and repair is an important goal in regenerative medicine. Tooth bioengineering is of great interest, because dental decay and tooth loss constitute an important public health issue. Additionally, tooth anomalies are common in many craniofacial syndromes, and the easy accessibility of the oral cavity makes teeth an excellent test case for organ replacement. A thorough understanding of the molecular and cellular processes that drive tooth renewal and regeneration will be crucial to efforts to build and repair teeth. We are using the mouse incisor as a model for understanding the mechanisms that underlie the ability of stem cells to contribute to renewal of dental tissues, because this remarkable tooth grows continuously due to the presence of adult stem cells. Our preliminary data indicate that the Hippo pathway, via its two effectors, Yes-associated Protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), regulates the incisor stem cell niche. Importantly, our initial studies suggest that YAP/TAZ in the incisor are regulated by mechanical cues; this important regulatory interaction has been understudied in adult stem cell biology. Thus, the rodent incisor provides a unique in vivo platform to study how the physical environment regulates YAP/TAZ, which will not only move the field of Hippo signaling forward but also lay the foundation for developing stem cell-based therapies in the treatment of dental diseases. In this application, we propose to test the hypothesis that mechanical signals, mediated by small Rho GTPases, can modulate YAP/TAZ activity, which in turn control key processes in stem cell biology. In the first Aim, we will determine the effect of Yap/Taz and Lats1/2 loss-of-function in the mouse incisor. Aim 2 will address how mechanical forces and small Rho GTPases regulate YAP/TAZ activity. A pilot RNAi screen will also be carried out to identify novel upstream mechano-regulators of YAP/TAZ. Aim 3 will address the role of the mTor pathway downstream of YAP/TAZ in the incisor stem cells. Together, these experiments will elucidate a linear mechanistic pathway that will help us to understand how Hippo signaling and YAP/TAZ mediate mechanical signals to regulate adult incisor stem cells in mice, and they will yield findings that will be of general interest to both dental researchers and to the stem cell and regenerative medicine communities. The results of these studies will advance our understanding of how nature normally uses stem cells in dental regeneration and enhance our ability to modulate both the physical and biochemical environments in order to generate dental stem cells for translational purposes.

 描述（由申请人提供）：利用成体干细胞进行器官更新和修复是再生医学的一个重要目标。牙齿生物工程是非常感兴趣的，因为蛀牙和牙齿脱落构成了一个重要的公共卫生问题。此外，牙齿异常在许多颅面综合征中很常见，口腔的易接近性使牙齿成为器官置换的绝佳测试案例。彻底了解驱动牙齿更新和再生的分子和细胞过程对于构建和修复牙齿的努力至关重要。我们正在使用小鼠切牙作为模型，以了解干细胞促进牙齿组织更新的能力的机制，因为由于成体干细胞的存在，这种显着的牙齿会持续生长。我们的初步数据表明，Hippo途径，通过其两个效应，是相关蛋白（雅普）和转录辅激活与PDZ结合基序（TAZ），调节切牙干细胞生态位。重要的是，我们的初步研究表明，门牙中的雅普/TAZ受到机械信号的调节;这种重要的调节相互作用在成人干细胞生物学中尚未得到充分研究。因此，啮齿动物切牙提供了一个独特的体内平台来研究物理环境如何调节雅普/TAZ，这不仅将推动Hippo信号传导领域的发展，而且还为开发基于干细胞的治疗牙科疾病的疗法奠定了基础。在本申请中，我们提出测试的假设，即机械信号，介导的小Rho GTP酶，可以调节雅普/TAZ的活动，这反过来又控制在干细胞生物学的关键过程。在第一个目标中，我们将确定雅普/Taz和Lats 1/2对小鼠切牙功能丧失的影响。目的2将阐述机械力和小Rho GTP酶如何调节雅普/TAZ活性。还将进行中试RNAi筛选以鉴定雅普/TAZ的新型上游机械调节剂。目的3将阐明mTor通路下游的雅普/TAZ在切牙干细胞中的作用。总之，这些实验将阐明一个线性的机制途径，这将有助于我们了解海马信号和雅普/TAZ如何介导机械信号来调节小鼠的成年切牙干细胞，并且它们将产生牙科研究人员以及干细胞和再生医学界普遍感兴趣的发现。这些研究的结果将促进我们对自然界通常如何在牙齿再生中使用干细胞的理解，并增强我们调节物理和生化环境的能力，以产生用于翻译目的的牙齿干细胞。