Regulation of Runx2 Function by Twist-1 in Tooth Development

Twist-1 在牙齿发育中对 Runx2 功能的调节

• 批准号: 7837315
• 负责人: Rena N. D'Souza
• 金额: $ 5.19万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7837315
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Address; Affect; Apoptosis; Award; Biochemical; Biological Assay; Boxing; Cell Differentiation process; Cell Line; Cell Proliferation; Cell Survival; Chotzen Syndrome; Classification; Cleidocranial Dysplasia; Craniosynostosis; DNA Binding Domain; Data; Defect; Dental; Dental Papilla; Dental Pulp; Dental Pulp Calcification; Dentin; Dentition; Development; Epithelial; Epithelium; Genes; Genetic; Goals; Hereditary Disease; Heterozygote; Homeostasis; Human; Human Genetics; Incisor; Laboratories; Learning; Mediating; Mesenchymal; Mesenchyme; Messenger RNA; Molecular; Molecular Genetics; Morphogenesis; Mus; Mutation; Nature; Nuclear Extract; Nuclear Protein; Nuclear Proteins; Odontoblasts; Organ; Pathogenesis; Pathway interactions; Pattern; Phenotype; Process; Proteins; Regulation; Research; Research Personnel; Role; Signal Transduction; Staging; Structure of fontanel of skull; Supernumerary Tooth; Testing; Tooth structure; Work; bHLH Domain; base; calcification; gain of function; genetic regulatory protein; interest; loss of function; molecular marker; osteoblast differentiation; programs; protein expression; protein protein interaction; research study

In this competing renewal, we propose to continue our research on the role of Runx2 in tooth development. Data from our work in the previous award period indicated key roles for Runx2 in directing the fate of dental epithelium during morphogenesis as well as in controlling the onset of odontoblast differentiation. Our studies point to the critical need to learn how Runx2 activities are precisely regulated during tooth morphogenesis and cell differentiation and whether its role in these processes is modulated through interactions with other molecules. The nuclear protein Twist-1 is of particular interest as a regulatory protein partner for Runx2. Our rationale for studying if Runx2, a cell differentiation factor, interacts with Twist-1, a cell survival factor, is derived from studies in our and other laboratories that suggest that these interactions between Runx2 and Twist-1 occur at the protein level. Our experiments will directly test the hypothesis that Runx2's key functions in odontoblast differentiation are regulated by Twist-1 at the level of protein- protein interactions that are functionally antagonistic in nature. The selective and transient blocking of Runx2 function by Twist-1 provides a means to restrain odontoblast differentiation until morphogenesis is complete. We further propose that interactions between Runx2 and Twist-1 are not mutually antagonistic as Twist-1 can mediate cell proliferation during morphogenesis via FGF-mediated epithelial mesenchymal signaling. Hence, the presence of supernumerary teeth in human CCD and accessory buds in Runx2(-/-) mice likely reflect increased activity of Twist-1 rather than a direct effect of decreased levels of Runx2. Aim 1 will determine if the patterns of Runx2 and Twist-1 (mRNA and protein) expression are compatible with their proposed partnership during tooth development and will correlate these patterns with the expression of molecular markers of tooth morphogenesis and odontoblast differentiation. Aim 2 will assess with mouse genetic loss-of-function and gain-of-function approaches whether alterations in Twist-1 expression affects tooth morphogenesis and odontoblast differentiation. Aim 3 will study the molecular basis of Runx2 - Twist-1 protein interactions in dental mesenchyme and the functional consequences of this interaction on Runx2 functions in odontoblast differentiation, and Aim 4 will test whether the bHLH domain of Twist-lean mediate tooth morphogenesis via FGF-signaling that is independent of its interactions with Runx2. These studies will increase our understanding of how Runx2 achieves its selective functions in tooth development through its partnership with Twist-1. Importantly, they will explain how supernumerary teeth form and if odontoblast differentiation is determined by the release of an inhibition. Such data will also provide a framework for understanding the pathogenesis of Cleidocranial Dysplasia and Saethre-Chotzen Syndrome, two human genetic disorders that threaten dentition.

在这次竞争更新中，我们建议继续研究 Runx2 在牙齿发育中的作用。数据来自 我们在上一个奖项期间的工作表明 Runx2 在指导牙上皮细胞命运方面发挥着关键作用 形态发生以及控制成牙本质细胞分化的开始。我们的研究表明迫切需要 了解 Runx2 活性在牙齿形态发生和细胞分化过程中如何精确调节，以及它是否 在这些过程中的作用是通过与其他分子的相互作用来调节的。核蛋白 Twist-1 是 作为 Runx2 的调节蛋白合作伙伴特别感兴趣。我们研究细胞分化 Runx2 的理由 因子与 Twist-1（一种细胞存活因子）相互作用，源自我们和其他实验室的研究，表明 Runx2 和 Twist-1 之间的这些相互作用发生在蛋白质水平。我们的实验将直接测试 假设 Runx2 在成牙本质细胞分化中的关键功能受到 Twist-1 在蛋白质水平上的调节 本质上是功能拮抗的蛋白质相互作用。 Runx2功能的选择性和瞬时阻断 Twist-1 提供了一种抑制成牙本质细胞分化直至形态发生完成的方法。我们进一步 认为 Runx2 和 Twist-1 之间的相互作用并不相互拮抗，因为 Twist-1 可以介导细胞 形态发生过程中通过 FGF 介导的上皮间质信号传导进行增殖。因此，存在 人类 CCD 中的多生牙和 Runx2(-/-) 小鼠中的副芽可能反映了 Twist-1 活性的增加 而不是 Runx2 水平降低的直接影响。目标 1 将确定 Runx2 和 Twist-1 的模式是否一致 （mRNA 和蛋白质）表达与其在牙齿发育过程中提议的伙伴关系相一致，并将 将这些模式与牙齿形态发生和成牙本质细胞分子标记的表达相关联 差异化。目标 2 将通过小鼠遗传功能丧失和功能获得方法评估是否 Twist-1 表达的改变会影响牙齿形态发生和成牙本质细胞分化。目标 3 将研究 牙齿间充质中 Runx2 - Twist-1 蛋白相互作用的分子基础及其功能后果 Runx2 上的这种相互作用在成牙本质细胞分化中发挥作用，目标 4 将测试 bHLH 结构域是否 Twist-lean 通过 FGF 信号介导牙齿形态发生，该信号独立于其与 Runx2 的相互作用。这些 研究将加深我们对 Runx2 如何通过其在牙齿发育中实现选择性功能的理解 与 Twist-1 合作。重要的是，他们将解释多生牙是如何形成的以及成牙本质细胞分化是否是 由抑制的释放来确定。这些数据还将为理解发病机制提供一个框架 锁骨颅骨发育不良和 Saethre-Chotzen 综合征是两种威胁牙列的人类遗传性疾病。

项目成果

Molecular studies on the roles of Runx2 and Twist1 in regulating FGF signaling.

Runx2 和 Twist1 在调节 FGF 信号传导中作用的分子研究。

• 发表时间: 2012-11
• 作者: Lu, Yongbo;Li, Yucheng;Cavender, Adriana C;Wang, Suzhen;Mansukhani, Alka;D'Souza, Rena N
• 通讯作者: D'Souza, Rena N

Runx2 mediates FGF signaling from epithelium to mesenchyme during tooth morphogenesis.

Runx2 在牙齿形态发生过程中介导从上皮到间充质的 FGF 信号传导。

• DOI: 10.1016/j.ydbio.2004.02.012
• 发表时间: 2004-06-01
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: T. Åberg;Xiu;Jung;T. Yamashiro;M. Bei;R. Rice;H. Ryoo;I. Thesleff
• 通讯作者: I. Thesleff

Identification of tooth-specific downstream targets of Runx2.

Runx2 牙齿特异性下游靶标的识别。

• DOI: 10.1016/s0378-1119(01)00759-4
• 发表时间: 2001-11-14
• 期刊: Gene
• 影响因子: 3.5
• 作者: J. Gaikwad;A. Cavender;R. D'Souza
• 通讯作者: R. D'Souza

TWIST1 promotes the odontoblast-like differentiation of dental stem cells.

TWIST1 促进牙齿干细胞的成牙本质细胞样分化。

• 发表时间: 2011-07
• 作者: Li, Y;Lu, Y;Maciejewska, I;Galler, K M;Cavender, A;D'Souza, R N
• 通讯作者: D'Souza, R N

Cell-specific patterns of Cbfa1 mRNA and protein expression in postnatal murine dental tissues.

出生后小鼠牙组织中 Cbfa1 mRNA 和蛋白表达的细胞特异性模式。

• 发表时间: 2001-03
• 影响因子: 2.6
• 作者: Bronckers, A L;Engelse, M A;Cavender, A;Gaikwad, J;D'Souza, R N
• 通讯作者: D'Souza, R N