DMP1 Mutations: Defects in Odontogenesis

描述（由申请人提供）：牙齿发育需要细胞生长、分化和矿化过程的精确时空协调。为了寻找正常牙齿形态发生所需的基因，我们研究了牙本质基质蛋白1 (DMP1)，这是一种在牙髓/成牙细胞中高度表达的非胶原基质蛋白，使用了体内功能丧失和功能获得的方法。dmp1缺失小鼠表现出牙髓腔增大、牙本质前区宽度增大、矿化程度低、第三磨牙形成延迟等严重牙齿异常。意识到常染色体隐性低磷血症佝偻病（ARHR）患者表现出与DMP1 - ko小鼠非常相似的表型变化，我们最近在这些患者中发现了两个DMP1突变。我们已经得出结论，这些突变是导致表型类似于Dmp1-KO小鼠的原因，尽管存在微小差异。在寻找DMP1控制成牙形成的机制时，我们意外地观察到无牙髓/成牙细胞中成骨的急剧减少。Osterix是一种转录因子，对成骨至关重要，但其在牙形成中的作用尚不清楚。DMP1在DMP1 - ko牙髓/成牙细胞中的靶向再表达可恢复成骨表达并修复牙齿形成缺陷。值得注意的是，我们在Dmp1-KO和osterix（常规或条件）KO小鼠中发现了类似的牙本质异常。因此，我们认为DMP1突变是ARHR患者牙本质缺陷的原因，并且DMP1在牙齿发育早期调控骨细胞的表达，在牙形成中起着关键作用。为了验证这一假设，我们将通过创建具有DMP1突变的小鼠模型来研究DMP1突变的分子遗传学和病理生理学。我们还将确定DMP1通过Osterix通过直接作用于细胞核和/或MAPK途径调节牙形成的机制。这些研究的成功完成将导致1)产生模拟这些人类突变的动物模型；2)通过细胞核水平和/或MAPK信号传导，了解DMP1通过骨组织直接控制牙形成的机制；3)鉴定DMP1的生物活性片段，这些片段最终可用于转化应用，通过提供与基因改变相对应的治疗方法，使公众受益，从而改善牙齿/口腔健康。公共卫生相关性：常染色体隐性低磷血症佝偻病（ARHR）患者表现出与我们在Dmp1-KO小鼠中观察到的牙本质缺陷非常相似的表型变化。我们在这些患者中发现的DMP1突变导致了当前研究计划的制定。这些研究的成功完成将提供牙本质形成的机理细节，并最终阐明牙本质结构缺陷的预防。此外，从这些研究中产生的小鼠模型可用于将个体基因突变与适当的临床干预联系起来。

DESCRIPTION (provided by applicant): Tooth development requires the precise spatial and temporal coordination of programs for cell growth, differentiation, and mineralization. In a search for genes required for normal tooth morphogenesis we have studied Dentin matrix protein1 (DMP1), a non- collagenous matrix protein highly expressed in pulp/odontoblast cells, using in vivo loss- and gain-of-function approaches. Dmp1-null mice display profound tooth abnormalities with enlarged pulp chambers, increased width of the predentin zone, hypomineralization, and delayed 3rd molar formation. Realizing that autosomal recessive hypophosphatemic rickets (ARHR) patients manifest phenotypic changes very similar to those observed in Dmp1-KO mice, we recently discovered two DMP1 mutations in these patients. We have concluded that these mutations are causes of the phenotypes resembling those of Dmp1-KO mice, although minor differences exist. In a search for mechanisms by which DMP1 controls odontogenesis, we unexpectedly observed a sharp reduction of osterix in the null pulp/odontoblasts. Osterix is a transcriptional factor that is essential for osteogenesis but its role in odontogenesis is unknown. Targeted re-expression of DMP1 in Dmp1-KO pulp/odontoblast cells restores osterix expression and rescues the defects in tooth formation. Notably, we found similar dentin abnormalities in Dmp1-KO, and osterix (conventional or conditional) KO mice. Therefore, we propose that DMP1 mutations are the cause of dentin defects in ARHR patients, and that DMP1 regulates osterix expression at an early stage of tooth development, which plays a critical role for odontogenesis. To test this hypothesis we will study the molecular genetics and pathophysiology of DMP1 mutations through creation of a mouse model with DMP1 mutations. We will also determine the mechanisms by which DMP1 modulates odontogenesis through Osterix via direct action at the nucleus and/or MAPK pathways. The successful completion of these studies will lead to 1) generation of an animal model mimicking these human mutations; 2) understanding the mechanism by which DMP1 controls odontogenesis through osterix via a direct mechanism within the nucleus level and/or MAPK signaling; and 3) identification of bioactive fragment(s) of DMP1 which can ultimately be used in translational applications that will benefit the public by providing therapeutic approaches corresponding to genetic alterations, leading to improved dental/oral health. PUBLIC HEALTH RELEVANCE: Patients with autosomal recessive hypophosphatemic rickets (ARHR) patients manifest phenotypic changes very similar to dentin defects we observed in Dmp1-KO mice. Our discovery of DMP1 mutations in these patients led to the formulation of the current research proposal. Our successful completion of these studies will provide mechanistic details about dentin formation, and will ultimately shed light on the prevention of structural defects in dentin. In addition, the mouse models which to be generated from these studies can be used to correlate individual gene mutations with the appropriate clinical intervention.