Redox and Ca2+ signaling regulation of enamel mineralization

牙釉质矿化的氧化还原和 Ca2 信号传导调节

基本信息

批准号：
10586833
负责人：
Rodrigo S. Lacruz
金额：
$ 48.4万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-10 至 2028-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10586833
关键词：
Acquired Dental Fluorosis Address Adult Affect Ameloblasts Amelogenesis Anabolism Apatites Biogenesis Biological Assay Brain Calcium Cell Line Cell physiology Cells Chromosome 21 Complex Confocal Microscopy Crystal Formation Data Defect Dental Enamel Dental caries Development Disease Down Syndrome Electron Transport Enamel Formation Exposure to Fluorides Foundations Functional disorder Funding Generations Genes Genetic Diseases Genus Hippocampus Glycolysis Goals Growth Heterozygote Histology Homeostasis Human Chromosomes Image Impairment In Vitro Incisor Individual Intake Knockout Mice Label Maturation-Stage Ameloblast Measures Mechanics Membrane Potentials Metabolic Minerals Mitochondria Mitochondrial DNA Mitochondrial Matrix Modeling Morphology Mus Muscle NADH Nutrient Oral health Organelles Oxidation-Reduction Oxygen Consumption Pancreas Patients Play Process Production Property Protein Biosynthesis Proteins Reactive Oxygen Species Recommendation Regulation Reporter Reporting Risk Rodent Role Safety Signal Transduction Sorting Structure Testing Tissues Tooth structure Trisomy bone calcification dosage drinking water fluorosis high risk human disease improved in vivo induced pluripotent stem cell mineralization mitochondrial dysfunction mitochondrial membrane mitochondrial metabolism mouse model overexpression response segregation single cell analysis single-cell RNA sequencing skeletal transcriptome transcriptome sequencing transcriptomic profiling uptake

项目摘要

Project Summary: Proper mineralization of dental enamel protects against bacterial attack that causes tooth decay (caries). The ameloblasts cells are responsible for producing and secreting an abundance of proteins (laying a foundation for enamel growth) as well as engaging in active mineral transport (calcifying the enamel). These functions are stage dependent with amelogenesis being divided into the secretory (protein synthesis) and maturation stage (mineralization). However, the mechanisms providing metabolic support for these processes and how mitochondrial dysfunction might alter them is poorly understood. Mitochondria are organelles within the cells that convert nutrients into energy via the production of ATP, and deficiency in mitochondrial function results in human diseases. Mitochondria play an important role in enamel as evidenced by defects in mitochondrial DNA causing abnormal enamel development. Therefore, the goal in this competing renewal proposal is to investigate the interplay between mitochondrial function and enamel formation during environmental insults and determine whether this is altered in patients with genetic disorders that effect mitochondrial function. We will specifically address this in the context of Down syndrome (DS), or Trisomy 21, and during dental fluorosis, a process in which exposure to excess of fluoride during development weakens tooth enamel. DS patients present with enamel defects including hypocalcification and hypoplasia, both caused by developmental defects in enamel formation. Mitochondrial dysfunction is widely reported in DS. The overarching hypothesis of this proposal is that altered mitochondrial function in DS ameloblasts alters enamel crystal formation and impacts sensitivity to fluorosis. In strong support, our preliminary data show that Dp- 16 mice (an established mouse model of DS) have mechanically weak and morphologically abnormal enamel. Moreover, overexpression of RCAN1 (a gene associated with DS pathophysiology that is expressed in ameloblasts) in enamel cell lines, significantly impaired mitochondrial function. We have also reported that fluoride exposure of enamel cells significantly affected the biosynthesis of the proteins of the electron transport chain (ETC) responsible for maintaining ATP production, but not in other cells tested, suggesting unique sensitivity of enamel cells to fluoride, possibly associated with higher ROS levels. In the proposed studies we will use DS mouse models (Dp16 mice, Rcan1-KO mice, Dp16 x Rcan1-KO mice) to address the role of mitochondria in the ameloblasts of these mice. We will also use recently developed reporter mice expressing fluorescently labelled secretory and maturation stage ameloblasts to induce fluoride and investigate mitochondrial defects using single cell RNASeq and bulk RNAseq to compare ameloblasts with other tissues. To address if ameloblasts of DS models are more sensitive to fluoride, we will treat the cells with fluoride and analyze mitochondrial function.

项目摘要：牙齿搪瓷的适当矿化可防止引起的细菌攻击蛀牙（龋齿）。成熟细胞细胞负责产生和分泌丰富的蛋白质（为搪瓷生长奠定基础）以及进行主动矿物运输（钙化搪瓷）。这些功能取决于阶段，而没有分泌分泌（蛋白质合成）和成熟阶段（矿化）。但是，提供代谢的机制对这些过程的支持以及线粒体功能障碍可能会改变它们，这是对它们的理解很少的。线粒体是细胞内的细胞器，可通过ATP的产生将养分转化为能量线粒体功能的缺乏会导致人类疾病。线粒体起重要作用在搪瓷中，线粒体DNA缺陷所证明，导致异常牙釉质发育。因此，该竞争续约提案的目标是调查环境侮辱期间的线粒体功能和搪瓷形成，并确定是否是影响线粒体功能的遗传疾病患者的改变。我们将专门解决这是在唐氏综合症（DS）或三体疾病的背景下，以及牙齿氟中毒期间发育过程中暴露于氟化物过量会削弱牙齿搪瓷。 DS患者出现牙釉质缺陷，包括低钙化和发育不全，都由发育缺陷引起搪瓷形成。线粒体功能障碍在DS中广泛报道。总体假设该提议是DS成成木中的线粒体功能改变了搪瓷晶体形成并影响对氟中毒的敏感性。为了有很大的支持，我们的初步数据表明DP- 16只小鼠（已建立的DS小鼠模型）在机械上弱且形态异常搪瓷。此外，RCAN1的过表达（与DS病理生理相关的基因是在牙釉质细胞系中以木材细胞表达）明显受损的线粒体功能。我们有还报道了搪瓷细胞的氟化物暴露显着影响蛋白质的生物合成负责维持ATP生产的电子传输链（ETC）的经过测试，表明搪瓷细胞对氟化物的独特敏感性，可能与较高的ROS有关水平。在拟议的研究中，我们将使用DS小鼠模型（DP16小鼠，RCAN1-KO小鼠，DP16 x RCAN1-KO小鼠）解决线粒体在这些小鼠的成成布中的作用。我们还将使用最近开发的记者小鼠表达荧光标记为分泌和成熟阶段木材细胞诱导氟化物并使用单细胞RNASEQ和大量研究线粒体缺陷 RNASEQ将成成木细胞与其他组织进行比较。解决DS模型的成成木是否更多对氟化物敏感，我们将用氟化物处理细胞并分析线粒体功能。