Characterization of sodium dependent phosphate transporter 2 signaling in hard tissue mineralization

硬组织矿化中钠依赖性磷酸盐转运蛋白 2 信号传导的表征

基本信息

批准号：
10661673
负责人：
Philip Adam Walczak
金额：
$ 4.81万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661673
关键词：
Affect Ameloblasts Amelogenesis American Animal Model Animals Apoptosis Award Basal Ganglia Biomedical Engineering Bone Density Bone Development Bone Growth CRISPR/Cas technology Calcium Cell Differentiation process Cell physiology Cells Chondrocytes Cloning Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Communities Complex Compressive Strength Congenital Abnormality Critical Thinking Data Data Analyses Dental Enamel Dentin Dentin Formation Dentinogenesis Dentistry Development Disease Doctor of Philosophy Education Enamel Formation Engineering Experimental Designs Faculty Fracture Frequencies Future Genes Grant Growth Health Sciences Histology Homeostasis Human Hydroxyapatites Impaired healing Impairment Incisor Injury Inorganic Phosphate Transporter Knock-out Knockout Mice Laboratories Leadership Learning Measures Mentors Mentorship Methods Molecular Morphogenesis Mus Mutation Odontoblasts Odontogenesis Oral health Osteoblasts Osteoclasts Osteogenesis Osteopenia Osteoporosis Patients Phosphorus Physiologic calcification Pilot Projects Positioning Attribute Process Proliferating Protocols documentation Publications Reading Regulation Reporting Research Resources Role Signal Transduction Skeletal Development Skeleton Sodium Specificity Techniques Technology Time Tissues Tooth structure Training Transgenic Mice Transmission Electron Microscopy Universities Vascular calcification Vesicle Washington Work Writing biomineralization bone calcification calcium phosphate career cell type experimental analysis experimental study extracellular fetal improved in vivo inorganic phosphate insight microCT mineralization mouse model multidisciplinary mutant novel therapeutics osteoblast differentiation repaired sensor skeletogenesis skills spatiotemporal tenure track therapeutic development therapeutic target

项目摘要

Project Summary Growth and homeostasis of bones require calcium and inorganic phosphorus (Pi). With millions of Americans suffering from bone mineralization disorders, there exists a need for understanding processes governing hard tissue mineralization. Pi is implicated in not only forming hydroxyapatite, the main crystal giving bone its compressive strength, but also regulating osteoblast and chondrocyte differentiation and function. PiT-2 (gene: Slc20a2) is the major form of sodium-dependent Pi transporters in mineralized tissue. Our laboratory is one of the first to have identified its potent role in regulating skeleton development and mineralization. Knockout of the PiT-2 gene in mice resulted in tooth and bone mineralization abnormalities, as evidenced by reduced osteoblast numbers, bone density and volume, and impaired incisor development and amelogenesis. The focus of this research is to explore further the role of PiT-2 in cellular and extracellular processes governing skeletogenesis, dentinogenesis, and amelogenesis. Specifically, Aim 1 will confirm and extend our preliminary findings to determine whether PiT-2 is required for regulation of odontoblast and ameloblast numbers, differentiation, and/or mineralizing activity with deficiencies leading to impaired dentinogenesis and amelogenesis. A role of PiT-2 in dentin repair and tertiary dentin formation will also be studied using a molar injury mouse model. In Specific Aim 2, we will define the mechanisms of action of PiT-2 in mineralizing cells using osteoblasts as an example. A Pi transport and sensing/signaling function of PiT-2 will be dissected through engineered PiT-2 transport deficient mutants. We expect these studies to provide a better understanding of how Pi influences functions of hard tissue forming cells, serving as a basis to develop novel therapeutics for patients suffering from mineralization disorders, such as osteoporosis and osteopenia. This project will take place at the University of Washington with collaborations between the Departments of Bioengineering and Oral Health Sciences during DDS-PhD training of the PI. The PI will develop critical thinking skills, learn state-of-the-art cellular and molecular techniques, in vivo transgenic mouse models, and data analysis that will prepare him for independent research. Ultimately, a strong mentorship team, multidisciplinary collaborative effort, and resources at the University of Washington with support through this award will prepare the trainee for a future tenure-track faculty position to study biomineralization processes.

项目摘要骨骼的生长和稳态需要钙和无机磷（PI）。有数百万美国人患有骨骼矿化障碍，需要了解努力的过程组织矿化。 PI不仅与形成羟基磷灰石有关，主要晶体使骨抗压强度，但还调节成骨细胞和软骨细胞分化和功能。 pit-2（基因： SLC20A2）是矿化组织中钠依赖性PI转运蛋白的主要形式。我们的实验室是第一个确定其在调节骨骼发育和矿化方面的有效作用的人。淘汰小鼠中的PIT-2基因导致牙齿和骨矿化异常，这是成骨细胞减少的证明数量，骨密度和体积，切牙发育和降解性受损。重点研究是为了进一步探索PIT-2在骨骼生成的细胞和细胞外过程中的作用，齿状生成和障碍发生。具体来说，AIM 1将确认并将我们的初步发现扩展到确定是否需要调节ODONTOBLAST和AMELEBLAST数字，分化和/或/或矿化活性的缺陷导致牙齿生成和休闲发生受损。 Pit-2在还将使用摩尔损伤小鼠模型研究牙本质修复和三级牙本质形成。在特定目标中 2，我们将使用成骨细胞来定义PIT-2在矿化细胞中的作用机理。 pi PIT-2的运输和传感/信号传导功能将通过工程pit-2传输不足来解剖突变体。我们希望这些研究能够更好地了解PI如何影响硬组织的功能形成细胞，作为开发矿化患者的新型治疗剂的基础疾病，例如骨质疏松和骨质减少症。该项目将在华盛顿大学举行在DDS-PHD培训期间，生物工程和口腔健康科学系之间的合作 pi。 PI将发展批判性思维技能，学习最先进的细胞和分子技术，体内转基因小鼠模型和数据分析将使他为独立研究做好准备。最终，一个强大的指导团队，多学科合作努力以及华盛顿大学的资源通过该奖项的支持将使受训者为未来的终身教师职位做好准备生物矿化过程。