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.

项目概要骨骼的生长和体内平衡需要钙和无机磷 (Pi)。与数百万美国人一起患有骨矿化障碍，需要了解控制硬骨的过程组织矿化。 Pi 不仅参与羟基磷灰石的形成，羟基磷灰石是骨骼的主要晶体抗压强度，还调节成骨细胞和软骨细胞的分化和功能。 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。我们的实验室是其中之一第一个发现其在调节骨骼发育和矿化方面的强大作用。淘汰赛的小鼠中的 PiT-2 基因导致牙齿和骨骼矿化异常，成骨细胞减少证明了这一点数量、骨密度和体积，以及门牙发育和釉质生成受损。本次的重点研究旨在进一步探索 PiT-2 在控制骨骼发生的细胞和细胞外过程中的作用，牙本质发生和釉质发生。具体来说，目标 1 将确认并扩展我们的初步发现确定 PiT-2 是否是调节成牙本质细胞和成釉细胞数量、分化和/或矿化活性不足，导致牙本质生成和釉质生成受损。 PiT-2 的作用牙本质修复和三级牙本质形成也将使用磨牙损伤小鼠模型进行研究。特定目标在图2中，我们将以成骨细胞为例定义PiT-2在矿化细胞中的作用机制。阿皮 PiT-2 的传输和传感/信号功能将通过工程化的 PiT-2 传输缺陷进行剖析突变体。我们希望这些研究能够更好地理解 Pi 如何影响硬组织的功能形成细胞，作为开发针对矿化患者的新型疗法的基础疾病，例如骨质疏松症和骨质减少。该项目将在华盛顿大学进行生物工程系和口腔健康科学系在 DDS-PhD 培训期间的合作 PI 的。 PI 将培养批判性思维技能，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为他的独立研究做好准备。最终，一个华盛顿大学强大的导师团队、多学科协作努力和资源通过该奖项的支持将为受训者做好未来终身教授职位的学习准备生物矿化过程。